|
1
|
Mendoza-Guevara CC, Martinez-Escobar A, Ramos-Godínez MDP, Muñoz-Medina JE, Ramon-Gallegos E. Efficient Delivery of SARS-CoV-2 Plasmid DNA in HEK-293T Cells Using Chitosan Nanoparticles. Pharmaceuticals (Basel) 2025; 18:683. [PMID: 40430502 PMCID: PMC12114681 DOI: 10.3390/ph18050683] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/04/2025] [Revised: 04/24/2025] [Accepted: 05/01/2025] [Indexed: 05/29/2025] Open
Abstract
Background/Objectives: Gene therapy has emerged as a promising strategy for treating a wide range of diseases. However, a major challenge remains in developing efficient and safe delivery systems for genetic material. Nanoparticles, particularly chitosan nanoparticles (CNPs), have gained significant attention as a potential solution. This study focuses on designing a SARS-CoV-2 plasmid DNA (pDNA) conjugated with CNPs and evaluating its in vitro delivery efficiency. Methods: The Omicron Spike DNA sequence was inserted into the pIRES2-eGFP expression vector, and CNPs were synthesized with optimized physicochemical properties to enhance stability, cellular uptake, and transfection efficiency. The conjugate was characterized using UV-Vis, FT-IR, DLS, and TEM techniques. Transfection efficiency was assessed and compared to the commercially available TurboFect reagent as a control. Results: CNPs-pDNA polyplexes with an average size of 159.0 ± 33.1 nm (TEM), a zeta potential of +19.7 ± 0.3 mV, and 100% ± 0.0 encapsulation efficiency were developed as a non-viral delivery system. CNPs efficiently serve as a delivery vehicle for the constructed pDNA without altering cell morphology, achieving transfection efficiencies of 62-74%, compared to 55-70% for TurboFect. Furthermore, RT-qPCR confirmed the expression of Spike mRNA, and Western blot assays validated the expression of Spike protein. Notably, Spike protein expression from CNPs was found to be two-fold higher than the control at 96 h post-transfection. Conclusions: These findings suggest that CNPs are a promising and versatile platform for delivering genetic material. Importantly, this study highlights the intrinsic properties of chitosan, without the use of additional ligands, as a key factor in achieving efficient gene delivery.
Collapse
Affiliation(s)
- Citlali Cecilia Mendoza-Guevara
- Laboratorio de Citopatología Ambiental, Departamento de Morfología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional (IPN), Ciudad de México 07738, Mexico; (C.C.M.-G.); (A.M.-E.)
| | - Alejandro Martinez-Escobar
- Laboratorio de Citopatología Ambiental, Departamento de Morfología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional (IPN), Ciudad de México 07738, Mexico; (C.C.M.-G.); (A.M.-E.)
| | | | - José Esteban Muñoz-Medina
- Banco de Muestras de la CCILE, Instituto Mexicano del Seguro Social (IMSS), Ciudad de México 07760, Mexico;
| | - Eva Ramon-Gallegos
- Laboratorio de Citopatología Ambiental, Departamento de Morfología, Escuela Nacional de Ciencias Biológicas, Instituto Politécnico Nacional (IPN), Ciudad de México 07738, Mexico; (C.C.M.-G.); (A.M.-E.)
| |
Collapse
|
|
2
|
Lin Z, Guha Ray P, Huang J, Buchmann P, Fussenegger M. Electromagnetic wireless remote control of mammalian transgene expression. NATURE NANOTECHNOLOGY 2025:10.1038/s41565-025-01929-w. [PMID: 40325210 DOI: 10.1038/s41565-025-01929-w] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Received: 07/25/2024] [Accepted: 04/02/2025] [Indexed: 05/07/2025]
Abstract
Communication between wireless field receivers and biological sensors remains a key constraint in the development of wireless electronic devices for minimally invasive medical monitoring and biomedical applications involving gene and cell therapies. Here we describe a nanoparticle-cell interface that enables electromagnetic programming of wireless expression regulation (EMPOWER) of transgenes via the generation of cellular reactive oxygen species (ROS) at a biosafe level. Multiferroic nanoparticles coated with chitosan to improve biocompatibility generate ROS in the cytoplasm of cells in response to a low-frequency (1-kHz) magnetic field. Overexpressed ROS-responsive KEAP1/NRF2 biosensors detect the generated ROS which is rewired to synthetic ROS-responsive promoters to drive transgene expression. In a proof-of-concept study, subcutaneously implanted alginate-microencapsulated cells stably expressing an EMPOWER-controlled insulin expression system normalized blood-glucose levels in a mouse model of type 1 diabetes in response to a weak magnetic field.
Collapse
Affiliation(s)
- Zhihua Lin
- Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland
| | - Preetam Guha Ray
- Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland
| | - Jinbo Huang
- Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland
| | - Peter Buchmann
- Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland
| | - Martin Fussenegger
- Department of Biosystems Science and Engineering, ETH Zurich, Basel, Switzerland.
- University of Basel, Faculty of Life Science, Basel, Switzerland.
| |
Collapse
|
|
3
|
Li TL, Zhou J, Gu JL, Zheng HW, Shen YX, Song MM. Oral pH- and inflammation-targeted delivery system with biodegradable multi-layer core-shell nanocapsules for the treatment of ulcerative colitis. NANOSCALE 2025; 17:10124-10141. [PMID: 40136045 DOI: 10.1039/d4nr04218b] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 03/27/2025]
Abstract
Most biologics require administration via parenteral routes; however, the pain and local allergic reaction brought about by injection usually lead to poor compliance, especially for chronic patients. Meanwhile, the oral delivery of biologics faces great challenges due to the complex physiological environment of the gastrointestinal tract. Herein, we developed a new formulation of multilayer core-shell nanocapsules composed of hyaluronan-modified silica nanocapsules, chitosan and alginate layers for the oral delivery of biologics. The mesencephalic astrocyte-derived neurotrophic factor (MANF) was selected as the model biologic for the treatment of ulcerative colitis (UC). MANF-loaded biodegradable silica (MBS) nanocapsules were first obtained simultaneously with the preparation. Then, MBS nanocapsules were surface-modified with hyaluronan (MBSH) for oral targeted delivery to the inflamed region via CD44-mediated endocytosis. To survive in the harsh gastrointestinal environment, MBSH was further modified using chitosan and alginate via polyelectrolyte interactions. With this delivery system, i.e., MBSH@CA, the cumulative release of MANF protein in the simulated gastric fluid (SGF) and simulated intestine fluid (SIF) was <10% of the total amount in MBSH@CA. Bio-distribution studies showed that the MBSH@CA nanocapsules were mainly distributed in the colon after 24 h treatment. Ex vivo imaging of the colons revealed a preferential accumulation of the MBSH@CA nanocapsules in the inflamed colons compared with the healthy colons. According to in vivo anti-inflammatory analysis, the oral MBSH@CA nanocapsules were effective in reducing related inflammatory symptoms caused by DSS-induced colitis. All of the above results suggested that the multilayer silica MBSH@CA nanocapsules could be employed for targeted drug delivery against UC.
Collapse
Affiliation(s)
- Tian-Le Li
- School of Basic Medical Sciences, Anhui Medical University, 81 Meishan Road, 230032 Hefei, Anhui, PR China.
| | - Jie Zhou
- School of Basic Medical Sciences, Anhui Medical University, 81 Meishan Road, 230032 Hefei, Anhui, PR China.
| | - Jin-Long Gu
- School of Basic Medical Sciences, Anhui Medical University, 81 Meishan Road, 230032 Hefei, Anhui, PR China.
| | - Han-Wen Zheng
- The Second School of Clinical Medicine, Anhui Medical University, 81 Meishan Road, 230032 Hefei, Anhui, PR China
| | - Yu-Xian Shen
- School of Basic Medical Sciences, Anhui Medical University, 81 Meishan Road, 230032 Hefei, Anhui, PR China.
| | - Meng-Meng Song
- School of Basic Medical Sciences, Anhui Medical University, 81 Meishan Road, 230032 Hefei, Anhui, PR China.
| |
Collapse
|
|
4
|
Deng D, Peng D, Lv J, Zhang W, Tian H, Wang T, Wu M, Zhao Y. Double-Network Hydrogel Based on Methacrylated Chitosan/Hyaluronic Acid Coacervate for Enhanced Wet-Tissue Adhesion. Biomacromolecules 2025; 26:2317-2330. [PMID: 40102053 DOI: 10.1021/acs.biomac.4c01645] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 03/20/2025]
Abstract
Developing robust wet tissue adhesives remains challenging due to interfacial water and irregular surfaces. While polyelectrolyte coacervates demonstrate promising hydrophobic/fluidic properties for wet adhesion, their low cohesion limits practical applications. Herein, a wet tissue bioadhesive based on coacervates formed from low- molecular-weight methacrylated chitosan (CSMA) and hyaluronic acid (HA) is reported. These homogeneous and transparent coacervates displayed high solid content (∼18.0%), fluidity (∼105 mPa·s), and tunable mechanical properties. Upon application to wet tissue surfaces, the coacervate can be photo-cross-linked to form a double-network hydrogel in situ, resulting in improved cohesion and durable adhesion. The resulting CSMA-HA hydrogel demonstrated robust adhesion to tissues, with a bursting pressure of 374 mmHg. Remarkably, the bursting pressure can be further enhanced (∼623 mmHg) after 24 h of PBS immersion due to dynamic bond reorganization and low swelling. The demonstrated stability under physiological conditions and robust wet adhesion position CSMA-HA coacervates as a transformative platform for tissue adhesive applications.
Collapse
Affiliation(s)
- Dafeng Deng
- Jihua Institute of Biomedical Engineering and Technology, Jihua Laboratory, Foshan 528200, China
- College of Sciences, Northeastern University, Shenyang 110819, China
| | - Deyi Peng
- Jihua Institute of Biomedical Engineering and Technology, Jihua Laboratory, Foshan 528200, China
- College of Sciences, Northeastern University, Shenyang 110819, China
| | - Jianhua Lv
- Jihua Institute of Biomedical Engineering and Technology, Jihua Laboratory, Foshan 528200, China
| | - Wenchang Zhang
- Jihua Institute of Biomedical Engineering and Technology, Jihua Laboratory, Foshan 528200, China
| | - Huaqin Tian
- Foshan Hospital of Traditional Chinese Medicine, Foshan, Guangdong 528000, China
| | - Tieqiang Wang
- College of Sciences, Northeastern University, Shenyang 110819, China
| | - Mi Wu
- Jihua Institute of Biomedical Engineering and Technology, Jihua Laboratory, Foshan 528200, China
| | - Yan Zhao
- Jihua Institute of Biomedical Engineering and Technology, Jihua Laboratory, Foshan 528200, China
| |
Collapse
|
|
5
|
Chaulagain B, Singh J. Penetratin and Mannose-Functionalized Cannabidiol Lipid Nanoparticles Encapsulating the BDNF Gene Reduce Amyloid-Induced Inflammation. Mol Pharm 2025; 22:154-167. [PMID: 39588752 PMCID: PMC11874068 DOI: 10.1021/acs.molpharmaceut.4c00811] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/27/2024]
Abstract
Inflammation is emerging as a critical player in the disease progression of Alzheimer's disease (AD) by its interaction with amyloid beta plaques in a feed-forward loop. There is also a decline in the nourishment and enriching neurotrophic factor, brain-derived neurotrophic factor (BDNF), in the brain. Therefore, supplementing the brain with BDNF by gene delivery and delivering the anti-inflammatory agent, cannabidiol (CBD) in this case, to mitigate inflammation-induced disease cascade offers an attractive treatment strategy. To achieve the brain localization of CBD and pBDNF, lipid nanoparticles (LNPs) functionalized with mannose and penetratin were utilized. CBD and pBDNF were successfully encapsulated in the LNPs (more than 80%) with a size less than 180 nm, polydispersity index less than 0.25, and zeta potential of 23 mV. CBD was released from the formulation over a period of a week. The dual-functionalized LNPs demonstrated higher cellular uptake of CBD and expressed a significantly higher amount of BDNF (p-value <0.05) after transfection than their nonmodified counterparts in four brain cell lines, i.e., brain endothelial cells (b.END3), immortalized microglia cells (IMGs), primary astrocytes, and primary neurons. Similarly, the permeation of CBD through the dual-modified LNPs across the in vitro coculture blood-brain barrier model was significantly higher (p-value <0.05) compared to free CBD or nonfunctionalized nanoparticles. The LNPs demonstrated anti-inflammatory activity against lipopolysaccharides and human amyloid beta1-42 oligomer induction as they reduced the protein and mRNA expression of pro-inflammatory cytokines TNF-α (p < 0.05) and IL-1β (p < 0.05) in IMG cells. In summary, the penetratin and mannose-functionalized LNPs encapsulating CBD and pBDNF could serve as a promising therapy in AD, requiring further validation in animal models.
Collapse
Affiliation(s)
- Bivek Chaulagain
- Department of Pharmaceutical Sciences, School of Pharmacy, College of Health and Human Sciences, North Dakota State University, Fargo, ND 58105, USA
| | - Jagdish Singh
- Department of Pharmaceutical Sciences, School of Pharmacy, College of Health and Human Sciences, North Dakota State University, Fargo, ND 58105, USA
| |
Collapse
|
|
6
|
Lou H, Shang X, Wang Z, Xiang S, Qiu Y, Wang C, Yu F. Chitosan-derived drug free "artificial beacon" simulating immunogenetic cell death cascade effector to initiate immune response for cancer therapy. Int J Biol Macromol 2025; 287:138434. [PMID: 39645118 DOI: 10.1016/j.ijbiomac.2024.138434] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/16/2024] [Revised: 12/02/2024] [Accepted: 12/04/2024] [Indexed: 12/09/2024]
Abstract
Immunogenetic cell death (ICD) is widely participated in tumor immune therapy. However, the stress responses triggered by individual ICD inducers are typically not strong enough to effectively kickstart an ICD effect and successful ICD necessitates a high level of ICD stimulus, which may be linked to dose-related toxicity. In this research, we developed a drug-free "artificial beacon" ATP/CSO@ECM that mimics the ICD cascade system to kickstart an immune response with cationic chitosan (CSO) as a bridge, which participated in integrating tumor antigens and functional damage-associated molecular patterns (DAMPs) into one effector by electrostatic interaction. This beacon is made up of ATP/CSO nanocomplexes covered by an engineered cell membrane (ECM), which is verified to enrich with high mobility group box 1 (HMGB1) and calreticulin (CRT). When exposed to the acidic tumor environment, the ATP/CSO@ECM underwent a morphological change by proton buffering capability of CSO. This resulted in the release of simulated DAMPs and the adjuvant CSO, all of which collaborated to activate dendritic cells and ultimately prolong the effectiveness of immunotherapy. This chitosan-derived "artificial beacon" ATP/CSO@ECM fully mobilizes the function of CSO and avoids the insufficient ICD effect by concentrating signaling molecules, providing a hopeful strategy for using the ICD process in targeted cancer therapy.
Collapse
Affiliation(s)
- Haiya Lou
- Department of Diagnostic Ultrasound and Echocardiography, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, China
| | - Xuwei Shang
- College of Pharmaceutical Science, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China
| | - Zixu Wang
- College of Pharmaceutical Science, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China
| | - Shanshan Xiang
- College of Pharmaceutical Science, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China
| | - Yihe Qiu
- College of Pharmaceutical Science, Zhejiang University, 866 Yuhangtang Road, Hangzhou 310058, China
| | - Cheng Wang
- School of Pharmacy, Changzhou University, Changzhou, Jiangsu, China.
| | - Fangying Yu
- Department of Diagnostic Ultrasound and Echocardiography, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, China.
| |
Collapse
|
|
7
|
Ma Y, Li S, Lin X, Chen Y. A perspective of lipid nanoparticles for RNA delivery. EXPLORATION (BEIJING, CHINA) 2024; 4:20230147. [PMID: 39713203 PMCID: PMC11655307 DOI: 10.1002/exp.20230147] [Citation(s) in RCA: 4] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 11/14/2023] [Accepted: 03/07/2024] [Indexed: 12/24/2024]
Abstract
Over the last two decades, lipid nanoparticles (LNPs) have evolved as an effective biocompatible and biodegradable RNA delivery platform in the fields of nanomedicine, biotechnology, and drug delivery. They are novel bionanomaterials that can be used to encapsulate a wide range of biomolecules, such as mRNA, as demonstrated by the current successes of COVID-19 mRNA vaccines. Therefore, it is important to provide a perspective on LNPs for RNA delivery, which further offers useful guidance for researchers who want to work in the RNA-based LNP field. This perspective first summarizes the approaches for the preparation of LNPs, followed by the introduction of the key characterization parameters. Then, the in vitro cell experiments to study LNP performance, including cell selection, cell viability, cellular association/uptake, endosomal escape, and their efficacy, were summarized. Finally, the in vivo animal experiments in the aspects of animal selection, administration, dosing and safety, and their therapeutic efficacy were discussed. The authors hope this perspective can offer valuable guidance to researchers who enter the field of RNA-based LNPs and help them understand the crucial parameters that RNA-based LNPs demand.
Collapse
Affiliation(s)
- Yutian Ma
- Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of PharmacyUniversity of North Carolina at Chapel HillChapel HillNorth CarolinaUSA
| | - Shiyao Li
- School of ScienceRMIT UniversityBundooraVictoriaAustralia
- ARC Centre of Excellence in Convergent Bio‐Nano Science and Technology, and the Department of Chemical EngineeringThe University of MelbourneParkvilleVictoriaAustralia
| | - Xin Lin
- Department of Cell BiologyDuke University Medical CenterDurhamNorth CarolinaUSA
| | - Yupeng Chen
- CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, CAS Center for Excellence in NanoscienceNational Center for Nanoscience and TechnologyBeijingChina
- University of Chinese Academy of SciencesBeijingChina
| |
Collapse
|
|
8
|
Yang J, Liu X, Yang H, Wang H, Xie B, Gao W, Xu M, Xu X, Liu B, Chen Z. Combination application of Pickering emulsion and BLS protein nanoparticles enhances vaccine efficacy. Int J Biol Macromol 2024; 283:137635. [PMID: 39547601 DOI: 10.1016/j.ijbiomac.2024.137635] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/19/2024] [Revised: 11/11/2024] [Accepted: 11/12/2024] [Indexed: 11/17/2024]
Abstract
The deficiency of recombinant protein immune response could be compensated for by using nanoparticle platforms or adding immune enhancers, however existing vaccines or adjuvants struggle to elicit durable cellular immune responses. In this work, a protein nanoscaffold, lumazine synthase isolated from Brucella (BLS) was optimally designed that could facilitate cellular uptake of displayed antigens and the maturation of bone marrow-derived dendritic cells (BMDCs), and enhancing humoral immune responses. To enhance cellular immune response, chitosan hydrochloride-stabilized Pickering emulsion (CHSPE) was evaluated as an adjuvant for the BLS nanoscaffold-based vaccine. CHSPE could enhance the recruitment and activation of DCs at the injection site and the uptake of antigen and activation of DCs in the draining lymph nodes (dLNs), compared with commercial adjuvant ISA 206. In addition, CHSPE induced antigen-specific antibody levels comparable to those of ISA 206 and increased the ratio of central memory T cells (TCM) and effector memory T cells (TEM), and promoted the secretion of Th1-type cytokines. Moreover, CHSPE-formulated vaccine provided a similar level of protection to the live attenuated vaccine and was superior to that of ISA 206. Taken together, the combination of the BLS nanoscaffold and CHSPE provides a feasible strategy for recombinant protein subunit vaccine development.
Collapse
Affiliation(s)
- Jianghua Yang
- Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University. Liaoning Province, Shenyang 110866, PR China
| | - Xiang Liu
- Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University. Liaoning Province, Shenyang 110866, PR China
| | - Haoyan Yang
- Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University. Liaoning Province, Shenyang 110866, PR China
| | - Hao Wang
- Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University. Liaoning Province, Shenyang 110866, PR China
| | - Bin Xie
- Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University. Liaoning Province, Shenyang 110866, PR China
| | - Weiyu Gao
- Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University. Liaoning Province, Shenyang 110866, PR China
| | - Man Xu
- Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University. Liaoning Province, Shenyang 110866, PR China
| | - Xinling Xu
- Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University. Liaoning Province, Shenyang 110866, PR China
| | - Baoshan Liu
- Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University. Liaoning Province, Shenyang 110866, PR China.
| | - Zeliang Chen
- Key Laboratory of Livestock Infectious Diseases in Northeast China, Ministry of Education, College of Animal Science and Veterinary Medicine, Shenyang Agricultural University. Liaoning Province, Shenyang 110866, PR China; NMPA Key Laboratory for Quality Monitoring and Evaluation of Vaccines and Biological Products, Key Laboratory of Tropical Diseases Control, School of public health, Sun Yat-sen University, Guangzhou 510275, PR China; Collaborative Innovation Center for Prevention and Control of Zoonoses, Jinzhou Medical University, Jinzhou 121001, PR China.
| |
Collapse
|
|
9
|
Wang Z, Zhitomirsky I. Strategies for alkali-free synthesis, surface modification, and electrophoretic deposition of composite films for biomedical applications. Colloids Surf A Physicochem Eng Asp 2024; 702:135150. [DOI: 10.1016/j.colsurfa.2024.135150] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 01/05/2025]
|
|
10
|
Li Y, Li XM, Wei LS, Ye JF. Advancements in mitochondrial-targeted nanotherapeutics: overcoming biological obstacles and optimizing drug delivery. Front Immunol 2024; 15:1451989. [PMID: 39483479 PMCID: PMC11524880 DOI: 10.3389/fimmu.2024.1451989] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/20/2024] [Accepted: 09/19/2024] [Indexed: 11/03/2024] Open
Abstract
In recent decades, nanotechnology has significantly advanced drug delivery systems, particularly in targeting subcellular organelles, thus opening new avenues for disease treatment. Mitochondria, critical for cellular energy and health, when dysfunctional, contribute to cancer, neurodegenerative diseases, and metabolic disorders. This has propelled the development of nanomedicines aimed at precise mitochondrial targeting to modulate their function, marking a research hotspot. This review delves into the recent advancements in mitochondrial-targeted nanotherapeutics, with a comprehensive focus on targeting strategies, nanocarrier designs, and their therapeutic applications. It emphasizes nanotechnology's role in enhancing drug delivery by overcoming biological barriers and optimizing drug design for specific mitochondrial targeting. Strategies exploiting mitochondrial membrane potential differences and specific targeting ligands improve the delivery and mitochondrial accumulation of nanomedicines. The use of diverse nanocarriers, including liposomes, polymer nanoparticles, and inorganic nanoparticles, tailored for effective mitochondrial targeting, shows promise in anti-tumor and neurodegenerative treatments. The review addresses the challenges and future directions in mitochondrial targeting nanotherapy, highlighting the need for precision, reduced toxicity, and clinical validation. Mitochondrial targeting nanotherapy stands at the forefront of therapeutic strategies, offering innovative treatment perspectives. Ongoing innovation and research are crucial for developing more precise and effective treatment modalities.
Collapse
Affiliation(s)
- Yang Li
- General Surgery Center, First Hospital of Jilin University, Changchun, China
- Department of Rehabilitation, School of Nursing, Jilin University, Changchun, China
| | - Xiao-meng Li
- Department of Rehabilitation, School of Nursing, Jilin University, Changchun, China
| | - Li-si Wei
- Department of Rehabilitation, School of Nursing, Jilin University, Changchun, China
| | - Jun-feng Ye
- General Surgery Center, First Hospital of Jilin University, Changchun, China
| |
Collapse
|
|
11
|
Guo X, Xu J, Lu X, Zheng X, Chen X, Sun Z, Shen B, Tang H, Duan Y, Zhou Z, Feng X, Chen Y, Wang J, Pang J, Jiang Q, Huang B, Gu N, Li J. Chenodeoxycholic Acid-Modified Polyethyleneimine Nano-Composites Deliver Low-Density Lipoprotein Receptor Genes for Lipid-Lowering Therapy by Targeting the Liver. Adv Healthc Mater 2024; 13:e2400254. [PMID: 38857027 DOI: 10.1002/adhm.202400254] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/25/2024] [Revised: 06/03/2024] [Indexed: 06/11/2024]
Abstract
Lipid-lowering drugs, especially statins, are extensively utilized in clinical settings for the prevention of hyperlipidemia. Nevertheless, prolonged usage of current lipid-lowering medications is associated with significant adverse reactions. Therefore, it is imperative to develop novel therapeutic agents for lipid-lowering therapy. In this study, a chenodeoxycholic acid and lactobionic acid double-modified polyethyleneimine (PDL) nanocomposite as a gene delivery vehicle for lipid-lowering therapy by targeting the liver, are synthesized. Results from the in vitro experiments demonstrate that PDL exhibits superior transfection efficiency compared to polyethyleneimine in alpha mouse liver 12 (AML12) cells and effectively carries plasmids. Moreover, PDL can be internalized by AML12 cells and rapidly escape lysosomal entrapment. Intravenous administration of cyanine5.5 (Cy5.5)-conjugated PDL nanocomposites reveals their preferential accumulation in the liver compared to polyethyleneimine counterparts. Systemic delivery of low-density lipoprotein receptor plasmid-loaded PDL nanocomposites into mice leads to reduced levels of low-density lipoprotein cholesterol (LDL-C) and triglycerides (TC) in the bloodstream without any observed adverse effects on mouse health or well-being. Collectively, these findings suggest that low-density lipoprotein receptor plasmid-loaded PDL nanocomposites hold promise as potential therapeutics for lipid-lowering therapy.
Collapse
Affiliation(s)
- Xiaotang Guo
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Jiming Xu
- School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, P. R. China
| | - Xiyuan Lu
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Xiaoyan Zheng
- School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, P. R. China
| | - Xi Chen
- The Affiliated Eye Hospital, Nanjing Medical University, Nanjing, 210029, P. R. China
- Department of Ophthalmology, Northern Jiangsu People's Hospital, Yangzhou, 225001, P. R. China
| | - Zhenning Sun
- School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, P. R. China
| | - Beilei Shen
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Hao Tang
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Yiman Duan
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Zhengwei Zhou
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Xu Feng
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Yang Chen
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Junjie Wang
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Jing Pang
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Qin Jiang
- The Affiliated Eye Hospital, Nanjing Medical University, Nanjing, 210029, P. R. China
| | - Bin Huang
- School of Biomedical Engineering and Informatics, Nanjing Medical University, Nanjing, 211166, P. R. China
| | - Ning Gu
- Medical School, Nanjing University, Nanjing, 210093, P. R. China
| | - Juxue Li
- State Key Laboratory of Reproductive Medicine and Offspring Health, Nanjing Medical University, Nanjing, 211166, P. R. China
- Key Laboratory of Human Functional Genomics of Jiangsu Province, Nanjing Medical University, Nanjing, 211166, P. R. China
- School of Biological Science and Medical Engineering, Southeast University, Nanjing, 210096, P. R. China
- The Affiliated Eye Hospital, Nanjing Medical University, Nanjing, 210029, P. R. China
- The Second Affiliated Hospital of Nanjing Medical University, Nanjing, 210011, P. R. China
| |
Collapse
|
|
12
|
Cojocaru E, Petriș OR, Cojocaru C. Nanoparticle-Based Drug Delivery Systems in Inhaled Therapy: Improving Respiratory Medicine. Pharmaceuticals (Basel) 2024; 17:1059. [PMID: 39204164 PMCID: PMC11357421 DOI: 10.3390/ph17081059] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/02/2024] [Revised: 08/03/2024] [Accepted: 08/09/2024] [Indexed: 09/03/2024] Open
Abstract
Inhaled nanoparticle (NP) therapy poses intricate challenges in clinical and pharmacodynamic realms. Recent strides have revolutionized NP technology by enabling the incorporation of diverse molecules, thus circumventing systemic clearance mechanisms and enhancing drug effectiveness while mitigating systemic side effects. Despite the established success of systemic NP delivery in oncology and other disciplines, the exploration of inhaled NP therapies remains relatively nascent. NPs loaded with bronchodilators or anti-inflammatory agents exhibit promising potential for precise distribution throughout the bronchial tree, offering targeted treatment for respiratory diseases. This article conducts a comprehensive review of NP applications in respiratory medicine, highlighting their merits, ranging from heightened stability to exacting lung-specific delivery. It also explores cutting-edge technologies optimizing NP-loaded aerosol systems, complemented by insights gleaned from clinical trials. Furthermore, the review examines the current challenges and future prospects in NP-based therapies. By synthesizing current data and perspectives, the article underscores the transformative promise of NP-mediated drug delivery in addressing chronic conditions such as chronic obstructive pulmonary disease, a pressing global health concern ranked third in mortality rates. This overview illuminates the evolving landscape of NP inhalation therapies, presenting optimistic avenues for advancing respiratory medicine and improving patient outcomes.
Collapse
Affiliation(s)
- Elena Cojocaru
- Morpho-Functional Sciences II Department, Faculty of Medicine, “Grigore T. Popa” University of Medicine and Pharmacy, 700115 Iasi, Romania;
| | - Ovidiu Rusalim Petriș
- Medical II Department, Faculty of Medicine, “Grigore T. Popa” University of Medicine and Pharmacy, 700115 Iasi, Romania
| | - Cristian Cojocaru
- Medical III Department, Faculty of Medicine, “Grigore T. Popa” University of Medicine and Pharmacy, 700115 Iasi, Romania;
| |
Collapse
|
|
13
|
Dubrovskii AV, Kim AL, Tikhonenko SA. The Buffer Capacity of Polyelectrolyte Microcapsules Depends on the Type of Template. Polymers (Basel) 2024; 16:2261. [PMID: 39204481 PMCID: PMC11359720 DOI: 10.3390/polym16162261] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/04/2024] [Revised: 08/02/2024] [Accepted: 08/08/2024] [Indexed: 09/04/2024] Open
Abstract
One of the key physicochemical parameters of polyelectrolyte microcapsules (PMCs) is their buffer capacity (BC). The BC of the microcapsules allows for an assessment of the change in protonation state across the entire polyelectrolyte system, which directly impacts the buffer barrier of PMCs, as well as the stability and physical properties of their shell. However, the buffer capacity of PMCs and their behavior under changes in ionic strength and temperature can differ depending on the type of core used to form the microcapsules. As part of this study, we revealed the buffer capacity (BC) of polyelectrolyte microcapsules formed on polystyrene cores (PMCPs) and studied the influence of ionic strength and environmental temperature on the BC of these capsules. We found that the buffer capacity of PMCPs differs from the BC of water at a pH above 8; the addition of sodium chloride leads to an increase in buffer capacity in alkaline conditions, and conversely, thermal treatment leads to its decrease at a pH of 9. The results obtained are different from the BC of polyelectrolyte microcapsules formed on CaCO3 cores, which suggests a difference in the physicochemical properties of these types of capsules. The buffer capacity of polyelectrolyte microcapsules depends on the type of template used.
Collapse
Affiliation(s)
| | | | - Sergey A. Tikhonenko
- Institute of Theoretical and Experimental Biophysics Russian Academy of Science, 3, Institutskaya Str., 142290 Puschino, Moscow Region, Russia; (A.V.D.); (A.L.K.)
| |
Collapse
|
|
14
|
Gu XR, Tai YF, Liu Z, Zhang XY, Liu K, Zhou LY, Yin WJ, Deng YX, Kong DL, Midgley AC, Zuo XC. Layer-by-Layer Assembly of Renal-Targeted Polymeric Nanoparticles for Robust Arginase-2 Knockdown and Contrast-Induced Acute Kidney Injury Prevention. Adv Healthc Mater 2024; 13:e2304675. [PMID: 38688026 DOI: 10.1002/adhm.202304675] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/31/2023] [Revised: 04/27/2024] [Indexed: 05/02/2024]
Abstract
The mitochondrial enzyme arginase-2 (Arg-2) is implicated in the pathophysiology of contrast-induced acute kidney injury (CI-AKI). Therefore, Arg-2 represents a candid target for CI-AKI prevention. Here, layer-by-layer (LbL) assembled renal-targeting polymeric nanoparticles are developed to efficiently deliver small interfering RNA (siRNA), knockdown Arg-2 expression in renal tubules, and prevention of CI-AKI is evaluated. First, near-infrared dye-loaded poly(lactic-co-glycolic acid) (PLGA) anionic cores are electrostatically coated with cationic chitosan (CS) to facilitate the adsorption and stabilization of Arg-2 siRNA. Next, nanoparticles are coated with anionic hyaluronan (HA) to provide protection against siRNA leakage and shielding against early clearance. Sequential electrostatic layering of CS and HA improves loading capacity of Arg-2 siRNA and yields LbL-assembled nanoparticles. Renal targeting and accumulation is enhanced by modifying the outermost layer of HA with a kidney targeting peptide (HA-KTP). The resultant kidney-targeting and siRNA loaded nanoparticles (PLGA/CS/HA-KTP siRNA) exhibit proprietary accumulation in kidneys and proximal tubular cells at 24 h post-tail vein injection. In iohexol-induced in vitro and in vivo CI-AKI models, PLGA/CS/HA-KTP siRNA delivery alleviates oxidative and nitrification stress, and rescues mitochondrial dysfunction while reducing apoptosis, thereby demonstrating a robust and satisfactory therapeutic effect. Thus, PLGA/CS/HA-KTP siRNA nanoparticles offer a promising candidate therapy to protect against CI-AKI.
Collapse
Affiliation(s)
- Xu-Rui Gu
- Department of Pharmacy, Third Xiangya Hospital, Central South University, Changsha, Hunan, 410013, China
| | - Yi-Fan Tai
- Key Laboratory of Bioactive Materials for the Ministry of Education and State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, China
| | - Zhen Liu
- Key Laboratory of Bioactive Materials for the Ministry of Education and State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, China
| | - Xin-Yan Zhang
- Key Laboratory of Bioactive Materials for the Ministry of Education and State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, China
| | - Kun Liu
- Department of Pharmacy, Third Xiangya Hospital, Central South University, Changsha, Hunan, 410013, China
| | - Ling-Yun Zhou
- Department of Pharmacy, Third Xiangya Hospital, Central South University, Changsha, Hunan, 410013, China
| | - Wen-Jun Yin
- Department of Pharmacy, Third Xiangya Hospital, Central South University, Changsha, Hunan, 410013, China
| | - Yi-Xuan Deng
- Department of Pharmacy, Third Xiangya Hospital, Central South University, Changsha, Hunan, 410013, China
| | - De-Ling Kong
- Key Laboratory of Bioactive Materials for the Ministry of Education and State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, China
| | - Adam C Midgley
- Key Laboratory of Bioactive Materials for the Ministry of Education and State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin, 300071, China
| | - Xiao-Cong Zuo
- Department of Pharmacy, Third Xiangya Hospital, Central South University, Changsha, Hunan, 410013, China
| |
Collapse
|
|
15
|
Li J, Tian Y, Qin Q, Ding Z, Zhao X, Tan W. Near-Infrared Light-Triggered NO Nanogenerator for Gas-Enhanced Photodynamic Therapy and Low-Temperature Photothermal Therapy to Eliminate Biofilms. Int J Nanomedicine 2024; 19:5763-5780. [PMID: 38882537 PMCID: PMC11180465 DOI: 10.2147/ijn.s454762] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2023] [Accepted: 05/25/2024] [Indexed: 06/18/2024] Open
Abstract
Purpose Owing to its noninvasive nature, broad-spectrum effectiveness, minimal bacterial resistance, and high efficiency, phototherapy has significant potential for antibiotic-free antibacterial interventions and combating antibacterial biofilms. However, finding effective strategies to mitigate the detrimental effects of excessive temperature and elevated concentrations of reactive oxygen species (ROS) remains a pressing issue that requires immediate attention. Methods In this study, we designed a pH-responsive cationic polymer sodium nitroside dihydrate/branched polyethylenimine-indocyanine green@polyethylene glycol (SNP/PEI-ICG@PEG) nanoplatform using the electrostatic adsorption method and Schiff's base reaction. Relevant testing techniques were applied to characterize and analyze SNP/PEI-ICG@PEG, proving the successful synthesis of the nanomaterials. In vivo and in vitro experiments were performed to evaluate the antimicrobial properties of SNP/PEI-ICG@PEG. Results The morphology and particle size of SNP/PEI-ICG@PEG were observed via TEM. The zeta potential and UV-visible (UV-vis) results indicated the synthesis of the nanomaterials. The negligible cytotoxicity of up to 1 mg/mL of SNP/PEI-ICG@PEG in the presence or absence of light demonstrated its biosafety. Systematic in vivo and in vitro antimicrobial assays confirmed that SNP/PEI-ICG@PEG had good water solubility and biosafety and could be activated by near-infrared (NIR) light and synergistically treated using four therapeutic modes, photodynamic therapy (PDT), gaseous therapy (GT), mild photothermal therapy (PTT, 46 °C), and cation. Ultimately, the development of Gram-positive (G+) Staphylococcus aureus (S. aureus) and Gram-negative (G-) Escherichia coli (E. coli) were both completely killed in the free state, and the biofilm that had formed was eliminated. Conclusion SNP/PEI-ICG@PEG demonstrated remarkable efficacy in achieving controlled multimodal synergistic antibacterial activity and biofilm infection treatment. The nanoplatform thus holds promise for future clinical applications.
Collapse
Affiliation(s)
- Junjuan Li
- Department of Respiratory Medicine, The First Affiliated Hospital of Weifang Medical University/Weifang People's Hospital, Weifang, 261000, People's Republic of China
| | - Yue Tian
- School of Clinical Medicine, Weifang Medical University, Weifang, 261053, People's Republic of China
| | - Qi Qin
- School of Clinical Medicine, Weifang Medical University, Weifang, 261053, People's Republic of China
| | - Zhaolei Ding
- Department of Respiratory Medicine, The First Affiliated Hospital of Weifang Medical University/Weifang People's Hospital, Weifang, 261000, People's Republic of China
| | - Xue Zhao
- Department of Laboratory Medicine, The First Affiliated Hospital of Weifang Medical University/Weifang People's Hospital, Weifang, 261000, People's Republic of China
| | - Wei Tan
- Department of Respiratory Medicine, The First Affiliated Hospital of Weifang Medical University/Weifang People's Hospital, Weifang, 261000, People's Republic of China
| |
Collapse
|
|
16
|
Genedy HH, Humbert P, Laoulaou B, Le Moal B, Fusellier M, Passirani C, Le Visage C, Guicheux J, Lepeltier É, Clouet J. MicroRNA-targeting nanomedicines for the treatment of intervertebral disc degeneration. Adv Drug Deliv Rev 2024; 207:115214. [PMID: 38395361 DOI: 10.1016/j.addr.2024.115214] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2023] [Revised: 02/09/2024] [Accepted: 02/14/2024] [Indexed: 02/25/2024]
Abstract
Low back pain stands as a pervasive global health concern, afflicting almost 80% of adults at some point in their lives with nearly 40% attributable to intervertebral disc degeneration (IVDD). As only symptomatic relief can be offered to patients there is a dire need for innovative treatments.Given the accumulating evidence that multiple microRNAs (miRs) are dysregulated during IVDD, they could have a huge potential against this debilitating condition. The way miRs can profoundly modulate signaling pathways and influence several cellular processes at once is particularly exciting to tackle this multifaceted disorder. However, miR delivery encounters extracellular and intracellular biological barriers. A promising technology to address this challenge is the vectorization of miRs within nanoparticles, providing both protection and enhancing their uptake within the scarce target cells of the degenerated IVD. This comprehensive review presents the diverse spectrum of miRs' connection with IVDD and demonstrates their therapeutic potential when vectorized in nanomedicines.
Collapse
Affiliation(s)
- Hussein H Genedy
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France; Univ Angers, INSERM, CNRS, MINT, SFR ICAT, F-49000 Angers, France
| | - Paul Humbert
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France
| | - Bilel Laoulaou
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France; Univ Angers, INSERM, CNRS, MINT, SFR ICAT, F-49000 Angers, France
| | - Brian Le Moal
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France; Univ Angers, INSERM, CNRS, MINT, SFR ICAT, F-49000 Angers, France
| | - Marion Fusellier
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France; Department of Diagnostic Imaging, CRIP, ONIRIS, College of Veterinary Medicine, Food Science and Engineering, Nantes F-44307, France
| | | | - Catherine Le Visage
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France
| | - Jérôme Guicheux
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France
| | - Élise Lepeltier
- Univ Angers, INSERM, CNRS, MINT, SFR ICAT, F-49000 Angers, France; Institut Universitaire de France (IUF), France.
| | - Johann Clouet
- Nantes Université, Oniris, CHU Nantes, INSERM, Regenerative Medicine and Skeleton, RMeS, UMR1229, Nantes, France
| |
Collapse
|
|
17
|
Wu X, Yuan R, Xu Y, Wang K, Yuan H, Meng T, Hu F. Functionalized lipid nanoparticles modulate the blood-brain barrier and eliminate α-synuclein to repair dopamine neurons. Asian J Pharm Sci 2024; 19:100904. [PMID: 38601010 PMCID: PMC11004078 DOI: 10.1016/j.ajps.2024.100904] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/09/2023] [Revised: 01/16/2024] [Accepted: 02/17/2024] [Indexed: 04/12/2024] Open
Abstract
The challenge in the clinical treatment of Parkinson's disease lies in the lack of disease-modifying therapies that can halt or slow down the progression. Peptide drugs, such as exenatide (Exe), with potential disease-modifying efficacy, have difficulty in crossing the blood-brain barrier (BBB) due to their large molecular weight. Herein, we fabricate multi-functionalized lipid nanoparticles (LNP) Lpc-BoSA/CSO with BBB targeting, permeability-increasing and responsive release functions. Borneol is chemically bonded with stearic acid and, as one of the components of Lpc-BoSA/CSO, is used to increase BBB permeability. Immunofluorescence results of brain tissue of 15-month-old C57BL/6 mice show that Lpc-BoSA/CSO disperses across the BBB into brain parenchyma, and the amount is 4.21 times greater than that of conventional LNP. Motor symptoms of mice in Lpc-BoSA/CSO-Exe group are significantly improved, and the content of dopamine is 1.85 times (substantia nigra compacta) and 1.49 times (striatum) that of PD mice. α-Synuclein expression and Lewy bodies deposition are reduced to 51.85% and 44.72% of PD mice, respectively. Immunohistochemical mechanism studies show AKT expression in Lpc-BoSA/CSO-Exe is 4.23 times that of PD mice and GSK-3β expression is reduced to 18.41%. Lpc-BoSA/CSO-Exe could reduce the production of α-synuclein and Lewy bodies through AKT/GSK-3β pathway, and effectively prevent the progressive deterioration of Parkinson's disease. In summary, Lpc-BoSA/CSO-Exe increases the entry of exenatide into brain and promotes its clinical application for Parkinson's disease therapy.
Collapse
Affiliation(s)
- Xiaomei Wu
- Department of Pharmacy, Ruijin Hospital, School of Medicine, Shanghai Jiaotong University, Shanghai 200000, China
| | - Renxiang Yuan
- College of Pharmaceutical Science, Zhejiang University, Hangzhou 310058, China
| | - Yichong Xu
- College of Pharmaceutical Science, Zhejiang University, Hangzhou 310058, China
| | - Kai Wang
- College of Pharmaceutical Science, Zhejiang University, Hangzhou 310058, China
| | - Hong Yuan
- College of Pharmaceutical Science, Zhejiang University, Hangzhou 310058, China
- Jinhua Institute of Zhejiang University, Jinhua 321299, China
| | - Tingting Meng
- College of Pharmaceutical Science, Zhejiang University, Hangzhou 310058, China
- Jinhua Institute of Zhejiang University, Jinhua 321299, China
| | - Fuqiang Hu
- College of Pharmaceutical Science, Zhejiang University, Hangzhou 310058, China
- Jinhua Institute of Zhejiang University, Jinhua 321299, China
| |
Collapse
|
|
18
|
Gholap AD, Kapare HS, Pagar S, Kamandar P, Bhowmik D, Vishwakarma N, Raikwar S, Garkal A, Mehta TA, Rojekar S, Hatvate N, Mohanto S. Exploring modified chitosan-based gene delivery technologies for therapeutic advancements. Int J Biol Macromol 2024; 260:129581. [PMID: 38266848 DOI: 10.1016/j.ijbiomac.2024.129581] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/09/2023] [Revised: 12/26/2023] [Accepted: 01/06/2024] [Indexed: 01/26/2024]
Abstract
One of the critical steps in gene therapy is the successful delivery of the genes. Immunogenicity and toxicity are major issues for viral gene delivery systems. Thus, non-viral vectors are explored. A cationic polysaccharide like chitosan could be used as a nonviral gene delivery vector owing to its significant interaction with negatively charged nucleic acid and biomembrane, providing effective cellular uptake. However, the native chitosan has issues of targetability, unpacking ability, and solubility along with poor buffer capability, hence requiring modifications for effective use in gene delivery. Modified chitosan has shown that the "proton sponge effect" involved in buffering the endosomal pH results in osmotic swelling owing to the accumulation of a greater amount of proton and chloride along with water. The major challenges include limited exploration of chitosan as a gene carrier, the availability of high-purity chitosan for toxicity reduction, and its immunogenicity. The genetic drugs are in their infancy phase and require further exploration for effective delivery of nucleic acid molecules as FDA-approved marketed formulations soon.
Collapse
Affiliation(s)
- Amol D Gholap
- Department of Pharmaceutics, St. John Institute of Pharmacy and Research, Palghar 401404, Maharashtra, India
| | - Harshad S Kapare
- Department of Pharmaceutics, Dr. D. Y. Patil Institute of Pharmaceutical Sciences and Research, Pune 411018, Maharashtra, India
| | - Sakshi Pagar
- Department of Pharmaceutical Sciences and Technology, Institute of Chemical Technology, Mumbai 400019, India
| | - Pallavi Kamandar
- Institute of Chemical Technology, Mumbai, Marathwada Campus, Jalna 431203, India
| | - Deblina Bhowmik
- Institute of Chemical Technology, Mumbai, Marathwada Campus, Jalna 431203, India
| | - Nikhar Vishwakarma
- Department of Pharmacy, Gyan Ganga Institute of Technology and Sciences, Jabalpur 482003, Madhya Pradesh, India
| | - Sarjana Raikwar
- Department of Pharmaceutical Sciences, Dr. Harisingh Gour Central University, Sagar 470003, Madhya Pradesh, India
| | - Atul Garkal
- Department of Pharmaceutics, Institute of Pharmacy, Nirma University, Ahmedabad 382481, Gujrat, India
| | - Tejal A Mehta
- Department of Pharmaceutics, Institute of Pharmacy, Nirma University, Ahmedabad 382481, Gujrat, India
| | - Satish Rojekar
- Department of Pharmacological Sciences, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
| | - Navnath Hatvate
- Institute of Chemical Technology, Mumbai, Marathwada Campus, Jalna 431203, India.
| | - Sourav Mohanto
- Department of Pharmaceutics, Yenepoya Pharmacy College & Research Centre, Yenepoya (Deemed to be University), Mangaluru, Karnataka 575018, India
| |
Collapse
|
|
19
|
Li B, Ashrafizadeh M, Jiao T. Biomedical application of metal-organic frameworks (MOFs) in cancer therapy: Stimuli-responsive and biomimetic nanocomposites in targeted delivery, phototherapy and diagnosis. Int J Biol Macromol 2024; 260:129391. [PMID: 38242413 DOI: 10.1016/j.ijbiomac.2024.129391] [Citation(s) in RCA: 29] [Impact Index Per Article: 29.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/09/2023] [Revised: 12/12/2023] [Accepted: 01/08/2024] [Indexed: 01/21/2024]
Abstract
The nanotechnology is an interdisciplinary field that has become a hot topic in cancer therapy. Metal-organic frameworks (MOFs) are porous materials and hybrid composites consisted of organic linkers and metal cations. Despite the wide application of MOFs in other fields, the potential of MOFs for purpose of cancer therapy has been revealed by the recent studies. High surface area and porosity, significant drug loading and encapsulation efficiency are among the benefits of using MOFs in drug delivery. MOFs can deliver genes/drugs with selective targeting of tumor cells that can be achieved through functionalization with ligands. The photosensitizers and photo-responsive nanostructures including carbon dots and gold nanoparticles can be loaded in/on MOFs to cause phototherapy-mediated tumor ablation. The immunogenic cell death induction and increased infiltration of cytotoxic CD8+ and CD4+ T cells can be accelerated by MOF platforms in providing immunotherapy of tumor cells. The stimuli-responsive MOF platforms responsive to pH, redox, enzyme and ion can accelerate release of therapeutics in tumor site. Moreover, MOF nanocomposites can be modified ligands and green polymers to improve their selectivity and biocompatibility for cancer therapy. The application of MOFs for the detection of cancer-related biomarkers can participate in the early diagnosis of patients.
Collapse
Affiliation(s)
- Beixu Li
- School of Policing Studies, Shanghai University of Political Science and Law, Shanghai 201701, China; Shanghai Fenglin Forensic Center, Shanghai 200231, China; State Key Laboratory of Medical Neurobiology, School of Basic Medical Sciences, Fudan University, Shanghai 200032, China; Department of Pathology, University of Maryland, Baltimore, MD 21201, USA
| | - Milad Ashrafizadeh
- Department of General Surgery, Institute of Precision Diagnosis and Treatment of Digestive System Tumors, Carson International Cancer Center, Shenzhen University General Hospital, Shenzhen University, Shenzhen, Guangdong 518055, China; International Association for Diagnosis and Treatment of Cancer, Shenzhen, Guangdong 518055, China; Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China; Department of Radiation Oncology, Shandong Cancer Hospital and Institute, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan, Shandong 250000, China.
| | - Taiwei Jiao
- Department of Gastroenterology and Endoscopy, The First Hospital of China Medical University, 155 North Nanjing St, Shenyang 110001, China.
| |
Collapse
|
|
20
|
Huete-Carrasco J, Lynch RI, Ward RW, Lavelle EC. Rational design of polymer-based particulate vaccine adjuvants. Eur J Immunol 2024; 54:e2350512. [PMID: 37994660 DOI: 10.1002/eji.202350512] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/28/2023] [Revised: 11/21/2023] [Accepted: 11/22/2023] [Indexed: 11/24/2023]
Abstract
Vaccination is considered one of the major milestones in modern medicine, facilitating the control and eradication of life-threatening infectious diseases. Vaccine adjuvants are a key component of many vaccines, serving to steer antigen-specific immune responses and increase their magnitude. Despite major advances in the field of adjuvant research over recent decades, our understanding of their mechanism of action remains incomplete. This hinders our capacity to further improve these adjuvant technologies, so addressing how adjuvants induce and control the induction of innate and adaptive immunity is a priority. Investigating how adjuvant physicochemical properties, such as size and charge, exert immunomodulatory effects can provide valuable insights and serve as the foundation for the rational design of vaccine adjuvants. Most clinically applied adjuvants are particulate in nature and polymeric particulate adjuvants present advantages due to stability, biocompatibility profiles, and flexibility in terms of formulation. These properties can impact on antigen release kinetics and biodistribution, cellular uptake and targeting, and drainage to the lymphatics, consequently dictating the induction of innate, cellular, and humoral adaptive immunity. A current focus is to apply rational design principles to the development of adjuvants capable of eliciting robust cellular immune responses including CD8+ cytotoxic T-cell and Th1-biased CD4+ T-cell responses, which are required for vaccines against intracellular pathogens and cancer. This review highlights recent advances in our understanding of how particulate adjuvants, especially polymer-based particulates, modulate immune responses and how this can be used as a guide for improved adjuvant design.
Collapse
Affiliation(s)
- Jorge Huete-Carrasco
- Adjuvant Research Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland
| | - Roisin I Lynch
- Adjuvant Research Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland
- Centre for Research on Adaptive Nanostructures and Nanodevices (CRANN) & Advanced Materials Bio-Engineering Research Centre (AMBER), Trinity College Dublin, Dublin, Ireland
| | - Ross W Ward
- Adjuvant Research Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland
| | - Ed C Lavelle
- Adjuvant Research Group, School of Biochemistry and Immunology, Trinity Biomedical Sciences Institute, Trinity College Dublin, Dublin, Ireland
- Centre for Research on Adaptive Nanostructures and Nanodevices (CRANN) & Advanced Materials Bio-Engineering Research Centre (AMBER), Trinity College Dublin, Dublin, Ireland
| |
Collapse
|
|
21
|
Valenti GE, Marengo B, Milanese M, Zuccari G, Brullo C, Domenicotti C, Alfei S. Imidazo-Pyrazole-Loaded Palmitic Acid and Polystyrene-Based Nanoparticles: Synthesis, Characterization and Antiproliferative Activity on Chemo-Resistant Human Neuroblastoma Cells. Int J Mol Sci 2023; 24:15027. [PMID: 37834475 PMCID: PMC10573130 DOI: 10.3390/ijms241915027] [Citation(s) in RCA: 2] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/08/2023] [Revised: 09/25/2023] [Accepted: 09/26/2023] [Indexed: 10/15/2023] Open
Abstract
Neuroblastoma (NB) is a childhood cancer, commonly treated with drugs, such as etoposide (ETO), whose efficacy is limited by the onset of resistance. Here, aiming at identifying new treatments for chemo-resistant NB, the effects of two synthesized imidazo-pyrazoles (IMPs) (4G and 4I) were investigated on ETO-sensitive (HTLA-230) and ETO-resistant (HTLA-ER) NB cells, detecting 4I as the more promising compound, that demonstrated IC50 values lower than those of ETO on HTLA ER. Therefore, to further improve the activity of 4I, we developed 4I-loaded palmitic acid (PA) and polystyrene-based (P5) cationic nanoparticles (P5PA-4I NPs) with high drug loading (21%) and encapsulation efficiency (97%), by a single oil-in-water emulsification technique. Biocompatible PA was adopted as an emulsion stabilizer, while synthesized P5 acted as an encapsulating agent, solubilizer and hydrophilic-lipophilic balance (HLB) improver. Optic microscopy and cytofluorimetric analyses were performed to investigate the micromorphology, size and complexity distributions of P5PA-4I NPs, which were also structurally characterized by chemometric-assisted Fourier transform infrared spectroscopy (FTIR). Potentiometric titrations allowed us to estimate the milliequivalents of PA and basic nitrogen atoms present in NPs. P5PA-4I NPs afforded dispersions in water with excellent buffer capacity, essential to escape lysosomal degradation and promote long residence time inside cells. They were chemically stable in an aqueous medium for at least 40 days, while in dynamic light scattering (DLS) analyses, P5PA-4I showed a mean hydrodynamic diameter of 541 nm, small polydispersity (0.194), and low positive zeta potentials (+8.39 mV), assuring low haemolytic toxicity. Biological experiments on NB cells, demonstrated that P5PA-4I NPs induced ROS-dependent cytotoxic effects significantly higher than those of pristine 4I, showing a major efficacy compared to ETO in reducing cell viability in HTLA-ER cells. Collectively, this 4I-based nano-formulation could represent a new promising macromolecular platform to develop a new delivery system able to increase the cytotoxicity of the anticancer drugs.
Collapse
Affiliation(s)
- Giulia Elda Valenti
- Department of Experimental Medicine (DIMES), University of Genova, Via Alberti L.B., 16132 Genoa, Italy; (G.E.V.); (B.M.)
| | - Barbara Marengo
- Department of Experimental Medicine (DIMES), University of Genova, Via Alberti L.B., 16132 Genoa, Italy; (G.E.V.); (B.M.)
| | - Marco Milanese
- Department of Pharmacy, Section of Chemistry and Pharmaceutical and Food Technologies, University of Genoa, Viale Cembrano, 4, 16148 Genoa, Italy;
| | - Guendalina Zuccari
- Department of Pharmacy (DIFAR), Section of Medicinal Chemistry and Cosmetic Product, University of Genoa, Viale Benedetto XV, 3, 16132 Genoa, Italy; (G.Z.); (C.B.)
| | - Chiara Brullo
- Department of Pharmacy (DIFAR), Section of Medicinal Chemistry and Cosmetic Product, University of Genoa, Viale Benedetto XV, 3, 16132 Genoa, Italy; (G.Z.); (C.B.)
| | - Cinzia Domenicotti
- Department of Experimental Medicine (DIMES), University of Genova, Via Alberti L.B., 16132 Genoa, Italy; (G.E.V.); (B.M.)
| | - Silvana Alfei
- Department of Pharmacy, Section of Chemistry and Pharmaceutical and Food Technologies, University of Genoa, Viale Cembrano, 4, 16148 Genoa, Italy;
| |
Collapse
|
|
22
|
Molinar C, Tannous M, Meloni D, Cavalli R, Scomparin A. Current Status and Trends in Nucleic Acids for Cancer Therapy: A Focus on Polysaccharide-Based Nanomedicines. Macromol Biosci 2023; 23:e2300102. [PMID: 37212473 DOI: 10.1002/mabi.202300102] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/10/2023] [Revised: 04/28/2023] [Indexed: 05/23/2023]
Abstract
The efficacious delivery of therapeutic nucleic acids to cancer still remains an open issue. Through the years, several strategies are developed for the encapsulation of genetic molecules exploiting different materials, such as viral vectors, lipid nanoparticles (LNPs), and polymeric nanoparticles (NPs). Indeed, the rapid approval by regulatory authorities and the wide use of LNPs complexing the mRNA coding for the spark protein for COVID-19 vaccination paved the way for the initiation of several clinical trials exploiting lipid nanoparticles for cancer therapy. Nevertheless, polymers still represent a valuable alternative to lipid-based formulations, due to the low cost and the chemical flexibility that allows for the conjugation of targeting ligands. This review will analyze the status of the ongoing clinical trials for cancer therapy, including vaccination and immunotherapy approaches, exploiting polymeric materials. Among those nanosized carriers, sugar-based backbones are an interesting category. A cyclodextrin-based carrier (CALAA-01) is the first polymeric material to enter a clinical trial complexed with siRNA for cancer therapy, and chitosan is one of the most characterized non-viral vectors able to complex genetic material. Finally, the recent advances in the use of sugar-based polymers (oligo- and polysaccharides) for the complexation of nucleic acids in advanced preclinical stage will be discussed.
Collapse
Affiliation(s)
- Chiara Molinar
- Department of Drug Science and Technology, University of Turin, Via P. Giuria 9, Torino, 10125, Italy
| | - Maria Tannous
- Department of Drug Science and Technology, University of Turin, Via P. Giuria 9, Torino, 10125, Italy
- Department of Chemistry, University of Turin, Via P. Giuria 7, Torino, 10125, Italy
| | - Domitilla Meloni
- Department of Drug Science and Technology, University of Turin, Via P. Giuria 9, Torino, 10125, Italy
| | - Roberta Cavalli
- Department of Drug Science and Technology, University of Turin, Via P. Giuria 9, Torino, 10125, Italy
| | - Anna Scomparin
- Department of Drug Science and Technology, University of Turin, Via P. Giuria 9, Torino, 10125, Italy
| |
Collapse
|
|
23
|
Gothwal A, Lamptey RNL, Trivedi R, Chaulagain B, Singh J. Non-Invasive Intranasal Delivery of pApoE2: Effect of Multiple Dosing on the ApoE2 Expression in Mice Brain. Int J Mol Sci 2023; 24:13019. [PMID: 37629200 PMCID: PMC10456017 DOI: 10.3390/ijms241613019] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/18/2023] [Revised: 08/16/2023] [Accepted: 08/19/2023] [Indexed: 08/27/2023] Open
Abstract
Chitosan-based polymeric micelles are promising non-viral nanocarriers for safe and targeted gene delivery. Multi-functionalized chitosan polymeric micelles were prepared by grafting fatty acid, cell-penetrating peptide, and mannose on the chitosan backbone. The polymeric micelles were subjected to surface morphology and surface topography using scanning electron microscopy and atomic force microscopy, respectively. The hemotoxic profile of the prepared polymeric micelles was established against erythrocytes and was found to be <5% hemotoxic up to the concentration of 600 µg/mL. In vitro ApoE2 expression in primary astrocytes and neurons was analyzed. Multi-functionalized polymeric micelles produced greater (p < 0.05) transfection in astrocytes and neurons in comparison to mono-functionalized micelles. Intranasal administration of polymeric micelles/pApoE2 polyplex led to significantly higher (p < 0.05) in vivo pApoE2 expression than chitosan and unfunctionalized polymeric micelles-treated mice groups. The outcomes of this study predict that the developed multi-functionalized polymeric micelles could be an effective and safe gene delivery platform to the brain through the intranasal route.
Collapse
Affiliation(s)
| | | | | | | | - Jagdish Singh
- Department of Pharmaceutical Sciences, School of Pharmacy, College of Health Professions, North Dakota State University, Fargo, ND 58105, USA; (A.G.); (R.N.L.L.); (R.T.); (B.C.)
| |
Collapse
|
|
24
|
Huang J, Lin G, Juenke T, Chung S, Lai N, Zhang T, Zhang T, Zhang M. Iron Oxide Nanoparticle-Mediated mRNA Delivery to Hard-to-Transfect Cancer Cells. Pharmaceutics 2023; 15:1946. [PMID: 37514132 PMCID: PMC10384052 DOI: 10.3390/pharmaceutics15071946] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/31/2023] [Revised: 07/05/2023] [Accepted: 07/10/2023] [Indexed: 07/30/2023] Open
Abstract
mRNA-based therapeutics have emerged as a promising strategy for cancer treatment. However, the effective delivery of mRNA into hard-to-transfect cancer cells remains a significant challenge. This study introduces a novel approach that utilizes iron oxide nanoparticles (NPs) synthesized through a layer-by-layer (LbL) method for safe and efficient mRNA delivery. The developed NPs consist of an iron oxide core modified with a thin charge-bearing layer, an mRNA middle layer, and an outer layer composed of perfluorinated polyethyleneimine with heparin (PPH), which facilitates efficient mRNA delivery. Through a comparative analysis of four nanoparticle delivery formulations, we investigated the effects of the iron oxide core's surface chemistry and surface charge on mRNA complexation, cellular uptake, and mRNA release. We identified an optimal and effective mRNA delivery platform, namely, (IOCCP)-mRNA-PPH, capable of transporting mRNA into various hard-to-transfect cancer cell lines in vitro. The (IOCCP)-mRNA-PPH formulation demonstrated significant enhancements in cellular internalization of mRNA, facilitated endosomal escape, enabled easy mRNA release, and exhibited minimal cytotoxicity. These findings suggest that (IOCCP)-mRNA-PPH holds great promise as a solution for mRNA therapy against hard-to-transfect cancers.
Collapse
Affiliation(s)
- Jianxi Huang
- Department of Materials Science and Engineering, University of Washington, Seattle, WA 98195, USA
| | - Guanyou Lin
- Department of Materials Science and Engineering, University of Washington, Seattle, WA 98195, USA
| | - Taylor Juenke
- Department of Materials Science and Engineering, University of Washington, Seattle, WA 98195, USA
| | - Seokhwan Chung
- Department of Materials Science and Engineering, University of Washington, Seattle, WA 98195, USA
| | - Nicholas Lai
- Department of Chemical Engineering, University of Washington, Seattle, WA 98195, USA
| | - Tianxin Zhang
- Department of Biology, University of Washington, Seattle, WA 98195, USA
| | - Tianyi Zhang
- Department of Biology, University of Washington, Seattle, WA 98195, USA
| | - Miqin Zhang
- Department of Materials Science and Engineering, University of Washington, Seattle, WA 98195, USA
| |
Collapse
|
|
25
|
Zou Y, Liu X, Chen Q, Oku H, Ma G, Wu J. Acid-Responsive Immune-Enhancing Chitosan Formulation Capable of Transforming from Particle Stabilization to Polymer Chain Stabilization. ACS APPLIED MATERIALS & INTERFACES 2023; 15:11403-11415. [PMID: 36825996 DOI: 10.1021/acsami.2c17505] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/18/2023]
Abstract
Chitosan with pH sensitivity and biocompatibility was selected to prepare chitosan nanoparticle-stabilized Pickering emulsion (CSPE). The flexibility of CSPE enables stress deformation when in contact with cell membranes, thereby mimicking the deformability of natural pathogens and facilitating their efficient uptake by cells. In the acidic environment of lysosomes, the amino groups of chitosan molecules are protonated, and the water solubility increases. CSPE transforms from particle-stabilized to polymer chain-stabilized, its subsequent swelling and proton accumulation lead to lysosome rupture. The experimental results evaluating CSPE as an adjuvant shows that CSPE could efficiently load antigens, promote endocytosis and antigen cross-presentation, recruit antigen-presenting cells at the injection site, boost T-cell activation, and enhance both humoral and cellular immune responses. In the prophylactic and therapeutic tumor models of E.G7-OVA lymphoma and B16-MUC1 melanoma, CSPE significantly inhibited tumor growth and prolonged the survival of mice. In summary, antigenic lysosomal escape resulted from the chitosan molecular state transition is the key to the enhancement of cellular immunity by CSPE, and CSPE is a promising vaccine adjuvant.
Collapse
Affiliation(s)
- Yongjuan Zou
- State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, P. R. China
| | - Xiaoxuan Liu
- State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, P. R. China
- Division of Molecular Science, Faculty of Science and Technology, Gunma University, Ota, Gunma 373-0057, Japan
| | - Qiuting Chen
- State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, P. R. China
- School of Biomedical Sciences and Engineering, South China University of Technology, Guangzhou 510006, P. R. China
| | - Hiroyuki Oku
- Division of Molecular Science, Faculty of Science and Technology, Gunma University, Ota, Gunma 373-0057, Japan
| | - Guanghui Ma
- State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, P. R. China
| | - Jie Wu
- State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, P. R. China
| |
Collapse
|
|
26
|
Parmaksız S, Gül A, Erkunt Alak S, Karakavuk M, Can H, Gül C, Karakavuk T, López-Macías C, Puralı N, Döşkaya M, Şenel S. Development of multistage recombinant protein vaccine formulations against toxoplasmosis using a new chitosan and porin based adjuvant system. Int J Pharm 2022; 626:122199. [PMID: 36115468 DOI: 10.1016/j.ijpharm.2022.122199] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/01/2022] [Revised: 09/07/2022] [Accepted: 09/09/2022] [Indexed: 10/14/2022]
Abstract
Toxoplasmosis is a global health problem affecting both human and animal populations. The lack of effective treatment makes the development of a vaccine against toxoplasmosis one of the main goals in the management of this disease. In our study, vaccine formulations containing the multistage recombinant antigens, rBAG1 + rGRA1 were developed with a combined adjuvant system consisting of chitosan and Salmonella Typhi porins in micro (MicroAS) and nanoparticulate (NanoAS) forms. BALB/c mice were immunized intraperitoneally with vaccine formulations two times at three-week intervals. Three weeks after the second vaccination, mice were challenged with 7-8 live tissue cysts of the virulent T. gondii PRU strain by oral gavage. Higher cellular uptake by macrophages and enhanced cellular (IFN-γ and I-4 in stimulated spleen cells) and humoral (IgG, IgG1, IgG2a) responses were obtained with the adjuvanted formulation, higher with microsystem when compared to that of nanosystem. Microsystem was found to stimulate Th1-polarized immune responses, whereasnon-adjuvanted antigens stimulated Th2-polarized immune response. The highest survival rate and reduction in cysts numbers and T. gondii DNA were obtained with the adjuvanted antigens.Our study showed that adjuvanted multistage recombinant vaccine systems increase theimmune response with strong protection againstT. gondii, more profoundly in microparticulate form.
Collapse
Affiliation(s)
- Selin Parmaksız
- Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical Technology, Ankara 06100, Turkey
| | - Aytül Gül
- Ege University, Faculty of Engineering, Department of Bioengineering, Bornova, Izmir 35040, Turkey; Ege University Vaccine Development, Application and Research Center, Izmir 35100, Turkey
| | - Sedef Erkunt Alak
- Ege University Vaccine Development, Application and Research Center, Izmir 35100, Turkey; Ege University, Faculty of Science, Department of Biology, Molecular Biology Section, Bornova, Izmir 35040, Turkey
| | - Muhammet Karakavuk
- Ege University Vaccine Development, Application and Research Center, Izmir 35100, Turkey; Ege University, Vocational School, Odemis, Izmir 35750, Turkey
| | - Hüseyin Can
- Ege University Vaccine Development, Application and Research Center, Izmir 35100, Turkey; Ege University, Faculty of Science, Department of Biology, Molecular Biology Section, Bornova, Izmir 35040, Turkey
| | - Ceren Gül
- Ege University Vaccine Development, Application and Research Center, Izmir 35100, Turkey; Ege University Institute of Science, Department of Biotechnology, Bornova, Izmir 35040, Turkey
| | - Tuğba Karakavuk
- Ege University Vaccine Development, Application and Research Center, Izmir 35100, Turkey; Ege University Institute of Science, Department of Biotechnology, Bornova, Izmir 35040, Turkey
| | - Constantino López-Macías
- Medical Research Unit on Immunochemistry, Specialties Hospital of the National Medical Centre ''Siglo XXI'', Mexican Institute for Social Security, Mexico City, Mexico
| | - Nuhan Puralı
- Hacettepe University, Faculty of Medicine, Department of Biophysics,06100 Ankara, Turkey
| | - Mert Döşkaya
- Ege University Vaccine Development, Application and Research Center, Izmir 35100, Turkey; Ege University Faculty of Medicine, Department of Parasitology, Bornova, Izmir 35100, Turkey
| | - Sevda Şenel
- Hacettepe University, Faculty of Pharmacy, Department of Pharmaceutical Technology, Ankara 06100, Turkey.
| |
Collapse
|
|
27
|
Kubiatowicz LJ, Mohapatra A, Krishnan N, Fang RH, Zhang L. mRNA nanomedicine: Design and recent applications. EXPLORATION (BEIJING, CHINA) 2022; 2:20210217. [PMID: 36249890 PMCID: PMC9539018 DOI: 10.1002/exp.20210217] [Citation(s) in RCA: 65] [Impact Index Per Article: 21.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Subscribe] [Scholar Register] [Received: 04/17/2022] [Accepted: 08/11/2022] [Indexed: 11/06/2022]
Abstract
The rational design and application of mRNA-based medicine have recently yielded some key successes in the clinical management of human diseases. mRNA technology allows for the facile and direct production of proteins in vivo, thus circumventing the need for lengthy drug development cycles and complex production workflows. As such, mRNA formulations can significantly improve upon the biological therapies that have become commonplace in modern medicine. Despite its many advantages, mRNA is inherently fragile and has specific delivery requirements. Leveraging the engineering flexibility of nanobiotechnology, mRNA payloads can be incorporated into nanoformulations such that they do not invoke unwanted immune responses, are targeted to tissues of interest, and can be delivered to the cytosol, resulting in improved safety while enhancing bioactivity. With the rapidly evolving landscape of nanomedicine, novel technologies that are under development have the potential to further improve the clinical utility of mRNA medicine. This review covers the design principles relevant to engineering mRNA-based nanomedicine platforms. It also details the current research on mRNA nanoformulations for addressing viral infections, cancers, and genetic diseases. Given the trends in the field, future mRNA-based nanomedicines have the potential to change how many types of diseases are managed in the clinic.
Collapse
Affiliation(s)
- Luke J. Kubiatowicz
- Department of NanoEngineering, Chemical Engineering Program, and Moores Cancer CenterUniversity of California San DiegoLa JollaCaliforniaUSA
| | - Animesh Mohapatra
- Department of NanoEngineering, Chemical Engineering Program, and Moores Cancer CenterUniversity of California San DiegoLa JollaCaliforniaUSA
| | - Nishta Krishnan
- Department of NanoEngineering, Chemical Engineering Program, and Moores Cancer CenterUniversity of California San DiegoLa JollaCaliforniaUSA
| | - Ronnie H. Fang
- Department of NanoEngineering, Chemical Engineering Program, and Moores Cancer CenterUniversity of California San DiegoLa JollaCaliforniaUSA
| | - Liangfang Zhang
- Department of NanoEngineering, Chemical Engineering Program, and Moores Cancer CenterUniversity of California San DiegoLa JollaCaliforniaUSA
| |
Collapse
|
|
28
|
Musin EV, Dubrovskii AV, Kim AL, Tikhonenko SA. A Study of the Buffer Capacity of Polyelectrolyte Microcapsules Depending on Their Concentration and the Number of Layers of the Polyelectrolyte Shell. Int J Mol Sci 2022; 23:ijms23179917. [PMID: 36077315 PMCID: PMC9456183 DOI: 10.3390/ijms23179917] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/29/2022] [Revised: 08/12/2022] [Accepted: 08/29/2022] [Indexed: 11/16/2022] Open
Abstract
Polyelectrolyte microcapsules are used in the development of new forms of targeted delivery systems, self-healing materials, sensors, and smart materials. Nevertheless, their buffer capacity has not been practically studied, although that characteristic makes it possible to estimate the change in the state of protonation of the entire polyelectrolyte system. This is necessary both for creating a buffer barrier system for pH-sensitive compounds (metals, enzymes, polyelectrolytes, drugs) and for the correct interpretation of the results of research and studying of the PMC structure. The buffer capacity of a PMC can be affected by the concentration of microcapsules in solution and the number of shell layers since the listed parameters affect other physicochemical properties of the PMC shell. This includes, for example, the electrical conductivity, permeability (of ions), osmotic pressure, charge density, etc. In this regard, we studied the change in the buffer capacity of polyelectrolyte microcapsules depending on their concentration and the number of shell layers. As a result, it was found that with an increasing concentration of microcapsules, the buffering capacity of the PMC increases, but at the same time, in the pH range from 4 to 5.5, the calculated buffering capacity of 1 billion capsules decreases with increasing their concentration. This effect may be associated with a decrease in the available -NH2 groups of the PMC’s shell. In addition, it was found that the main contribution to the buffer capacity of a PMC is made by the entire shell of the microcapsule and not just its surface. At the same time, the buffer capacity of the capsules has non-linear growth with an increase in the number of PMC shell layers. It is presumably associated either with a decrease in the polyelectrolyte layer with an increase in their number or with a decrease in the permeability of hydrogen protons.
Collapse
|
|
29
|
Hausig-Punke F, Richter F, Hoernke M, Brendel JC, Traeger A. Tracking the Endosomal Escape: A Closer Look at Calcein and Related Reporters. Macromol Biosci 2022; 22:e2200167. [PMID: 35933579 DOI: 10.1002/mabi.202200167] [Citation(s) in RCA: 16] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/27/2022] [Revised: 07/19/2022] [Indexed: 11/11/2022]
Abstract
Crossing the cellular membrane and delivering active pharmaceuticals or biologicals into the cytosol of cells is an essential step in the development of nanomedicines. One of the most important intracellular processes regarding the cellular uptake of biologicals is the endolysosomal pathway. Sophisticated nanocarriers have been developed overcoming a major hurdle, the endosomal entrapment, and delivering their cargo to the required site of action. In parallel, in vitro assays have been established analyzing the performance of these nanocarriers. Among them, the release of the membrane-impermeable dye calcein has become a popular and straightforward method. It is accessible for most researchers worldwide, allows for rapid conclusions about the release potential, and enables the study of release mechanisms. This review is intended to provide an overview and guidance for scientists applying the calcein release assay. It comprises a survey of several applications in the study of endosomal escape, considerations of potential pitfalls, challenges and limitations of the assay, and a brief summary of complementary methods. Based on this review, we hope to encourage further research groups to take advantage of the calcein release assay for their own purposes and help to create a database for more efficient cross-correlations between nanocarriers. This article is protected by copyright. All rights reserved.
Collapse
Affiliation(s)
- Franziska Hausig-Punke
- Laboratory of Organic and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Humboldtstrasse 10, 07743, Jena, Germany.,Jena Center for Soft Matter (JCSM), Friedrich Schiller University Jena, Philosophenweg 7, 07743, Jena, Germany
| | - Friederike Richter
- Laboratory of Organic and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Humboldtstrasse 10, 07743, Jena, Germany.,Jena Center for Soft Matter (JCSM), Friedrich Schiller University Jena, Philosophenweg 7, 07743, Jena, Germany
| | - Maria Hoernke
- Chemistry and Pharmacy, Albert-Ludwigs-Universität Freiburg, Hermann-Herder-Str. 9, 79104, Freiburg i.Br., Germany
| | - Johannes C Brendel
- Laboratory of Organic and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Humboldtstrasse 10, 07743, Jena, Germany.,Jena Center for Soft Matter (JCSM), Friedrich Schiller University Jena, Philosophenweg 7, 07743, Jena, Germany
| | - Anja Traeger
- Laboratory of Organic and Macromolecular Chemistry (IOMC), Friedrich Schiller University Jena, Humboldtstrasse 10, 07743, Jena, Germany.,Jena Center for Soft Matter (JCSM), Friedrich Schiller University Jena, Philosophenweg 7, 07743, Jena, Germany
| |
Collapse
|
|
30
|
Yang E, Jung HS, Chang PS. Preparation and Characterization of pH-Sensitive Capsosomes for Oral Delivery of Therapeutic Proteins. LANGMUIR : THE ACS JOURNAL OF SURFACES AND COLLOIDS 2022; 38:9294-9300. [PMID: 35863074 DOI: 10.1021/acs.langmuir.2c01089] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/15/2023]
Abstract
Oral administration of therapeutic proteins is very challenging because of gastrointestinal instability and decomposition. In this study, we developed a system for oral delivery of superoxide dismutase (SOD) as one of the therapeutic proteins. SOD-loaded capsosomes (SOD-C) were formed by the assembly of chitosan-coated solid lipid nanoparticles and SOD-loaded liposomes (SOD-L). Unlike raw SOD activity decreases to 19.41% in SGF and 13.70% in SIF, the SOD-C in SGF (89.30%) condition retained its initial catalytic activity and decreased but exhibited a three-fold higher raw SOD activity even after incubation in SIF (41.63%). TEM analysis indicated that after intestinal digestion, the residual amount of intact liposomes affected the higher catalytic activity of SOD-C compared to raw SOD and SOD-L. Based on these results, significantly higher cellular uptake of SOD-C was observed compared to raw SOD. Also, SOD-C remarkably suppressed the cellular malondialdehyde (MDA) concentration by maintaining the antioxidative capacity of SOD to remove MDA produced in the oxidative stress-induced cells, thereby contributing to a significant five-fold difference with SOD-R (p < 0.05). This delivery system can facilitate the oral application of other therapeutic proteins, improving gastrointestinal stability.
Collapse
Affiliation(s)
- Eunhye Yang
- Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea
| | - Ho-Sup Jung
- Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea
| | - Pahn-Shick Chang
- Department of Agricultural Biotechnology, Seoul National University, Seoul 08826, Republic of Korea
- Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea
- Center for Agricultural Microorganism and Enzyme, Seoul National University, Seoul 08826, Republic of Korea
- Research Institute of Agriculture and Life Sciences, Seoul National University, Seoul 08826, Republic of Korea
| |
Collapse
|
|
31
|
Fang T, Cao X, Ibnat M, Chen G. Stimuli-responsive nanoformulations for CRISPR-Cas9 genome editing. J Nanobiotechnology 2022; 20:354. [PMID: 35918694 PMCID: PMC9344766 DOI: 10.1186/s12951-022-01570-y] [Citation(s) in RCA: 9] [Impact Index Per Article: 3.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/19/2022] [Accepted: 07/22/2022] [Indexed: 12/07/2022] Open
Abstract
The CRISPR-Cas9 technology has changed the landscape of genome editing and has demonstrated extraordinary potential for treating otherwise incurable diseases. Engineering strategies to enable efficient intracellular delivery of CRISPR-Cas9 components has been a central theme for broadening the impact of the CRISPR-Cas9 technology. Various non-viral delivery systems for CRISPR-Cas9 have been investigated given their favorable safety profiles over viral systems. Many recent efforts have been focused on the development of stimuli-responsive non-viral CRISPR-Cas9 delivery systems, with the goal of achieving efficient and precise genome editing. Stimuli-responsive nanoplatforms are capable of sensing and responding to particular triggers, such as innate biological cues and external stimuli, for controlled CRISPR-Cas9 genome editing. In this Review, we overview the recent advances in stimuli-responsive nanoformulations for CRISPR-Cas9 delivery, highlight the rationale of stimuli and formulation designs, and summarize their biomedical applications.
Collapse
Affiliation(s)
- Tianxu Fang
- Department of Biomedical Engineering, McGill University, Montreal, QC, H3G 0B1, Canada.,Rosalind & Morris Goodman Cancer Institute, McGill University, Montreal, QC, H3G 0B1, Canada
| | - Xiaona Cao
- Department of Biomedical Engineering, McGill University, Montreal, QC, H3G 0B1, Canada.,Rosalind & Morris Goodman Cancer Institute, McGill University, Montreal, QC, H3G 0B1, Canada.,School of Nursing, Tianjin Medical University, Tianjin, China
| | - Mysha Ibnat
- Department of Biomedical Engineering, McGill University, Montreal, QC, H3G 0B1, Canada.,Rosalind & Morris Goodman Cancer Institute, McGill University, Montreal, QC, H3G 0B1, Canada
| | - Guojun Chen
- Department of Biomedical Engineering, McGill University, Montreal, QC, H3G 0B1, Canada. .,Rosalind & Morris Goodman Cancer Institute, McGill University, Montreal, QC, H3G 0B1, Canada.
| |
Collapse
|
|
32
|
Functionalized chitosan as a promising platform for cancer immunotherapy: A review. Carbohydr Polym 2022; 290:119452. [DOI: 10.1016/j.carbpol.2022.119452] [Citation(s) in RCA: 40] [Impact Index Per Article: 13.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/10/2022] [Revised: 03/30/2022] [Accepted: 03/30/2022] [Indexed: 12/20/2022]
|
|
33
|
Zhang J, Liu Y, Liu Y, Liu W, Lu F, Yuan Z, Lu C. Gold Nanocluster-Encapsulated Hyperbranched Polyethyleneimine for Selective and Ratiometric Dopamine Analyses by Enhanced Self-Polymerization. Front Chem 2022; 10:928607. [PMID: 35873048 PMCID: PMC9307107 DOI: 10.3389/fchem.2022.928607] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/26/2022] [Accepted: 05/17/2022] [Indexed: 11/13/2022] Open
Abstract
The exploitation of selective and sensitive dopamine (DA) sensors is essential to more deeply understand its biological function and diagnosis of related diseases. In this study, gold nanocluster-encapsulated hyperbranched polyethyleneimine (hPEI-Au NCs) has been explored as the specific and ratiometric DA nanoprobe through hPEI-assisted DA self-polymerization reactions. The Au NCs encapsulation not only provides a fluorescent internal reference but also enhances the DA self-polymerization by weakening the proton sponge effect of the hPEI layer. Rapid and sensitive DA detection is realized through the proposed hPEI-Au NC nanoprobe with a limit of detection of 10 nM. The favorable selectivity over other possible interferents including amino acids, sugars, and salts is due to the specific self-polymerization reaction. The DA analysis in urine samples with small relative standard deviations has been accomplished with an hPEI-Au NC nanoprobe.
Collapse
Affiliation(s)
- Jing Zhang
- State Key Laboratory of Chemical Resource Engineering, College of Chemistry, Beijing University of Chemical Technology, Beijing, China
| | - Ying Liu
- State Key Laboratory of Chemical Resource Engineering, College of Chemistry, Beijing University of Chemical Technology, Beijing, China
| | - Yang Liu
- State Key Laboratory of Chemical Resource Engineering, College of Chemistry, Beijing University of Chemical Technology, Beijing, China
| | - Wencai Liu
- State Key Laboratory of Chemical Resource Engineering, College of Chemistry, Beijing University of Chemical Technology, Beijing, China
| | - Fengniu Lu
- Department of Chemistry and Chemical Engineering, Beijing Institute of Technology, Beijing, China
| | - Zhiqin Yuan
- State Key Laboratory of Chemical Resource Engineering, College of Chemistry, Beijing University of Chemical Technology, Beijing, China
- Beijing Key Laboratory of Plant Resources Research and Development, Beijing Technology and Business University, Beijing, China
- *Correspondence: Zhiqin Yuan, ; Chao Lu,
| | - Chao Lu
- State Key Laboratory of Chemical Resource Engineering, College of Chemistry, Beijing University of Chemical Technology, Beijing, China
- Green Catalysis Center, College of Chemistry, Zhengzhou University, Zhengzhou, China
- *Correspondence: Zhiqin Yuan, ; Chao Lu,
| |
Collapse
|
|
34
|
Sun L, Nie X, Lu W, Zhang Q, Fang W, Gao S, Chen S, Hu R. Mucus-Penetrating Alginate-Chitosan Nanoparticles Loaded with Berberine Hydrochloride for Oral Delivery to the Inflammation Site of Ulcerative Colitis. AAPS PharmSciTech 2022; 23:179. [PMID: 35761150 DOI: 10.1208/s12249-022-02327-4] [Citation(s) in RCA: 19] [Impact Index Per Article: 6.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/13/2022] [Accepted: 06/07/2022] [Indexed: 01/06/2023] Open
Abstract
The rectal enemas of berberine hydrochloride (BH) have emerged as one of the most effective strategies in the clinical treatment of ulcerative colitis (UC). However, oral dosages of BH exhibit a poor anti-inflammatory effect of UC, which may attribute to premature absorption of BH by the upper gastrointestinal tract. Moreover, the thick colonic mucus layer obstructs the penetration of the drug, resulting in low bioavailability to the inflammatory site of the colon. The aim of this study was to develop the mucus-penetrating sodium alginate-chitosan nanoparticles (SA-CS NPs) for oral delivery of BH to the site of colonic ulcer lesions. BH-loaded SA-CS NPs were developed through the ionic gelation method and analyzed for physicochemical characteristics, release performance, penetrability, site retention, and therapeutic efficacy. The results showed that the NPs have a particle size of 257 nm with a negative charge, presenting desired pH-dependent release behavior. The permeation studies elucidated that negatively charged SA-CS NPs had 2.9 times higher mucus penetration ability than positively charged CS NPs. An ex vivo retention study indicated the high retention of BH-SA-CS NPs at the colon site for more than 16 h. In vivo therapeutic effectiveness demonstrated that the prepared NPs could not only alleviate colonic injury by decreasing the disease activity index and colon mucosa damage index, but also improve the immunologic function by decreasing the spleen index. In conclusion, the BH-SA-CS NPs could enhance the mucus permeability and deliver drugs to the colonic inflammation site, providing new insights into improving the therapeutic effect of UC.
Collapse
Affiliation(s)
- Linfeng Sun
- Anhui Province Key Laboratory of Pharmaceutical Technology and Application, Key Laboratory of Xin'an Medicine Ministry of Education, Anhui Province Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine, Hefei, 230038, Anhui, China
| | - Xiangjiang Nie
- Anhui Province Key Laboratory of Pharmaceutical Technology and Application, Key Laboratory of Xin'an Medicine Ministry of Education, Anhui Province Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine, Hefei, 230038, Anhui, China
| | - Wenjie Lu
- School of Pharmacy, China Pharmaceutical University, Nanjing, 210009, Jiangsu, China
| | - Qing Zhang
- Department of Pharmacy, School of Pharmacy, Nanjing Medical University, Nanjing, 210009, Jiangsu, China
| | - Wenyou Fang
- Anhui Province Key Laboratory of Pharmaceutical Technology and Application, Key Laboratory of Xin'an Medicine Ministry of Education, Anhui Province Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine, Hefei, 230038, Anhui, China
| | - Song Gao
- Anhui Province Key Laboratory of Pharmaceutical Technology and Application, Key Laboratory of Xin'an Medicine Ministry of Education, Anhui Province Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine, Hefei, 230038, Anhui, China.
| | - Shengqi Chen
- Anhui Province Key Laboratory of Pharmaceutical Technology and Application, Key Laboratory of Xin'an Medicine Ministry of Education, Anhui Province Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine, Hefei, 230038, Anhui, China.
| | - Rongfeng Hu
- Anhui Province Key Laboratory of Pharmaceutical Technology and Application, Key Laboratory of Xin'an Medicine Ministry of Education, Anhui Province Key Laboratory of Chinese Medicinal Formula, Anhui University of Chinese Medicine, Hefei, 230038, Anhui, China.
| |
Collapse
|
|
35
|
A Study of the Buffer Capacity of Polyelectrolyte Microcapsules Depending on Their Ionic Environment and Incubation Temperature. Int J Mol Sci 2022; 23:ijms23126608. [PMID: 35743051 PMCID: PMC9223497 DOI: 10.3390/ijms23126608] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/27/2022] [Revised: 06/10/2022] [Accepted: 06/12/2022] [Indexed: 02/06/2023] Open
Abstract
Polyelectrolyte microcapsules (PMCs) are used in the development of new forms of drugs, coatings and diagnostic systems. Their buffer capacity, depending on the conditions of the medium, has not been practically studied, although it can affect the structure of both the capsule itself and the encapsulated agents. In this connection, we studied the buffer capacity of polyelectrolyte microcapsules of the composition (polystyrene sulfonate/polyallylamine)3 ((PSS/PAH)3) depending on the concentration and the type of salt in solution, as well as the microcapsule incubation temperature. It was found that the buffer capacity of microcapsules in the presence of mono- and di-valent salts of the same ionic strength did not differ practically. Increasing the NaCl concentration to 1 M led to an increase of buffer capacity of PMCs at pH ≥ 5, and an increase in NaCl concentration above 1 M did not change buffer capacity. The study of the buffer capacity of pre-heated PMCs showed that buffer capacity decreased with increasing incubation temperature, which was possibly due to the compaction of the PMCs and an increase in the number of compensated PAH sites. The addition of 1 M sodium chloride to heated PMCs presumably reversed the process described above, since an increase in the ionic strength of the solution led to an increase of the buffer capacity of the PMCs. The effects described above confirm the hypothesis put forward that the buffer properties of microcapsules are determined by uncompensated PAH regions in their composition.
Collapse
|
|
36
|
Interleukin-12 Plasmid DNA Delivery by N-[(2-Hydroxy-3-trimethylammonium)propyl]chitosan-Based Nanoparticles. Polymers (Basel) 2022; 14:polym14112176. [PMID: 35683849 PMCID: PMC9182864 DOI: 10.3390/polym14112176] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/20/2022] [Revised: 05/04/2022] [Accepted: 05/24/2022] [Indexed: 02/04/2023] Open
Abstract
Cationic polysaccharides are capable of forming polyplexes with nucleic acids and are considered promising polymeric gene carriers. The objective of this study was to evaluate the transfection efficiency and cytotoxicity of N-[(2-hydroxy-3-trimethylammonium)propyl] chitosan salt (HTCS), a quaternary ammonium derivative of chitosan (CS), which benefits from non-ionizable positive charges. In this work, HTCS with a full quaternization of amino groups and a molar mass of 130,000 g·mol−1 was synthesized to use for delivery of a plasmid encoding the interleukin-12 (IL-12) gene. Thus, a polyplex based on HTCS and the IL-12 plasmid was prepared and then was characterized in terms of particle size, zeta potential, plasmid condensation ability, and protection of the plasmid against enzymatic degradation. We showed that HTCS was able to condense the IL-12 plasmid by the formation of polyplexes in the range of 74.5 ± 0.75 nm. The level of hIL-12 production following the transfection of the cells with HTCS polyplexes at a C/P ratio of 8:1 was around 4.8- and 2.2-fold higher than with CS and polyethylenimine polyplexes, respectively. These findings highlight the role of HTCS in the formation of polyplexes for the efficient delivery of plasmid DNA.
Collapse
|
|
37
|
Butt AM, Abdullah N, Rani NNIM, Ahmad N, Amin MCIM. Endosomal Escape of Bioactives Deployed via Nanocarriers: Insights Into the Design of Polymeric Micelles. Pharm Res 2022; 39:1047-1064. [PMID: 35619043 DOI: 10.1007/s11095-022-03296-w] [Citation(s) in RCA: 10] [Impact Index Per Article: 3.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/30/2021] [Accepted: 05/17/2022] [Indexed: 10/18/2022]
Abstract
Cytoplasmic delivery of bioactives requires the use of strategies such as active transport, electroporation, or the use of nanocarriers such as polymeric nanoparticles, liposomes, micelles, and dendrimers. It is essential to deliver bioactive molecules in the cytoplasm to achieve targeted effects by enabling organelle targeting. One of the biggest bottlenecks in the successful cytoplasmic delivery of bioactives through nanocarriers is their sequestration in the endosomes that leads to the degradation of drugs by progressing to lysosomes. In this review, we discussed mechanisms by which nanocarriers are endocytosed, the mechanisms of endosomal escape, and more importantly, the strategies that can be and have been employed for their escape from the endosomes are summarized. Like other nanocarriers, polymeric micelles can be designed for endosomal escape, however, a careful control is needed in their design to balance between the possible toxicity and endosomal escape efficiency. Keeping this in view, polyion complex micelles, and polymers that have the ability to escape the endosome, are fully discussed. Finally, we provided some perspectives for designing the polymeric micelles for efficient cytoplasmic delivery of bioactive agents through endosomal escape.
Collapse
Affiliation(s)
- Adeel Masood Butt
- Institute of Pharmaceutical Sciences, University of Veterinary and Animal Sciences, Lahore, 54000, Pakistan.
| | - Nabiha Abdullah
- Department of Pharmacy, Quaid-i-Azam University, 45320, Islamabad, Pakistan.,Department of Chemistry, Quaid-i-Azam University, 45320, Islamabad, Pakistan
| | - Nur Najihah Izzati Mat Rani
- Faculty of Pharmacy and Health Sciences, Universiti Kuala Lumpur Royal College of Medicine Perak, 30450, Ipoh, Perak, Malaysia.,Centre for Drug Delivery Technology, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300, Kuala Lumpur, Malaysia
| | - Naveed Ahmad
- Department of Pharmaceutics, College of Pharmacy, Jouf University, Sakaka, 72388, Aljouf, Saudi Arabia
| | - Mohd Cairul Iqbal Mohd Amin
- Centre for Drug Delivery Technology, Faculty of Pharmacy, Universiti Kebangsaan Malaysia, Jalan Raja Muda Abdul Aziz, 50300, Kuala Lumpur, Malaysia.
| |
Collapse
|
|
38
|
Kermaniyan SS, Chen M, Zhang C, Smith SA, Johnston APR, Such C, Such GK. Understanding the Biological Interactions of pH Swellable Nanoparticles. Macromol Biosci 2022; 22:e2100445. [PMID: 35182032 DOI: 10.1002/mabi.202100445] [Citation(s) in RCA: 14] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/02/2021] [Revised: 02/02/2022] [Indexed: 11/07/2022]
Abstract
pH responsive nanoparticles have generated significant interest for use as drug delivery systems due to their potential for inducible release at low pH. The pH variation from the blood stream (pH 7.4) to intracellular compartments of cells called endosomes/lysosomes (pH < 5.0) has been of particular interest. However, one of the limitations with nanoparticle delivery systems is the ability to migrate out of these compartments to the cytosol or other organelles, via a process termed endosomal escape. Previous studies have postulated that pH responsive nanoparticles can facilitate endosomal escape through a range of mechanisms including membrane interaction, pH-induced swelling, and the proton-sponge effect. In this study we designed a series of pH swellable nanoparticles (85-100 nm) and investigated their impact on biological interactions, particularly endosomal escape. The particles exhibited tuneable pH-induced swelling (from 120% to 200%) and had good buffering capacity. Cellular association was studied using flow cytometry and endosomal escape was determined using a calcein leakage assay. Interestingly, we found no endosomal escape with all nanoparticle formulations, which suggests there are limitations with both the proton-sponge effect and pH-induced swelling mechanism as the primary methods for inducing endosomal escape. This article is protected by copyright. All rights reserved.
Collapse
Affiliation(s)
- Sarah S Kermaniyan
- School of Chemistry, The University of Melbourne, Parkville, Victoria, 3010, Australia
| | - Moore Chen
- Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, 3052, Australia
| | - Changhe Zhang
- School of Chemistry, The University of Melbourne, Parkville, Victoria, 3010, Australia
| | - Samuel A Smith
- School of Chemistry, The University of Melbourne, Parkville, Victoria, 3010, Australia
| | - Angus P R Johnston
- Monash Institute of Pharmaceutical Sciences, Monash University, Parkville, Victoria, 3052, Australia
| | - Chris Such
- School of Chemistry, The University of Melbourne, Parkville, Victoria, 3010, Australia
| | - Georgina K Such
- School of Chemistry, The University of Melbourne, Parkville, Victoria, 3010, Australia
| |
Collapse
|
|
39
|
Preparation and Characterization of Antibacterial Films with Eggshell-Membrane Biopolymers Incorporated with Chitosan and Plant Extracts. Polymers (Basel) 2022; 14:polym14030383. [PMID: 35160373 PMCID: PMC8839977 DOI: 10.3390/polym14030383] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/06/2021] [Revised: 01/07/2022] [Accepted: 01/17/2022] [Indexed: 11/16/2022] Open
Abstract
A series of films containing chitosan (CS), eggshell membrane (ESM), soluble eggshell membrane (SEP), and plant extracts from Thymus vulgaris and Origanum valgare were prepared with varying concentrations and compositions. These novel films were characterized extensively with respect to film thickness and uniformity, solution absorption, degradation, microenvironmental pH, and antibacterial properties. All the films were flexible with appropriate mechanical stability. After 48 h of soaking in a lysozyme solution, all the films degraded 64 ± 4%, which would be expected to allow for the release of the plant extracts. The plant extracts on their own showed a pH of approximately 4, with the blended films having microenvironmental pHs from approximately 6.4–7.0, which would be expected to promote wound healing. A CS-ESM-SEP film with 5% of each plant extract inhibited almost all E. coli growth in liquid cultures and had no detriments to fluid absorption. Fluid absorption was approximately 100–150% by weight for all the films. The incorporation of SEP and plant extracts to a CS-ESM film provides a promising and novel method for the incorporation of SEP and antibacterial agents in a film with no detriment to wound fluid absorption or film degradation.
Collapse
|
|
40
|
Sarwar S, Bashir S, Asim MH, Ikram F, Ahmed A, Omema U, Asif A, Chaudhry AA, Hu Y, Ustundag CB. In-depth drug delivery to tumoral soft tissues via pH responsive hydrogel. RSC Adv 2022; 12:31402-31411. [PMID: 36348995 PMCID: PMC9627957 DOI: 10.1039/d2ra05639a] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/07/2022] [Accepted: 10/26/2022] [Indexed: 11/07/2022] Open
Abstract
A pH responsive nanoparticle–hydrogel hybrid drug delivery system was investigated for in-depth anticancer drug delivery to solid tumours. It consists of acid susceptible polymer nanoparticles loaded in a chitosan hydrogel. The hybrid formulation was characterized by UV-visible spectroscopy, FTIR, SEM, TEM, particle size analysis, zeta potential measurement and viscosity measurement. Drug encapsulation and nanoparticle loading efficiencies were found to be 48% and 72% respectively which describes the efficient interaction of the chemical entities in this hybrid drug delivery system. The hydrogel exhibited pH responsive behaviour: minimal drug and nanoparticle release at physiological pH but an increase in viscosity under acidic conditions and fast nanoparticle and drug release. The cytotoxicity of the drug loaded hydrogel was investigated against the MCF-7 breast cancer cell line along with the drug and nanoparticles without hydrogel. The drug loaded hydrogel showed a better cytotoxic effect on MCF-7 cancer cells. Thus, drug loaded nanoparticles containing hydrogel could be a better option for maximum drug distribution in tumours. A pH responsive nanoparticle–hydrogel hybrid drug delivery system was investigated for in-depth anticancer drug delivery to solid tumours.![]()
Collapse
Affiliation(s)
- Shumaila Sarwar
- Interdisciplinary Research Centre in Biomedical Materials, COMSATS University Islamabad, Lahore, Pakistan
- College of Pharmacy, University of Sargodha, Sargodha, Pakistan
| | - Sajid Bashir
- College of Pharmacy, University of Sargodha, Sargodha, Pakistan
| | | | - Fakhera Ikram
- Interdisciplinary Research Centre in Biomedical Materials, COMSATS University Islamabad, Lahore, Pakistan
| | - Arsalan Ahmed
- Interdisciplinary Research Centre in Biomedical Materials, COMSATS University Islamabad, Lahore, Pakistan
| | - Ume Omema
- Interdisciplinary Research Centre in Biomedical Materials, COMSATS University Islamabad, Lahore, Pakistan
| | - Anila Asif
- Interdisciplinary Research Centre in Biomedical Materials, COMSATS University Islamabad, Lahore, Pakistan
| | - Aqif Anwar Chaudhry
- Interdisciplinary Research Centre in Biomedical Materials, COMSATS University Islamabad, Lahore, Pakistan
| | - Yong Hu
- Institute of Materials Engineering, National Laboratory of Solid State Microstructure, College of Engineering and Applied Sciences, Nanjing University, Nanjing, Jiangsu, China
| | - Cem Bulent Ustundag
- Department of Bioengineering, Faculty of Chemical and Metallurgical Engineering, Yildiz Technical University, Turkey
| |
Collapse
|
|
41
|
Development of erythrosine-based photodynamic therapy with a targeted drug delivery system to induce HepG2 cell apoptosis in vitro. Biochem Eng J 2022. [DOI: 10.1016/j.bej.2021.108267] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/13/2022]
|
|
42
|
Park SY, Yun YH, Park BJ, Seo HI, Chung I. Fabrication and Biological Activities of Plasmid DNA Gene Carrier Nanoparticles Based on Biodegradable l-Tyrosine Polyurethane. Pharmaceuticals (Basel) 2021; 15:ph15010017. [PMID: 35056074 PMCID: PMC8780858 DOI: 10.3390/ph15010017] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/02/2021] [Revised: 12/15/2021] [Accepted: 12/22/2021] [Indexed: 12/04/2022] Open
Abstract
Gene therapy is a suitable alternative to chemotherapy due to the complications of drug resistance and toxicity of drugs, and is also known to reduce the occurrence of cellular mutation through the use of gene carriers. In this study, gene carrier nanoparticles with minimal toxicity and high transfection efficiency were fabricated from a biocompatible and biodegradable polymer, l-tyrosine polyurethane (LTU), which was polymerized from presynthesized desaminotyrosyl tyrosine hexyl ester (DTH) and polyethylene glycol (PEG), by using double emulsion and solvent evaporation techniques, resulting in the formation of porous nanoparticles, and then used to evaluate their potential biological activities through molecular controlled release and transfection studies. To assess cellular uptake and transfection efficiency, two model drugs, fluorescently labeled bovine serum albumin (FITC-BSA) and plasmid DNA-linear polyethylenimine (LPEI) complex, were successfully encapsulated in nanoparticles, and their transfection properties and cytotoxicities were evaluated in LX2 as a normal cell and in HepG2 and MCF7 as cancer cells. The morphology and average diameter of the LTU nanoparticles were confirmed using light microscopy, transmission electron microscopy, and dynamic light scattering, while confocal microscopy was used to validate the cellular uptake of FITC-BSA-encapsulated LTU nanoparticles. Moreover, the successful cellular uptake of LTU nanoparticles encapsulated with pDNA-LPEI and the high transfection efficiency, confirmed by gel electrophoresis and X-gal assay transfection, indicated that LTU nanoparticles had excellent cell adsorption ability, facilitated gene encapsulation, and showed the sustained release tendency of genes through transfection experiments, with an optimal concentration ratio of pDNA and LPEI of 1:10. All the above characteristics are ideal for gene carriers designed to transport and release drugs into the cytoplasm, thus facilitating effective gene therapy.
Collapse
Affiliation(s)
- Soo-Yong Park
- Department of Polymer Science and Engineering, Pusan National University, Busan 46241, Korea;
| | - Yang H. Yun
- Department of Biomedical Engineering, College of Engineering, The University of Akron, Akron, OH 44325, USA;
| | - Bum-Joon Park
- Department of Molecular Biology, College of Natural Science, Pusan National University, Busan 46241, Korea;
| | - Hyung-Il Seo
- Department of Surgery, Biomedical Research Institute, Pusan National University Hospital, Busan 49241, Korea;
| | - Ildoo Chung
- Department of Polymer Science and Engineering, Pusan National University, Busan 46241, Korea;
- Correspondence:
| |
Collapse
|
|
43
|
Dubrovskii AV, Kim AL, Musin EV, Ramazanov BR, Tikhonenko SA. The Discovery of the Buffer Capacity of Various Types of Polyelectrolyte Microcapsules. Polymers (Basel) 2021; 13:4026. [PMID: 34833325 PMCID: PMC8620602 DOI: 10.3390/polym13224026] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/03/2021] [Revised: 11/17/2021] [Accepted: 11/19/2021] [Indexed: 11/17/2022] Open
Abstract
Polyelectrolyte microcapsules, which are obtained by the method of alternate adsorption of oppositely charged polyelectrolytes onto colloidal particles of micron size, are widely used in science and industry. Nevertheless, the properties of microcapsules are still poorly understood. In particular, there is no information in the literature on the buffer capacity. However, information on the presence of a buffer capacity and an understanding of its mechanisms can both simplify the use of microcapsules and expand the scope of their application. In this regard, the buffer capacity of various types of microcapsules was studied. It was found that polyelectrolyte microcapsules consisting of polyallylamine, and polystyrene sulfonate have a buffer capacity. In addition, in an acidic medium, the buffer capacity of microcapsules containing BSA is significantly greater than that of microcapsules without protein. This is due to the fact that BSA contributes to the buffering of microcapsules. Differences in the behaviour of the buffer capacity of microcapsules with the composition (PAH/PSS)3 and (PSS/PAH)3 were found. In addition, a hypothesis has been proposed that regions of unbound polyallylamine are responsible for the buffering properties of polyelectrolyte microcapsules. This hypothesis is confirmed by the fact that incubation of microcapsules in 0.5 M NaCl increases the amount of unbound polyallylamine, which leads to an increase in the buffer capacity of microcapsules at alkaline pH values higher than the buffer capacity of capsules in an aqueous solution.
Collapse
Affiliation(s)
- Alexey V. Dubrovskii
- Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya st., 3, 142290 Puschino, Russia; (A.V.D.); (A.L.K.); (E.V.M.)
| | - Aleksandr L. Kim
- Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya st., 3, 142290 Puschino, Russia; (A.V.D.); (A.L.K.); (E.V.M.)
| | - Egor V. Musin
- Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya st., 3, 142290 Puschino, Russia; (A.V.D.); (A.L.K.); (E.V.M.)
| | - Bulat R. Ramazanov
- Faculty of Biology, M. V. Lomonosov Moscow State University, 119991 Moscow, Russia;
| | - Sergey A. Tikhonenko
- Institute of Theoretical and Experimental Biophysics Russian Academy of Science, Institutskaya st., 3, 142290 Puschino, Russia; (A.V.D.); (A.L.K.); (E.V.M.)
| |
Collapse
|
|
44
|
Supramolecular Assembly and Reversible Transition and of Chitosan Fluorescent Micelles by Noncovalent Modulation. ADVANCES IN POLYMER TECHNOLOGY 2021. [DOI: 10.1155/2021/5175473] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 11/17/2022]
Abstract
Chitosan-based intelligent artificial systems have been of increasing interest for their biocompatibility, multifunctionality, biological activity, and low cost. Herein, we report the fabrication of supramolecular nanoparticles based on water-soluble chitosan (WCS) and 1,1
,1
,1
-(ethene-1,1,2,2-tetrayl)tetrakis(benzene-4,1-diyl) tetrakis(azanediyl)tetraacetic acid (TPE-(N-COOH)4), which is capable of reversible transition between polyion complexes (PICs) and hydrogen bonding complexes (HBCs) with tunable aggregation-induced emission driven by pH value. The PIC micelles could be formed via electrostatic interaction between ammonium cations and carboxylate anions under mild alkaline conditions. The formation of the micelles dramatically blocks the nonradiative pathway and enhances the fluorescence of TPE moieties, and the maximum fluorescence intensity was achieved near the isoelectric point due to the restriction of intramolecular motion. In addition, the fluorescence intensity and size of the PIC micelles exhibited a temperature response in the range from 20 to 80°C. Upon adjusting the solution pH to 2, the PIC micelles were reconstructed into hydrogen-bonding complexes while the hydrogen bonding interaction between the protonated carboxyl groups of TPE-(N-COOH)4 and chitosan. Moreover, the size of the micelles underwent a remarkable decrease, whereas the fluorescence emission was further enhanced by ~6.25-fold. The pH actuated micellar transition from PIC to HBC with tunable fluorescence performance is fully reversible. This study provides novel multifunctional materials that are of great importance for their potential application in the fields of optoelectronic devices and chemical and biomedical sensors.
Collapse
|
|
45
|
Racaniello GF, Laquintana V, Vergnaud J, Lopedota A, Cutrignelli A, Lopalco A, Leonetti F, Franco M, Fiume M, Pontrelli P, Gesualdo L, Fattal E, Denora N. Development of purified glycogen derivatives as siRNA nanovectors. Int J Pharm 2021; 608:121128. [PMID: 34560204 DOI: 10.1016/j.ijpharm.2021.121128] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/04/2021] [Revised: 09/17/2021] [Accepted: 09/18/2021] [Indexed: 10/20/2022]
Abstract
Purified Glycogen (PG) is a highly hyper branched carbohydrate, characterized by high water solubility and very moderate increase in viscosity. The dendrimeric structure of PG, appropriately functionalized, makes it an alternative to current synthetic gene delivery agents. The present study explores the preparation of purified glycogen polycationic derivatives (PGPDs), developed and characterized starting from a single step reaction between PG and N,N-dialkylamino alkyl halides. Subsequently PGPDs were used for the complexation of a model siRNA nucleic acid, a transfection reagent siRNA and a fluorescein-labelled dsRNA oligomer. PGPDs-siRNA complexes were fully characterized by agarose gel electrophoresis and their efficacy was assessed by both confocal microscopy and transfection assays on breast and renal cancer cells. Results proved that PGPDs-siRNA complexes were efficient and not cytotoxic, maintaining their spherical and dendrimeric structure and, particularly, were able to effectively transfect the target cells by releasing the siRNA.
Collapse
Affiliation(s)
| | - Valentino Laquintana
- Department of Pharmacy - Pharmaceutical Sciences, University of Bari "A. Moro", Orabona, St. 4, 70125 Bari, Italy
| | - Juliette Vergnaud
- Institut Galien Paris-Saclay, UMR CNRS 8612, Université Paris Saclay, Châtenay-Malabry, Paris, France
| | - Angela Lopedota
- Department of Pharmacy - Pharmaceutical Sciences, University of Bari "A. Moro", Orabona, St. 4, 70125 Bari, Italy
| | - Annalisa Cutrignelli
- Department of Pharmacy - Pharmaceutical Sciences, University of Bari "A. Moro", Orabona, St. 4, 70125 Bari, Italy
| | - Antonio Lopalco
- Department of Pharmacy - Pharmaceutical Sciences, University of Bari "A. Moro", Orabona, St. 4, 70125 Bari, Italy
| | - Francesco Leonetti
- Department of Pharmacy - Pharmaceutical Sciences, University of Bari "A. Moro", Orabona, St. 4, 70125 Bari, Italy
| | - Massimo Franco
- Department of Pharmacy - Pharmaceutical Sciences, University of Bari "A. Moro", Orabona, St. 4, 70125 Bari, Italy
| | - Mauro Fiume
- Department of Emergency and Organ Transplantation, University of Bari "A. Moro", Orabona, St. 4, 70125 Bari, Italy
| | - Paola Pontrelli
- Department of Emergency and Organ Transplantation, University of Bari "A. Moro", Orabona, St. 4, 70125 Bari, Italy
| | - Loreto Gesualdo
- Department of Emergency and Organ Transplantation, University of Bari "A. Moro", Orabona, St. 4, 70125 Bari, Italy
| | - Elias Fattal
- Institut Galien Paris-Saclay, UMR CNRS 8612, Université Paris Saclay, Châtenay-Malabry, Paris, France
| | - Nunzio Denora
- Department of Pharmacy - Pharmaceutical Sciences, University of Bari "A. Moro", Orabona, St. 4, 70125 Bari, Italy.
| |
Collapse
|
|
46
|
Reshad RAI, Jishan TA, Chowdhury NN. Chitosan and its Broad Applications: A Brief Review. JOURNAL OF CLINICAL AND EXPERIMENTAL INVESTIGATIONS 2021. [DOI: 10.29333/jcei/11268] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/20/2022] Open
|
|
47
|
Sun R, Fang L, Lv X, Fang J, Wang Y, Chen D, Wang L, Chen J, Qi Y, Tang Z, Zhang J, Tian Y. In vitro and in vivo evaluation of self-assembled chitosan nanoparticles selectively overcoming hepatocellular carcinoma via asialoglycoprotein receptor. Drug Deliv 2021; 28:2071-2084. [PMID: 34595970 PMCID: PMC8491732 DOI: 10.1080/10717544.2021.1983077] [Citation(s) in RCA: 19] [Impact Index Per Article: 4.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/27/2022] Open
Abstract
Hepatocellular carcinoma (HCC) is one of the major causes of cancer-related mortality worldwide. Nowadays, liver-targeting drug delivery system has been proven as a promising strategy for overcoming HCC. Asialoglycoprotein receptor (ASGPR) is an ideal receptor for liver targeting, which is mainly expressed on hepatocytes. In this study, we developed several novel liver-targeting chitosan nanoparticles to selectively overcome HCC via ASGPR. Chitosan nanoparticles (Gly-CS-VE, Gal-Gly-CS-VE, Gly-CS-DCA, and Gal-Gly-CS-DCA) were prepared by grafting hydrophilic group (glycidol, Gly), hydrophobic group (deoxycholic acid, DCA or vitamin E succinate, VE), and ASGPR recognizing group (galactose, Gal). Subsequently, their characterizations were measured by 1H NMR, FT-IR, TEM, and DLS. Doxorubicin (DOX) was loaded in nanoparticles and released out in a pH-dependent manner. Most importantly, the galactosylated Gal-Gly-CS-VE and Gal-Gly-CS-DCA nanoparticles exhibited significantly stronger in vitro cell internalization, cytotoxicity, anti-migration capabilities and in vivo anticancer efficacies than the corresponding Gly-CS-VE and Gly-CS-DCA nanoparticles, as well as free DOX. Finally, the four chitosan nanoparticles exhibited good biocompatibility without causing any obvious histological damage to the major organs. Overall, the galactosylated chitosan nanoparticles were proven to be promising pharmaceutical formulations for selectively overcoming HCC, with great potential for clinical applications.
Collapse
Affiliation(s)
- Rensong Sun
- Collage of Pharmacy, Dalian Medical University, Dalian, China
| | - Linlin Fang
- Collage of Pharmacy, Dalian Medical University, Dalian, China
| | - Xia Lv
- Collage of Integrative Medicine, Dalian Medical University, Dalian, China
| | - Jiani Fang
- Collage of Pharmacy, Dalian Medical University, Dalian, China
| | - Yuting Wang
- Collage of Pharmacy, Dalian Medical University, Dalian, China
| | - Dapeng Chen
- Laboratory Animal Center, Dalian Medical University, Dalian, China
| | - Liang Wang
- Laboratory Animal Center, Dalian Medical University, Dalian, China
| | - Jun Chen
- Laboratory Animal Center, Dalian Medical University, Dalian, China
| | - Yan Qi
- Collage of Pharmacy, Dalian Medical University, Dalian, China
| | - Zeyao Tang
- Collage of Pharmacy, Dalian Medical University, Dalian, China
| | - Jianbin Zhang
- Collage of Pharmacy, Dalian Medical University, Dalian, China
| | - Yan Tian
- Collage of Pharmacy, Dalian Medical University, Dalian, China
| |
Collapse
|
|
48
|
Sharifiaghdam M, Shaabani E, Sharifiaghdam Z, De Keersmaecker H, Lucas B, Lammens J, Ghanbari H, Teimoori-Toolabi L, Vervaet C, De Beer T, Faridi-Majidi R, De Smedt SC, Braeckmans K, Fraire JC. Macrophage reprogramming into a pro-healing phenotype by siRNA delivered with LBL assembled nanocomplexes for wound healing applications. NANOSCALE 2021; 13:15445-15463. [PMID: 34505619 DOI: 10.1039/d1nr03830c] [Citation(s) in RCA: 14] [Impact Index Per Article: 3.5] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 06/13/2023]
Abstract
Excessive inflammatory responses in wounds are characterized by the presence of high levels of pro-inflammatory M1 macrophages rather than pro-healing M2 macrophages, which leads to delayed wound healing. Macrophage reprogramming from the M1 to M2 phenotype through knockdown of interferon regulatory factor 5 (irf5) has emerged as a possible therapeutic strategy. While downregulation of irf5 could be achieved by siRNA, it very much depends on successful intracellular delivery by suitable siRNA carriers. Here, we report on highly stable selenium-based layer-by-layer (LBL) nanocomplexes (NCs) for siRNA delivery with polyethyleneimine (PEI-LBL-NCs) as the final polymer layer. PEI-LBL-NCs showed good protection of siRNA with only 40% siRNA release in a buffer of pH = 8.5 after 72 h or in simulated wound fluid after 4 h. PEI-LBL-NCs also proved to be able to transfect RAW 264.7 cells with irf5-siRNA, resulting in successful reprogramming to the M2 phenotype as evidenced by a 3.4 and 2.6 times decrease in NOS-2 and TNF-α mRNA expression levels, respectively. Moreover, irf5-siRNA transfected cells exhibited a 2.5 times increase of the healing mediator Arg-1 and a 64% increase in expression of the M2 cell surface marker CD206+. Incubation of fibroblast cells with conditioned medium isolated from irf5-siRNA transfected RAW 264.7 cells resulted in accelerated wound healing in an in vitro scratch assay. These results show that irf5-siRNA loaded PEI-LBL-NCs are a promising therapeutic approach to tune macrophage polarization for improved wound healing.
Collapse
Affiliation(s)
- Maryam Sharifiaghdam
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ghent, B-9000, Belgium.
- Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
| | - Elnaz Shaabani
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ghent, B-9000, Belgium.
- Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
| | - Zeynab Sharifiaghdam
- Department of Physiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
| | - Herlinde De Keersmaecker
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ghent, B-9000, Belgium.
- Center for Advanced Light Microscopy, Ghent University, 9000 Ghent, Belgium
| | - Bart Lucas
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ghent, B-9000, Belgium.
| | - Joris Lammens
- Laboratory of Pharmaceutical Technology, Department of Pharmaceutics, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium
| | - Hossein Ghanbari
- Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
| | | | - Chris Vervaet
- Laboratory of Pharmaceutical Technology, Department of Pharmaceutics, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium
| | - Thomas De Beer
- Laboratory of Pharmaceutical Process Analytical Technology (LPPAT), Department of Pharmaceutical Analysis, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium
| | - Reza Faridi-Majidi
- Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.
| | - Stefaan C De Smedt
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ghent, B-9000, Belgium.
| | - Kevin Braeckmans
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ghent, B-9000, Belgium.
- Center for Advanced Light Microscopy, Ghent University, 9000 Ghent, Belgium
| | - Juan C Fraire
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ghent, B-9000, Belgium.
| |
Collapse
|
|
49
|
Ture N, Desai D, Shende P. Polyplexes of retinoic acid: an in vitro study of complex nanostructures against colorectal cancer cell line (HCT-15). JOURNAL OF MATERIALS SCIENCE. MATERIALS IN MEDICINE 2021; 32:122. [PMID: 34519890 PMCID: PMC8440295 DOI: 10.1007/s10856-021-06571-1] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Figures] [Subscribe] [Scholar Register] [Received: 11/20/2020] [Accepted: 06/07/2021] [Indexed: 06/13/2023]
Abstract
Despite recent advances in the treatment of human colon cancer, the chemotherapeutic efficacy against colon cancer is still unsatisfactory. The complexity in colorectal cancer treatment leads to new research in combination therapy to overcome multidrug resistance in cancer and increase apoptosis. The objective of the present research work was to develop polyplexes for co-delivery of plasmid DNA with retinoic acid against colorectal cancer cell line (HCT-15). Plain polyplexes were prepared using chitosan and hyaluronic acid solution (0.1% w/v), whereas retinoic acid polyplexes were prepared using ethanol: water (1:9 v/v) system. The particle size was observed in the order of chitosan solution > blank polyplex > retinoic acid-loaded polyplex. Encapsulation efficiency of retinoic acid was found to be 81.51 ± 4.33% for retinoic acid-loaded polyplex formulation. The drug release was observed to be in a controlled pattern with 72.23 ± 1.32% release of retenoic acid from polyplex formulation. Cell line studies of the formulation displayed better cell inhibition and low cytotoxicity for the retinoic acid-loaded polyplexes in comparison to pure retinoic acid, thus demonstrating better potential action against colorectal cancer cell line HCT-15. Retinoic acid-loaded polyplexes indicated higher potential for the delivery of the active whereas the cell line studies displayed the efficacy of the formulation against colorectal cancer cell line HCT-15.
Collapse
Affiliation(s)
- Narayan Ture
- Shobhaben Pratapbhai Patel School of Pharmacy and Technology Management, SVKM'S NMIMS, V. L. Mehta Road, Vile Parle (W), Mumbai, India
| | - Drashti Desai
- Shobhaben Pratapbhai Patel School of Pharmacy and Technology Management, SVKM'S NMIMS, V. L. Mehta Road, Vile Parle (W), Mumbai, India
| | - Pravin Shende
- Shobhaben Pratapbhai Patel School of Pharmacy and Technology Management, SVKM'S NMIMS, V. L. Mehta Road, Vile Parle (W), Mumbai, India.
| |
Collapse
|
|
50
|
Shaabani E, Sharifiaghdam M, Lammens J, De Keersmaecker H, Vervaet C, De Beer T, Motevaseli E, Ghahremani MH, Mansouri P, De Smedt S, Raemdonck K, Faridi-Majidi R, Braeckmans K, Fraire JC. Increasing Angiogenesis Factors in Hypoxic Diabetic Wound Conditions by siRNA Delivery: Additive Effect of LbL-Gold Nanocarriers and Desloratadine-Induced Lysosomal Escape. Int J Mol Sci 2021; 22:9216. [PMID: 34502144 PMCID: PMC8431033 DOI: 10.3390/ijms22179216] [Citation(s) in RCA: 24] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 07/30/2021] [Revised: 08/16/2021] [Accepted: 08/24/2021] [Indexed: 12/23/2022] Open
Abstract
Impaired wound healing in people with diabetes has multifactorial causes, with insufficient neovascularization being one of the most important. Hypoxia-inducible factor-1 (HIF-1) plays a central role in the hypoxia-induced response by activating angiogenesis factors. As its activity is under precise regulatory control of prolyl-hydroxylase domain 2 (PHD-2), downregulation of PHD-2 by small interfering RNA (siRNA) could stabilize HIF-1α and, therefore, upregulate the expression of pro-angiogenic factors as well. Intracellular delivery of siRNA can be achieved with nanocarriers that must fulfill several requirements, including high stability, low toxicity, and high transfection efficiency. Here, we designed and compared the performance of layer-by-layer self-assembled siRNA-loaded gold nanoparticles with two different outer layers-Chitosan (AuNP@CS) and Poly L-arginine (AuNP@PLA). Although both formulations have exactly the same core, we find that a PLA outer layer improves the endosomal escape of siRNA, and therefore, transfection efficiency, after endocytic uptake in NIH-3T3 cells. Furthermore, we found that endosomal escape of AuNP@PLA could be improved further when cells were additionally treated with desloratadine, thus outperforming commercial reagents such as Lipofectamine® and jetPRIME®. AuNP@PLA in combination with desloratadine was proven to induce PHD-2 silencing in fibroblasts, allowing upregulation of pro-angiogenic pathways. This finding in an in vitro context constitutes a first step towards improving diabetic wound healing with siRNA therapy.
Collapse
Affiliation(s)
- Elnaz Shaabani
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium; (E.S.); (M.S.); (H.D.K.); (S.D.S.); (K.R.); (J.C.F.)
- Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
| | - Maryam Sharifiaghdam
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium; (E.S.); (M.S.); (H.D.K.); (S.D.S.); (K.R.); (J.C.F.)
- Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
| | - Joris Lammens
- Laboratory of Pharmaceutical Technology, Department of Pharmaceutics, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium; (J.L.); (C.V.)
| | - Herlinde De Keersmaecker
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium; (E.S.); (M.S.); (H.D.K.); (S.D.S.); (K.R.); (J.C.F.)
- Center for Advanced Light Microscopy, Ghent University, 9000 Ghent, Belgium
| | - Chris Vervaet
- Laboratory of Pharmaceutical Technology, Department of Pharmaceutics, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium; (J.L.); (C.V.)
| | - Thomas De Beer
- Laboratory of Pharmaceutical Process Analytical Technology (LPPAT), Department of Pharmaceutical Analysis, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium;
| | - Elahe Motevaseli
- Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran;
| | - Mohammad Hossein Ghahremani
- Department of Toxicology and Pharmacology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran;
| | - Parvin Mansouri
- Skin and Stem Cell Research Center, Tehran University of Medical Sciences, Tehran, Iran;
| | - Stefaan De Smedt
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium; (E.S.); (M.S.); (H.D.K.); (S.D.S.); (K.R.); (J.C.F.)
- Center for Advanced Light Microscopy, Ghent University, 9000 Ghent, Belgium
| | - Koen Raemdonck
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium; (E.S.); (M.S.); (H.D.K.); (S.D.S.); (K.R.); (J.C.F.)
| | - Reza Faridi-Majidi
- Department of Medical Nanotechnology, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran
| | - Kevin Braeckmans
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium; (E.S.); (M.S.); (H.D.K.); (S.D.S.); (K.R.); (J.C.F.)
- Center for Advanced Light Microscopy, Ghent University, 9000 Ghent, Belgium
| | - Juan C. Fraire
- Laboratory of General Biochemistry and Physical Pharmacy, Faculty of Pharmacy, Ghent University, Ottergemsesteenweg 460, 9000 Ghent, Belgium; (E.S.); (M.S.); (H.D.K.); (S.D.S.); (K.R.); (J.C.F.)
| |
Collapse
|