Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Dec 26, 2020; 12(12): 1667-1690
Published online Dec 26, 2020. doi: 10.4252/wjsc.v12.i12.1667
Effects of normobaric cyclic hypoxia exposure on mesenchymal stem-cell differentiation–pilot study on bone parameters in elderly
Marta Camacho-Cardenosa, José Manuel Quesada-Gómez, Alba Camacho-Cardenosa, Alejo Leal, Gabriel Dorado, Bárbara Torrecillas-Baena, Antonio Casado-Díaz
Marta Camacho-Cardenosa, Alba Camacho-Cardenosa, Facultad Ciencias del Deporte, Universidad De Extremadura, Cáceres 10003, Spain
José Manuel Quesada-Gómez, Bárbara Torrecillas-Baena, Antonio Casado-Díaz, CIBER De Fragilidad Y Envejecimiento Saludable (CIBERFES), Unidad De Gestión Clínica De Endocrinología Y Nutrición, Instituto Maimónides De Investigación Biomédica De Córdoba, Hospital Universitario Reina Sofía, Córdoba 14004, Spain
Alejo Leal, Servicio de Traumatología, Hospital de Cáceres, Cáceres 10004, Spain
Gabriel Dorado, Departamento Bioquímica y Biología Molecular, Campus Rabanales C6-1-E17, Campus de Excelencia Internacional Agroalimentario (ceiA3), Universidad de Córdoba-CIBERFES, 14071 Córdoba, Spain
Author contributions: Camacho-Cardenosa M, Casado-Díaz A and Quesada-Gómez JM conceived and designed the experiments; Camacho-Cardenosa M, Torrecillas-Baena B and Casado-Díaz A performed the experiments in vitro; Camacho-Cardenosa M, Camacho-Cardenosa A and Leal A performed the studies with the elderly volunteers; Camacho-Cardenosa M, Casado-Díaz A, Dorado G and Quesada-Gómez JM wrote the paper. All authors analyzed and interpreted the data.
Supported by Government of Extremadura GAEDAF Research Group, No. GR18003; Ministerio de Educación, Cultura y Deporte, No. FPU15/00452; and Instituto de Salud Carlos III, No. PI15/01857 and No. PI18/01659.
Institutional review board statement: The study was approved by the Bioethical and Biosecurity Commission of the University of Extremadura (17/2016).
Conflict-of-interest statement: The authors have nothing to disclose.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Marta Camacho-Cardenosa, PhD, Postdoc, Facultad Ciencias Del Deporte, Universidad De Extremadura, Av. Universidad, s/n, Cáceres 10003, Spain. mcamachocardenosa@unex.es
Received: September 3, 2020
Peer-review started: September 3, 2020
First decision: September 15, 2020
Revised: September 30, 2020
Accepted: October 20, 2020
Article in press: October 20, 2020
Published online: December 26, 2020
Processing time: 114 Days and 8.9 Hours
Abstract
BACKGROUND

Mesenchymal stem cells (MSC) of bone marrow are the progenitor of osteoblasts and adipocytes. MSC tend to differentiate into adipocytes, instead of osteoblasts, with aging. This favors the loss of bone mass and development of osteoporosis. Hypoxia induces hypoxia inducible factor 1α gene encoding transcription factor, which regulates the expression of genes related to energy metabolism and angiogenesis. That allows a better adaptation to low O2 conditions. Sustained hypoxia has negative effects on bone metabolism, favoring bone resorption. Yet, surprisingly, cyclic hypoxia (CH), short times of hypoxia followed by long times in normoxia, can modulate MSC differentiation and improve bone health in aging.

AIM

To evaluate the CH effect on MSC differentiation, and whether it improves bone mineral density in elderly.

METHODS

MSC cultures were induced to differentiate into osteoblasts or adipocytes, in CH (3% O2 for 1, 2 or 4 h, 4 d a week). Extracellular-matrix mineralization and lipid-droplet formation were studied in MSC induced to differentiate into osteoblast or adipocytes, respectively. In addition, gene expression of marker genes, for osteogenesis or adipogenesis, have been quantified by quantitative real time polymerase chain reaction. The in vivo studies with elderly (> 75 years old; n = 10) were carried out in a hypoxia chamber, simulating an altitude of 2500 m above sea level, or in normoxia, for 18 wk (36 CH sessions of 16 min each). Percentages of fat mass and bone mineral density from whole body, trunk and right proximal femur (femoral, femoral neck and trochanter) were assessed, using dual-energy X-ray absorptiometry.

RESULTS

CH (4 h of hypoxic exposure) inhibited extracellular matrix mineralization and lipid-droplet formation in MSC induced to differentiate into osteoblasts or adipocytes, respectively. However, both parameters were not significantly affected by the other shorter hypoxia times assessed. The longest periods of hypoxia downregulated the expression of genes related to extracellular matrix formation, in MSC induced to differentiate into osteoblasts. Interestingly, osteocalcin (associated to energy metabolism) was upregulated. Vascular endothelial growth factor an expression and low-density lipoprotein receptor related protein 5/6/dickkopf Wnt signaling pathway inhibitor 1 (associated to Wnt/β-catenin pathway activation) increased in osteoblasts. Yet, they decreased in adipocytes after CH treatments, mainly with the longest hypoxia times. However, the same CH treatments increased the osteoprotegerin/receptor activator for nuclear factor kappa B ligand ratio in both cell types. An increase in total bone mineral density was observed in elderly people exposed to CH, but not in specific regions. The percentage of fat did not vary between groups.

CONCLUSION

CH may have positive effects on bone health in the elderly, due to its possible inhibitory effect on bone resorption, by increasing the osteoprotegerin / receptor activator for nuclear factor kappa B ligand ratio.

Keywords: Normobaric cyclic hypoxia; Mesenchymal stem cells; Cellular differentiation; Osteoblasts; Adipocytes; Bone health

Core Tip: Cyclic hypoxia (CH) can improve bone health in aging. Mesenchymal stem cells were induced to differentiate into osteoblasts or adipocytes, in hypoxia (3% O2) for 1, 2 or 4 h, 4 d a week. Older people were treated with 36 CH sessions of 16 min (simulating an altitude of 2500 m), for 18 wk. Exposure to longer periods of hypoxia inhibited mesenchymal stem cells differentiation, but increased osteoprotegerin/ receptor activator for nuclear factor kappa B ligand expression in both cell types. CH increased total bone mineral density, but not fat content, in elderly. CH may improve bone health in elderly, due to its inhibitory effect on bone resorption, by increasing the osteoprotegerin/receptor activator for nuclear factor kappa B ligand ratio.