Osteomyelitis is a severe infection of bone tissue that can lead to bone loss and even osteonecrosis. This condition is mostly caused by Gram-positive bacteria, with Staphylococcus aureus being the most common etiological agent. Among the pathophysiological mechanisms involved in osteomyelitis, the ability of S. aureus to be internalized by osteoblasts or osteocytes and to survive within these cells, is particularly noteworthy. Infected osteoblasts and osteocytes not only serve as reservoirs in chronic cases of osteomyelitis but also play an active role in the osteoimmunology process, notably by producing mediators that promote the bone resorption activity of osteoclasts, thereby disrupting bone homeostasis. The present review explores both historical and recent literature on the internalization of S. aureus by osteoblasts and osteocytes, its intracellular behavior following internalization, and its mechanisms for inducing cell death. Additionally, it examines how S. aureus affects bone formation activity and promotes the production of inflammatory and pro-osteoclastic mediators. This review aims to highlight the limitations of current findings and outline key questions for future investigations.

By 2050, the global aging population is predicted to reach 1.5 billion, highlighting the need to enhance the quality of life of the elderly population. Osteoporotic fractures are projected to affect one in three women and one in five men over age 50. Initial treatments for osteoporosis in postmenopausal women include antiresorptive agents such as bisphosphonates, strontium ranelate, estrogen replacement therapy (ERT) and selective estrogen receptor modulators (SERMs). However, these do not rebuild bone, limiting their effectiveness. Denosumab, an FDA-approved antiresorptive monoclonal antibody, also has drawbacks including high costs, biannual subcutaneous injections, slow healing, impaired bone growth and side effects like eczema, flatulence, cellulitis, osteonecrosis of the jaw (ONJ) and an increased risk of spinal fractures after discontinuation of treatment. Nutraceuticals, particularly phytoestrogens, are gaining attention for their health benefits and safety in osteoporosis prevention, management and treatment. Phytoestrogens are plant metabolites similar to mammalian estrogens and include isoflavones, coumestans, lignans, stilbenes, and flavonoids. They interact with estrogen receptor isoforms ERα and ERβ, acting as agonists or antagonists based on concentration and bioavailability. Their tissue-selective activities are particularly significant: anti-estrogenic effects in reproductive tissues may lower the risk of hormone-related cancers (such as ovarian, uterine, breast and prostate), while estrogenic effects on bone could contribute to the preservation of bone mineral density.Phytoestrogens are, thus, used in managing breast and prostate cancers, cardiovascular diseases, menopause and osteoporosis. The present review focuses on the botanical origin, classification, sources and mechanism(s) of action of major phytoestrogens, their potential in prevention and management of osteoporosis and the requirement for additional clinical trials to achieve more definitive outcomes in order to confirm their efficacy and dosage safety.

Cardiovascular disease and osteoporosis are clinically associated. Bone adapts to mechanical forces by altering its overall structure and mass. In response to mechanical strain bone cells release signaling molecules and activate the nervous system. Bone also exhibits endocrine functions that modulate a number of tissues including the heart. We hypothesized that bone mechanical loading acutely alters cardiac function via neural and/or endocrine mechanisms. To test this hypothesis, we performed in vivo tibia mechanical loading in anesthetized mice while monitoring heart parameters using electrocardiogram (ECG). An immediate, transient reduction in resting heart rate was observed during tibial loading in both adult male and female mice (p < 0.01) with concurrent increases in heart rate variability (HRV) (p < 0.01). ECG intervals, PR, QRS and QTc were unaffected with loading. In further studies, we found that at least 3 N of load was necessary to elicit this heart response in adult mice. With aging to 11-12 months the responsiveness of the heart to loading was blunted, suggesting this bone-heart connection may weaken with age. Administration of lidocaine around the tibia significantly diminished the heart rate response to bone loading (p < 0.05). Moreover, pre-treatment with sympathetic antagonist propranolol inhibited this heart rate response to loading (p < 0.05), while parasympathetic antagonist atropine did not (p > 0.05). This suggests that a neuronal afferent pathway in the hindlimb and reduction in efferent sympathetic tone mediate this bone-neuro-heart reflex. In conclusion, the findings that tibia bone loading age-dependently modulates heart function support the concept of physiological coupling of the skeletal and cardiovascular systems.

This review explores the role of cell communication network (CCN) proteins in regulating skeletal physiology, aging, and disease, particularly within the context of balanced bone remodeling.

Recent conceptualization of paracrine and endocrine networks in bone marrow as a form of osteoimmunological crosstalk suggests a significant role for matricellular signaling in regulating bone homeostasis. As multifunctional adapters of cell-matrix interactions, CCNs are emerging as a focal point for parathyroid hormone (PTH) signaling and regulation of the RANKL/RANK/OPG axis in skeletal aging. Altered bone marrow CCN expression creates a permissive environment for accelerated postmenopausal bone loss and may contribute to the pathogenesis of osteoporosis and other diseases related to skeletal aging. CCNs modulate fundamental signaling mechanisms in bone development, homeostasis and repair. During aging, dysregulation of CCNs may negatively affect skeletal health and contribute to disease progression. As a result, CCNs may constitute promising therapeutic targets for improving and maintaining aging bone health.

Osteoarthritis (OA) is a multifactorial degenerative disease involved subchondral bone remodeling, cartilage destruction and synovium inflammation. While receptor activator of nuclear factor-κB ligand (RANKL), a tumor necrosis factor (TNF) superfamily protein, is the critical regulator in bone metabolism associated with subchondral bone resorption, TNF-α is also an important inflammatory factor involved in the OA inflammation and cartilage destruction. Based on previous compound Y1599, we identified a novel tetrahydro-β-carboline derivative Y2641 with both RANKL and TNF-α inhibition in this study. Y2641 exhibited potent RANKL-induced osteoclastogenic inhibition (IC50 = 109.1 nM), and had anti-inflammatory and cartilage destruction inhibiting effects at 10 μM with low cytotoxicity. SPR assays demonstrated the binding affinity of Y2641 to RANKL (Kd = 3.984 μM) and TNF-α (Kd = 18.59 μM). In vivo assay further revealed the disease-modifying effects of Y2641 in OA rats, establishing Y2641 as a promising lead compound for the development of disease-modifying osteoarthritis drugs.

Osteoporosis, characterized by excessive osteoclast activation, is mediated through the RANKL/RANK/OPG signaling axis. While flavonoids from Eucommia ulmoides (EU) have demonstrated anti-osteoclastogenic activity, their atomic-level mechanisms remain elusive. Here, we investigated six EU-derived flavonoids (cyrtominetin, quercetin, syringetin, genistein, ombuin, and kaempferol) targeting RANKL using integrated computational approaches. Molecular docking revealed strong binding affinities (Total_Score > 4.0) for all compounds, with cyrtominetin exhibiting the highest affinity (-50.205 kJ/mol via MM-PBSA), primarily through hydrogen bonds with Gly178, His180, Lys181, and Asn295. Moreover, most flavonoids interacted with RANKL by forming strong hydrogen bonds with Gly178 and Asn295, exhibiting higher binding affinity that was identified as essential for the activity. All-atom molecular dynamics simulations (100 ns) confirmed complex stability, demonstrating: low RMSD fluctuations (< 4.0 Å) and compact Rg values (16.0-17.0 Å). Notably, binding free energy decomposition identified both electrostatic and van der Waals contributions as critical for stabilization. These results identify cyrtominetin as a promising lead compound for RANKL inhibition, providing structural insights for designing flavonoid-based therapeutics against osteoporosis.

Aging is typically associated with decreased musculoskeletal function, leading to reduced mobility and increased frailty. As a hallmark of aging, cellular senescence plays a crucial role in various age-related musculoskeletal diseases, including osteoporosis, osteoarthritis, intervertebral disc degeneration, and sarcopenia. The detrimental effects of senescence are primarily due to impaired regenerative capacity of stem cells and the pro-inflammatory environment created by accumulated senescent cells. The secreted senescence-associated secretory phenotype (SASP) can induce senescence in neighboring cells, further amplifying senescent signals. Although the removal of senescent cells and the suppression of SASP factors have shown promise in alleviating disease progression and restoring musculoskeletal health in mouse models, clinical trials have yet to demonstrate significant efficacy. This review summarizes the mechanisms of cellular senescence in age-related musculoskeletal diseases and discusses potential therapeutic strategies targeting cellular senescence.

A large number of studies have shown that osteoporosis is closely related to bone immunology. The purpose of this study is to conduct bibliometrics and visual analysis of the fields related to osteoimmunology and osteoporosis from 2013 to 2022 and to summarize the research hotspots and trends in this field.

We searched the Web of Science core collection database for articles on osteoimmunology and osteoporosis published between 2013 and 2022. Vosviewer 1.6.18 and CiteSpace.6.2. R4 were used to analyze the retrieved data.

A total of 3218 articles on osteoimmunology and osteoporosis were included in this study. A total of 76 countries, 347 institutions, and 502 authors were included in the articles examined in this study. The main research countries were China, the United States, and South Korea. Shanghai Jiaotong University, Harvard University, and the University of California system were the main research institutions. The author who published the most papers was Xu, Jiake.

This study is the first to summarize the global research trends in the field of osteoimmunology and osteoporosis from 2013 to 2022. That helps researchers quickly understand the research hotspots and directions in this field.

Osteoporosis is the most common skeletal disease worldwide, characterized by low bone mineral density, resulting in weaker bones, and an increased risk of fragility fractures. The maintenance of bone mass relies on the precise balance between bone synthesis and resorption. The close relationship between the immune and skeletal systems, called "osteoimmunology", was coined to identify these overlapping "scientific worlds", and its function resides in the evaluation of the mutual effects of the skeletal and immune systems at the molecular and cellular levels, in both physiological and pathological states. Lipids play an essential role in skeletal metabolism and bone health. Indeed, bone marrow and its skeletal components demand a dramatic amount of daily energy to control hematopoietic turnover, acquire and maintain bone mass, and actively being involved in whole-body metabolism. Statins, the main therapeutic agents in lowering plasma cholesterol levels, are able to promote osteoblastogenesis and inhibit osteoclastogenesis. This review is meant to provide an updated overview of the pathophysiology of bone remodelling cycle, focusing on the interplay between bone, immune system and lipids. Novel therapeutic strategies for the management of osteoporosis are also discussed.

Cadmium (Cd) is a toxic heavy metal with a long biological half-life, exerting adverse effects on most tissues and organs in the human body. Inhalation, ingestion, and skin contact are the main ways of exposure to Cd. Bone is one of the target organs of Cd. The aging of the population has been considered as the reason for the high incidence rate of osteoporosis, but recent studies have emphasized that the risk of osteoporosis is related to Cd exposure. With the widespread use of Cd-containing materials in industrial and agricultural activities, the risk of Cd exposure is worrying. This review covers the epidemiological, in vivo, and in vitro studies on Cd exposure and osteoporosis. Epidemiological evidence has emphasized a positive association between Cd exposure and the occurrence rates of osteoporosis and fractures. Experimental studies have demonstrated that Cd induces osteoporosis through both direct and indirect pathways. The indirect pathway encompasses inducing renal dysfunction to impair calcium and phosphorus metabolism, whereas the direct pathway consists of directly influencing bone cells. This review aims to emphasize that Cd exposure may be an overlooked risk factor for osteoporosis and to elucidate the direct and indirect molecular mechanisms by which Cd induces osteoporosis. Understanding the pathogenesis of Cd-induced osteoporosis is crucial for the development of preventive and therapeutic strategies.

Bone has long been acknowledged as a fundamental structural entity that provides support and protection to the body's organs. However, emerging research indicates that bone plays a crucial role in the regulation of systemic metabolism. This is achieved through the secretion of a variety of hormones, cytokines, metal ions, extracellular vesicles, and other proteins/peptides, collectively referred to as bone-derived factors (BDFs). BDFs act as a medium through which bones can exert targeted regulatory functions upon various organs, thereby underscoring the profound and concrete implications of bone in human physiology. Nevertheless, there remains a pressing need for further investigations to elucidate the underlying mechanisms that inform the effects of bone on other body systems. This review aims to summarize the current findings related to the roles of these significant modulators across different organs and metabolic contexts by regulating critical genes and signaling pathways in vivo. It also addresses their involvement in the pathogenesis of various diseases affecting the musculoskeletal system, circulatory system, glucose and lipid metabolism, central nervous system, urinary system, and reproductive system. The insights gained from this review may contribute to the development of innovative therapeutic strategies through a focused approach to bone secretomes. Continued research into BDFs is expected to enhance our understanding of bone as a multifunctional organ with diverse regulatory roles in human health.

Osteoarthritis, a progressive and degenerative joint disease, disrupts the integrity of the entire joint structure, underscoring the urgency of identifying more effective therapeutic strategies and innovative targets. Among these, exercise therapy is considered a key component in the early management of osteoarthritis, functioning by stimulating the secretion of myokines from the skeletal muscle system. Irisin, a myokine predominantly secreted by skeletal muscle during exercise and encoded by the FNDC5 gene, has garnered attention for its regulatory effects on bone health. Emerging evidence suggests that irisin may play a protective role in osteoarthritis by promoting tissue homeostasis, enhancing subchondral bone density and microstructure, and inhibiting chondrocyte apoptosis. By improving chondrocyte viability, preserving extracellular matrix integrity, and maintaining homeostasis in osteoblasts, osteoclasts, and osteocytes, irisin emerges as a promising therapeutic target for osteoarthritis. This review delves into the role of irisin in osteoarthritis pathogenesis, highlighting its influence on cartilage and bone metabolism as well as its dynamic relationship with exercise. Additionally, this review suggests that further exploration on its specific molecular mechanisms, optimization of drug delivery systems, and strategic utilization of exercise-induced benefits will be pivotal in unlocking the full potential of irisin as a novel intervention for osteoarthritis.

Osteoporosis, a common metabolic condition that affects the bones, increases the risk of fractures, thereby diminishing one's quality of life and, in severe cases, can even result in life-threatening conditions. Osteoporosis is becoming increasingly prevalent worldwide as the population ages. Previous research on osteoporosis has focused on skeletal cellular components such as osteoblasts and osteoclasts. The emerging field of "osteoimmunology" has recently been introduced through new research. The concept highlights the critical impact of bone-immune system interactions on osteoporosis progression. The pathogenesis of osteoporosis is significantly influenced by T cells, particularly cytotoxic and helper T cells, which modulate osteoclast differentiation and activity. A crucial aspect of understanding osteoporosis is how T lymphocytes interact with osteoclasts. However, the precise mechanisms underlying T cell-osteoclast crosstalk remain poorly understood. This review systematically examines T cell and osteoclast involvement in osteoimmunology, with a particular focus on their involvement in osteoporosis. It seeks to elucidate the immune mechanisms driving the progression of osteoporosis and identify key molecules involved in T cell-osteoclast interactions. This aims to discover novel molecular targets and intervention strategies to improve early diagnosis and management of osteoporosis. Furthermore, this article will explore the potential of intervening in T cell-osteoclast interactions using conventional therapies, traditional Chinese medicine, immunomodulatory agents, and nanomaterial-based treatments, providing new perspectives for future osteoporosis management.

Aims: Periprosthetic osteolysis (PPO), a leading cause of aseptic loosening in joint replacement, arose from complex interactions among osteoblasts, osteoclasts, and osteocytes. Given the pivotal role of connexin 43 (Cx43) in osteocyte communication and bone remodeling, investigating its function was essential for understanding the mechanisms of osteolysis. Our previous studies showed that titanium (Ti) particles increased Cx43 expression in osteocytes. However, the role of Cx43 in osteolysis remained unclear. This study investigated the role of Cx43-mediated regulation of osteocytes on osteoclastogenesis in wear debris-induced osteolysis. Results: Using Dmp1-cre conditional Cx43 knockout mice and the MLO-Y4 osteocyte cell line, we demonstrated that Cx43 deficiency reduced bone resorption and osteoclastogenesis, thereby improving bone remodeling in a Ti particle-induced osteolysis model. Sequencing analysis revealed that Cx43 gene expression changes might be linked to oxidative stress and the Janus Kinase (JAK)-STAT pathway. Elevated Cx43 expression in osteocytes stimulated by Ti particles increased STAT1 protein phosphorylation, induced oxidative stress, elevated the Receptor Activator of Nuclear Factor Kappa-Β Ligand (RANKL)/Osteoprotegerin (OPG) ratio, and promoted osteoclast activation and bone resorption. Conversely, Cx43 gene knockout decreased STAT1 protein phosphorylation and enhanced Nuclear Factor Erythroid 2-Related Factor 2 (NrF2) protein expression. Blocking the JAK-STAT signaling pathway activated by Cx43 increased NrF2 expression, reduced reactive oxygen species levels, and subsequently decreased the RANKL/OPG ratio. Innovation and Conclusions: This study identified a novel mechanism where Cx43 in osteocytes promoted osteoclastogenesis through JAK-STAT pathway activation and oxidative stress in wear debris-induced osteolysis. These findings highlighted the critical role of Cx43 in bone resorption and suggested targeting Cx43 or the JAK-STAT pathway as potential therapeutic strategies to mitigate osteolysis and improve implant longevity. Antioxid. Redox Signal. 00, 000-000.

Background: Our previous study demonstrated that miR-140-3p induced osteocalcin expression in osteoblastic MC3T3-E1 cells. In this study, we investigated the direct effects of miR-140-3p on bone turnover in senescence-accelerated mice. Methods: In order to evaluate the effects of miR-140-3p, we formulated lipid nanoparticles (LNPs) containing miR-140-3p (100 μg/mL), with or without flotillin-2 (Flo2), a microvesicle marker excreted by osteoblasts. LNP was administered into the right tibia of the P6 strain of senescence-accelerated mice (SAMP6). Four-week-old SAMP6 males were divided into three groups: control, LNP, and LNP + Flo2. LNPs were administered five times, once every three days. No gait abnormalities were observed in any group. Two days after the last administration of LNPs, blood and urine samples were collected to measure bone turnover markers and blood chemistry and to perform urinalysis. Bone histomorphometry was performed on the left femur, contralateral to the administration site. The pancreas was removed for insulin staining of the Langerhans islets. Results: The LNP + Flo2 group showed greater bone volume, trabecular thickness, and osteoid thickness in bone histomorphometry. Carboxylated osteocalcin, a bone formation marker, was also higher in the LNP + Flo2 group, indicating that LNP + Flo2 activated osteoblastic function. Insulin levels in the islets of Langerhans did not differ across the groups, consistent with under-carboxylated osteocalcin levels. Conclusions: LNP + Flo2 effectively improved bone metabolism.

Diabetes mellitus (DM) is associated with increased bone fragility despite normal or elevated bone mineral density, partially due to the accumulation of advanced glycation end products (AGEs) in bone tissue. AGEs, such as carboxymethyl lysine (CML), impair osteocyte function by activating the receptor for advanced glycation end products (RAGE), triggering oxidative stress and inflammatory responses. This study aimed to investigate the effects of high glucose (HG) and CML on bone remodeling, glycation, inflammatory markers, and cellular functions in osteocytes. Using the murine osteocyte cell line OCY454-12H, we treated cells with HG (30 mM glucose) or 3 μM CML to simulate diabetic conditions. We assessed the expression of bone remodeling markers (SOST, RANKL, OPG, CTsK), glycation markers (RAGE, AGER1), inflammatory cytokines (IL-6, TNF-α), and cellular functions, including proliferation, viability, and apoptosis, using quantitative PCR and functional assays. HG treatment resulted in a 10-fold increase in SOST expression (9.3 vs. 0.9, p ≤ 0.0001) and a 2.4-fold increase in RANKL expression (2.75 vs. 1.15, p ≤ 0.0001), with a concurrent 2-fold increase in OPG (2.60 vs. 1.04, p ≤ 0.0001). The RANKL/OPG ratio remained unchanged (p = 0.15). HG also significantly increased RAGE expression by 3.67-fold (4.20 vs. 1.15, p ≤ 0.0001) and AGER1 by 1.65-fold (1.94 vs. 1.15, p ≤ 0.0001), along with a 2.02-fold increase in IL-6 (2.32 vs. 1.12, p ≤ 0.001) and a 7.35-fold increase in TNF-α (7.04 vs. 1.04, p ≤ 0.0001). Cell viability and proliferation were significantly higher under HG, accompanied by increased caspase-3 activity, indicating enhanced apoptosis. In contrast, CML exposure significantly upregulated RAGE (3.18 vs. 1.15, p ≤ 0.0001) and AGER1 (2.10 vs. 1.14, p = 0.028) but had no significant effects on bone remodeling markers, inflammatory cytokines, or cellular functions at physiological concentrations. Our findings demonstrate that HG disrupts osteocyte function by altering bone remodeling, glycation, and inflammatory pathways, while CML at physiological levels selectively activates glycation markers without inducing broader cellular dysfunction. These results underscore the role of the AGE-RAGE axis in diabetic bone fragility and highlight the need for future in vivo studies to explore therapeutic strategies targeting AGE accumulation and RAGE signaling in bone.

Multiple myeloma (MM) is a clonal plasma cell proliferative malignancy characterized by a debilitating bone disease. Osteolytic destruction, a hallmark of MM, is driven by increased osteoclast number and exacerbated bone resorption, primarily fueled by the excessive production of RANKL, the master regulator of osteoclast formation, within the tumor niche. We previously reported that osteocytes, the most abundant cells in the bone niche, promote tumor progression and support MM bone disease by overproducing RANKL. However, the molecular mechanisms underlying RANKL dysregulation in osteocytes in the context of MM bone disease are not entirely understood. Here, we present evidence that MM-derived CCL3 induces upregulation of RANKL expression in both human and murine osteocytes. Through a combination of in vitro, ex vivo, and in vivo models and clinical data, we demonstrate that genetic or pharmacologic inhibition of CCL3 prevents RANKL upregulation in osteocytes and attenuates the bone loss induced by MM cells. Mechanistic studies revealed that MM-derived CCL3 triggers the secretion of HMGB1 by osteocytes, a process required for osteocytic RANKL upregulation by MM cells. These findings identify a previously unknown CCL3-HMGB1 signaling axis in the MM tumor niche that drives bone resorption by promoting RANKL overproduction in osteocytes.

Drug treatments against osteoporosis are commonly divided into anti-catabolic and anabolic. Anti-catabolic drugs reduce bone turnover and increase bone mass mainly through mineralization of the existing bone matrix. Anabolic drugs, on the other hand, enhance osteoblastic activity, resulting in new bone formation. Treatments are often limited to a few years due to reported side effects, which increases fracture risk upon discontinuation. Switching to a different drug is a common strategy. However, it is not clear what is the best combination of a dual-drug therapy, the lapse between treatments and other parameters defining the combination. In this study, we conducted in silico trials to assess the efficacy of two drugs: denosumab (anti-catabolic) and romosozumab (anabolic and anti-catabolic). Our simulations indicate that starting treatment with romosozumab leads to greater bone mass gain. This is because anti-catabolic treatments reduce bone rate and, due to osteoblast-osteoclast coupling, the number of osteoblast precursors. Romosozumab increases the proliferation of these precursors, so their population should be maximised for optimal efficacy. Therefore, prior administration of an anti-catabolic drug may be counterproductive to the effectiveness of romosozumab. We also found that a rest period between treatments does not benefit bone mass gain. Furthermore, concurrent administration of romosozumab and denosumab results in greater bone mass gain and might be worth investigating in future clinical trials. Finally, we showed that reduction of fracture risk in patients undergoing sequential treatments is dose dependent and consequently, dosage could be optimised in a patient-specific manner.

Musculoskeletal health is strongly influenced by regulatory interactions of bone and muscle. Recent discoveries have identified a number of key mechanisms through which soluble factors released during exercise by bone exert positive effects on muscle and by muscle on bone. Although exercise can delay the negative effects of aging, these beneficial effects are diminished with aging. The limited response of aged muscle and bone tissue to exercise are accompanied by a failure in bone and muscle communication. Here, we propose that exercise induced beneficial factors must battle changes in circulating endocrine and inflammatory factors that occur with aging. Furthermore, sedentary behavior results in the release of negative factors impacting the ability of bone and muscle to respond to physical activity especially with aging. In this review we report on exercise responsive factors and evidence of modification occurring with aging.

Bone resorption by osteoclasts is a critical step in bone remodeling, a process important for maintaining bone homeostasis and repairing injured bone. We previously identified a bone marrow mesenchymal subpopulation, marrow adipogenic lineage precursors (MALPs), and showed that its production of RANKL stimulates bone resorption in young mice using Adipoq-Cre. To exclude developmental defects and to investigate the role of MALPs-derived RANKL in adult bone, we generated inducible reporter mice (Adipoq-CreER Tomato) and RANKL deficient mice (Adipoq-CreER RANKLflox/flox, iCKO). Single cell-RNA sequencing data analysis and lineage tracing revealed that Adipoq+ cells contain not only MALPs but also some mesenchymal progenitors capable of osteogenic differentiation. In situ hybridization showed that RANKL mRNA is only detected in MALPs, but not in osteogenic cells. RANKL deficiency in MALPs induced at 3 months of age rapidly increased trabecular bone mass in long bones as well as vertebrae due to diminished bone resorption but had no effect on the cortical bone. Ovariectomy (OVX) induced trabecular bone loss at both sites. RANKL depletion either before OVX or at 6 weeks post OVX protected and restored trabecular bone mass. Furthermore, bone healing after drill-hole injury was delayed in iCKO mice. Together, our findings demonstrate that MALPs play a dominant role in controlling trabecular bone resorption and that RANKL from MALPs is essential for trabecular bone turnover in adult bone homeostasis, postmenopausal bone loss, and injury repair.

Osseous echinococcosis, caused by Echinococcus granulosus infection, is characterized by progressive bone destruction driven by abnormal osteoclast activation. Dual-specificity phosphatase 4 (DUSP4), a key negative regulator of the MAPK pathway, inhibits osteoclast differentiation and bone resorption. This study aimed to elucidate the role of DUSP4 in E. granulosus-induced bone loss.

In vitro, a co-culture system of E. granulosus protoscoleces (PSCs) and bone marrow-derived macrophages (BMMs) was established. Osteoclast differentiation and bone resorption were assessed using TRAP staining and F-actin immunofluorescence. Transcriptome sequencing identified DUSP4 as a key regulator. DUSP4 overexpression was performed to evaluate its effects on osteoclast markers and MAPK signaling (ERK, JNK, p38). In vivo, a mouse model of osseous echinococcosis was developed, and DUSP4 overexpression was achieved via lentiviral transduction. Bone destruction was analyzed using X-ray, micro-CT, and histology.

PSCs significantly enhanced osteoclast differentiation and bone resorption, upregulated osteoclast markers (CTSK, NFATc1), and activated MAPK signaling. DUSP4 overexpression reversed these effects, reducing osteoclast activity and MAPK phosphorylation. In vivo, PSC infection caused severe bone destruction, which was mitigated by DUSP4 overexpression.

This study reveals the molecular mechanism by which Echinococcus granulosus drives abnormal osteoclast activation through the DUSP4-MAPK signaling axis. Parasitic infection suppresses DUSP4 expression, relieving its negative regulation of the MAPK pathway and leading to excessive osteoclast differentiation. Restoring DUSP4 expression effectively reverses abnormal MAPK pathway activation, reducing osteoclast bone resorption activity to physiological levels. These findings not only provide new insights into the pathological mechanisms of bone destruction in osseous echinococcosis but also establish DUSP4 as a critical therapeutic target for pathological bone resorption, laying the groundwork for host-directed treatment strategies for parasitic bone diseases.

Bone remodeling is a dynamic and continuous process involving three components: bone formation mediated by osteoblasts, bone resorption mediated by osteoclasts, and bone formation-resorption balancing regulated by osteocytes. Excessive osteocyte death is found in various bone diseases, such as postmenopausal osteoporosis (PMOP), and osteoclasts are found increased and activated at osteocyte death sites. Currently, apart from apoptosis and necrosis as previously established, more forms of cell death are reported, including necroptosis, ferroptosis and pyroptosis. These forms of cell death play important role in the development of inflammatory diseases and bone diseases. Increasing studies have revealed that various forms of osteocyte death promote osteoclast formation via different mechanism, including actively secreting pro-inflammatory and pro-osteoclastogenic cytokines, such as tumor necrosis factor alpha (TNF-α) and receptor activator of nuclear factor-kappa B ligand (RANKL), or passively releasing pro-inflammatory damage associated molecule patterns (DAMPs), such as high mobility group box 1 (HMGB1). This review summarizes the established and potential mechanisms by which various forms of osteocyte death regulate osteoclast formation, aiming to provide better understanding of bone disease development and therapeutic target.

Little is known about how tissues mediate the ability to selectively form or resorb bone, as required during orthodontic tooth movement (OTM), facial growth, continued tooth eruption and for healing after fractures, maxillofacial surgical repositioning or implant dentistry. OTM has the unique ability to selectively cause apposition, resorption or a combination of both at the alveolar periosteal surface and therefore, provides an optimal process to study the regulation of bone physiology at a tissue level. Our aim was to elucidate the mechanisms and signaling pathways of the bone remodeling regulatory system (BRRS) as well as to investigate its clinical applications in osteoporosis treatment, orthopedic surgery, fracture management and orthodontic treatment. OTM is restricted to a specific range in which the BRRS permits remodeling; however, surpassing this limit may lead to bone dehiscence. Low-intensity pulsed ultrasound, vibration or photobiomodulation with low-level laser therapy have the potential to modify BRRS with the aim of reducing bone dehiscence and apical root resorption or accelerating OTM. Unloading of bone and periodontal compression promotes resorption via receptor activator of nuclear factor κB-ligand, monocyte chemotactic protein-1, parathyroid hormone-related protein (PTHrP), and suppression of anti-resorptive mediators. Furthermore, proinflammatory cytokines, such as interleukin-1 (IL-1), IL-6, IL-8, tumor necrosis factor-α, and prostaglandins exert a synergistic effect on bone resorption. While proinflammatory cytokines are associated with periodontal sequelae such as bone dehiscence and gingival recessions, they are not essential for OTM. Integrins mediate mechanotransduction by converting extracellular biomechanical signals into cellular responses leading to bone apposition. Active Wnt signaling allows β-catenin to translocate into the nucleus and to stimulate bone formation, consequently converging with integrin-mediated mechanotransductive signals. During OTM, periodontal fibroblasts secrete PTHrP, which inhibits sclerostin secretion in neighboring osteocytes via the PTH/PTHrP type 1 receptor interaction. The ensuing sclerostin-depleted region may enhance stem cell differentiation into osteoblasts and subperiosteal osteoid formation. OTM-mediated BRRS modulation suggests that administering sclerostin-inhibiting antibodies in combination with PTHrP may have a synergistic bone-inductive effect. This approach holds promise for enhancing osseous wound healing, treating osteoporosis, bone grafting and addressing orthodontic treatments that are linked to periodontal complications.

Over the past decade, advancements in cell line development and genetic tools have led to a wealth of new insights into the effects of parathyroid hormone (PTH), particularly on osteocytes-cells deeply embedded within the bone's mineral matrix. These cells were once believed to be inactive bystanders, with little, if any, role in skeletal and mineral homeostasis. This concept of passive osteocytes has been challenged in recent years and osteocytes are now recognized for their crucial functions in skeletal mechanotransduction, bone modeling and remodeling, mineral ion regulation, and hematopoiesis. Moreover, osteocytes are key targets of PTH, and studies utilizing genetically modified mice, in which the PTH receptor is either deleted or overexpressed, have shed light on the hormone's complex effects on these cells. Several signaling molecules, including salt kinase inhibitors and histone deacetylases, have been identified as part of PTH's intracellular signaling cascade. In addition to its effects on bone metabolism, PTH has also been implicated in regulating bone energy metabolism, skeletal aging, and hematopoiesis. This review summarizes both classical and emerging effects of PTH on osteocytes, highlights the limitations of current research, and offers perspectives for future investigations in the field.

Bone disorders affect more than half of the adult population worldwide who may have a poor quality of life and physical independence worldwide. Multi-omic techniques are increasingly adopted and applied to determine the molecular mechanisms of bone tissue repair, providing perspective towards personalized medical intervention. Data from genomics, epigenomics, transcriptomics, proteomics, glycomics, and lipidomics were combined to elucidate dynamic processes in bone repair. In this narrative review, the key role of genetic and epigenetic factors in regulating injured cellular responses is highlighted, and changes in RNA and protein expression during the healing phase, as well as glucolipid metabolism adaptation, are described in detail how the repair process is affected. In a word, the integration of multi-omic techniques in this review not only benefits the comprehensive identification of new biomarkers, but also facilitates the development of personalized treatment strategies of bone disorders to revolutionize regenerative medicine.

Our study elucidates the crucial role of the liver in bone homeostasis through the p53-miR17 family (miR17-miR20/miR20-miR106/miR93-miR106)-Rankl axis. We demonstrate the enhanced hepatocyte Rankl expression in inflammaging conditions, such as aging, ovariectomized (OVX) mice, and elderly humans. Mice with hepatocyte-specific Rankl deletion exhibit significant resistance to bone mass loss associated with aging, lipopolysaccharide (LPS)-induced inflammation, or estrogen deficiency, compared with controls. Our study highlights hepatocytes as the primary source of Rankl in the liver and serum under these conditions. We identify the p53-miR17 family axis as a crucial regulator for hepatocyte Rankl expression, with p53 inhibiting the miR17 family transcription. Through bioinformatics analysis and in vitro validation, we identify Rankl mRNA as a direct target of the miR17 family. Targeting this axis via CasRx-mediated mRNA editing or miRNA interference significantly attenuates bone mass loss in mice. Our investigation underscores the pivotal significance and therapeutic potential of modulating the p53-miR17 family-Rankl axis in the treatment of inflammaging-associated osteoporosis.

Mechanical and thermal cell damage can occur due to invasive procedures related to drilling, the insertion of dental implants, and periodontal treatments. Necrotic cells release the content of their cytoplasm and membrane fragments, thereby signaling the need for repair, which includes bone resorption by osteoclasts and inflammation. Here we screened lysates from human gingival fibroblasts, HSC2 and TR146 oral squamous carcinoma cell lines, as well as murine IDG-SW3 osteocytic and RAW264.7 macrophage cell lines for their potential to modulate in vitro osteoclastogenesis in murine bone marrow cultures. We also tested the impact of necrotic lysates on modulating the expression of inflammatory cues in murine ST2 bone marrow stromal cells. We report here that independent of human or murine origin, all cell lysates significantly reduced in vitro osteoclastogenesis in bone marrow cultures, as indicated by the expression of the osteoclast marker genes cathepsin K and tartrate-resistant acid phosphatase and the respective histochemical staining in multinucleated cells. We also found that lysates from HSC2 and TR146 cells significantly pushed the expression of CCL2, CCL5, CXCL1, IL1, and IL6 in ST2 cells. These findings suggest that oral cell lysates reduce in vitro osteoclastogenesis, but only damaged oral squamous carcinoma cells can force murine stromal cells to produce an inflammatory environment.

Bone is a unique organ crucial for locomotion, mineral metabolism, and hematopoiesis. It maintains homeostasis through a balance between bone formation by osteoblasts and bone resorption by osteoclasts, which is regulated by the basic multicellular unit (BMU). Abnormal bone metabolism arises from an imbalance in the BMU. Osteoclasts, derived from the monocyte-macrophage lineage, are regulated by the RANKL-RANK-OPG system, which is a key factor in osteoclast differentiation. RANKL activates osteoclasts through its receptor RANK, while OPG acts as a decoy receptor that inhibits RANKL. In trabecular bone, high turnover involves rapid bone formation and resorption, influenced by conditions such as malignancy and inflammatory cytokines that increase RANKL expression. Cortical bone remodeling, regulated by aged osteocytes expressing RANKL, is less understood, despite ongoing research into how Rett syndrome, characterized by MeCP2 abnormalities, affects RANKL expression. Balancing trabecular and cortical bone involves mechanisms that preserve cortical bone, despite overall bone mass reduction due to aging or oxidative stress. Research into genes like sFRP4, which modulates bone mass, highlights the complex regulation by BMUs. The roles of the RANKL-RANK-OPG system extend beyond bone, affecting processes such as aortic valve formation and temperature regulation, which highlight the interconnected nature of biological research.

Bone remodeling is regulated by the interaction between receptor activator of nuclear factor kappa-B ligand (RANKL) and its receptor RANK on osteoblasts and osteoclasts, respectively. Osteoprotegerin (OPG) is secreted from osteoblasts and inhibits osteoclast differentiation by acting as a decoy receptor for RANKL. Despite its importance, the mechanism underlying the secretion of OPG remains poorly understood. Here, we applied a method of video-rate bioluminescence imaging using a fusion protein with Gaussia luciferase (GLase) and visualized the secretion of OPG from living mouse osteoblastic MC3T3-E1 cells. The bioluminescence imaging revealed that the secretion of OPG fused to GLase (OPG-GLase) occurred frequently and widely across the cell surface. Notably, co-expression of RANKL significantly reduced the secretion of OPG-GLase, indicating an inhibitory role of RANKL on OPG secretion within cells. Further imaging and biochemical analyses using deletion mutants of OPG and RANKL, as well as RANKL mutants that cause autosomal recessive osteopetrosis, demonstrated the essential role of protein-protein interaction between OPG and RANKL in the inhibition of OPG secretion. Treatment with proteasome inhibitors resulted in increased levels of OPG in both culture medium and cell lysates. However, the fold-increase of OPG was similar regardless of the presence or absence of RANKL, suggesting that the regulation of OPG secretion by RANKL is independent of proteasome activity. This report visualized the secretion of OPG from living cells and provided evidence for a novel intracellular inhibitory effect of RANKL on OPG secretion.

In this study, we developed a hydrostatic pressurizing chamber capable of applying hydrostatic pressure to osteocytic spheroids derived from mouse osteoblastic MC3T3-E1 cells. Our results demonstrate that a 4-h exposure to 200 kPa of hydrostatic pressure did not alter the apparent morphology of the spheroids. However, gene expression analysis revealed a significant up-regulation of Sost, a marker of late-stage osteocyte differentiation.

Tumor necrosis factor-alpha (TNF-α) is a significant cytokine that regulates bone resorption under inflammatory conditions. However, its mechanism of action in osteocytes remains unclear. In this study, highly purified osteocytes were isolated from dentin matrix protein 1 (DMP1)-Topaz mice using cell sorter. RNA sequencing (RNA-seq) revealed that TNF-α stimulation increased C-X-C motif chemokine ligand 10 (CXCL10) gene expression in osteocytes. Although CXCL10 did not affect osteoclast differentiation in vitro, it enhanced the migration of osteoclast precursors. Additionally, in the transwell co-culture system, TNF-α induced the migration of osteoclast precursors. However, this effect was attenuated by a CXCL10-neutralizing antibody. In vivo, mice were administered supracalvarial injections of TNF-α with or without the CXCL10-neutralizing antibody for 5 days. The percentage of CXCL10-positive osteocytes increased after TNF-α administration. Additionally, osteoclast formation and bone resorption were assessed. CXCL10-neutralizing antibody-treated calvariae exhibited a significantly lower number of osteoclasts and bone resorption than those treated with TNF-α alone. These results indicated that TNF-α-induced CXCL10, which affects the migration of osteocyte-derived osteoclast precursors, may enhance TNF-α-triggered osteoclast formation and bone resorption in vivo.

Craniometaphyseal dysplasia (CMD), a rare craniotubular disorder, occurs in an autosomal dominant (AD) or autosomal recessive (AR) form. CMD is characterized by hyperostosis of craniofacial bones and metaphyseal flaring of long bones. Many patients with CMD suffer from neurological symptoms. The pathogenesis of CMD is not fully understood. Treatment is limited to craniofacial surgery. Here, we report a knock in (KI) mouse model for AR CMD carrying a Cx43R239Q mutation. Cx43KI/KI mice replicate typical features of AR CMD, including thickening of craniofacial bones, club-shaped femurs, and widened diaphyseal cortical bones. Female Cx43KI/KI mice display remarkably more bone overgrowth than male Cx43KI/KI mice as they age. In contrast to Cx43+/+ littermates, Cx43KI/KI mice exhibit periosteal bone deposition and increased osteoclast (OC) numbers in the endosteum of long bones. Although formation of resting OCs in Cx43+/+ and Cx43KI/KI mice is comparable, the actively resorbing Cx43KI/KI OCs have reduced resorption on bone chips. Cx43KI/KI mice display reduced osteocyte dendrites. RNA from Cx43KI/KI femoral cortical bones show reduced expression levels of Sost, Tnf-α, IL-1β, Esr1, Esr2, and a lower Rankl/Opg ratio. Moreover, the Cx43R239Q mutation results in altered spatial expression of Cx43 protein and mild reduction of gap junction and hemichannel activity. The distinct phenotype seen in Cx43KI/KI mice but not in Cx43 ablation models suggests that Cx43 loss-of-function is unlikely the main cause of AR CMD. Additional studies are required to investigate new roles of CMD-mutant Cx43.

The significance of biomedical applications of bio-functional niobium (Nb)-based metallic biomaterials is underscored by their potential utilization in implant application. Nb-based metallic materials present reliable physicomechanical and biological properties, thus represent materials highly suitable for implant application. This review provides an overview on the advances of pure niobium and Nb-based metallic materials as implant materials over the past 20 years, and highlights the advantages of Nb-based metallic biomaterials for implant application in terms of their physicomechanical properties, corrosion resistance in biological media, magnetic resonance imaging (MRI) compatibility, cell compatibility, blood compatibility, osteogenesis, and bioactivity. An introduction is provided for the production and processing techniques for Nb-based metallic biomaterials, including traditional melting processes like vacuum arc remelting, additive manufacturing like selective laser melting (SLM), electron beam melting (EBM), spark plasma sintering (SPS), and severe plastic deformation like equal channel angular pressing (ECAP), multi-axial forging (MAF), high pressure torsion (HPT), as well as their physicomechanical properties and implant application. Also suggested are the critical issues, challenges, and prospects in the further development of Nb-based metallic biomaterials for implant applications. STATEMENT OF SIGNIFICANCE: Nb-based biomaterials have gained significant interest for bioimplantable scaffolds because of their appropriate mechanical characteristics and biocompatibility. No prior work has been published specifically reviewing bio-functional Nb-based biomaterials for exploring their physicomechanical properties, biological significance, and implant applications. This review provides an overview on the advances of niobium and Nb-based materials as implant materials over the past 20 years, and highlights the advantages of Nb-based biomaterials for implant application. An introduction is provided for the production and processing techniques for Nb-based biomaterials, as well as their physicomechanical properties and implant application. Also suggested are the critical issues, challenges, and prospects in the further development of Nb-based biomaterials for implant applications.

This study assessed the novel concept that osteoclast-derived Grem1 has regulatory functions in the skeletal response to calcium stress using an osteoclastic Grem1 conditional knockout (cKO) mouse model. The calcium stress was initiated by feeding cKO mutants and wildtype (WT) littermates a calcium-deficient diet for 2 weeks. Deletion of Grem1 in mature osteoclasts did not affect developmental bone growth nor basal bone turnover. In response to calcium depletion, male cKO mutants showed greater increases in osteoclastic resorption and trabecular bone loss than male WT littermates, indicating an enhanced skeletal sensitivity to calcium depletion in male mutants. The enhanced sensitivity to calcium depletion was sex-dependent, as female cKO mutants showed lower increases in osteoclastic resorption and bone loss than female WT littermates as well as male cKO mutants. The sex disparity in osteoclastic resorption response to calcium stress was intrinsic to osteoclasts since osteoclasts of male but not female cKO mutants showed greater in vitro bone resorption activity than osteoclasts of WT littermates of respective sex. Male cKO mutants displayed smaller bone formation response to calcium depletion than male WT littermates, while female mutants showed bigger bone formation response than female WT littermates, indicating that cKO mutants also displayed sex disparity in bone formation response. The sex disparity in bone formation response was not caused by intrinsic differences in osteoblasts but might be due to sex-dependent differential osteoclastic release of osteogenic factors. In summary, osteoclast-derived gremlin-1 has complicated and sex-dependent regulatory roles in skeletal response to calcium stress.

Bone remodeling maintains the robustness of the bone tissue by balancing bone resorption by osteoclasts and bone formation by osteoblasts. Although these cells together play a crucial role in bone remodeling, only a few reports are available on the common factors involved in the differentiation of the two types of cells. Here, we show family with sequence similarity 102 member A (Fam102a) as a bone-remodeling factor that positively regulates both osteoclast and osteoblast differentiation. Fam102a regulates osteoblast differentiation by controlling recombination signal binding protein for immunoglobulin κ J region-like (Rbpjl). The Fam102a-Rbpjl axis promotes the nuclear translocation of transcription factors and enhances the expression of Osterix, a transcription factor essential for osteoblast differentiation. The deletion of Fam102a or a functional mutation in Rbpjl leads to osteopenia accompanied by reduced osteoblastic bone formation. Thus, the Fam102a-Rbpjl axis plays an important role in osteoblasts and this finding provides insights into bone remodeling.

Osteocytes reside within the bone matrix and produce both paracrine and endocrine factors that influence the skeleton and other tissues. Despite their abundance and physiological importance, osteocytes have been challenging to study in vitro because they are difficult to extract and purify and do not retain their phenotype in standard culture conditions. New techniques for this purpose are emerging. This chapter will describe two methods and adaptations we use to study osteocytes: (1) isolating and purifying primary osteocytes from murine bone, with and without hematopoietic lineage depletion and (2) differentiating cultured osteoblasts or osteoblast-lineage cell lines (including cell lines termed "osteocytic") until they reach a stage of osteocytic gene expression. We will also discuss the limitations of these methods and possible directions for future improvements.

Iron overload leads to apoptosis and increased expression of receptor activator of nuclear factor kappa-Β ligand (RANKL) in osteocytes, which in turn accelerates osteoclastogenesis. Since osteocytes are the main RANKL producers, we hypothesized that apoptotic osteocytes increase RANKL expression in osteocytes, which in turn stimulates osteoclastogenesis and bone resorption. In this study, alendronate or IG9402, a bisphosphonate (BP) analog which does not inhibit bone resorption, inhibited iron overload-induced osteocyte apoptosis and increased RANKL expression. Both BPs also prevented osteoblast apoptosis but did not inhibit the increase in osteoblastic RANKL. Alendronate, but not IG9402, prevented the increase in osteoclastogenesis and serum levels of the bone resorption marker C-telopeptide of type I collagen (CTX) in iron-overloaded mice. Alendronate also prevented the iron overload-induced reduction in femoral bone mineral density, disruption of bone microstructure, and weakness of bone strength. Although IG9402 did not prevent bone loss due to iron overload, it partially prevented reduction of strength, suggesting that osteocyte viability contributes to the maintenance of bone strength. In conclusion, although osteocyte apoptosis in the presence of iron overload leads to an increase in osteocytic RANKL production. However, blocking these events was not sufficient to inhibit iron overload-induced osteoclastogenesis and bone loss.

During reproduction, multiple species such as insects and all mammals undergo extensive physiological and morphological adaptions to ensure health and survival of the mother and optimal development of the offspring. Here we report that the intestinal epithelium undergoes expansion during pregnancy and lactation in mammals. This enlargement of the intestinal surface area results in a novel geometry of expanded villi. Receptor activator of nuclear factor-κΒ (RANK, encoded by TNFRSF11A) and its ligand RANKL were identified as a molecular pathway involved in this villous expansion of the small intestine in vivo in mice and in intestinal mouse and human organoids. Mechanistically, RANK-RANKL protects gut epithelial cells from cell death and controls the intestinal stem cell niche through BMP receptor signalling, resulting in the elongation of villi and a prominent increase in the intestinal surface. As a transgenerational consequence, babies born to female mice that lack Rank in the intestinal epithelium show reduced weight and develop glucose intolerance after metabolic stress. Whereas gut epithelial remodelling in pregnancy/lactation is reversible, constitutive expression of an active form of RANK is sufficient to drive intestinal expansion followed by loss of villi and stem cells, and prevents the formation of Apcmin-driven small intestinal stem cell tumours. These data identify RANK-RANKL as a pathway that drives intestinal epithelial expansion in pregnancy/lactation, one of the most elusive and fundamental tissue remodelling events in mammalian life history and evolution.