Aims. Currently, the effect of drug treatment for osteoporosis is relatively poor, and the side effects are numerous and serious. Melatonin is a potential drug to improve bone mass in postmenopausal women. Unfortunately, the mechanism by which melatonin improves bone metabolism remains unclear. The aim of this study was to further investigate the potential mechanism of melatonin in the treatment of osteoporosis. Methods. The effects of melatonin on mitochondrial apoptosis protein, bmal1 gene, and related pathway proteins of RAW264.7 (mouse mononuclear macrophage leukaemia cells) were analyzed by western blot. Cell Counting Kit-8 was used to evaluate the effect of melatonin on cell viability. Flow cytometry was used to evaluate the effect of melatonin on the apoptosis of RAW264.7 cells and mitochondrial membrane potential. A reactive oxygen species (ROS) detection kit was used to evaluate the level of ROS in osteoclast precursors. We used bmal1-small interfering RNAs (siRNAs) to downregulate the Bmal1 gene. We established a postmenopausal mouse model and verified the effect of melatonin on the bone mass of postmenopausal osteoporosis in mice via micro-CT. Bmal1 lentiviral activation particles were used to establish an in vitro model of overexpression of the bmal1 gene. Results. Melatonin promoted apoptosis of RAW264.7 cells and increased the expression of BMAL1 to inhibit the activation of ROS and phosphorylation of mitogen-activated protein kinase (MAPK)-p38. Silencing the bmal1 gene weakened the above effects of melatonin. After that, we used dehydrocorydaline (DHC) to enhance the activation of MAPK-p38, and the effects of melatonin on reducing ROS levels and promoting apoptosis of RAW264.7 cells were also blocked. Then, we constructed a mouse model of postmenopausal osteoporosis and administered melatonin. The results showed that melatonin improves bone loss in ovariectomized mice. Finally, we established a model of overexpression of the bmal1 gene, and these results suggest that the bmal1 gene can regulate ROS activity and change the level of the MAPK-p38 signalling pathway. Conclusion. Our study confirmed that melatonin promotes the apoptosis of RAW264.7 cells through BMAL1/ROS/MAPK-p38, and revealed the therapeutic effect and mechanism of melatonin in postmenopausal osteoporosis. This finding enriches BMAL1 as a potential target for the treatment of osteoporosis and the pathogenesis of postmenopausal osteoporosis. Cite this article: Bone Joint Res 2023;12(11):677–690

Aims. We aimed to develop a gene signature that predicts the occurrence of postmenopausal osteoporosis (PMOP) by studying its genetic mechanism. Methods. Five datasets were obtained from the Gene Expression Omnibus database. Unsupervised consensus cluster analysis was used to determine new PMOP subtypes. To determine the central genes and the core modules related to PMOP, the weighted gene co-expression network analysis (WCGNA) was applied. Gene Ontology enrichment analysis was used to explore the biological processes underlying key genes. Logistic regression univariate analysis was used to screen for statistically significant variables. Two algorithms were used to select important PMOP-related genes. A logistic regression model was used to construct the PMOP-related gene profile. The receiver operating characteristic area under the curve, Harrell’s concordance index, a calibration chart, and decision curve analysis were used to characterize PMOP-related genes. Then, quantitative real-time polymerase chain reaction (qRT-PCR) was used to verify the expression of the PMOP-related genes in the gene signature. Results. We identified three PMOP-related subtypes and four core modules. The muscle system process, muscle contraction, and actin filament-based movement were more active in the hub genes. We obtained five feature genes related to PMOP. Our analysis verified that the gene signature had good predictive power and applicability. The outcomes of the GSE56815 cohort were found to be consistent with the results of the earlier studies. qRT-PCR results showed that RAB2A and FYCO1 were amplified in clinical samples. Conclusion. The PMOP-related gene signature we developed and verified can accurately predict the risk of PMOP in patients. These results can elucidate the molecular mechanism of RAB2A and FYCO1 underlying PMOP, and yield new and improved treatment strategies, ultimately helping PMOP monitoring. Cite this article: Bone Joint Res 2022;11(8):548–560

Aims. This study explored the shared genetic traits and molecular interactions between postmenopausal osteoporosis (POMP) and sarcopenia, both of which substantially degrade elderly health and quality of life. We hypothesized that these motor system diseases overlap in pathophysiology and regulatory mechanisms. Methods. We analyzed microarray data from the Gene Expression Omnibus (GEO) database using weighted gene co-expression network analysis (WGCNA), machine learning, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis to identify common genetic factors between POMP and sarcopenia. Further validation was done via differential gene expression in a new cohort. Single-cell analysis identified high expression cell subsets, with mononuclear macrophages in osteoporosis and muscle stem cells in sarcopenia, among others. A competitive endogenous RNA network suggested regulatory elements for these genes. Results. Signal transducer and activator of transcription 3 (STAT3) was notably expressed in both conditions. Single-cell analysis pinpointed specific cells with high STAT3 expression, and microRNA (miRNA)-125a-5p emerged as a potential regulator. Experiments confirmed the crucial role of STAT3 in osteoclast differentiation and muscle proliferation. Conclusion. STAT3 has emerged as a key gene in both POMP and sarcopenia. This insight positions STAT3 as a potential common therapeutic target, possibly improving management strategies for these age-related diseases. Cite this article: Bone Joint Res 2024;13(8):411–426

Objectives

Insufficient protein ingestion may affect muscle and bone mass, increasing the risk of osteoporotic fractures in the elderly, and especially in postmenopausal women. We evaluated how a low-protein diet affects bone parameters under gonadal hormone deficiency and the improvement led by hormone replacement therapy (HRT) with 17β-oestradiol.

Aims

This study examined whether systemic administration of melatonin would have different effects on osseointegration in ovariectomized (OVX) rats, depending on whether this was administered during the day or night.

Purpose of the study: Oestrogen depletion leads to osteoclastic hyperactivity and subsequent postmenopausal osteoporosis. Little is known about interactions with bone absorption induced by wear particles from joint bearings. The purpose of this study was to evaluate bone response to polyethylene (PE) particles in a mouse model of oestrogen deficiency. Material and methods: Particles of PE were implanted in the calvaria of seven non-ovariectomised mice and in seven ovariectomised mice (OVX). Fourteen mice were operated on without implantation of the particles (7 non-OVX and 7 OVX, control groups). The mice were sacrificed at two weeks. The crania were studied under a microscanner and histologically without decalcification. Results: The microscanner showed that particles of PE induced a significant decrease in bone thickness in non-OVX mice (p=0.04), while the thickness remained unchanged in OVX mice who had received the particles (p=0.40). After implantation of the PE particles, the number of osteoclasts per mm of bone perimeter was 2.84±1.6 in the non-OVX mice and 1.74±1.3 in OVX mice (p=0.004). Compared with controls, the mean loss of bone was 12±10% in the non-OVX mice versus 4.7±0.9%in the OVX mice (p=0.004). Discussion: The volume of osteolysis induced by PE particles was smaller in OVX mice compared with non-OVX mice. Conclusion: These results suggest that a deficit in oestrogens has a protective effect against bone adsorption induced by PE particles

Aims

Transcription factor nuclear factor kappa B (NF-κB) plays a major role in the pathogenesis of chronic inflammatory diseases in all organ systems. Despite its importance, NF-κB targeted drug therapy to mitigate chronic inflammation has had limited success in preclinical studies. We hypothesized that sex differences affect the response to NF-κB treatment during chronic inflammation in bone. This study investigated the therapeutic effects of NF-κB decoy oligodeoxynucleotides (ODN) during chronic inflammation in male and female mice.

Aims: Compelling evidences suggest that increased production of osteoclastogenic cytokines by activated T cells plays a relevant role in the bone loss induced by estrogen (E) deficiency in the mouse. However, little information is available on the role of T cells in post-menopausal bone loss in humans. Methods: To investigate this issue we have assessed the production of cytokines involved in osteoclasts (OCs) formation (RANKL, TNFα and OPG), in vitro Ocs formation in pre and postmenopausal women, the latter with or without osteoporosis. We also evaluate OC precursors in peripheral blood and the ability of peripheral blood mononuclear cells to produce TNFα in both basal and stimulated condition by flow cytometry in these subjects. Results: Our data demonstrate that E enhances the production of the pro-osteoclastogenetic cytokines TNF alpha and RANKL and increases the number of circulating OCs precursors. Furthermore we show that T cells and monocytes from women with osteoporosis exhibit a higher production of TNF α then those from the other two groups. Conclusions: Our findings suggest that E deficiency stimulates OCs formation both by increasing the production of TNF and RANKL and increasing the number of OCs precursors. Women with postmenopausal osteoporosis have a higher T cells activity than healthy postmenopausal subjects, T cells thus contribute to the bone loss induced by E deficiency in humans as they do in the mouse

Aims

Osteoporosis (OP) is a metabolic bone disease, characterized by a decrease in bone mineral density (BMD). However, the research of regulatory variants has been limited for BMD. In this study, we aimed to explore novel regulatory genetic variants associated with BMD.

Bone regeneration and repair are crucial to ambulation and quality of life. Factors such as poor general health, serious medical comorbidities, chronic inflammation, and ageing can lead to delayed healing and nonunion of fractures, and persistent bone defects. Bioengineering strategies to heal bone often involve grafting of autologous bone marrow aspirate concentrate (BMAC) or mesenchymal stem cells (MSCs) with biocompatible scaffolds. While BMAC shows promise, variability in its efficacy exists due to discrepancies in MSC concentration and robustness, and immune cell composition. Understanding the mechanisms by which macrophages and lymphocytes – the main cellular components in BMAC – interact with MSCs could suggest novel strategies to enhance bone healing. Macrophages are polarized into pro-inflammatory (M1) or anti-inflammatory (M2) phenotypes, and influence cell metabolism and tissue regeneration via the secretion of cytokines and other factors. T cells, especially helper T1 (Th1) and Th17, promote inflammation and osteoclastogenesis, whereas Th2 and regulatory T (Treg) cells have anti-inflammatory pro-reconstructive effects, thereby supporting osteogenesis. Crosstalk among macrophages, T cells, and MSCs affects the bone microenvironment and regulates the local immune response. Manipulating the proportion and interactions of these cells presents an opportunity to alter the local regenerative capacity of bone, which potentially could enhance clinical outcomes.

Cite this article: Bone Joint Res 2024;13(9):462–473.

Aims

Insufficient treatment response in rheumatoid arthritis (RA) patients requires novel treatment strategies to halt disease progression. The potential benefit of combination of cytokine-inhibitors in RA is still unclear and needs further investigation. To explore the impact of combined deficiency of two major cytokines, namely interleukin (IL)-1 and IL-6, in this study double deficient mice for IL-1αβ and IL-6 were investigated in different tumour necrosis factor (TNF)-driven inflammatory bone disorders, namely peripheral arthritis and sacroiliitis, as well as systemic bone loss.

Aims

The association of auraptene (AUR), a 7-geranyloxycoumarin, on osteoporosis and its potential pathway was predicted by network pharmacology and confirmed in experimental osteoporotic mice.

Aims

The role of N,N-dimethylformamide (DMF) in diabetes-induced osteoporosis (DM-OS) progression remains unclear. Here, we aimed to explore the effect of DMF on DM-OS development.

Introduction: The evidence of genetic background as an important causative factor in disc degeneration and osteoporosis is increasing. Defects in the COL2A1 gene coding for type II collagen are known to lead to disturbed chondrogenesis and ossification. Retardation of growth, abnormal shape of vertebral bodies and intervertebral discs and occult spina bifida have been described in young mice with the defect. How the gene defect is manifested later in life has not been described. Purpose of the study: The purpose of this study was to describe, at the microscopic level, the structure of intervertebral discs of transgenic Del1 mice carrying a deletion mutation in the Col2a1 gene, and the effect of the gene defect on the structural properties of bone. In addition, we wanted to see how the gene defect manifests in disc tissue and skeletal bone later in life and if there were differences between sexes. Materials and methods: The study material consisted of transgenic male (n=27) and female (n=21) mice and their age-matched littermate controls (n=22 and 21, respectively). The transgenic mice were offspring of the transgenic founder mouse Del1 harbouring six copies of a mouse type II collagen transgene with a 150-bp deletion. The mice were divided into two age groups, the younger group being 3 to 13 months and the older 15 to 21 months of age. The two major macromolecules of the intervertebral discs, proteoglycans (PGs) and collagen, were studied. The PG concentration of the intervertebral discs’ nucleus pulposus, annulus fibrosus, and the vertebral bodies and end plates was measured from Safranin-O-stained sections using digital densitometry. Collagen orientation of these structures was evaluated using quantitative polarised light microscopy. Bone mineral density (BMD) was measured with dual energy x ray absorptiometry (DXA), and the breaking force of the femoral bone with three point bending test only for nine 14-month-old females (four control mice and five with gene defect) and fourteen 14-month-old male mice (six control mice and eight with gene defect). Results: In the young mice, there were no changes in the measured parameters in the intervertebral discs due to the gene defect. However, Safranin-O density and thus PG concentration of the vertebral trabecular bone was 47 % lower in the young transgenic female mice than in the controls (p< 0.001). Ageing had a significant effect on the measured parameters. The Safranin-O density in the nucleus pulposus of the old transgenic male mice was 35 % higher than in the age-matched controls (p< 0.05). In the females, however, Safranin-O density in the nucleus pulposus was 53 % (p< 0.01) and in the vertebral bone 68 % (p< 0.01) lower in the transgenic mice than in the controls. The Safranin-O density in the annulus fibrosus of the transgenic female mice was not changed as compared to the controls. The collagen orientation in the nucleus pulposus of old transgenic male mice was 27 % higher than in the age-matched controls (p< 0.05). In the old females there was no difference in the collagen orientation of the nucleus pulposus between the transgenic mice and controls but in the annulus fibrosus the orientation was 41 % (p< 0.01) and in the vertebral bone 70 % (p< 0.05) lower in the transgenic mice than in the controls. There was no difference in the BMD and the breaking force of the femurs of 14-month-old male mice as compared with the age-matched controls. However, in the old transgenic female mice, the femoral BMD was 14 % (p=0.05) and the breaking force 27 % (p=0.09) lower than in the controls. Conclusions: The transgene of the Col2a1 gene caused a decrease in the nucleus pulposus PG concentration and in the annulus fibrosus collagen orientation in the old female mice. These features can compromise the structural and load-bearing properties of the discs and thus predispose to disc degeneration. Interestingly enough, the male mice seemed to benefit from the genetic defect in this respect. In addition, in the old transgenic female mice, the PG concentration and the collagen orientation of the vertebral trabecular bone were decreased which contributed to the loss of BMD and breaking force of bone seen in these mice. The fact, that these differences in the bone were not seen in the male mice suggests that this animal model could possibly be used in studies of postmenopausal osteoporosis

Aims

Osteoarthritis (OA) is prevalent among the elderly and incurable. Intra-articular parathyroid hormone (PTH) ameliorated OA in papain-induced and anterior cruciate ligament transection-induced OA models; therefore, we hypothesized that PTH improved OA in a preclinical age-related OA model.

Aims

The effect of the gut microbiota (GM) and its metabolite on bone health is termed the gut-bone axis. Multiple studies have elucidated the mechanisms but findings vary greatly. A systematic review was performed to analyze current animal models and explore the effect of GM on bone.

Aims

Cigarette smoking has a negative impact on the skeletal system, causes a decrease in bone mass in both young and old patients, and is considered a risk factor for the development of osteoporosis. In addition, it disturbs the bone healing process and prolongs the healing time after fractures. The mechanisms by which cigarette smoking impairs fracture healing are not fully understood. There are few studies reporting the effects of cigarette smoking on new blood vessel formation during the early stage of fracture healing. We tested the hypothesis that cigarette smoke inhalation may suppress angiogenesis and delay fracture healing.

A balanced inflammatory response is important for successful fracture healing. The response of osteoporotic fracture healing is deranged and an altered inflammatory response can be one underlying cause. The objectives of this review were to compare the inflammatory responses between normal and osteoporotic fractures and to examine the potential effects on different healing outcomes. A systematic literature search was conducted with relevant keywords in PubMed, Embase, and Web of Science independently. Original preclinical studies and clinical studies involving the investigation of inflammatory response in fracture healing in ovariectomized (OVX) animals or osteoporotic/elderly patients with available full text and written in English were included. In total, 14 articles were selected. Various inflammatory factors were reported; of those tumour necrosis factor-α (TNF-α) and interleukin (IL)-6 are two commonly studied markers. Preclinical studies showed that OVX animals generally demonstrated higher systemic inflammatory response and poorer healing outcomes compared to normal controls (SHAM). However, it is inconclusive if the local inflammatory response is higher or lower in OVX animals. As for clinical studies, they mainly examine the temporal changes of the inflammatory stage or perform comparison between osteoporotic/fragility fracture patients and normal subjects without fracture. Our review of these studies emphasizes the lack of understanding that inflammation plays in the altered fracture healing response of osteoporotic/elderly patients. Taken together, it is clear that additional studies, preclinical and clinical, are required to dissect the regulatory role of inflammatory response in osteoporotic fracture healing.

Cite this article: Bone Joint Res 2020;9(7):368–385.

Objectives

This study aimed to assess the effect of age and osteoporosis on the proliferative and differentiating capacity of bone-marrow-derived mesenchymal stem cells (MSCs) in female rats. We also discuss the role of these factors on expression and migration of cells along the C-X-C chemokine receptor type 4 (CXCR-4) / stromal derived factor 1 (SDF-1) axis.

Long non-coding RNAs (lncRNAs) are transcripts longer than 200 nucleotides with limited coding potential, which have emerged as novel regulators in many biological and pathological processes, including growth, development, and oncogenesis. Accumulating evidence suggests that lncRNAs have a special role in the osteogenic differentiation of various types of cell, including stem cells from different sources such as embryo, bone marrow, adipose tissue and periodontal ligaments, and induced pluripotent stem cells. Involved in complex mechanisms, lncRNAs regulate osteogenic markers and key regulators and pathways in osteogenic differentiation. In this review, we provide insights into the functions and molecular mechanisms of lncRNAs in osteogenesis and highlight their emerging roles and clinical value in regenerative medicine and osteogenesis-related diseases.

Cite this article: J. Zhang, X. Hao, M. Yin, T. Xu, F. Guo. Long non-coding RNA in osteogenesis: A new world to be explored. Bone Joint Res 2019;8:73–80. DOI: 10.1302/2046-3758.82.BJR-2018-0074.R1.