Cellular alterations and crosstalk in the osteochondral joint in osteoarthritis and promising therapeutic strategies.

Osteoarthritis (OA) is a joint disorder involving cartilage degeneration and subchondral bone sclerosis. The bone-cartilage interface is implicated in OA pathogenesis due to its susceptibility to mechanical and biological factors. The crosstalk between cartilage and the underlying subchondral bone is elevated in OA due to multiple factors, such as increased vascularization, porosity, microcracks and fissures. Changes in the osteochondral joint are traceable to alterations in chondrocytes and bone cells (osteoblasts, osteocytes and osteoclasts). The phenotypes of these cells can change with the progression of OA. Aberrant intercellular communications among bone cell-bone cell and bone cell-chondrocyte are of great importance and might be the factors promoting OA development. An appreciation of cellular phenotypic changes in OA and the mechanisms by which these cells communicate would be expected to lead to the development of targeted drugs with fewer side effects.

[Fluid-solid coupling numerical simulation on ideal porous structure of rat alveolar bone].

Fluid shear stress (FSS) caused by interstitial fluid flow within trabecular bone cavities under mechanical loading is the key factor of stimulating biological response of bone cells. Therefore, to investigate the FSS distribution within cancellous bone is important for understanding the transduction process of mechanical forces within alveolar bone and the regulatory mechanism at cell level during tooth development and orthodontics. In the present study, the orthodontic tooth movement experiment on rats was first performed. Finite element model of tooth-periodontal ligament-alveolar bone based on micro computed tomography (micro-CT) images was established and the strain field in alveolar bone was analyzed. An ideal model was constructed mimicking the porous structure of actual rat alveolar bone. Fluid flow in bone was predicted by using fluid-solid coupling numerical simulation. Dynamic occlusal loading with orthodontic tension loading or compression loading was applied on the ideal model. The results showed that FSS on the surface of the trabeculae along occlusal direction was higher than that along perpendicular to occlusal direction, and orthodontic force has little effect on FSS within alveolar bone. This study suggests that the orientation of occlusal loading can be changed clinically by adjusting the shape of occlusal surface, then FSS with different level could be produced on trabecular surface, which further activates the biological response of bone cells and finally regulates the remodeling of alveolar bone.

Corrigendum: Nitazoxanide, an antiprotozoal drug, reduces bone loss in ovariectomized mice by inhibition of RANKL-induced osteoclastogenesis.

[This corrects the article DOI: 10.3389/fphar.2021.781640.].

Distinct pathological changes of osteochondral units in early OVX-OA involving TGF-ß signaling.

Introduction: Different opinions exist about the role of subchondral bone in osteoarthritis (OA), probably because subchondral bone has different effects on cartilage degeneration in OA induced by different pathologies. Animal studies to illustrate the role of subchondral bone in cartilage degeneration were mostly based on post-traumatic OA (PT-OA). Postmenopausal women experience a much higher occurrence of OA than similar-aged men. The physiological changes and pathogenesis of the osteochondral unit in ovariectomy-induced OA (OVX-OA) might be distinct from other types of OA. Methods: The osteochondral alterations of post-traumatic OA (PT-OA) and OVX-OA at week 9 after surgery were compared. Then the alterations of osteochondral units in OVX-OA rats were tracked over time for the designed groups: Sham, OVX and OVX rats treated with estrogen (OVX+E). DXA, micro-CT, and histochemical staining were performed to observe alterations in osteochondral units. Results: Rapid cartilage degeneration and increased bone formation were observed in PT-OA, while only mild cartilage erosion and significant bone loss were observed in OVX-OA at week 9 after surgery. Subchondral bone degradation preceded cartilage degeneration by 6 weeks in OVX-OA. TGF-ß expression was downregulated in the osteochondral unit of OVX rats. Estrogen supplementation inhibited subchondral bone loss, cartilage degradation and TGF-ß expression decrease. Discussion: This research demonstrated the distinct behaviors of the osteochondral unit and the critical role of subchondral bone in early OVX-OA compared with PT-OA. Inhibiting subchondral bone catabolism at the early stage of OVX-OA could be an effective treatment for post-menopausal OA. Based on the results, estrogen supplementation and TGF-ß modulation at the early stage are both potential therapies for post-menopausal OA.

Nitazoxanide, an Antiprotozoal Drug, Reduces Bone Loss in Ovariectomized Mice by Inhibition of RANKL-Induced Osteoclastogenesis.

Nitazoxanide (NTZ) is an FDA-approved anti-parasitic drug with broad-spectrum anti-infective, anti-inflammatory, and antineoplastic potential. However, its regulatory effects on osteoclastogenesis and the underlying mechanisms remain unclear. The present study found that NTZ potently inhibited osteoclast formation at the early stage of receptor activator of NF-κB ligand-induced osteoclastogenesis in a concentration-dependent manner at a non-growth inhibitory concentration. NTZ suppressed actin ring formation and decreased osteoclast marker gene expression, including TRAP, MMP9, and cathepsin K. NTZ significantly impaired the bone resorption activity of osteoclasts. In vivo, ovariectomized mice were treated with 50, 100 and 200 mg/kg/d NTZ for 3 months. NTZ (100 mg/kg/d) administration markedly reduced ovariectomy-induced bone loss by suppressing osteoclast activity. Mechanistically, osteoclastogenesis blockade elicited by NTZ resulted from inhibition of STAT3 phosphorylation, and reduction of the Ca2+ fluorescence intensity and NFATc1 expression. NTZ weakened the binding between STAT3 and the NFATc1 promoter region. Furthermore, enforced NFATc1 expression partly rescued the impaired osteoclast differentiation in NTZ-treated RAW264.7 cells. In summary, NTZ could inhibit osteoclastogenesis and bone loss through modulation of the receptor activator of NF-κB ligand-induced STAT3-NFATc1 signaling pathway, which might be a potential alternative treatment regimen against bone destruction-related diseases including osteoporosis.

Phenotype changes of subchondral plate osteoblasts based on a rat model of ovariectomy-induced osteoarthritis.

BACKGROUND: Osteoarthritis (OA) is prevalent in postmenopausal women. Subchondral bone in ovariectomized (OVX) rats might play a more important role in cartilage degeneration compared with other types of OA. How subchondral osteoblast changes in OVX rats is still unclear. Understanding of osteoblast changes obtained from OVX subchondral bone might be helpful to clarify pathogenesis of OVX-OA. METHODS: Female Sprague-Dawley rats were randomly divided into two groups: Sham (n=20) and OVX (n=20). Serum levels of Alkaline phosphatase (ALP) and C-telopeptide of type I collagen (CTX-I) were measured every one or two weeks. All rats were executed at week 9 post surgery. The weight of rats and the wet weight of uterus were assessed. Micro-computed Tomography (micro-CT) was used to analyze the knee microstructure, and toluidine blue staining was employed to evaluate cartilage erosion. Subchondral osteoblast proliferation ability by cell counting kit-8 assay, osteogenic genes expressions by reverse transcription polymerase chain reaction (RT-PCR), differentiation and mineralization ability by ALP staining and alizarin red staining were evaluated and compared between Sham and OVX. RESULTS: Ovariectomy induced significant increases of serum ALP and CTX-I as early as at week 2. At week 9 after surgery, the body weight of OVX rats was significantly increased, and uterus weight of OVX rats was remarkably decreased. OVX rats demonstrated significant subchondral bone change and cartilage erosion compared with Sham rats. mRNA levels of early markers of osteogenic differentiation (ALP, type I collagen, Runx2) were enhanced in OVX rats, but the late marker (osteocalcin) was not significantly different. ALP activity of osteoblasts increased, but the mineralization capacity decreased in OVX rats. CONCLUSIONS: Subchondral osteoblasts in OVX rats exhibited different proliferation, differentiation and mineralization abilities from normal counterparts.

Identification of Candidate Genetic Markers and a Novel 4-genes Diagnostic Model in Osteoarthritis through Integrating Multiple Microarray Data.

BACKGROUND: Osteoarthritis (OA) is a joint disease that leads to a high disability rate and a low quality of life. With the development of modern molecular biology techniques, some key genes and diagnostic markers have been reported. However, the etiology and pathogenesis of OA are still unknown. OBJECTIVE: To develop a gene signature in OA. METHOD: In this study, five microarray data sets were integrated to conduct a comprehensive network and pathway analysis of the biological functions of OA related genes, which can provide valuable information and further explore the etiology and pathogenesis of OA. RESULTS AND DISCUSSION: Differential expression analysis identified 180 genes with significantly expressed expression in OA. Functional enrichment analysis showed that the up-regulated genes were associated with rheumatoid arthritis (p < 0.01). Down-regulated genes regulate the biological processes of negative regulation of kinase activity and some signaling pathways such as MAPK signaling pathway (p < 0.001) and IL-17 signaling pathway (p < 0.001). In addition, the OA specific protein-protein interaction (PPI) network was constructed based on the differentially expressed genes. The analysis of network topological attributes showed that differentially upregulated VEGFA, MYC, ATF3 and JUN genes were hub genes of the network, which may influence the occurrence and development of OA through regulating cell cycle or apoptosis, and were potential biomarkers of OA. Finally, the support vector machine (SVM) method was used to establish the diagnosis model of OA, which not only had excellent predictive power in internal and external data sets (AUC > 0.9), but also had high predictive performance in different chip platforms (AUC > 0.9) and also had effective ability in blood samples (AUC > 0.8). CONCLUSION: The 4-genes diagnostic model may be of great help to the early diagnosis and prediction of OA.

Efficient regeneration of rat calvarial defect with gelatin-hydroxyapatite composite cryogel.

To induce bone regeneration efficiently, a properly designed organic-inorganic composite scaffold is necessary and important. Gelatin-hydroxyapatite (HA) composite is a suitable choice for the purpose because it can resemble the chemical composition of natural bone tissue. The gelatin-HA composite can be implanted into bone defects as a hydrogel or cryogel, however, it is interesting to know the effect of their different morphology on inducing osteogenesis in vivo. Herein, HA nanowire (HANW) reinforced photocrosslinkable methacrylated gelatin (GelMA) cryogel and hydrogel are prepared and comparatively investigated by being implanted into rat calvarial defects. The cryogel acts as a kind of sponge with interconnected macropores, allowing cell infiltration, as well as, displaying rapid shape recovery and excellent mechanical stability under cyclic compression loading. Conversely, the hydrogel is rigid and easily crushed during the first compression test, showing no shape recovery ability, instead inhibiting cell migration and spreading. Accordingly, the GelMA/HANW composite cryogel is able to promote osteogenesis significantly more in comparison with the corresponding hydrogel at six and 12 weeks post-implantation, as revealed by comprehensive evaluations using radiographic examination, histochemical and immunohistochemical staining methods. Neo-bone tissues have grown into the macroporous cryogel six and 12 weeks after the implantation, while the dense hydrogel prevents the tissue ingrowth, causing the newly formed sparse bone tissue to only elongate into the gaps between cracked hydrogel blocks. In summary, organic-inorganic macroporous cryogels demonstrate superiority for in vivo applications to induce bone regeneration.

Macroporous scaffolds developed from CaSiO3 nanofibers regulating bone regeneration via controlled calcination.

Calcium silicate (CS) is envisioned as a good substrate for bone tissue engineering applications because it can provide bioactive ions like Ca2+ and Si4+ to promote bone regeneration. Calcination temperature is a critical factor in determining the crystallinity of CS ceramic, which subsequently influences its degradation and ion release behaviors. To investigate the effect of calcination temperature on the capacity of CS in inducing bone regeneration, CS nanofibers were fabricated via electrospinning of precursor sol-gel and subsequent sintering at 800 °C, 1000 °C or 1200 °C. As the calcination temperature was increased, the obtained CS nanofibers displayed higher crystallinity and slower degradation rate. The CS nanofibers calcined at 800 °C (800 m) would like to cause high pH (>9) in cell culture medium due to its rapid ion release rate, displaying adverse effect on cell viability. Among all the preparations, it was found the CS nanofibers calcined at 1000 °C (1000 m) demonstrated the strongest promotion effect on the osteogenic differentiation of bone marrow mesenchymal stromal cells. To facilitate in vivo implantation, the CS nanofibers were shaped into three-dimensional macroporous scaffolds and coated with gelatin to improve their mechanical stability. By implanting the scaffolds into rat calvarial defects, it was confirmed the scaffold made of CS nanofibers calcined at 1000 °C was able to enhance new bone formation more efficiently than the scaffolds made of CS nanofibers calcined at 800 °C or 1200 °C. To summarize, calcination temperature could be an effective and useful tool applied to produce CS bioceramic substrates with improved potential in enhancing osteogenesis by regulating their degradation and bioactive ion release behaviors.