Low-intensity pulsed ultrasound inhibits IL-6 in subchondral bone of temporomandibular joint osteoarthritis by suppressing the TGF-ß1/Smad3 pathway.

OBJECTIVE: This study aimed to provide further information on the exact mechanisms involved in the anti-inflammatory effect of low-intensity pulsed ultrasound (LIPUS) on rabbit temporomandibular joint osteoarthritis (TMJOA) on interleukin-6 (IL-6) production in subchondral bone, IL-6 production in IL-1ß stimulated via inhibition of the TGF-ß1/Smad3 pathway in mouse embryo osteoblast precursor (MC3T3-E1) cells. DESIGN: Bilateral joints were injected with type II collagenase to establish TMJOA models in two male and four female rabbits. The left joint was continuously stimulated by LIPUS, while the right joint was treated with the power off in this model. One male and two female rabbits were used as normal healthy controls without treatment. The histological features of subchondral bone were examined by Safranin-O/Fast staining. Immunohistochemistry was conducted to evaluate IL-6 expression. Then, cells were stimulated by LIPUS with IL-1ß. IL-6 expression and activity of the TGF-ß1/Smad3 pathway were evaluated by Enzyme-linked immunosorbent assay (ELISA), Immunofluorescence and Western blotting, respectively. Specific inhibition of the TGF-ß1/Smad3 pathway was conducted by transfecting with small interfering RNA (siRNA) of type II receptor (siTßRII). RESULTS: LIPUS significantly ameliorated the production of IL-6 in vitro and in vivo. Its inhibitory effect on the production of IL-6 induced by IL-1ß in MC3T3-E1 cells was partly reversed by siTßRII knockdown. CONCLUSIONS: LIPUS inhibited IL-6 production by suppressing the TGF-ß1/Smad3 pathway of subchondral bone in TMJOA. These data revealed the part of the pathways involved in the anti-inflammatory effect of LIPUS and provided a possible treatment strategy for TMJOA patients and other inflammatory diseases.

Overexpression of HIF-1alpha in Bone Marrow Mesenchymal Stem Cells Promote the Repair of Mandibular Condylar Osteochondral Defect in a Rabbit Model.

PURPOSE: The self-repair ability of temporomandibular joint (TMJ) cartilage is limited. Hypoxia-inducible factor-1 alpha (HIF-1alpha) may induce stem cells to promote chondrogenic repair. The purpose of this study was to systematically evaluate the effect of HIF-1alpha overexpression in bone marrow mesenchymal stem cells (BMSCs) combined with collagen scaffolds on the repair of TMJ condylar osteochondral defects in a rabbit model. METHODS: Osteochondral defects of 3-mm diameter × 2-mm depth were created at the right side of the mandibular condyle in 40 New Zealand white rabbits. The defect sites were treated with simple empty, collagen scaffolds (COL), BMSCs/COL, and HIF-1alpha overexpression BMSCs/COL groups. The histomorphologic features of condylar cartilage were monitored by gross examination, safranin O-fast green staining (Solarbio, Beijing, China), and immunohistochemical staining. The changes in subchondral bone were examined by microcomputed tomography. Immunofluorescence staining was used to trace the transplanted BMSCs in vivo. RESULTS: At 12 weeks postimplantation, histologic staining showed that the osteochondral defects in the simple empty and COL groups were mainly filled with fibrous tissue, whereas the BMSCs/COL and HIF-1alpha overexpression BMSCs/COL groups repaired the defect with fibrocartilage. Furthermore, the cartilage was better organized in the HIF-1alpha overexpression BMSCs/COL group compared with the BMSCs/COL group. Microcomputed tomography showed that osteochondral defects can cause abnormal hyperosteogeny in subchondral bone, and the transplantation of BMSCs, especially HIF-1alpha overexpression BMSCs, may alleviate osteosclerosis. Immunofluorescence staining showed that HIF-1alpha overexpression can promote the survival of transplanted BMSCs. CONCLUSIONS: The transplantation of HIF-1alpha overexpression BMSCs combined with a COL scaffold promotes cartilaginous repair of condylar cartilage and inhibits subchondral bone sclerosis in TMJ condylar osteochondral defect rabbits.