Experimental Study of Dentin Matrix Protein-1 in Interleukin-1β induced Rat Chondrocyte Inflammatory Lesion
Department of Orthopaedics，The First People's Hospital of Wuhu
Department of Orthopaedics，Harbin First Hospital
Department of Spinal Surgery，The First Affiliated Hospital of Harbin Medical University
Objective To explore the biological effect of Dentin Matrix Protein-1 (DMP-1) on chondrocytes in osteoarthritis (OA).Methods The expression of DMP-1 in cartilage cells of OA rat model was analyzed with Affymetrix microarray.The chondrocyte was isolated，cultured and transfected with DMP-1 gene.Subsequently，the effect of DMP-1 on the proliferation of chondrocyte was examined with CCK-8 assay.ELISA assay was performed for detection of chondrocyte glycosaminoglycan (GAG) secretion and proinflammatory cytokines (IL-6 and TNF-α).The expression of Col2a1 was observed by immunofluorescence.RT-qPCR was to evaluate the expression levels of DMP-1，Sox9，Acan，and Col2a1 genes.Western Blot assay was conducted for DMP-1，Sox9，Acan，Col2a1 expressions.Flow cytometry and Calcein AM/PI staining assays were conducted for detection of the anti-apoptosis ability of chondrocyte treated with DMP-1.Results The Microarray analysis showed that the expression level of DMP-1 in OA group was lower than that in Control group in vivo (P＜0.05)；The expression level of DMP-1 in chondrocytes of OA group was also lower than that in Control group in vitro (P＜0.001).After transfection with DMP-1 gene，the expression level of DMP-1 in chondrocytes and chondrocytes induced by IL-1β was higher than that in the non-transfected group.The viability of chondrocytes induced by IL-1β was higher in transfected DMP-1 chondrocyte than that in the non-transfected group (P＜0.001).The expression of GAGs and immunofluorescence of Col2a1 in the transfected DMP-1 chondrocytes induced by IL-1β was significantly higher than that in the untransfected group，as well as the expression levels of Sox9，Acan and Col2a1 were significantly higher than that in the untransfected group (P＜0.001).The levels of IL-6 and TNF-α and apoptosis of transfected DMP-1 chondrocytes induced by IL-1β were significantly lower than those of non-transfected group (P＜0.001). Conclusion DMP-1 can be used to transfect chondrocytes and reduce the progression of OA.
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