Abstract
Purpose: In this study, we performed the gene transfection by introducing sox9/sox6 and ANGOTL4 genes into DEX-conjugated PEI for the treatment of osteoarthritis. Methods: Three genes SOX6, SOX9, and ANGPTL4 were fabricated in a single plasmid and this multicistronic plasmid was complexed with the DEX-conjugated PEI (DEX-PEI). The surface chemistries of the synthesized DEX-PEI were characterized by FTIR and 1H NMR. SEM and TEM analysis were conducted to observe the morphology of DEX-PEI complexed with the multicistronic plasmid (DEX-PEI-SOX9/SOX6/shANGPTL4). Transfection efficiency of DEX-PEI-SOX9/SOX6/shANGPTL4 complexes in hADSCs was evaluated by flow cytometry analysis and in vitro chondrogenesis of hADSCs transfected by DEX-PEI-SOX9/SOX6/shANGPTL4 complexes (hADSCSOX9/SOX6/shANG) was evaluated by RT-qPCR, western-blot and immunohistochemistry. In vivo study, the effects of intra-articular (IA) administered hADSCSOX9/SOX6/shANG were evaluated in a surgically-induced OA rat model. Results: Covalently crosslinked DEX-PEI had a diameter of 190.1±86.2 nm. The uptake of DEX-PEI-SOX9/SOX6/shANGPTL4 in hADSCs was 52.5%. Thereafter, overexpression of SOX6 and SOX9 and down-regulation of shANGPTL4 was observed. pMC_SOX9/SOX6 expressed more than control, and pMC_SOX9/SOX6/shANGPTL4 was more than pMC_SOX9/SOX6. The expression of Col2α1 and Aggrecan in hADSCSOX9SOX/6/shANG showed significantly higher than those of DEX-PEI-SOX9/SOX6 complexes-transfected hADSCSOX9/SOX6. After Safranin O staining and Immunofluorescense, it was identified that RDbSOX9 / SOX6 / shANGPTL4 expressed more than control and TGF-BMP. The OARSI and Mankin scores of hADSCSOX/9SOX/6/shANG-injected rats were significantly lower than those of vehicle and vector control. Conclusions: In conclusion, the effectiveness of DEX-PEI complexes with SOX9/SOX6/shANGPTL4 genes for the transfection of hADSCs to osteoarthritis treatment is suggested.
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