Modified Hif1α Gene Promotes Osteogenic Differentiation In Diabetic Rat ADSCs

To improve the osteogenic differentiation ability of T-ADSCs through HIF-1α gene modification, and to provide reliable strategies and insights for the clinical treatment of diabetic patients. Flow cytometry, alizarin red staining, and oil red O staining were used for isolation, identification, and culture of ADSCs from T2DM and normal sources. Transcriptomic sequencing (RNA-Seq) was performed to discover gene expression in ADSCs. CCK-8 analysis, scratch assay, Transwell assay, RT-qPCR and Western Blot analysis were used to detect the proliferation, migration, cell adhesion, and osseo-related gene expressions. Alkaline phosphatase (ALP) staining, Alizarin red (ARS), and AKP activity assay were performed to evaluate early osteogenic activity. Statistical analyses were performed using GraphPad Prism 5.0 (GraphPad Software, San Diego, CA, USA). Images were analyzed using ImageJ and Image Pro. A T2DM rat model was successfully established, and ADSCs were isolated from the fat of T2DM and normal rats. T-ADSCs were constructed and transfected with the HIF-1α plasmid. Overexpression of HIF-1α significantly improved cell proliferation, adhesion, migration, and osteogenic ability of T-ADSCs. The expression levels of osteogenesis-related proteins in T-ADSCs were significantly upregulated. 1. It was verified that HIF-1α mRNA expression in T-ADSCs was significantly decreased. 2. It was established that HIF-1α gene modification promoted cell proliferation, adhesion, migration, and osteogenic differentiation in T-ADSCs.