Effects of oral implants with miR-122-modified cell sheets on rat bone marrow mesenchymal stem cells

Abstract

The aim of the present study was to regulate the transformation of bone marrow mesenchymal stem cells (BMMSCs) to osteoblasts to promote bone formation and osseointegration surrounding oral implants. BMMSCs were cultured using the whole bone marrow adherence method. Cell surface markers were detected by flow cytometry, and multi-lineage differentiation potential was detected by osteogenic and adipogenic tests. miR-122-modified cell sheets were prepared by non-viral transfection and complexed with micro-arc titanium oxide implants to construct a gene-modified tissue-engineered implant, with its surface morphology observed by scanning electron microscopy (SEM). In vitro osteogenic activity of the implant was determined by alkaline phosphatase (ALP), Sirius Red, alizarin red staining, polymerase chain reaction and western blot analysis. The BMMSCs were spindle- or triangular-shaped. Surface markers, cluster of differentiation 29(CD29), CD90 and CD105 were positively expressed, whereas blood cell markers CD34, CD45 and CD31 were negatively expressed. Osteogenic staining exhibited deposition of calcified nodules, while adipogenic staining demonstrated the formation of lipid droplets. miR-122 modification significantly enhanced the in vitro osteogenic activity of the sheets. On day 3 of osteogenic induction, runt-related transcription factor 2, osterix, osteocalcin, collagen I, ALP and bone morphogenetic protein 2 expression levels of the experimental group were 2.0, 3.1, 4.6, 3.2, 10.5 and 4.5 times those of the blank control group, respectively. SEM imaging of the modified sheet demonstrated close adhesion and fitting between abundant cellular and extracellular matrices, and the porous surface of the implant. In vitro osteogenesis of the complex was promoted and accelerated. Thus, miR-122 effectively promoted osteogenic differentiation of the BMMSC sheet. Therefore, it is feasible to construct gene-modified tissue-engineered implants by complexing miR-122-modified sheets with micro-arc titanium oxide implants.