Abstract:
Background Exosomes derived from gingival mesenchymal stem cells (GMSCs) have similar biological functions to GMSCs, which is a promising treatment for tissue repair. However, the research of GMSCs derived exosomes on bone regeneration is rarely reported.
Objective To investigate the effects of GMSCs-derived exosomes on the proliferation and osteogenic differentiation of osteoblast-like cells MC3T3-E1 in vitro.
Methods GMSCs were cultured, and exosomes were extracted by ultracentrifugation from supernatants. Uptake of exosomes by MC3T3-E1 was observed under an inverted fluorescence microscope. CCK-8 assay and scratch test were used to detect the changes of proliferation and migration of MC3T3-E1 with different concentrations of exosomes. After exosomes acted on MC3T3-E1, the activity of alkaline phosphatase (ALP) was detected by ALP staining, and the formation of calcium nodules was detected by alizarin red staining and quantitative experiments. RT-qPCR method was used to detect the mRNA expression levels of osteogenesis related genes ALP, COL1A1 (Collagen Type I Alpha 1) and RUNX2 (RUNX Family Transcription Factor 2).
Results The extracted exosomes derived from GMSCs presented a round membranous structure with a size of about 100 nm and expressed CD63 and Tsg101 marker proteins. GMSCs derived exosomes could promote the proliferation and migration of MC3T3-E1 (P＜0.05) in a dose-dependent manner. The ALP activity of MC3T3-E1 and the calcium nodule Nodal formation increased under the action of exosomes, and the mRNA expression levels of osteogenic related genes ALP, COL1A1 and RUNX2 were significantly up-regulated (all P＜0.05).
Conclusion GMSCs derived exosomes can promote the proliferation and osteogenic differentiation of MC3T3-E1, which lays the foundation for the application of GMSCs derived exosomes to bone regeneration treatment.
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