NASA-Approved Rotary Bioreactor Enhances Proliferation and Osteogenesis of Human Periodontal Ligament Stem Cells

Previous studies have suggested that periodontal ligament stem cells (PDLSCs) play crucial role in regeneration of periodontal defects, and recently tissue engineering based on PDLSCs to enhance periodontal regeneration has been the focus of periodontal research. A theoretical way to achieve this goal would be to provide a "stimulatory'' environment to rapidly expand PDLSCs in vitro to expedite tissue engineering of periodontium. We hypothesize that three-dimensional (3D) dynamic simulated microgravity (SMG) culture system have effect on periodontal stem cells, and would benefit periodontal stem cells proliferation and differentiation, but up to now, there are no related reports on this aspect. In this study, we investigated the biological effect of three-dimensional dynamic SMG induced by rotary cell culture system (RCCS) on human periodontal ligament stem cells (hPDLSCs) in vitro. hPDLSCs were isolated from surgically extracted human teeth and enriched by collecting multiple colonies. hPDLSCs were inoculated on Cytodex 3 microcarriers and cultured in RCCS. The results showed that SMG affected the biology of hPDLSCs as indicated by promotion of proliferation and viability, alterations of morphology, and disorganization of microfilament system. Besides, SMG-treated hPDLSCs presented increased matrix mineralization and up-regulated expression of mineralization associated genes after incubation in osteogenic medium. For it is the first time to investigate effects of SMG on PDLSCs, the research may lend insight into variations of cell response in 3D environment, and contribute to achievement of desirable periodontal regeneration utilizing PDLSCs-based tissue engineering approaches.