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  • Title: The effect of the coumarin-like derivative osthole on the osteogenic properties of human periodontal ligament and jaw bone marrow mesenchymal stem cell sheets.
    Author: Gao LN, An Y, Lei M, Li B, Yang H, Lu H, Chen FM, Jin Y.
    Journal: Biomaterials; 2013 Dec; 34(38):9937-51. PubMed ID: 24095254.
    Abstract:
    Cell sheet engineering is a scaffold-free delivery concept that has been shown to improve mesenchymal stem cell-mediated regeneration of injured or pathologically damaged periodontal tissues in preclinical studies and several clinical trials. However, the best strategy for cell sheet production remains to be identified. The aim of this study was to investigate the biological effects of osthole, a coumarin-like derivative extracted from Chinese herbs, on the cell sheet formation and osteogenic properties of human periodontal ligament stem cells (PDLSCs) and jaw bone marrow mesenchymal stem cells (JBMMSCs). Patient-matched PDLSCs and JBMMSCs were isolated, and an appropriate concentration of osthole for cell culture was screened for both cell types in terms of cell proliferation and alkaline phosphatase (ALP) activity. Next, the best mode of osthole stimulation for inducing the formation of sheets by each cell type was selected by evaluating the amount of their extracellular matrix (ECM) protein production as well as osteogenic-related gene expression. Furthermore, both PDLSC and JBMMSC sheets obtained from each optimized technique were transplanted subcutaneously into nude mice to evaluate their capacity for ectopic bone regeneration. The results revealed that 10(-5) m/L osthole significantly enhanced the proliferation of both PDLSCs and JBMMSCs (P < 0.05), although for JBMMSCs, there was no concentration-related change among the four established osthole groups (P > 0.05). In addition, 10(-5) m/L osthole was the best concentration to promote the ALP activities of both cells (P < 0.01). Based on both the production of ECM proteins (collagen type I, integrin β1, and fibronectin) and the expression of osteogenic genes (ALP, Runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN)), the provision of 10(-5) m/L osthole throughout the entire culture stage (10 days) for PDLSCs or at the early stage (first 3 days) for JBMMSCs was the most effective osthole administration mode for cell sheet formation (P < 0.05). The results of in vivo transplantation showed that osthole-mediated PDLSC and JBMMSC sheets formed more new bone than those obtained without osthole intervention (P < 0.001). Our data suggest that a suitable concentration and mode of osthole stimulation may enhance ECM production and positively affect cell behavior in cell sheet engineering.
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