Recombinant growth/differentiation factor-5 enhances cell proliferation and tendogenic differentiation of human periodontal ligament stem cells in vitro.

Abstract

e22191 Background: To investigate the function of growth/differentiation factor-5 (GDF-5) on cell proliferation and tendogenic differentiation of human periodontal ligment stem cells (PDLSCs) in vitro. Methods: PDLSCs were obtained from human healthy periodontal ligaments of wisdom teeth. Immunofluorescence cytochemistry was used to detect the characters of mesenchymal stem cells (MSCs) in PDLSCs. Recombinant mouse GDF-5 (rmGDF-5) protein was used to stimulate the PDLSCs. Tetrazolium reduction assay was carried out for cell proliferation. Alkaline phosphatase (ALP) activity was measured by ALP activity assay and ALP staining. Real-time RT-PCR was performed to detect the gene expressions of aggrecan (ACAN), collagen, type III (COL3), scleraxis (SCX), decorin (DCN), tenomodulin (TNMD) and tenascin C (TNC) in PDLSCs after stimulated by rmGDF-5 protein. Results: 5.6% STRO-1 positive cells was detected in primary PDLSCs. The cell proliferation of PDLSCs was enhanced by rmGDF-5 in a dose-dependent manner at 0, 20, 100 and 500 ng/ml of concentration. The special markers of tendogenic differention, ACAN, COL3, SCX and TNMD were up-regulated at mRNA level after rmGDF-5 treatment for 1, 2 or 3 weeks, and the 20 ng/ml was the more effective than 100 ng/ml and 500 ng/ml for rmGDF-5 in PDLSCs. The expressions of TNC and DCN were increased at 4 or 8 hours after 20, 50 and 100 ng/m rmGDF-5 treatment. The ALP activity results showed that ALP activities were decreased paralleled with increased rmGDF-5 concentration. Conclusions: The rmGDF-5 could promote the cell proliferation and tendogenic differentiation of PDLSCs in vitro.