Effects of CAP on the biological activity of BMSCs

Objectives: The purpose of this study is to investigate the influence of Cementum attachment protein (CAP) on the attachment, spreading, proliferation and mineralization of cultured Rhesus bone marrow stromal cells (BMSCs) in vitro. Methods: BMSCs were obtained from Rhesus bone marrow aspiration and cultured in DMEM medium with 10% FBS. The third passage cells were exposed to various concentrations of CAP (0.125µg, 0.25µg, 0.5µg, 1µg and 2µg /ml). Controls of BMSCs were cultured in DMEM medium without CAP. Attachment ability of BMSCs was detected by counting cell number. Cell spreading rates were performed at culture time of 1h, 4h and 7h by analysis of micrographs taken at predetermined sites of each wells. BMSCs proliferation rates and alkaline phosphatase (ALP) synthesis were assessed by an MTT assay. A parametric one- way analysis of variance(ANOVA) and multiple comparisons were used to test the treatments based on the Stument- Newman- Keuls test. Results: It was shown that BMSCs well attached and spread on CAP - coated substrata, The attachment of BMSCs was significantly stimulated by CAP at a concentration of 0.5 and 1µg/ml(P<0.01) than Control group. whereas there was no significant change between the control group and experimental groups in cell spreading rate. The proliferation of BMSCs was significantly stimulated by CAP at a concentration of 0.5 to 2µg/ml(P<0.05) than Control group. whereas there was no significant change between the control group and experimental groups in alkaline phosphatase (ALP) synthesis.

Conclusions: CAP could promote the attachment and the proliferating ability of Rhesus BMSCs in vitro, but have no effects on its spreading and alkaline phosphatase (ALP) synthesis.