pentosan polysulfate promotes proliferation and chondrogenic differentiation of adult human bone marrow-derived mesenchymal precursor cells戊聚糖polysulfate促进成人骨骨髓来源间充质干细胞的增殖和chondrogenic前体细胞.pdf

Ghosh et al. Arthritis Research Therapy 2010, 12:R28 /content/12/1/R28 RESEARCH ARTICLE Open Access Pentosan polysulfate promotes proliferation and chondrogenic differentiation of adult human bone marrow-derived mesenchymal precursor cells 1,2* 1 1 3 4 2 Peter Ghosh , Jiehua Wu , Susan Shimmon , Andrew CW Zannettino , Stan Gronthos , Silviu Itescu Abstract Introduction: This study was undertaken to determine whether the anti-osteoarthritis drug pentosan polysulfate (PPS) influenced mesenchymal precursor cell (MPC) proliferation and differentiation. Methods: Human MPCs were maintained in monolayer, pellet or micromass cultures (MMC) for up to 10 days with PPS at concentrations of 0 to 20 μg/ml. MPC viability and proliferation was assessed using the WST-1 assay and 3H-thymidine incorporation into DNA, while apoptosis was monitored by flow cytometry. Proteoglycan (PG) biosynthesis was determined by 35SO42- incorporation and staining with Alcian blue. Proteoglycan and collagen type I and collagen type II deposition in pellet cultures was also examined by Toluidine blue and immunohistochemical staining, respectively. The production of hyaluronan (HA) by MPCs in MMC was assessed by ELISA. The relative outcome of PPS, HA, heparin or dextran sulfate (DS) on PG synthesis was compared in 5-day MMC. Gene expression of MPCs in 7-day and 10-day MMC was examined using real-time PCR. MPC differentiation was investigated by co-culturing with PPS in osteogenic or adipogenic inductive culture media for 28 days. Results: Significant MPC proliferation was evident by day 3 at PPS concentrations of 1 to 5 μg/ml (P 0.01). In the presence of 1 to 10 μg/ml PPS, a 38% reduction in IL-4/IFNg-induced MPC apoptosis was observe