Pentosan polysulfate promotes proliferation and chondrogenic differentiation of adult human bone marrow-derived mesenchymal precursor cells.
Pentosan polysulfate promotes proliferation and chondrogenic differentiation of adult human bone marrow-derived mesenchymal precursor cells.
复制标题
DOI：
10.1186/ar2935
复制
发表时间：
2010
影响因子：
4.9
通讯作者：
Itescu S
中科院分区：
文献类型：
--
作者：
Ghosh P;Wu J;Shimmon S;Zannettino AC;Gronthos S;Itescu S
本研究旨在确定抗骨关节炎药物聚硫酸戊聚糖（PPS）是否影响间充质前细胞（MPC）的增殖和分化。在PPS浓度为0 ~ 20 μg/ml的条件下，将人MPCs在单层、颗粒或微团培养（MMC）中维持10天。采用WST-1检测和3h -胸腺嘧啶并入DNA检测MPC的活力和增殖，流式细胞术检测细胞凋亡。蛋白多糖（PG）的生物合成采用35SO42掺入法和阿利新蓝染色法。用甲苯胺蓝和免疫组织化学染色分别检测微球培养中蛋白多糖、ⅰ型胶原和ⅱ型胶原沉积。ELISA法测定MMC中MPCs产生透明质酸（HA）的情况。比较PPS、HA、肝素和硫酸葡聚糖（DS）在5 d MMC中对PG合成的相对影响。采用实时荧光定量PCR检测7天和10天MMC中MPCs的基因表达情况。在成骨或成脂诱导培养基中与PPS共培养28天，观察MPC的分化情况。PPS浓度为1 ~ 5 μg/ml时，第3 d MPC增殖显著（P < 0.01）。PPS浓度为1 ~ 10 μg/ml时，IL-4/ ifn γ-诱导的MPC细胞凋亡减少38%。在5 d MMC中，2.5 μg/ml PPS对PG合成的刺激达到130% (P < 0.0001)，而5.0 μg/ml PPS对7 d和10 d的刺激最大（P < 0.05）。≥5 μg/ml的HA和DS抑制了5 d MMC中PG的合成（P < 0.05）。≥0.5 μg/ml PPS时，MMC显著沉积ⅱ型胶原（P < 0.001 ~ 0.05）。在MPC-PPS颗粒培养中，与对照组相比，PG、II型胶原沉积较多，而I型胶原沉积较少。Real-time PCR结果与蛋白质数据一致。5、10 μg/ml PPS抑制MPC成骨分化（P < 0.01）。本研究首次证实PPS促进MPC增殖和软骨形成，为骨关节炎关节的软骨再生和修复提供了新的策略。
This study was undertaken to determine whether the anti-osteoarthritis drug pentosan polysulfate (PPS) influenced mesenchymal precursor cell (MPC) proliferation and differentiation. 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. 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/IFNγ-induced MPC apoptosis was observed. In 5-day MMC, 130% stimulation of PG synthesis occurred at 2.5 μg/ml PPS (P < 0.0001), while 5.0 μg/ml PPS achieved maximal stimulation in the 7-day and 10-day cultures (P < 0.05). HA and DS at ≥ 5 μg/ml inhibited PG synthesis (P < 0.05) in 5-day MMC. Collagen type II deposition by MMC was significant at ≥ 0.5 μg/ml PPS (P < 0.001 to 0.05). In MPC-PPS pellet cultures, more PG, collagen type II but less collagen type I was deposited than in controls. Real-time PCR results were consistent with the protein data. At 5 and 10 μg/ml PPS, MPC osteogenic differentiation was suppressed (P < 0.01). This is the first study to demonstrate that PPS promotes MPC proliferation and chondrogenesis, offering new strategies for cartilage regeneration and repair in osteoarthritic joints.
登录
查看更多内容
影响因子：
4.9
作者：
Chen FH;Tuan RS
通讯作者：
Tuan RS
影响因子：
4
作者：
Gronthos, Stan;McCarty, Rosa;Zannettino, Andrew C. W.
通讯作者：
Zannettino, Andrew C. W.
影响因子：
4
作者：
Gronthos, S;Zannettino, ACW;Simmons, PJ
通讯作者：
Simmons, PJ
影响因子：
4.9
作者：
Grogan SP;Miyaki S;Asahara H;D'Lima DD;Lotz MK
通讯作者：
Lotz MK
影响因子：
2.9
作者：
Denker, AE;Haas, AR;Tuan, RS
通讯作者：
Tuan, RS