Proliferation and collagen synthesis of human anterior cruciate ligament cells in vitro:: Effects of ascorbate-2-phosphate, dexamethasone and oxygen tension
Proliferation and collagen synthesis of human anterior cruciate ligament cells in vitro:: Effects of ascorbate-2-phosphate, dexamethasone and oxygen tension
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DOI：
10.1006/cbir.1998.0302
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发表时间：
1998-01-01
影响因子：
3.9
通讯作者：
Gage, J
中科院分区：
文献类型：
--
作者：
Fermor, B;Urban, J;Gage, J
临床和实验研究表明，损伤的前交叉韧带（ACL）通常不能愈合，用于修复ACL的自体移植物在早期迅速减弱，需要很长时间才能恢复强度。本研究的目的是建立一个体外培养系统，在该系统中，环境和生化因素的影响，增殖和基质合成的细胞来自人前交叉韧带可以studied. Human ACL细胞的原代培养获得的生长从正常ACL的外植体在Dulbecco的最低必需培养基（DMEM）中获得的膝关节置换骨关节炎。在培养4、8和12天后，评估添加剂100 μ M L-抗坏血酸-2-磷酸盐（Asc-2-P）和10 nM地塞米松（dex）对增殖和胶原合成的影响。韧带细胞在100 μ M asc-2-P和10 nM dex存在下，在0、5、10和21%pO（2）下生长。培养3、6和12天后，使用Hoechst染料法评估DNA含量，并通过掺入5 mCi/ml [H-3]脯氨酸来评估胶原合成。在21%pO（2）下，asc-2-P和dex的存在显着增加（P
Clinical and experimental studies demonstrate that injured anterior cruciate ligaments (ACL) do not usually heal and that autografts used to repair the ACL rapidly weaken in the early period and take a long time to regain strength. The aim of this study was to develop an in vitro culture system in which environmental and biochemical factors influencing the proliferation and matrix synthesis of cells derived from human anterior cruciate ligaments can be studied.Primary cultures of human ACL cells were obtained by outgrowth from explants of normal ACL obtained at knee replacement for osteoarthritis in Dulbecco's minimum essential medium (DMEM). The effects of the additives 100 mu M L-ascorbic acid-2-phosphate (Asc-2-P) and 10 nM dexamethasone (dex) on proliferation and collagen synthesis were assessed after 4, 8 and 12 days in culture. Ligament cells were grown at 0, 5, 10 and 21% pO(2) in the presence of 100 mu M asc-2-P and 10 nM dex. DNA content was assessed using the Hoechst dye method and collagen synthesis by the incorporation of 5 mCi/ml [H-3]proline after 3, 6 and 12 days in culture.At 21% pO(2), the presence of asc-2-P and dex induced significantly greater (P