Development of an in vitro model to investigate joint ochronosis in alkaptonuria
Development of an in vitro model to investigate joint ochronosis in alkaptonuria
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开发体外模型，研究碱蛋白尿的关节chronosis问题
DOI：
10.1093/rheumatology/keq246
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发表时间：
2011-02-01
期刊：
影响因子：
5.5
通讯作者：
Gallagher, James A.
中科院分区：
文献类型：
--
作者：
Tinti, Laura;Taylor, Adam M.;Gallagher, James A.
方法：研究方法。骨肉瘤细胞株MG63、SAOS-2和TE85在含HGA的0.1 mM~1 mM的培养液中培养。培养物进行光镜和电子显微镜检查，并用Schmorl‘s染色检测体外色素沉积，观察到AKU组织中的同时性色素。测定HGA对细胞生长和胶原合成的影响。从33um M到0.33 mm HGA，细胞和相关基质中的色素沉积与剂量相关。体外的色素沉着比体内的要快得多，表明在原位组织中存在保护机制。色素沉积依赖于细胞的存在，并在无毒的HGA浓度下观察到。0.33 mM HGA对细胞生长有抑制作用，对I型胶原合成有刺激作用，但1 mM HGA对细胞毒性较大。我们已经建立了一个延时过长的体外模型，这应该有助于理解AKU的关节破坏和骨关节炎的病因学。
Methods. Osteosarcoma cell lines MG63, SaOS-2 and TE85 were cultured in medium containing HGA from 0.1 mu M to 1 mM. Cultures were examined by light microscopy and transmission electron microscopy, and Schmorl's stain was used to detect pigment deposits in vitro, following the observation that this stain identifies ochronotic pigment in AKU tissues. The effects of HGA on cell growth and collagen synthesis were also determined.Results. There was a dose-related deposition of pigment in cells and associated matrix from 33 mu M to 0.33 mM HGA. Pigmentation in vitro was much more rapid than in vivo, indicating that protective mechanisms exist in tissues in situ. Pigment deposition was dependent on the presence of cells and was observed at HGA concentrations that were not toxic. There was an inhibition of cell growth and a stimulation of type I collagen synthesis up to 0.33 mM HGA, but severe cell toxicity at 1 mM HGA.Conclusion. We have developed an in vitro model of ochronosis that should contribute to understanding joint destruction in AKU and to the aetiology of OA.