CCN family 2/connective tissue growth factor (CCN2/CTGF) stimulates proliferation and differentiation of auricular chondrocytes
CCN family 2/connective tissue growth factor (CCN2/CTGF) stimulates proliferation and differentiation of auricular chondrocytes
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DOI：
10.1016/j.joca.2007.11.001
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发表时间：
2008-07-01
影响因子：
7
通讯作者：
Takigawa, M.
中科院分区：
文献类型：
--
作者：
Fujisawa, T.;Hattori, T.;Takigawa, M.
目的：CCN 家族 2/结缔组织生长因子 (CCN2/CTGF) 是生长板软骨细胞的非典型生长因子。它在体外增殖和分化中发挥重要作用，但不刺激关节软骨细胞肥大或钙化。我们首次报道CCN2/CTGF在体外和体内促进耳廓软骨细胞的生长和分化，并维持其分子表型。方法：通过胰蛋白酶-胶原酶处理从兔耳廓软骨中分离耳廓软骨细胞，并用人重组CCN2/CTGF处理或用携带ccn2/ctgf基因的腺病毒感染。通过[3H]胸苷掺入和MTS测定来测量细胞增殖，并通过实时聚合酶链反应、Northern杂交和组织学分析来监测耳廓软骨细胞标记物基因表达的变化。对于体内研究，耳廓软骨细胞以颗粒形式培养，并在重组人 CCN2/CTGF 处理后植入皮下。对异位形成的软骨进行组织学分析。通过原位TUNEL分析监测细胞死亡。结果：CCN2/CTGF以剂量依赖性方式刺激兔原代耳廓软骨细胞的增殖、分化和弹性蛋白和蛋白聚糖的合成。与对照相比，CCN2/CTGF 导致弹性蛋白表达增加 2.5 倍，导致耳廓软骨细胞单层培养物中弹性蛋白纤维的沉积增强。未诱导矿化；相反，CCN2/CTGF 刺激基质 gla 蛋白的表达，已知该蛋白会损害矿化。此外，用CCN2/CTGF预处理耳廓软骨细胞颗粒并皮下植入显着增强了表达耳廓软骨细胞表型标记物（包括11型和X胶原）的异位耳廓软骨片的生长。值得注意的是，CCN2/CTGF 会损害软骨细胞凋亡。结论：这些发现表明，CCN2/CTGF 可能是促进耳廓软骨细胞分化和生长、同时防止矿化和凋亡的合适试剂，并表明 CCN2/CTGF 可能有助于弹性软骨的修复或重建。 (C) 2007 年国际骨关节炎研究协会。由爱思唯尔有限公司出版。保留所有权利。
Objectives: CCN family 2/connective tissue growth factor (CCN2/CTGF) is an atypical growth factor for growth plate chondrocytes. It plays an important role in their proliferation and differentiation in vitro, but does not stimulate hypertrophy or calcification of articular chondrocytes. We herein report for the first time that CCN2/CTGF promotes growth and differentiation of auricular chondrocytes and maintains their molecular phenotype in vitro and in vivo.Methods: Auricular chondrocytes were isolated from rabbit auricular cartilage by trypsin-collagenase treatment, and treated with human recombinant CCN2/CTGF or infected with adenovirus harboring the ccn2/ctgf gene. Cell proliferation was measured by [3 H] thymidine incorporation and MTS assay, and changes in gene expression of auricular chondrocyte markers were monitored by real-time polymerase chain reaction, Northern hybridization, and histological analysis. For in vivo studies, auricular chondrocytes were cultured as pellets and implanted subcutaneously after treatment of recombinant human CCN2/CTGF. Ectopically formed cartilage was subjected to histological analysis. Cell death was monitored by in situ TUNEL analysis.Results: CCN2/CTGF stimulated proliferation, differentiation and synthesis of elastin and proteoglycans of rabbit primary auricular chondrocytes in a dose-dependent manner. CCN2/CTGF caused a 2.5-fold increase in the expression of elastin in comparison to the control, resulting in enhanced deposition of elastin fibers in a monolayer culture of auricular chondrocytes. Mineralization was not induced; in contrast, CCN2/CTGF stimulated expression of matrix gla protein which is known to impair mineralization. Furthermore, pretreatment of pellets of auricular chondrocytes with CCN2/CTGF and subcutaneous implantation significantly enhanced the growth of ectopic auricular cartilage pieces expressing phenotypic markers of auricular chondrocytes including type 11 and X collagen. Notably, chondrocyte apoptosis was impaired by CCN2/CTGF.Conclusions: These findings show that CCN2/CTGF may be a suitable agent for promoting differentiation and growth of auricular chondrocytes, while preventing mineralization and apoptosis, and suggests that CCN2/CTGF may be useful for the repair or reconstruction Of Elastic cartilage. (C) 2007 Ostecarthritis Research Society International. Published by Elsevier Ltd. All rights reserved.