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A silanized hydroxypropyl methylcellulose hydrogel for the three-dimensional culture of chondrocytes

机译:用于软骨细胞三维培养的硅烷化羟丙基甲基纤维素水凝胶

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Articular cartilage has limited intrinsic repair capacity. In order to promote cartilage repair, the amplification and transfer of autologous chondrocytes using three-dimensional scaffolds have been proposed. We have developed an injectable and self-setting hydrogel consisting of hydroxypropyl methylcellulose grafted with silanol groups (Si-HPMC). The aim of the present work is to assess both the in vitro cytocompatibility of this hydrogel and its ability to maintain a chondrocytc-specific phenotype. Primary chondrocytes isolated from rabbit articular cartilage (RAC) and two human chondrocytic cell lines (SW1353 and C28/12) were cultured into the hydrogel. Methyl tetrazolium salt (NITS) assay and cell counting indicated that Si-HPMC hydrogel did not affect respectively chondrocyte viability and proliferation. Fluorescent microscopic observations of RAC and C28/12 chondrocytes double-labeled with cell tracker green and ethidium homodimer-1 revealed that chondrocytes proliferated within Si-HPMC. Phenotypic analysis (RT-PCR and Alcian blue staining) indicates that chondrocytes, when three-dimensionnally cultured within Si-HPMC, expressed transcripts encoding type 11 collagen and aggrecan and produced sulfated glycosaminoglycans. These results show that Si-HPMC allows the growth of differentiated chondrocytes. Si-HPMC therefore appears as a potential scaffold for three-dimensional amplification and transfer of chondrocytes in cartilage tissue engineering. (c) 2005 Elsevier Ltd. All rights reserved.
机译:关节软骨具有有限的固有修复能力。为了促进软骨修复,已经提出了使用三维支架来扩增和转移自体软骨细胞的方法。我们已经开发了一种由羟丙基甲基纤维素与硅烷醇基团(Si-HPMC)接枝的可注射的自凝水凝胶。本工作的目的是评估该水凝胶的体外细胞相容性及其维持软骨细胞特异性表型的能力。从兔关节软骨(RAC)和两种人软骨细胞系(SW1353和C28 / 12)中分离出的原代软骨细胞被培养到水凝胶中。甲基四唑盐(NITS)测定和细胞计数表明,Si-HPMC水凝胶不会分别影响软骨细胞的活力和增殖。 RAC和C28 / 12软骨细胞用细胞追踪绿色和乙二胺均二聚体1双重标记的荧光显微镜观察表明,软骨细胞在Si-HPMC中增殖。表型分析(RT-PCR和Alcian蓝染色)表明,当软骨细胞在Si-HPMC中三维培养时,其表达的转录本编码11型胶原蛋白和聚集蛋白聚糖,并产生硫酸化的糖胺聚糖。这些结果表明Si-HPMC允许分化的软骨细胞生长。因此,Si-HPMC似乎是软骨组织工程中软骨细胞三维扩增和转移的潜在支架。 (c)2005 Elsevier Ltd.保留所有权利。

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