Vinatier C, Magne D, Weiss P, Trojani C, Rochet N, Carle G F, Vignes-Colombeix C, Chadjichristos C, Galera P, Daculsi G, Guicheux J
INSERM EM 9903, Research Center on Materials with Biological Interest, School of Dental Surgery, University of Nantes, 1-Place Alexis Ricordeau 44042, Nantes Cedex 1, France.
Biomaterials. 2005 Nov;26(33):6643-51. doi: 10.1016/j.biomaterials.2005.04.057.
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 chondrocyte-specific phenotype. Primary chondrocytes isolated from rabbit articular cartilage (RAC) and two human chondrocytic cell lines (SW1353 and C28/I2) were cultured into the hydrogel. Methyl tetrazolium salt (MTS) assay and cell counting indicated that Si-HPMC hydrogel did not affect respectively chondrocyte viability and proliferation. Fluorescent microscopic observations of RAC and C28/I2 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 II 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.
关节软骨的内在修复能力有限。为了促进软骨修复，人们提出了使用三维支架扩增和移植自体软骨细胞的方法。我们开发了一种由接枝有硅醇基团的羟丙基甲基纤维素（Si-HPMC）组成的可注射自固化水凝胶。本研究的目的是评估这种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。将从兔关节软骨（RAC）分离的原代软骨细胞以及两个人软骨细胞系（SW1353和C28/I2）培养于该水凝胶中。甲基四氮唑盐（MTS）测定和细胞计数表明，Si-HPMC水凝胶分别不影响软骨细胞的活力和增殖。用细胞追踪绿和乙锭同二聚体-1对RAC和C28/I2软骨细胞进行双重标记后的荧光显微镜观察显示，软骨细胞在Si-HPMC内增殖。表型分析（RT-PCR和阿尔新蓝染色）表明，当在Si-HPMC内进行三维培养时，软骨细胞表达编码II型胶原蛋白和聚集蛋白聚糖的转录本，并产生硫酸化糖胺聚糖。这些结果表明，Si-HPMC能促进分化软骨细胞的生长。因此，Si-HPMC似乎是软骨组织工程中软骨细胞三维扩增和移植的潜在支架。
