Deponti Daniela, Di Giancamillo Alessia, Gervaso Francesca, Domenicucci Marco, Domeneghini Cinzia, Sannino Alessandro, Peretti Giuseppe Michele
1 IRCCS Istituto Ortopedico Galeazzi , Milan, Italy .
Tissue Eng Part A. 2014 Mar;20(5-6):1113-26. doi: 10.1089/ten.TEA.2013.0171. Epub 2013 Dec 13.
This study (i) developed a scaffold made of collagen I designed for hosting the autologous chondrocytes, (ii) focused on the optimization of chondrocytes seeding by the addition of the fibrin glue, and (iii) investigated the culture time for the ideal scaffold maturation in vitro. In the first part of the study, fresh chondrocytes were isolated from infant swine articular cartilage, and immediately seeded onto the collagen sponges either in medium or in fibrinogen in order to show the contribute of fibrin glue in cell seeding and survival into the scaffold. In the second part of the study, chondrocytes were first expanded in vitro and then resuspended in fibrinogen, seeded in collagen sponges, and cultured for 1, 3, and 5 weeks in order to identify the optimal time for the rescue of cell phenotype and for the scaffold maturation into a tissue with chondral properties. The histological and immunohistochemical data from the first part of the study (study with primary chondrocytes) demonstrated that the presence of fibrin glue ameliorated cell distribution and survival into the chondral composites. The second part of this work (study with dedifferentiated chondrocytes) showed that the prolongation of the culture to 3 weeks promoted a significant restoration of the cell phenotype, resulting in a composite with proper morphological features, biochemical composition, and mechanical integrity. In conclusion, this study developed a collagenic-fibrin glue scaffold that was able to support chondrocyte survival and synthetic activity in a static culture; in particular, this model was able to turn the engineered samples into a tissue with chondral-like properties when cultured in vitro for at least 3 weeks.
(i) 开发了一种由I型胶原蛋白制成的支架，用于容纳自体软骨细胞；(ii) 重点通过添加纤维蛋白胶来优化软骨细胞接种；(iii) 研究体外理想支架成熟的培养时间。在研究的第一部分，从幼猪关节软骨中分离出新鲜软骨细胞，并立即将其接种到培养基或纤维蛋白原中的胶原海绵上，以显示纤维蛋白胶在细胞接种和在支架内存活中的作用。在研究的第二部分，先将软骨细胞进行体外扩增，然后重悬于纤维蛋白原中，接种到胶原海绵上，并培养1、3和5周，以确定挽救细胞表型和使支架成熟为具有软骨特性组织的最佳时间。研究第一部分（原代软骨细胞研究）的组织学和免疫组化数据表明，纤维蛋白胶的存在改善了细胞在软骨复合材料中的分布和存活。这项工作的第二部分（去分化软骨细胞研究）表明，将培养时间延长至3周可显著促进细胞表型的恢复，从而形成具有适当形态特征、生化组成和机械完整性的复合材料。总之，本研究开发了一种胶原-纤维蛋白胶支架，能够在静态培养中支持软骨细胞存活和合成活性；特别是，该模型在体外培养至少3周时能够将工程化样本转化为具有软骨样特性的组织。
