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Decellularized cartilage matrix as a novel biomatrix for cartilage tissue-engineering applications

机译:去细胞软骨基质作为用于软骨组织工程应用的新型生物基质

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Damage of cartilage structures in the head and neck region as well as in orthopedic sites are frequently caused by trauma, tumor resection, or congenital defects. Despite a high demand in many clinical fields, until today, no adequate cartilage replacement matrix is available for these fields of application. Materials that are clinically applied for joint cartilage repair still need optimization due to difficult intraoperative handling and risk of early mechanical damage. We have developed and applied a novel chemical process to completely decellularize and sterilize human and porcine cartilage tissues (meniscus cartilage and nasal septum) to generate a new type of bioimplant matrix. To characterize this matrix and to determine the effect of the decellularization process, the content of denatured collagen (w D) and the content of glycosaminoglycans (GAGs) (w G) were determined. Possible cytotoxic effects and cellular compatibility of the matrix in vitro have been examined by seeding processed cartilage biomatrices with human primary chondrocytes as well as murine fibroblasts (L929). Vitality and state of metabolism of cells were measured using MTS assays. Both cell types adhered to scaffold surfaces and proliferated. No areas of growth inhibition or cytotoxic effects were detected. New synthesis of cartilage-specific extracellular matrix was observed. By histological staining, electron microscopy, and μCT analysis, an increase of matrix porosity, complete cell elimination, and high GAG removal were demonstrated. Being from natural-origin, processed xenogenic and allogeneic cartilage biomatrices are highly versatile with regard to shape, size, and biomechanics, making them promising candidates for various biomedical applications.
机译:头部,颈部以及整形外科部位的软骨结构损坏通常是由外伤,肿瘤切除或先天性缺陷引起的。尽管在许多临床领域都有很高的需求,但是直到今天,对于这些应用领域还没有足够的软骨替代基质。由于术中操作困难和早期机械损伤的风险,临床上用于关节软骨修复的材料仍需要优化。我们已经开发并应用了一种新颖的化学方法,可以对人和猪的软骨组织(半月板软骨和鼻中隔)进行完全脱细胞和灭菌,以生成新型的生物植入物基质。为了表征该基质并确定脱细胞过程的效果,测定了变性胶原蛋白的含量(w D)和糖胺聚糖(GAGs)的含量(w G)。通过在处理过的软骨生物基质中接种人原代软骨细胞和鼠成纤维细胞(L929),已经检查了体外基质的可能的细胞毒性作用和细胞相容性。使用MTS测定法测量细胞的活力和代谢状态。两种细胞类型均粘附在支架表面并增殖。没有检测到生长抑制或细胞毒性作用的区域。观察到了软骨特异性细胞外基质的新合成。通过组织学染色,电子显微镜和μCT分析,证明了基质孔隙率的增加,细胞的完全清除和高GAG去除率。经过加工的异种和同种异体软骨生物基质来自天然,在形状,大小和生物力学方面具有很高的通用性,使其成为各种生物医学应用的有希望的候选者。

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