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脱细胞软骨基质作为一种新型的生物基质在软骨组织工程应用中。

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

机构信息

Department of Otorhinolaryngology, Ulm University Medical Center, Ulm, Germany.

出版信息

Tissue Eng Part A. 2012 Nov;18(21-22):2195-209. doi: 10.1089/ten.TEA.2011.0705. Epub 2012 Jul 20.

DOI:10.1089/ten.TEA.2011.0705
PMID:22690787
Abstract

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.

摘要

头颈部和骨科部位的软骨结构损伤通常由创伤、肿瘤切除或先天缺陷引起。尽管许多临床领域对此有很高的需求,但直到今天,还没有一种合适的软骨替代基质可以应用于这些领域。由于术中处理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进一步优化。我们已经开发并应用了一种新的化学处理方法,对人源和猪源软骨组织(半月板软骨和鼻中隔)进行完全脱细胞和灭菌处理,以生成一种新型的生物植入基质。为了对该基质进行表征并确定脱细胞过程的效果,我们测定了变性胶原(w(D))和糖胺聚糖(GAGs)(w(G))的含量。我们通过将处理过的软骨生物基质与原代人软骨细胞和小鼠成纤维细胞(L929)共培养,在体外研究了该基质的潜在细胞毒性和细胞相容性。使用 MTS 检测法测量细胞活力和代谢状态。两种细胞类型均黏附在支架表面并增殖。未检测到生长抑制或细胞毒性的区域。观察到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,证明了基质孔隙率增加、细胞完全消除和 GAG 大量去除。经过处理的异种和同种异体软骨生物基质来源于天然组织,具有形状、尺寸和生物力学的多样性,是各种生物医学应用的有前途的候选材料。

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