Matrix-assisted autologous chondrocyte transplantation for remodeling and repair of chondral defects in a rabbit model.
Matrix-assisted autologous chondrocyte transplantation for remodeling and repair of chondral defects in a rabbit model.
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DOI：
10.3791/4422
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发表时间：
2013-05
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通讯作者：
M. Berninger;Gabriele Wexel;E. Rummeny;A. Imhoff;M. Anton;T. Henning;S. Vogt
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文献类型：
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作者：
M. Berninger;Gabriele Wexel;E. Rummeny;A. Imhoff;M. Anton;T. Henning;S. Vogt
关节软骨缺陷被认为是一个主要的健康问题，因为关节软骨的自我再生能力有限 (1)。未经治疗的软骨损伤会导致持续疼痛，对生活质量产生负面影响，并容易患骨关节炎。在过去的几十年中，已经开发了几种手术技术来治疗此类病变。然而，到目前为止，还无法实现用透明关节软骨覆盖缺损或提供令人满意的长期恢复的完全修复 (2-4)。因此，关节软骨损伤仍然是组织工程等再生技术的主要目标。与其他通常导致纤维或纤维软骨组织形成的外科技术不同，组织工程旨在利用移植细胞的软骨形成潜力，完全恢复原始关节软骨的复杂结构和特性。最近的发展为再生软骨疗法开辟了有希望的可能性。 Lars Peterson 和 Mats Brittberg 于 1994 年首次采用基于细胞的方法治疗全层软骨或骨软骨损伤，他们率先进行了临床自体软骨细胞植入 (ACI) (5)。如今，该技术在临床上已成熟用于治疗膝关节大面积透明软骨缺损，即使在植入后 10 至 20 年也能保持良好的临床效果 (6)。近年来，自体软骨细胞的植入发展迅速。使用人工三维胶原蛋白基质（随后将细胞重新移植到其上）变得越来越流行 (7-9)。 MACT 包括两个外科手术：首先，为了收集软骨细胞，需要从膝关节的非承重软骨区域进行软骨活检。然后，软骨细胞被提取、纯化并在体外扩增至足够的细胞数量。然后将软骨细胞接种到三维矩阵上，随后可以重新植入。在制备组织工程植入物时，增殖率和分化能力对于组织再生的成功至关重要 (10)。使用三维矩阵作为细胞载体被认为支持这些细胞特性 (11)。以下协议将总结并演示从软骨活检中分离软骨细胞、体外增殖以及接种到 3D 基质上的技术（Chondro-Gide，Geistlich Biomaterials，Wollhusen，瑞士）。最后，将描述将细胞基质构建体植入兔膝关节的人工软骨缺损中。该技术可用作软骨修复进一步实验的实验设置。
Articular cartilage defects are considered a major health problem because articular cartilage has a limited capacity for self-regeneration (1). Untreated cartilage lesions lead to ongoing pain, negatively affect the quality of life and predispose for osteoarthritis. During the last decades, several surgical techniques have been developed to treat such lesions. However, until now it was not possible to achieve a full repair in terms of covering the defect with hyaline articular cartilage or of providing satisfactory long-term recovery (2-4). Therefore, articular cartilage injuries remain a prime target for regenerative techniques such as Tissue Engineering. In contrast to other surgical techniques, which often lead to the formation of fibrous or fibrocartilaginous tissue, Tissue Engineering aims at fully restoring the complex structure and properties of the original articular cartilage by using the chondrogenic potential of transplanted cells. Recent developments opened up promising possibilities for regenerative cartilage therapies. The first cell based approach for the treatment of full-thickness cartilage or osteochondral lesions was performed in 1994 by Lars Peterson and Mats Brittberg who pioneered clinical autologous chondrocyte implantation (ACI) (5). Today, the technique is clinically well-established for the treatment of large hyaline cartilage defects of the knee, maintaining good clinical results even 10 to 20 years after implantation (6). In recent years, the implantation of autologous chondrocytes underwent a rapid progression. The use of an artificial three-dimensional collagen-matrix on which cells are subsequently replanted became more and more popular (7-9). MACT comprises of two surgical procedures: First, in order to collect chondrocytes, a cartilage biopsy needs to be performed from a non weight-bearing cartilage area of the knee joint. Then, chondrocytes are being extracted, purified and expanded to a sufficient cell number in vitro. Chondrocytes are then seeded onto a three-dimensional matrix and can subsequently be re-implanted. When preparing a tissue-engineered implant, proliferation rate and differentiation capacity are crucial for a successful tissue regeneration (10). The use of a three-dimensional matrix as a cell carrier is thought to support these cellular characteristics (11). The following protocol will summarize and demonstrate a technique for the isolation of chondrocytes from cartilage biopsies, their proliferation in vitro and their seeding onto a 3D-matrix (Chondro-Gide, Geistlich Biomaterials, Wollhusen, Switzerland). Finally, the implantation of the cell-matrix-constructs into artificially created chondral defects of a rabbit's knee joint will be described. This technique can be used as an experimental setting for further experiments of cartilage repair.