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基质辅助自体软骨细胞移植用于兔模型关节软骨缺损的重塑与修复

Matrix-assisted autologous chondrocyte transplantation for remodeling and repair of chondral defects in a rabbit model.

作者信息

Berninger Markus T, Wexel Gabriele, Rummeny Ernst J, Imhoff Andreas B, Anton Martina, Henning Tobias D, Vogt Stephan

机构信息

Department of Orthopaedic Sports Medicine, Klinikum rechts der Isar der Technischen Universität München.

出版信息

J Vis Exp. 2013 May 21(75):e4422. doi: 10.3791/4422.

Abstract

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.

摘要

关节软骨缺损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,并易引发骨关节炎。在过去几十年中,已经开发出几种外科技术来治疗此类损伤。然而,到目前为止,无论是用透明关节软骨覆盖缺损,还是实现令人满意的长期恢复,都无法做到完全修复(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。与其他常常导致纤维组织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生成潜力,完全恢复原始关节软骨的复杂结构和特性。最近的进展为再生软骨治疗开辟了有前景的可能性。1994年,拉尔斯·彼得森和马茨·布里特伯格率先进行了临床自体软骨细胞植入(ACI),这是第一种基于细胞治疗全层软骨或骨软骨损伤的方法(5)。如今,该技术在临床上已被广泛用于治疗膝关节的大面积透明软骨缺损,即使在植入10至20年后仍能保持良好的临床效果(6)。近年来,自体软骨细胞植入技术发展迅速。使用人工三维胶原基质,随后将细胞重新种植在上面,这种方法越来越受欢迎(7-9)。基质辅助自体软骨细胞移植(MACT)包括两个外科手术步骤:首先,为了收集软骨细胞,需要从膝关节非负重软骨区域进行软骨活检。然后,提取、纯化软骨细胞,并在体外扩增到足够的细胞数量。接着将软骨细胞接种到三维基质上,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成功的组织再生至关重要(10)。使用三维基质作为细胞载体被认为有助于这些细胞特性(11)。以下方案将总结并演示一种从软骨活检中分离软骨细胞、在体外增殖并接种到三维基质(Chondro-Gide, Geistlich Biomaterials, Wollhusen,瑞士)上的技术。最后,将描述将细胞-基质构建体植入兔膝关节人工制造的软骨缺损中的过程。该技术可作为软骨修复进一步实验的实验设置。

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Cartilage repair: generations of autologous chondrocyte transplantation.软骨修复:历代自体软骨细胞移植
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本文引用的文献

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Autologous chondrocyte implantation: a long-term follow-up.自体软骨细胞移植:长期随访。
Am J Sports Med. 2010 Jun;38(6):1117-24. doi: 10.1177/0363546509357915. Epub 2010 Feb 24.

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