Extraction techniques for the decellularization of tissue engineered articular cartilage constructs.
Extraction techniques for the decellularization of tissue engineered articular cartilage constructs.
复制标题
DOI：
10.1016/j.biomaterials.2009.03.050
复制
发表时间：
2009-08
期刊：
影响因子：
14
通讯作者：
Athanasiou, Kyriacos A.
中科院分区：
文献类型：
--
作者：
Elder, Benjamin D.;Eleswarapu, Sriram V.;Athanasiou, Kyriacos A.
一些先前的研究已经进行，以确定组织脱细胞的可行性，以创建一个非免疫原性异种组织替代膀胱，血管，心脏瓣膜，膝关节半月板，颞下颌关节盘，韧带和肌腱。然而，有限的工作已经完成了关节软骨，并没有研究检查组织工程结构的脱细胞。本研究的目的是评估不同脱细胞处理对关节软骨构建的影响，采用无支架的方法，经过4周的培养，采用两阶段的方法。在第一阶段，研究了五种不同的处理方法：1)1% SDS, 2) 2% SDS, 3) 2%磷酸三丁酯，4)2% Triton X-100, 5)低渗后高渗溶液。这些处理分别应用1小时或8小时，然后在PBS中洗涤2小时。清洗后，对构建体进行组织学、生物化学的细胞度、GAG和胶原含量评估，以及生物力学的压缩和拉伸性能评估。在II期，为了优化施用时间，将I期的最佳处理分别施用1、2、4、6或8 h。用2% SDS处理1小时或2小时显著降低了组织的DNA含量，同时保持了组织的生化和生物力学性能。另一方面，2% SDS作用6 h或8 h可导致组织学上完全脱细胞，组织学染色上细胞核完全消失，但GAG含量和压缩性能显著降低。总的来说，2%的SDS，作用1或2小时，似乎是软骨脱细胞最有效的药物，因为它在保持功能特性的同时导致脱细胞。这项研究的结果是令人兴奋的，因为它们表明了创造非免疫原性的工程软骨作为替代组织的可行性。
Several prior studies have been performed to determine the feasibility of tissue decellularization to create a non-immunogenic xenogenic tissue replacement for bladder, vasculature, heart valves, knee meniscus, temporomandibular joint disc, ligament, and tendon. However, limited work has been performed with articular cartilage, and no studies have examined the decellularization of tissue engineered constructs. The objective of this study was to assess the effects of different decellularization treatments on articular cartilage constructs, engineered using a scaffoldless approach, after 4 wks of culture, using a two-phased approach. In the first phase, five different treatments were examined: 1) 1% SDS, 2) 2% SDS, 3) 2% Tributyl phosphate, 4) 2% Triton X-100, and 5) Hypotonic followed by hypertonic solution. These treatments were applied for either 1 h or 8 h, followed by a 2 h wash in PBS. Following this wash, the constructs were assessed histologically, biochemically for cellularity, GAG, and collagen content, and biomechanically for compressive and tensile properties. In phase II, the best treatment from phase I was applied for 1, 2, 4, 6, or 8 h in order to optimize the application time. Treatment with 2% SDS for 1 h or 2 h significantly reduced the DNA content of the tissue, while maintaining the biochemical and biomechanical properties. On the other hand, 2% SDS for 6 h or 8 h resulted in complete histological decellularization, with complete elimination of cell nuclei on histological staining, although GAG content and compressive properties were significantly decreased. Overall, 2% SDS, for 1 or 2 h, appeared to be the most effective agent for cartilage decellularization, as it resulted in decellularization while maintaining the functional properties. The results of this study are exciting as they indicate the feasibility of creating engineered cartilage that may be non-immunogenic as a replacement tissue.
登录
查看更多内容
DOI：
10.1016/j.jtcvs.2005.03.021
发表时间：
2005-08-01
影响因子：
6
作者：
Meyer, SR;Nagendran, J;Ross, DB
通讯作者：
Ross, DB
影响因子：
1.7
作者：
Kasimir, MT;Rieder, E;Simon, P
通讯作者：
Simon, P
影响因子：
4.9
作者：
Meyer, Steven R.;Chiu, Brian;Ross, David B.
通讯作者：
Ross, David B.
影响因子：
2.8
作者：
ATHANASIOU, KA;AGARWAL, A;DZIDA, FJ
通讯作者：
DZIDA, FJ
DOI：
10.2519/jospt.1998.28.4.192
发表时间：
1998-10-01
影响因子：
6.1
作者：
Buckwalter, JA
通讯作者：
Buckwalter, JA