Quantitative outcome measures of cartilage repair in patients treated by tissue engineering
Quantitative outcome measures of cartilage repair in patients treated by tissue engineering
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DOI：
10.1089/ten.2005.11.277
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发表时间：
2005-01-01
影响因子：
--
通讯作者：
Hollander, AP
中科院分区：
文献类型：
--
作者：
Dickinson, SC;Sims, TJ;Hollander, AP
可靠和可重复的结果测量对于评估竞争性和新的组织工程技术的有效性是必不可少的。这项研究的目的是比较传统的组织学分析和新开发的定量生化结果指标在关节软骨修复中的作用。介绍了一种新的抗多肽抗体的制备，以及一种新的I型胶原蛋白提取和免疫分析方法的开发和验证。该分析结合现有的II型胶原和蛋白多糖的分析方法，用于测量从接受新的组织工程治疗的患者的修复组织活检中获得的基质成分的水平。从相同的活检组织中切下的冰冻切片用藏红花O进行蛋白多糖染色，并使用免疫组织化学分析来评估I型和II型胶原的染色。尽管染色的程度和三种基质成分的数量基本一致，但根据低、中或丰富染色的组织学分类，活检组织的生化含量有很大程度的重叠。结果表明，用基质蛋白丰度的组织学分级将修复软骨分为透明软骨或纤维软骨往往具有误导性。此外，我们首次证明了在修复组织活检中测量胶原交联物的能力，并表明它可以用作组织成熟度的替代标记。我们的一系列新的生化技术提供了一种标准化的方法来评估体外生产的工程软骨和修复体内植入后的组织活检的质量。
Reliable and reproducible outcome measures are essential to assess the efficacy of competing and novel tissue-engineering techniques. The aim of this study was to compare traditional histological analyses with newly developed quantitative biochemical outcome measures for the repair of articular cartilage. The production of a new anti-peptide antibody and the development and validation of a novel method for the extraction and immunoassay of type I collagen are described. The assay was used, in conjunction with existing assays for type II collagen and proteoglycans, to measure levels of the matrix components in repair tissue biopsies obtained from patients treated with the new tissue-engineering therapy Hyalograft(R) C. Frozen sections cut from the same biopsies were stained for proteoglycans, using safranin O, and immunohistochemical analysis was used to assess type I and II collagen staining. Although there was general agreement between the extent of staining and the amounts of the three matrix components, there was a large degree of overlap in biochemical content between biopsies classified histologically on the basis of low, moderate, or abundant staining. The results demonstrate that histological grading of matrix protein abundance to classify repair cartilage as hyaline or fibrocartilagenous is often misleading. In addition, we demonstrate for the first time the ability to measure collagen cross-links in repair tissue biopsies and show that it can be used as a surrogate marker for tissue maturity. Our new range of biochemical techniques provides a standardized method to assess the quality of both engineered cartilage produced in vitro and repair tissue biopsies obtained after in vivo implantation.