中文摘要：

目的应用比较蛋白质组学分析方法筛选组织工程化软骨成熟过程中变化显著的细胞外基质蛋白，通过线性相关分析，研究与软骨力学性能相关的蛋白。方法分离人胎儿软骨细胞培养并扩增，接种到聚羟基乙酸／聚乳酸（PGA／PLA）支架材料，体外培养4周后随机分为3组，每组6例，A组继续体外培养6周、B组回植裸鼠体内培养6周、c组回植裸鼠体内培养12周。取材后进行大体观察、湿重及体积测定、组织学及生物力学等相关检测，比较体内外构建软骨与天然软骨形成质量上的差异。同时进行蛋白质组学分析，选择细胞外基质中比值大于2倍的蛋白与相应各组杨氏模量做相关性分析，根据相关系数筛选与软骨力学性能最相关的蛋白。结果3组均能在体内外形成透明软骨样组织，c组和B组软骨外观近似天然成人软骨，组织学显示大量成熟软骨陷窝，而A组形成的软骨组织结构较松散，组织学可见不规则肥大软骨陷窝。c组湿重和生物力学检测分别为（372．5±35．4）mg和（8．68±2．65）MPa，明显高于B组的（346±34．5）mg和（3．25±1．24）MPa（P〈0．01），而A组为（184．4±12．28）mg和（0．7±0．23）MPa。应用蛋白质组学技术检测到细胞外基质中共44个差异蛋白，选择胞外基质中C组和A组之间差异最大的6个蛋白，与相应各组杨氏模量进行线性相关分析，发现核心蛋白聚糖（Decorin）、软骨粘附素（Chondroadhefin）和纤调蛋白（Fibromodulin）与软骨力学性能变化显著相关（P〈0．05）。结论蛋白质组学技术能够提供大量的蛋白质信息，组织构建与比较蛋白质组学技术相结合，有利于进一步分析组织工程化软骨胞外基质蛋白与力学性能的关系。

英文摘要：

Objective To study proteins correlated with the mechanical properties of engineered cartilage by screening significantly changed proteins during cartilage formation by comparative proteomic analysis. Methods Human chondrocyte, cultured and expanded, were seeded onto a polyglycolic acid/ polylactic acid （PGA/PLA） scaffolds. After 4 weeks of culture in vitro, the constructs were divided into three groups. There were 6 specimens in each group. For the regular in vitro culture group （ A）, the constructs were kept in culture at the original condition for an additional 6 weeks. For in vivo groups, the constructs were implanted subcutaneously into nude mice for either 6 weeks （B） or 12 weeks （C）. All specimens were harvested for gross observation, average wet weight and volume measurement, histology, immunohistochemistry and biomechanics to evaluate the results. Meanwhile, comparative proteomic analysis was performed for each group, and those proteins involved in extracellular matrix with at least 2 folds up-regulation were choosen for further exploration. The correlations between Young＇ s modulus and the relative content of the selected proteins were analyzed by Pearson correlation coefficient. Results All these samples in the three groups eventually formed hyaline-like cartilage structure. Specimens in C and B groups were similar with adult articular cartilage in appearance, and had multiple mature lacuna in histology. However, those specimens in A group had loose texture with irregular hypertrophy lucuna. Specimens implanted for 12 weeks in vivo had better wet weight （372.5 ± 35.4） mg and Young＇ s modulus （8.68 ± 2. 65） MPa than those cultured in vivo for 6 weeks （346 ± 34.5） mg, （3.25 ± 1.24） MPa （P 〈 0.01）. In group A, they were （184.4 ± 12.28） mgand （0.7 ± 0.23） MPa. This study had detected 44 proteins in ECM by comparative proteomic analysis, then chosing the greatest ratio of 6 up-regulation proteins compared between C and A groups. The correlation results