组织工程化软骨分泌蛋白的初步蛋白质组学分析

Preliminary Proteomic Analysis of Proteins Secreted by Tissue-engineered Cartilage

目的通过蛋白质组学筛选组织工程化软骨分泌的可溶性蛋白中具有软骨诱导作用的因子。方法分别培养人软骨细胞与皮肤成纤维细胞,接种到PGA上,两种细胞-材料复合物培养2周后,移至无血清培养基中培养6d,并收集所有培养液,用于差异蛋白质组学分析。结果蛋白组学分析显示有21个差异点。对其中差异最大的点作质谱分析,显示该蛋白与Col2A1异构体2最为匹配。结论用蛋白质组学方法 ,可以高通量筛选软骨细胞分泌的,与软骨分化相关的重要蛋白;Col2A1异构体2可能与软骨分化有关。

Objective To screen the chondrogenic factors in soluble proteins secreted by the engineered cartilage using proteomic approach. Methods Human chondrocytes and dermal fibroblasts were collected, and then seeded onto the PGA scaffolds. At 2 weeks, the two types of cell-scaffold constructs were cultured in serum-free media for 6 days. All the serum-free media were collected for proteomic analysis. Results The proteome maps showed 21 difference spots. On mass spectrometry （MS）, the protein showing the most significant difference of expression matched best with alpha 1 type Ⅱ collagen isoform 2. Conclusion Proteomic approach is a high throughout method to screen the important chondrogenic factors secreted by the tissue-engineered cartilage. Alpha 1 type Ⅱ collagen isoform 2 may be related to chondrogenic differentiation.