目的探究鸡蛋清制备多孔支架用于组织工程软骨构建的可行性。方法将鸡蛋清与去离子水以不同体积比(1:0、1:1和1:2)混合,通过冷冻干燥制备成多孔支架,行大体观、孔隙率及力学性能检测。取兔耳软骨细胞接种于不同体积比支架,于体外培养1 d、4 d、7 d后进行细胞染色和细胞增殖实验;体外培养至3周后,行组织学检测,观察其成软骨情况。结果三种不同体积比的蛋清均能制备成多孔结构的支架材料。随着去离子水比例的增加,其孔径大小明显提高,但是力学强度却呈下降趋势。活死细胞染色及细胞增殖实验证明,细胞可以在支架上黏附、增殖。延长体外培养时间至3周,HE染色显示三组蛋清支架上均有稚嫩的新生软骨基质形成,证实蛋清支架体外构建软骨的可行性。值得注意的是,随着去离子水比例的增加,残余的支架材料减少,新生软骨基质成分增多。结论基于鸡蛋清制备的多孔支架适合用于组织工程软骨构建。
Objective To explore the feasibility of porous scaffold based on egg white for tissue-engineered cartilage regeneration. Methods Egg white was mixed with deionized water in different volume ratios(1:0, 1:1 and 1:2) to fabricate porous scaffolds after freeze-drying. Afterwards, the three kinds of scaffolds were conducted with gross observation, porous size and mechanical property examination. Chondrocytes derived from rabbit ear in the second passage were seeded into the scaffolds with different volume ratios and cultured in vitro. The examination of Living & dead staining and cell proliferation assay were conducted after in vitro cultivation for 1, 4 and 7 days. Moreover, the chondrogenesis was observed by histological examination after in vitro culture for 3 weeks. Results Porous scaffolds could be fabricated by all the three kinds of egg white with different volume ratios. The pore size presented significantly increasing trend with the increase of deionized water content, but the mechanical property presented the reversed trend. The examination of both living & dead staining and cell proliferation assay showed that the chondrocytes could attach and proliferate on all the three kinds of scaffold. When extended the in vitro culture time to 3 weeks, HE staining showed that egg white scaffolds of all three groups had immature neocartilage matrix formation, which confirmed the feasibility of constructing cartilage based on egg white scaffold in vitro.Notably, with the increase of deionized water ratio, the amounts of residual scaffolds decreased but the component of neocartilage matrix increased. Conclusion Porous scaffolds based on egg white are suitable for constructing tissue engineered cartilage.
