Kang Ning, Liu Xia, Cao Yilin, Xiao Ran
Zhonghua Zheng Xing Wai Ke Za Zhi. 2014 Jan;30(1):33-40.
To compare the tissue engineered cartilage constructed with chondrocytes derived from auricular and articular cartilage.
Chondrocytes were isolated from porcine auricular and articular cartilage, and BMSCs were obtained from bone marrow aspirate and cultured. Each kind of chondrocytes were resuspended alone or mixed with BMSCs at a ratio of 1:1, and seeded onto PGA/PLA scaffolds to construct tissue engineered cartilage (n = 4). The constructs were cultured for 8 weeks in vitro and then subcutaneously implanted into nude mice for 6 weeks. The differences between chondrocytes monoculture from articular and auricular cartilage or between each of them co-cultured with BMSCs were evaluated by gross view, measurement of thickness and wet weight, histological examinations including H&E, Safranin O, type II collagen, and Ponceau's & Victoria blue staining, and gene expression analysis of cartilage related genes.
No obvious differences were found histologically among the complexes constructed in vitro 8 weeks except for few elastic fibers secreted in the auricular chondrocytes + BMSCs co-culture group. Neo-cartilage is thicker in the groups of articular chondrocytes (38. 1% than the group of auricular chondrocytes, P < 0.05) and articular chondrocytes + BMSCs co-culture (19.3% than the group of auricular chondrocytes + BMSCs, P < 0.05). However, after 6 weeks in vivo the elastic fibers were found positive in the complexes constructed by auricular chondrocytes, and its staining was even stronger and more homogenous in the group of auricular chondrocytes + BMSCs co-culture. The tissues generated by articular chondrocytes alone and co-cultured with BMSCs both formed the characteristic features of three-layer structure of hyaline cartilage and ossified in vivo with significant up-regulation of COL10A1 and MMP-13. To summarize, auricular chondrocytes formed the elastic cartilage while articular chondrocytes formed the hyaline cartilage during the development of tissue engineered cartilage either by monoculture or the co-culture with BMSCs.
The chondrogenic response of chondrocytes from different cartilage origins demonstrates that an initial chondrocyte and cartilage type recapitulates the same in later tissue-engineered development.
比较用耳软骨和关节软骨来源的软骨细胞构建的组织工程软骨。
从猪的耳软骨和关节软骨中分离软骨细胞，从骨髓抽吸物中获取骨髓间充质干细胞（BMSCs）并进行培养。将每种软骨细胞单独重悬或与BMSCs按1:1比例混合，接种到聚乙醇酸/聚乳酸（PGA/PLA）支架上构建组织工程软骨（n = 4）。构建物在体外培养8周，然后皮下植入裸鼠体内6周。通过大体观察、厚度和湿重测量、包括苏木精-伊红（H&E）、番红O、II型胶原以及丽春红和维多利亚蓝染色的组织学检查，以及软骨相关基因的基因表达分析，评估关节软骨和耳软骨的软骨细胞单培养之间或它们各自与BMSCs共培养之间的差异。
体外培养8周构建的复合物在组织学上未发现明显差异，除了耳软骨细胞+BMSCs共培养组分泌少量弹性纤维。关节软骨细胞组和关节软骨细胞+BMSCs共培养组的新生软骨更厚（分别比耳软骨细胞组厚38.1%，P < 0.05；比耳软骨细胞+BMSCs组厚19.3%，P < 0.05）。然而，在体内6周后，发现耳软骨细胞构建的复合物中弹性纤维呈阳性，且在耳软骨细胞+BMSCs共培养组中其染色更强且更均匀。单独的关节软骨细胞以及与BMSCs共培养产生的组织均形成了透明软骨三层结构的特征，并在体内发生骨化，伴有COL10A1和基质金属蛋白酶-13（MMP-13）的显著上调。总之，在组织工程软骨的发育过程中，无论是单培养还是与BMSCs共培养，耳软骨细胞形成弹性软骨，而关节软骨细胞形成透明软骨。
不同软骨来源的软骨细胞的软骨形成反应表明，初始的软骨细胞和软骨类型在后期的组织工程发育中会重现相同的情况。
