Abstract
Objective To investigate the tissue structure, chondrocyte characteristics, and the differential expression of related genes and cell surface markers of auricular cartilage of patients in different ages, in order to provide a basis for the age selection of tissue engineered cartilage repair defects. Methods The auricular cartilage tissue was obtained from 22 patients with microtia in the Plastic Surgery Hospital, Chinese Academy of Medical Science, ranged from 6 to 28 years old, and divided into the child group (6-12 years old), the adolescent group (13-18 years old) and the adult group (21-28 years old). The proliferation and differentiation features of chondrocytes which from different-aged patients were detected. Furthermore, quantitative real-time PCR was used to detect the differences in the expression of genes related to cell proliferation and chondrocyte extracellular matrix. Flow cytometry, immunofluorescence and immunohistochemistry were used to detect the differences in the expression of mesenchymal stem cell markers CD90, CD44, CD73 and CD105 in chondrocytes. SPSS Statistics 21.0 software was used to process statistics. Results The proliferation capability of auricular chondrocytes of children was stronger than adolescents and adults, the child group vs the adult group P<0.05, the child group vs the adolescent group P<0.01. The expression of cartilage extracellular matrix related gene COL2A1 increased with age, the child group vs the adult group P<0.01, the adolescent group vs the adult group P<0.01. While the capability of cell osteogenic differentiation decreased with age(P<0.05). However, there was no significant difference in the capability of adipogenic differentiation when considering the ages of patients. The results of both flow cytometry and real-time PCR showed that the expression of mesenchymal stem cell markers decreased with age, with the most significant decrease in CD90(P<0.01). Conclusions The biological characteristics and stem cell content of cells derived from auricular cartilage tissue was influenced by the patients′age. Key words: Age; Auricular cartilage; Cell proliferation; Cell differentiation; Cartilage stem/progenitor cells
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