Bonello John-Peter, Tse M Yat, Robinson Trevor J G, Bardana Davide D, Waldman Stephen D, Pang Stephen C
Department of Biomedical and Molecular Sciences, Queen's University, Kingston, ON, Canada.
Division of Surgery, Kingston General Hospital, Kingston, ON, Canada.
Cartilage. 2024 Apr 14:19476035241241930. doi: 10.1177/19476035241241930.
While substantial progress has been made in engineering cartilaginous constructs for animal models, further research is needed to translate these methodologies for human applications. Evidence suggests that cultured autologous chondrocytes undergo changes in phenotype and gene expression, thereby affecting their proliferation and differentiation capacity. This study was designed to evaluate the expression of chondrogenic markers in cultured human articular chondrocytes from passages 3 (P3) and 7 (P7), beyond the current clinical recommendation of P3.
Cultured autologous chondrocytes were passaged from P3 up to P7, and quantitative polymerase chain reaction (qPCR) was used to assess mRNA expression of chondrogenic markers, including collagen type I (COLI), collagen type II (COLII), aggrecan (AGG), bone morphogenetic protein 4 (BMP4), transcription factor SOX-9 (SOX9), proteoglycan 4 (PGR4), and transformation-related protein 53 (p53), between P3 and P7.
Except for AGG, no significant differences were found in the expression of markers between passages, suggesting the maintenance of chondrogenic potential in cultured chondrocytes. Differential expression identified between SOX9 and PGR4, as well as between COLI and SOX9, indicates that differences in chondrogenic markers are present between age groups and sexes, respectively.
Overall, expression profiles of younger and male chondrocytes exhibit conversion of mature cartilage characteristics compared to their counterparts, with signs of dedifferentiation and loss of phenotype within-group passaging. These results may have implications in guiding the use of higher passaged chondrocytes for engineering constructs and provide a foundation for clinical recommendations surrounding the repair and treatment of articular cartilage pathology in both sexes.
虽然在为动物模型构建软骨组织方面已取得重大进展，但仍需进一步研究将这些方法转化用于人类。有证据表明，培养的自体软骨细胞会发生表型和基因表达变化，从而影响其增殖和分化能力。本研究旨在评估传代3次（P3）和7次（P7）的培养人关节软骨细胞中软骨生成标志物的表达情况，超出了目前临床推荐的P3代。
将培养的自体软骨细胞从P3代传至P7代，采用定量聚合酶链反应（qPCR）评估P3代和P7代之间软骨生成标志物的mRNA表达，包括I型胶原（COLI）、II型胶原（COLII）、聚集蛋白聚糖（AGG）、骨形态发生蛋白4（BMP4）、转录因子SOX - 9（SOX9）、蛋白聚糖4（PGR4）和转化相关蛋白53（p53）。
除AGG外，各传代之间标志物表达未发现显著差异，表明培养的软骨细胞软骨生成潜能得以维持。SOX9和PGR4之间以及COLI和SOX9之间的差异表达分别表明，年龄组和性别之间软骨生成标志物存在差异。
总体而言，与年轻和男性软骨细胞的对应细胞相比，其表达谱显示出成熟软骨特征的转变，在组内传代过程中有去分化和表型丧失的迹象。这些结果可能对指导使用传代次数更高的软骨细胞构建工程组织具有意义，并为围绕两性关节软骨病理修复和治疗的临床建议提供基础。
