Zwickl Hannes, Niculescu-Morzsa Eugenia, Halbwirth Florian, Bauer Christoph, Jeyakumar Vivek, Reutterer Angelique, Berger Manuela, Nehrer Stefan
Center for Regenerative Medicine and Orthopaedics, Department for Clinical Medicine and Biotechnology, Danube University Krems, Krems, Austria.
Cartilage. 2016 Apr;7(2):185-92. doi: 10.1177/1947603515615388. Epub 2016 Mar 17.
Matrix-assisted autologous chondrocyte implantation is frequently applied to replace damaged cartilage in order to support tissue regeneration or repair and to prevent progressive cartilage degradation and osteoarthritis. Its application, however, is limited to primary defects and contraindicated in the case of osteoarthritis that is partially ascribed to dedifferentiation and phenotype alterations of chondrocytes obtainable from patients' biopsies. The differentiation state of chondrocytes is reflected at the level of structural gene (COL2A1, ACAN, COL1A1) and transcription factor (SOX9, 5, 6) expression.
METHODS/DESIGN: We determined the mRNA abundances of COL2A1, ACAN, and COL1A1as well as SOX9, -5, and -6 of freshly isolated and passaged collagen I implant-derived and osteoarthritic chondrocytes via reverse transcription-polymerase chain reaction. Moreover, we analyzed the correlation of structural and transcription factor gene expression. Thus, we were able to evaluate the impact of the mRNA levels of transcription factors on the expression of cartilage-specific structural genes.
Significant differences were obtained (1) for freshly isolated osteoarthritic versus collagen I implant-derived chondrocytes, (2) due to passaging of the respective cell sources, (3) for osteoarthritic versus nonosteoarthritic chondrocytes, and (4) for COL2A1 versus ACAN expression with respect to the coherence with SOX9, -5, and -6 transcript levels.
Our results might contribute to a better understanding of the transcriptional regulation of structural gene expression of chondrocytes with implications for their use in matrix-assisted autologous chondrocyte implantation.
基质辅助自体软骨细胞移植常用于替换受损软骨,以支持组织再生或修复,并预防软骨进行性退变和骨关节炎。然而,其应用仅限于原发性缺损,在骨关节炎情况下则为禁忌,部分原因是从患者活检获得的软骨细胞会发生去分化和表型改变。软骨细胞的分化状态反映在结构基因(COL2A1、ACAN、COL1A1)和转录因子(SOX9、5、6)的表达水平上。
方法/设计:我们通过逆转录-聚合酶链反应测定了新鲜分离的、传代后的胶原I植入物来源的软骨细胞以及骨关节炎软骨细胞中COL2A1、ACAN和COL1A1以及SOX9、-5和-6的mRNA丰度。此外,我们分析了结构基因和转录因子基因表达的相关性。因此,我们能够评估转录因子mRNA水平对软骨特异性结构基因表达的影响。
在以下方面获得了显著差异:(1)新鲜分离的骨关节炎软骨细胞与胶原I植入物来源的软骨细胞之间;(2)由于各自细胞来源的传代;(3)骨关节炎软骨细胞与非骨关节炎软骨细胞之间;(4)COL2A1与ACAN表达相对于与SOX9、-5和-6转录水平的一致性。
我们的结果可能有助于更好地理解软骨细胞结构基因表达的转录调控,这对其在基质辅助自体软骨细胞移植中的应用具有重要意义。