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原代纤维软骨细胞在创伤性和退行性半月板病变中的基因表达谱分析。

Gene expression profiling of primary fibrochondrocyte cultures in traumatic and degenerative meniscus lesions.

机构信息

Department of Orthopaedics and Traumatology, 64298Cerrahpasa Faculty of Medicine, Istanbul University-Cerrahpasa, Istanbul, Turkey.

Brain and Neurodegenerative Disorders Research Laboratory, Department of Medical Biology, 64298Cerrahpasa Faculty of Medicine, Istanbul University-Cerrahpasa, Istanbul, Turkey.

出版信息

J Orthop Surg (Hong Kong). 2021 Jan-Apr;29(1):23094990211000168. doi: 10.1177/23094990211000168.

Abstract

PURPOSE

This study aimed to investigate how fibroblastic and chondrocytic properties of human meniscal fibrochondrocytes are affected in culture conditions according to the type of meniscal pathology and localization, and to provide basic information for tissue-engineering studies.

METHODS

Primary fibrochondrocyte cultures were prepared from meniscus samples of patients who had either traumatic tear or degeneration due to osteoarthritis. Cultures were compared in terms of mRNA expression levels of COL1A1, COL2A1, COMP1, HIF1A, HIF2A, and SOX9 and secreted total collagen and sulfated sGAG levels according to the type of meniscal pathology, anatomical localization, and the number of subcultures.

RESULTS

mRNA expression levels of COL1A1, COMP1, HIF1A, HIF2A, and SOX9 were found to be increased in subsequent subcultures in all specimens. COL1A1 mRNA expression levels of both lateral and medial menisci of patients with traumatic tear were significantly higher than in patients with degenerative pathology, indicating a more fibroblastic character. P1 subculture of lateral and P3 or further subculture of medial meniscus showed more fibroblastic characteristics in patients with degenerative pathology. Furthermore, in patients with degenerative pathology, the subcultures of the lateral meniscus (especially on the inner part) presented more chondrocytic characteristics than did those of medial meniscus.

CONCLUSIONS

The mRNA expression levels of the cultures showed significant differences according to the anatomical localization and pathology of the meniscus, indicating distinct chondrocytic and fibroblastic features. This fundamental knowledge would help researchers to choose more efficient cell sources for cell-seeding of a meniscus scaffold, and to generate a construct resembling the original meniscus tissue.

摘要

目的

本研究旨在探讨人半月板纤维软骨细胞在不同半月板病变类型和定位的培养条件下,其成纤维细胞和软骨细胞特性的变化,并为组织工程研究提供基础信息。

方法

从创伤性撕裂或骨关节炎退变患者的半月板样本中制备原代纤维软骨细胞培养物。根据半月板病变类型、解剖定位和传代数,比较 COL1A1、COL2A1、COMP1、HIF1A、HIF2A 和 SOX9 的 mRNA 表达水平以及总胶原和硫酸软骨素 sGAG 的分泌水平。

结果

所有标本的 COL1A1、COMP1、HIF1A、HIF2A 和 SOX9 的 mRNA 表达水平在后续传代中均升高。创伤性撕裂患者的外侧和内侧半月板的 COL1A1 mRNA 表达水平明显高于退行性病变患者,表明其具有更强的成纤维细胞特征。退行性病变患者的外侧 P1 代和内侧 P3 代或进一步传代的培养物具有更强的成纤维细胞特征。此外,在退行性病变患者中,外侧半月板(尤其是内部分)的培养物比内侧半月板具有更强的软骨细胞特征。

结论

培养物的 mRNA 表达水平根据半月板的解剖定位和病变类型存在显著差异,表明其具有明显的成纤维细胞和软骨细胞特征。这些基础知识将有助于研究人员选择更有效的细胞来源用于半月板支架的细胞接种,并生成更接近原始半月板组织的构建体。

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