Qiao Yusen, Yi Dan, Reed David Andrew, Mercuri Louis G, Chen Di, Oh Chun-do
Department of Orthopedic Surgery, Rush University Medical Center, Chicago, USA.
Department of Oral Biology, University of Illinois Chicago, Chicago, USA.
J Dent Sci. 2022 Jul;17(3):1378-1386. doi: 10.1016/j.jds.2022.04.017. Epub 2022 May 12.
BACKGROUND/PURPOSE: The temporomandibular joint (TMJ) is a bi-arthrodial joint that is composed of the temporal bone glenoid fossa and the condylar head of the mandible both having fibrocartilaginous articular surfaces. Functional overloading of the TMJ is the main cause of TMJ osteoarthritis (TMJ OA) disease. The aim of this study was to establish immortalized TMJ fibrocartilage cell clones to provide enough cells to adequately investigate the molecular mechanisms studies of TMJ OA.
We have isolated temporomandibular condyle chondrocytes from adult Sprague Dawley rat. The cells were cultured and immortalized by treating with Y-27632, a well-characterized inhibitor of Rho-Associated Kinase (ROCK). Clones were characterized on the basis of cell morphology and analyses of marker gene expression through 45 passages.
Cells from the condylar fibrocartilage of the TMJ were successfully immortalized by ROCK inhibitor, retaining a consistent cuboidal cell morphology and the expression of several cell markers of polymorphic cell fate. In addition, they retained phenotype features similar to the primary parental TMJ fibrocartilage cells when the cells were challenged with different cytokines and growth factors.
These studies establish a novel immortalized cell line through ROCK inhibitor Y-27632, that retains the polymorphic phenotype of primary cell lines from TMJ fibrocartilage chondrocyte cell through a high number of passages, serving as a valuable preclinical resource for mechanistic assessment of TMJ health, disease, and regeneration.
背景/目的:颞下颌关节(TMJ)是一种双关节,由颞骨的关节窝和下颌骨的髁突组成,二者均具有纤维软骨关节面。颞下颌关节的功能过载是颞下颌关节骨关节炎(TMJ OA)疾病的主要原因。本研究的目的是建立永生化的颞下颌关节纤维软骨细胞克隆,以提供足够的细胞来充分研究颞下颌关节骨关节炎的分子机制。
我们从成年Sprague Dawley大鼠中分离出颞下颌髁突软骨细胞。通过用Y-27632(一种已充分表征的Rho相关激酶(ROCK)抑制剂)处理,对细胞进行培养并使其永生化。通过45代培养,根据细胞形态和标记基因表达分析对克隆进行表征。
通过ROCK抑制剂成功使颞下颌关节髁突纤维软骨细胞永生化,保持一致的立方形细胞形态以及几种多形性细胞命运的细胞标记物的表达。此外,当用不同的细胞因子和生长因子刺激细胞时,它们保留了与原代亲代颞下颌关节纤维软骨细胞相似的表型特征。
这些研究通过ROCK抑制剂Y-27632建立了一种新型永生化细胞系,该细胞系通过大量传代保留了来自颞下颌关节纤维软骨细胞原代细胞系的多形性表型,可作为评估颞下颌关节健康、疾病和再生机制的有价值的临床前资源。