山羊颞下颌关节盘细胞肌动蛋白骨架的研究

口腔医学研究 ›› 2015, Vol. 31 ›› Issue (10): 953-956.

• 基础研究论著 • 下一篇

山羊颞下颌关节盘细胞肌动蛋白骨架的研究

孔楠楠^1,2,张文霞²,康宏²,包广洁^1.2*

1. 西北民族大学口腔医学国家民委重点实验室甘肃兰州 730030;
2. 兰州大学口腔医学研究所甘肃兰州 730000

收稿日期:2015-04-20 出版日期:2015-10-28 发布日期:2016-05-04
通讯作者: 包广洁,电话:0931-2977518
作者简介:孔楠楠(1988~ ),女,甘肃人,硕士,主要从事口腔内科学临床研究,现就职于西安市儿童医院口腔科。
基金资助:
国家自然科学基金项目(编号:81160139);甘肃省科技支撑项目(编号:1104FKCA144)

Study on the Cytoskeleton of Cultured Goat Temporomandibular Joint Disc Cells

KONG Nan-nan^1,2, ZHANG Wen-xia¹, KANG Hong¹, BAO Guang-jie^1,2

1. Key Lab of Stomatology of State Ethnic Affairs Commission, Northwest University for Nationalities, Lanzhou 730030, China;
2. School of Stomatology, Lanzhou University, Lanzhou 730000, China

Received:2015-04-20 Online:2015-10-28 Published:2016-05-04

摘要/Abstract

摘要： 目的:通过激光扫描共聚焦显微镜(Laser Scanning Confocal Microscope,LSCM)研究原代及传代山羊颞下颌关节盘细胞骨架(Cytoskeleton,CSK)蛋白-肌动蛋白的形态及蛋白量的改变,以说明关节盘细胞随着传代的发生,细胞骨架蛋白-肌动蛋白的差异。方法:原代(P0)山羊颞下颌关节盘细胞及传代1、2、3、4(P1、P2、P3、P4)代的关节盘细胞,分别接种在有方形盖玻片的6孔板中制作细胞爬片。对各代关节盘细胞肌动蛋白骨架进行免疫荧光染色,利用激光共聚焦显微镜获取各代关节盘细胞肌动蛋白的形态,同时利用荧光强度测定软件对各代关节盘细胞肌动蛋白荧光量进行测定。结果:丝状的肌动蛋白主要沿着细胞膜外围均匀排列,随着传代丝状结构逐渐清晰,且逐渐增粗,P4代时形成较粗的纤维束。荧光强度整体趋势是随着传代荧光量表达是递增的,P3代以内没有统计学差异,而在P4代时统计学差异明显。结论:肌动蛋白骨架随着传代过程形态学上发生了改变,在P4代时可见骨架蛋白形成较粗的纤维束,荧光量表达同其他代细胞差异明显,这为以后工程化颞下颌关节盘种子细胞的选择提供了依据。

关键词: 关节盘细胞, 传代, 细胞骨架, 激光共聚焦显微镜

Abstract: Objective: To study the morphology and quantify the cytoskeleton (CSK)-actin in primary and passaged goat temporomandibular joint disc (TMJ) cells, and to illustrate the differences of actin protein during passaging using laser scanning confocal microscope (LCSM). Methods: The primary, 1st, 2nd, 3rd and 4th (P0, P1, P2, P3, P4) TMJ disc cells were inoculated into 6-well plates with square coverslips placed in each well. The CSK-actin of each generation of TMJ disc cells was stained by immunofluorescence staining and observed under the LCSM and later quantitatively analyzed by a fluorescence intensity software. Results: Filamentous actin mainly located uniformly at the outer periphery of cell membrane. The CSK-actin became thicker and clearer during cell passaging, and thick fiber bundles formed obviously in P4 generation. The fluorescence intensity also increased during passaging and was significantly higher in P4 than in P1, P2 and P3. Conclusion: The morphology of the CSK-actin changed during the passaging of TMJ disc cells. This finding might provide basis for seed cell choose in TMJ disc engineering.

Key words: TMJ disc cell, Passage, Cytoskeleton, Laser scanning confocal microscope

中图分类号:

R782.6

孔楠楠,张文霞,康宏,包广洁. 山羊颞下颌关节盘细胞肌动蛋白骨架的研究[J]. 口腔医学研究, 2015, 31(10): 953-956.

KONG Nan-nan, ZHANG Wen-xia, KANG Hong, BAO Guang-jie. Study on the Cytoskeleton of Cultured Goat Temporomandibular Joint Disc Cells[J]. Journal of Oral Science Research, 2015, 31(10): 953-956.

参考文献

[1] Allen KD, Athanasiou KA. Tissue Engineering of the TMJ Disc: An Review [J]. Tissue engineering,2006,12(5)∶1183-1196
[2] Allen KD, Athanasiou KA. Effect of passage and topography on gene expression of temporomandibular joint disc cells [J]. Tissue engineering, 2007, 13(1)∶101-110
[3] 李凯, 卫小春, 许刚, 等. 原代和传代兔关节软骨细胞糖胺多糖合成的动态观察[J].中国组织工程研究与临床康复,2009,13(37)∶7217-7220
[4] Heath JP, Holifield BF. Cell locomotion: New research tests old ldeas on membrane and cytoskeletal flow [J]. Cell motility and the cytoskeleton, 1991, 18(4)∶245-257
[5] Turner CE, Burridge K. Transmembrane molecular assemblies in cell-extracellular matrix interactions [J]. Current opinion in cell biology, 1991, 3(5)∶849-853
[6] Juliano RL, Haskill S. Signal transduction from the extracellular matrix [J]. Journal of Cell Biology, 1993, 120∶577-577
[7] Noriega S, Hasanova G, Subramanian A. The effect of ultrasound stimulation on the cytoskeletal organization of chondrocytes seeded in three-dimensional matrices [J]. Cells Tissues Organs, 2012, 197(1)∶14-26
[8] de Paz LEC. Image analysis software based on color segmentation for characterization of viability and physiological activity of biofilms [J]. Applied and environmental microbiology, 2009, 75(6)∶1734-1739
[9] Dziedzic K, Zalewski M, Gadek A, et al. Chondrocytes application in regenerative medicine [J]. Przeglad lekarski, 2013, 71(6)∶334-339
[10] Zeng L, Chen X, Zhang Q, et al. Redifferentiation of dedifferentiated chondrocytes in a novel three‐dimensional microcavitary hydrogel [J]. Journal of Biomedical Materials Research Part A, 2014
[11] Gurusinghe S, Strappe P. Gene Modification of Mesenchymal Stem Cells and Articular Chondrocytes to Enhance Chondrogenesis [J]. BioMed Research International, 2014
[12] 张艳,柴岗,刘伟,等.体外培养过程中去分化的人软骨细胞基因表达谱的变化[J].中华整形外科杂志,2007,23(4)∶331-334
[13] Vinall RL, Lo SH, Reddi AH. Regulation of articular chondrocyte phenotype by bone morphogenetic protein 7, interleukin 1, and cellular context is dependent on the cytoskeleton [J]. Experimental cell research, 2002, 272(1)∶32-44
[14] Berkovitz BKB, Becker D. Detailed morphology and distribution of gap junction protein associated with cells from the intra-articular disc of the rat temporomandibular joint [J]. Connective tissue research, 2003, 44(1)∶12-18
[15] Mallein-Gerin F, Garrone R, Van der Rest M. Proteoglycan and collagen synthesis are correlated with actin organization in dedifferentiating chondrocytes [J]. European journal of cell biology, 1991, 56(2)∶364-373
[16] Spiegelman BM, Ginty CA. Fibronectin modulation of cell shape and lipogenic gene expression in 3T3-adipocytes [J]. Cell, 1983, 35(3)∶657-666
[17] Yourek G, Hussain MA, Mao JJ. Cytoskeletal changes of mesenchymal stem cells during differentiation [J]. ASAIO.2007,53(2)∶218-228
[18] Haudenschild DR, Chen J, Steklov N, et al. Characterization of the chondrocyte actin cytoskeleton in living three-dimensional culture: response to anabolic and catabolic stimuli [J]. Molecular & cellular biomechanics: MCB, 2009, 6(3)∶135-144
[19] Rotsch C, Radmacher M. Drug-induced changes of cytoskeletal structure and mechanics in fibroblasts: an atomic force microscopy study [J]. Biophysical journal, 2000, 78(1)∶520-535
[20] Ohashi T, Ishii Y, Ishikawa Y, et al. Experimental and numerical analyses of local mechanical properties measured by atomic force microscopy for sheared endothelial cells [J]. Bio-medical materials and engineering, 2002, 12(3)∶319-327

山羊颞下颌关节盘细胞肌动蛋白骨架的研究

Study on the Cytoskeleton of Cultured Goat Temporomandibular Joint Disc Cells

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