EFFECTS OF HYDROSTATIC PRESSURE AND INSULIN-LIKE GROWTH FACTOR 1 ON EXPRESSION OF FILAMENTOUS ACTIN OF TEMPOROMANDIBULAR JOINT DISC CELLS IN GOATS_Chinese Journal of Reparative and Reconstructive Surgery

Authors：

ZHANG Chunyu ¹ , BAO Guangjie ² , KANG Hong ¹

1Department of Prosthodontics, Hospital of Stomatology, Lanzhou University, Lanzhou Gansu, 730000, P.R.China;;
2Key Laboratory of Stomatology of State Ethnic Affairs Commission, Northwest University for Nationalities. Corresponding author: KANG Hong, E-mail: kanghong@lzu.edu.cn;

Corresponding author：

Keywords：

Hydrostatic pressure Insulin-like growth factor 1; Temporomandibular joint disc cells; Filamentous actin; Goat

DOI：

10.7507/1002-1892.20130167

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Objective To investigate the effect of hydrostatic pressure and insulin-like growth factor 1 (IGF-1) on the expression of filamentous actin (F-actin) of temporomandibular joint disc cells in goats, and to analyze the F-actin changes of temporomandibular joint disc cells in vitro under hydrostatic pressure and IGF-1 stimulation. Methods The bilateral temporomandibular joint discs were harvested from 4 1-month-old goats, and temporomandibular joint disc cells were isolated with collagenase. Immunohistochemical staining for collagen type I and collagen type II was performed for identification. The cells at passages 2-3 were used; the experiment was divided into 4 groups according to different interventions: the cells were cultivated with complete medium in group A as control; the cells were intervened by hydrostatic pressure (0.2 MPa and 1 Hz for 3 hours) in group B, by complete medium containing IGF-1 (10 ng/mL) in group C, and by a combination of hydrostatic pressure (0.2 MPa and 1 Hz for 3 hours) and complete medium containing IGF-1 (10 ng/mL) in group D. The changes of F-actin at 24 and 72 hours after cultivation were observed by immunofluorescence staining. The cell fluorescence intensity was measured. Results The cultivated cells were identified to be temporomandibular joint disc cells by morphological observation and immunohistochemical staining. At 24 hours, fluorescence intensity of groups A and C was b and clear, with normal morphology of temporomandibular joint disc cells; F-actin arranged in disorder in group B, and F-actin was thinner with arrangement disorder in group D. At 72 hours, the F-actin arranged regularly in groups A and C; however, some F-actin became blurry with irregular arrangement, breakage, and pseudopodia in group B; and F-actin was thinner and ruptured formed in group D. With time passing, the fluorescence intensity of F-actin in groups A, B, and D had an increasing trend, showing significant differences between 24 hours and 72 hours (P lt; 0.05); but there was no significant difference between 24 and 72 hours in group C (t=0.284, P=0.781). At 24 hours, fluorescence intensity of F-actin was highest in group C and was lowest in group B, showing significant difference when compared with groups A and D (P lt; 0.05). At 72 hours, fluorescence intensity in groups B and D was significantly lower than that in groups A and C (P lt; 0.05), but there was no significant difference between groups B and D, and between groups A and C (P gt; 0.05). Conclusion Hydrostatic pressure may cause the F-actin breakage and rearrangement of temporomandibular joint disc cells, and IGF-1 can up-regulate the F-actin expression. Such effects may be correlated with the biological behavior of the temporomandibular joint disc cells.

Citation： ZHANG Chunyu,BAO Guangjie,KANG Hong. EFFECTS OF HYDROSTATIC PRESSURE AND INSULIN-LIKE GROWTH FACTOR 1 ON EXPRESSION OF FILAMENTOUS ACTIN OF TEMPOROMANDIBULAR JOINT DISC CELLS IN GOATS. Chinese Journal of Reparative and Reconstructive Surgery, 2013, 27(6): 755-759. doi: 10.7507/1002-1892.20130167 Copy

1.	Knight MM, Toyoda T, Lee DA, et al. Mechanical compression and hydrostatic pressure induce reversible changes in actin cytoskeletal organisation in chondrocytes in agarose. J Biomech, 2006, 39(8): 1547-1551.
2.	Deibler M, Spatz JP, Kemkemer R. Actin fusion proteins alter the dynamics of mechanically induced cytoskeleton rearrangement. PLoS One, 2011, 6(8): e22941.
3.	堵安庆, 王羽, 赵森, 等. 周期性张应力作用下成骨样细胞MG-63中c-fos基因和丝状肌动蛋白相互关系的研究. 华西口腔医学杂志, 2012, 30(4): 430-433, 438.
4.	朱庆党, 巢永烈, 陈新民, 等. 动态张、压应力刺激下人牙周膜成纤维细胞细胞骨架变化. 实用口腔医学杂志, 2008, 24(4): 559-562.
5.	Hu K, Ji L, Applegate KT, et al. Differential transmission of actin motion within focal adhesions. Science, 2007, 315(5808): 111-115.
6.	Jaworski J, Kapitein LC, Gouveia SM, et al. Dynamic microtubules regulate dendritic spine morphology and synaptic plasticity. Neuron, 2009, 61(1): 85-100.
7.	Okamoto K, Bosch M, Hayashi Y. The roles of CaMKII and F-actin in the structural plasticity of dendritic spines: a potential molecular identity of a synaptic tag? Physiology (Bethesda), 2009, 24: 357-366.
8.	Wu MJ, Gu ZY, Sun W. Effects of hydrostatic pressure on cytoskeleton and BMP-2, TGF-beta, SOX-9 production in rat temporomandibular synovial fibroblasts. Osteoarthritis Cartilage, 2008, 16(1): 41-47.
9.	康宏, 李振强, 闭艳妲. 自组装山羊颞下颌关节盘组织工程纤维软骨模型的构建. 华西口腔医学杂志, 2011, 29(3): 314-317.
10.	舒维娜, 康宏, 张卫平, 等. 山羊颞下颌关节盘细胞体外培养研究. 实用口腔医学杂志, 2010, 26(2): 165-168.
11.	Akisaka T, Yoshida H, Inoue S, et al. Organization of cytoskeletal F-actin, G-actin, and gelsolin in the adhesion structures in cultured osteoclast. J Bone Miner Res, 2001, 16(7): 1248-1255.
12.	Mouneimne G, Hansen SD, Selfors LM, et al. Differential remodeling of actin cytoskeleton architecture by profilin isoforms leads to distinct effects on cell migration and invasion. Cancer Cell, 2012, 22(5): 615-630.
13.	Gardel ML, Schneider IC, Aratyn-Schaus Y, et al. Mechanical integration of actin and adhesion dynamics in cell migration. Annu Rev Cell Dev Biol, 2010, 26: 315-333.
14.	Rivero F, Köppel B, Peracino B, et al. The role of the cortical cytoskeleton: F-actin crosslinking proteins protect against osmotic stress, ensure cell size, cell shape and motility, and contribute to phagocytosis and development. J Cell Sci, 1996, 109(Pt 11): 2679-2691.
15.	邓银栓, 唐康来, 谢美明, 等. 不同牵伸载荷对体外培养人肌腱细胞纤维型肌动蛋白的影响. 中华医学杂志, 2011, 91(25): 1780-1785.
16.	彭庭莉, 杨军, 周继祥, 等. 不同张应力对体外培养的人牙周膜成纤维细胞形态和纤维型肌动蛋白改变的影响. 第三军医大学学报, 2005, 27(9): 898-900.
17.	王彬, 闵苏. 机械牵拉对人肺微血管内皮细胞单层通透性、F-肌动蛋白变化的影响. 临床心血管病杂志, 2011, 27(1): 59-62.
18.	Wuertz K, Urban JP, Klasen J, et al. Influence of extracellular osmolarity and mechanical stimulation on gene expression of intervertebral disc cells. J Orthop Res, 2007, 25(11): 1513-1522.
19.	康宏, 闭艳妲, 李振强, 等. 两种生长因子对颞下颌关节盘细胞自组装基体合成影响的动物实验. 中华口腔医学杂志, 2011, 46(9): 541-546.
20.	Leipzig ND, Eleswarapu SV, Athanasiou KA. The effects of TGF-beta1 and IGF-I on the biomechanics and cytoskeleton of single chondrocytes. Osteoarthritis Cartilage, 2006, 14(12): 1227-1236.

1. Knight MM, Toyoda T, Lee DA, et al. Mechanical compression and hydrostatic pressure induce reversible changes in actin cytoskeletal organisation in chondrocytes in agarose. J Biomech, 2006, 39(8): 1547-1551.
2. Deibler M, Spatz JP, Kemkemer R. Actin fusion proteins alter the dynamics of mechanically induced cytoskeleton rearrangement. PLoS One, 2011, 6(8): e22941.
3. 堵安庆, 王羽, 赵森, 等. 周期性张应力作用下成骨样细胞MG-63中c-fos基因和丝状肌动蛋白相互关系的研究. 华西口腔医学杂志, 2012, 30(4): 430-433, 438.
4. 朱庆党, 巢永烈, 陈新民, 等. 动态张、压应力刺激下人牙周膜成纤维细胞细胞骨架变化. 实用口腔医学杂志, 2008, 24(4): 559-562.
5. Hu K, Ji L, Applegate KT, et al. Differential transmission of actin motion within focal adhesions. Science, 2007, 315(5808): 111-115.
6. Jaworski J, Kapitein LC, Gouveia SM, et al. Dynamic microtubules regulate dendritic spine morphology and synaptic plasticity. Neuron, 2009, 61(1): 85-100.
7. Okamoto K, Bosch M, Hayashi Y. The roles of CaMKII and F-actin in the structural plasticity of dendritic spines: a potential molecular identity of a synaptic tag? Physiology (Bethesda), 2009, 24: 357-366.
8. Wu MJ, Gu ZY, Sun W. Effects of hydrostatic pressure on cytoskeleton and BMP-2, TGF-beta, SOX-9 production in rat temporomandibular synovial fibroblasts. Osteoarthritis Cartilage, 2008, 16(1): 41-47.
9. 康宏, 李振强, 闭艳妲. 自组装山羊颞下颌关节盘组织工程纤维软骨模型的构建. 华西口腔医学杂志, 2011, 29(3): 314-317.
10. 舒维娜, 康宏, 张卫平, 等. 山羊颞下颌关节盘细胞体外培养研究. 实用口腔医学杂志, 2010, 26(2): 165-168.
11. Akisaka T, Yoshida H, Inoue S, et al. Organization of cytoskeletal F-actin, G-actin, and gelsolin in the adhesion structures in cultured osteoclast. J Bone Miner Res, 2001, 16(7): 1248-1255.
12. Mouneimne G, Hansen SD, Selfors LM, et al. Differential remodeling of actin cytoskeleton architecture by profilin isoforms leads to distinct effects on cell migration and invasion. Cancer Cell, 2012, 22(5): 615-630.
13. Gardel ML, Schneider IC, Aratyn-Schaus Y, et al. Mechanical integration of actin and adhesion dynamics in cell migration. Annu Rev Cell Dev Biol, 2010, 26: 315-333.
14. Rivero F, Köppel B, Peracino B, et al. The role of the cortical cytoskeleton: F-actin crosslinking proteins protect against osmotic stress, ensure cell size, cell shape and motility, and contribute to phagocytosis and development. J Cell Sci, 1996, 109(Pt 11): 2679-2691.
15. 邓银栓, 唐康来, 谢美明, 等. 不同牵伸载荷对体外培养人肌腱细胞纤维型肌动蛋白的影响. 中华医学杂志, 2011, 91(25): 1780-1785.
16. 彭庭莉, 杨军, 周继祥, 等. 不同张应力对体外培养的人牙周膜成纤维细胞形态和纤维型肌动蛋白改变的影响. 第三军医大学学报, 2005, 27(9): 898-900.
17. 王彬, 闵苏. 机械牵拉对人肺微血管内皮细胞单层通透性、F-肌动蛋白变化的影响. 临床心血管病杂志, 2011, 27(1): 59-62.
18. Wuertz K, Urban JP, Klasen J, et al. Influence of extracellular osmolarity and mechanical stimulation on gene expression of intervertebral disc cells. J Orthop Res, 2007, 25(11): 1513-1522.
19. 康宏, 闭艳妲, 李振强, 等. 两种生长因子对颞下颌关节盘细胞自组装基体合成影响的动物实验. 中华口腔医学杂志, 2011, 46(9): 541-546.
20. Leipzig ND, Eleswarapu SV, Athanasiou KA. The effects of TGF-beta1 and IGF-I on the biomechanics and cytoskeleton of single chondrocytes. Osteoarthritis Cartilage, 2006, 14(12): 1227-1236.

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Chinese Journal of Reparative and Reconstructive Surgery

EFFECTS OF HYDROSTATIC PRESSURE AND INSULIN-LIKE GROWTH FACTOR 1 ON EXPRESSION OF FILAMENTOUS ACTIN OF TEMPOROMANDIBULAR JOINT DISC CELLS IN GOATS

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