Effects of cytoskeleton reorganization inhibition on the proliferation and differentiation of osteoblasts under fluid shear stress

doi:10.3877/cma.j.issn.1674-1366.2011.01.007

Abstract

Abstract:

Objective

The study aimed at investigating the effects of cytoskeleton reorganization inhibition with RNA interference （RNAi） on cell proliferation and differentiation of osteoblasts under fluid shear stress （FSS）.

Methods

Mouse primary osteoblasts were cultured and treated with or without LIMK2 specific siRNA， and then loaded or unloaded under FSS. MTT assay， alkaline phosphatase （ALP） activity measurement and cell cycle analysis were conducted to evaluate their effects on cell proliferation and differentiation.

Results

Treated with RNAi only， osteoblasts of each group free of FSS showed no significant difference on the percentage of S-phase cells ［（3.600±0.447）% and （3.420±0.217）%， t=-0.810， P＞0.05］， the proliferation viability （0.208 ± 0.724 and 0.211± 0.044， t=0.251， P＞0.05） and the ALP activity （0.095 ± 0.001 and 0.090 ± 0.020，t=-1.857，P＞0.05）. Without effects of RNAi， FSS up-regulated the percentage of S-phase cells ［（11.110 ± 3.840）% ＞（3.420 ± 0.217）%， t=-4.460， P＜0.05］， the proliferation viability （0.336 ± 0.029 ＞0.211 ± 0.044， t=-13.878， P＜0.05） and the ALP activity （0.110 ±0.010 ＞0.090±0.012， t=-7.937， P＜0.05） of osteoblasts. And after the cytoskeleton reorganization inhibited with RNAi， the percentage of S-phase cells ［（25.140 ± 3.329）% ＞（3.600 ± 0.447）%，t=-14.339， P＜0.05］， the proliferation viability （0.480 ± 0.953 ＞0.208 ± 0.724， t=-13.454， P＜0.05） and the ALP activity（0.140±0.006 ＞0.095±0.001， t=-7.384， P＜0.05） of cells under FSS had significantly increased. There was a synergistic effect of cytoskeleton reorganization inhibition with RNA interference and fluid shear stress on the percentage of S-phase cells （F=240.125，P＜0.05）， the proliferation viability （F=44.550， P＜0.05）， and as well as the ALP activity （F=13.798， P＜0.05）.

Conclusion

Current findings illustrated that cytoskeleton reorganization inhibition with RNA interference promoted the proliferation and differentiation of osteoblasts under fluid shear stress.

Key words: Fluid shear stress, Cytoskeleton, Osteoblasts, RNA interference

Min-feng SHAO, Ya-juan XU, Ying-hui XIANG, Yourui LI, Yi-ping ZHANG, Rui CHEN, Qiang FU. Effects of cytoskeleton reorganization inhibition on the proliferation and differentiation of osteoblasts under fluid shear stress[J]. Chinese Journal of Stomatological Research(Electronic Edition), 2011, 5(01): 44-51.

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URL: https://zhkqyxyjzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-1366.2011.01.007

https://zhkqyxyjzz.cma-cmc.com.cn/EN/Y2011/V5/I01/44

Figures/Tables 7

References 20

1	Pavalko FM， Chen NX， Turner CH， et al. Fluid shear-induced mechanical signaling in MC3T3-E1 osteoblasts requires cytoskeleton-integrin interactions. Am J Physiol， 1998，275（6 Pt 1）：C1591-C1601.
2	Takai E， Costa KD， Shaheen A， et al. Osteoblast elastic modulus measured by atomic force microscopy is substrate dependent. Ann Biomed Eng， 2005，33（7）：963-971.
3	Fu Q， Wu C， Shen Y， et al. Effect of LIMK2 RNAi on reorganization of the actin cytoskeleton in osteoblasts induced by fluid shear stress.J Biomech， 2008，41（15）：3225-3228.
4	张艺平，覃峰，吴昌敬，等. 沉默LIMK2 基因对成骨细胞c-fos 基因力学敏感性的影响. 中华医学杂志， 2009，89（44）：3143-3146.
5	吴昌敬. 抑制细胞骨架改建对流体剪切力诱导成骨细胞cox-2 基因的影响. 广州：中山大学， 2009.
6	Kido S， Kuriwaka-Kido R， Imamura T， et al. Mechanical stress induces Interleukin-11 expression to stimulate osteoblast differentiation.Bone， 2009，45（6）：1125-1132.
7	McGarry JG， Klein-Nulend J， Mullender MG， et al. A comparison of strain and fluid shear stress in stimulating bone cell responses--a computational and experimental study. FASEB J， 2005，19（3）：482-484.
8	Takai E， Costa KD， Shaheen A， et al. Osteoblast elastic modulus measured by atomic force microscopy is substrate dependent. Ann Biomed Eng， 2005，33（7）：963-971.
9	Jaasma MJ， Jackson WM， Keaveny TM. The effects of morphology，confluency，and phenotype on whole-cell mechanical behavior. Ann Biomed Eng， 2006，34（5）：759-768.
10	Nomura S， Takano-Yamamoto T. Molecular events caused by mechanical stress in bone. Matrix Biol， 2000，19（2）：91-96.
11	Clarke B. Normal bone anatomy and physiology. Clin J Am Soc Nephrol， 2008，3（Suppl 3）：S131-S139.
12	Rosenberg N.The role of the cytoskeleton in mechanotransduction in human osteoblast-like cells. Hum Exp Toxicol， 2003，22（5）：271-274.
13	McGarry JG， Klein-Nulend J， Prendergast PJ. The effect of cytoskeletal disruption on pulsatile fluid flow-induced nitric oxide and prostaglandin E2 release in osteocytes and osteoblasts. Biochem Biophys Res Commun， 2005，330（1）：341-348.
14	Lee DY， Yeh CR， Chang SF， et al. Integrin-mediated expression of bone formation-related genes in osteoblast-like cells in response to fluid shear stress： roles of extracellular matrix， Shc， and mitogen-activated protein kinase. J Bone Miner Res， 2008，23（7）：1140-1149.
15	Forgacs G. On the possible role of cytoskeletal filamentous networks in intracellular signaling： an approach based on percolation. J Cell Sci，1995，108（Pt 6）：2131-2143.
16	Traub O， Monia BP， Dean NM， et al. PKC-epsilon is required for mechano-sensitive activation of ERK1/2 in endothelial cells. J Biol Chem， 1997，272（50）：31251-31257.
17	Zambuzzi WF， Bruni-Cardoso A， Granjeiro JM， et al. On the road to understanding of the osteoblast adhesion： cytoskeleton organization is rearranged by distinct signaling pathways. J Cell Biochem， 2009，108（1）：134-144.
18	Ingber DE. TensegrityⅡ. How structural networks influence cellular information processing networks. J Cell Sci， 2003，116 （Pt 8）：1397-1408.
19	Lee YC， Huang CF， Murshed M， et al. Src family kinase/abl inhibitor dasatinib suppresses proliferation and enhances differentiation of osteoblasts. Oncogene， 2010，29（22）：3196-3207.
20	Zhang J， Ryder KD， Bethel JA， et al. PTH-induced actin depolymerization increases mechanosensitive channel activity to enhance mechanically stimulated Ca²⁺ signaling in Osteoblasts. J Bone Mineral Res， 2006，21（11）：1729-1737.

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