Effect of tensile strain on perlecan expression of human periodontal ligament cells

Abstract

Abstract: PURPOSE: To investigate the perlecan expression of human periodontal ligament cells (hPDLCs) under cyclic tensile strain in vitro, and learn the molecular mechanism of periodontal remolding during tooth movement. METHODS: hPDLCs isolated by enzyme digestion were loaded with 12% elongation, 1 Hz of uniaxial tensile strain for 12, 24 and 48 h. The unloaded cells were used as control. Real-time PCR and enzyme-linked immune sorbent assay (ELISA) were applied to analyze the mRNA and protein expression of perlecen in each sample respectively. The data was analyzed with SPSS 19.0 software package. RESULTS: Within 12 h, mRNA expression was transiently elevated, but no significant difference was detected compared with the control. After 12 h, the mRNA expression was significantly decreased. It would decreased to (0.28±0.049) at lowest level of control at lowest level at 48 h (P<0.05). The protein expression of perlecan was time-dependently decreased. Specifically, it was downregulated from (14.03±0.71) pg/mL (control) to (11.06±0.15) pg/mL at lowest level at 48 h (P<0.05). CONCLUSIONS: Tensile strain time-dependently down-regulates perlecan expression, indicating perlecan may play a pivot role in PDLCS responding to mechanical loading in vitro.

Key words: Supported, by, Xin, Miao, Talent, Program, of, Zhejiang, Province, (2013R423044), and, Research, Project, of, Wenzhou, Science, and, Technology, Department, (Y20120228).

CLC Number:

R781.4

WANG Yi¹, SUN Chao-fan¹, CHEN Li-jiao¹, WANG Yan-liang², HU Rong-dang¹. Effect of tensile strain on perlecan expression of human periodontal ligament cells[J]. Shanghai Journal of Stomatology, 2014, 23(6): 675-680.

References

[1] Henneman S, Von den Hoff JW, Maltha JC. Mechanobiology of tooth movement [J]. Eur J Orthod, 2008, 30(3): 299-306.
[2] Magan A, Ripamonti U. Biological aspects of periodontal tissue regeneration: cementogenesis and the induction of Sharpey’s fibres [J]. SADJ, 2013, 68(7): 304-306, 308-312, 314 passim.
[3] Liu M, Dai J, Lin Y, et al. Effect of the cyclic stretch on the expression of osteogenesis genes in human periodontal ligament cells [J]. Gene, 2012, 491(2): 187-193.
[4] Whitelock JM, Melrose J, Iozzo RV. Diverse cell signaling events modulated by perlecan [J]. Biochemistry, 2008, 47(43): 11174-11183.
[5] Jha AK, Yang W, Kirn-Safran CB, et al. Perlecan domain I-conjugated, hyaluronic acid-based hydrogel particles for enhanced chondrogenic differentiation via BMP-2 release [J]. Biomaterials, 2009, 30(36): 6964-6975.
[6] Whitelock JM, Murdoch AD, Iozzo RV, et al. The degradation of human endothelial cell-derived perlecan and release of bound basic fibroblast growth factor by stromelysin, collagenase, plasmin, and heparanases[J]. J Biol Chem, 1996, 271(17): 10079-10086.
[7] Schmidtchen A, Carlstedt I, Malmström A, et al. Inventory of human skin fibroblast proteoglycans. Identification of multiple heparan and chondroitin/dermatan sulphate proteoglycans [J]. Biochem J, 1990, 265(1): 289-300.
[8] Larjava H, Hakkinen L, Rahemtulla F. A biochemical analysis of human periodontal tissue proteoglycans[J].Biochem J,1992,284 (Pt 1):267-274.
[9] 李玉珍, 刘秀华, 蔡莉蓉. 串珠素表达下调参与低氧对大鼠心肌微血管内皮细胞增殖的抑制作用 [J]. 生理学报, 2007, 59(2): 221-226.
[10] Baker AB, Ettenson DS, Jonas M, et al. Endothelial cells provide feedback control for vascular remodeling through a mechanosensitive autocrine TGF-beta signaling pathwayc [J]. Circ Res, 2008, 103(3): 289-297.
[11] Natali AN, Pavan PG, Scarpa C. Numerical analysis of tooth mobility: formulation of a non-linear constitutive law for the periodontal ligament [J]. Dent Mater, 2004, 20(7): 623-629.
[12] Lee RT, Yamamoto C, Feng Y, et al. Mechanical strain induces specific changes in the synthesis and organization of proteoglycans by vascular smooth muscle cells [J]. J Biol Chem, 2001, 276(17): 13847-13851.
[13] 李永明, 冯雪, 林珠, 等. 周期性牵张力对人牙周膜细胞MMP-3及TIMP-1表达的影响[J]. 临床口腔医学杂志, 2004, 20(4): 195-198.
[14] 李永明, 唐林, 张晓东, 等. 牵张应力对人牙周膜细胞MMP-13/TIMP-1表达的影响及其信号转导途径研究 [J]. 医用生物力学, 2010, 25(6): 422-427.
[15] Leonardi R, Talic NF, Loreto C. MMP-13 (collagenase 3) immunolocalisation during initial orthodontic tooth movement in rats [J]. Acta Histochem, 2007, 109(3): 215-220.
[16] Wang B, Lai X, Price C, et al. Perlecan-containing pericellular matrix regulates solute transport and mechanosensing within the osteocyte lacunar-canalicular system[J]. J Bone Miner Res, 2014, 29(4): 878-891.
[17] Lowe DA, Lepori-Bui N, Fomin PV, et al. Deficiency in perlecan/HSPG2 during bone development enhances osteogenesis and decreases quality of adult bone in mice [J]. Calcif Tissue Int, 2014, 95(1): 29-38.
[18] Reiland J, Kempf D, Roy M, et al. FGF2 binding, signaling, and angiogenesis are modulated by heparanase in metastatic melanoma cells [J]. Neoplasia, 2006, 8(7): 596-606.
[19] Elkin M, Ilan N, Ishai-Michaeli R, et al. Heparanase as mediator of angiogenesis: mode of action [J]. FASEB J, 2001, 15(9): 1661-1663.
[20] Takayama S, Murakami S, Nozaki T, et al. Expression of receptors for basic fibroblast growth factor on human periodontal ligament cells [J]. J Periodontal Res, 1998, 33(6): 315-322.