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Differential regulation of collagen, lysyl oxidase and MMP ‐2 in human periodontal ligament cells by low‐ and high‐level mechanical stretching
Author(s) -
Chen YJ.,
Jeng JH.,
Chang HH.,
Huang MY.,
Tsai FF.,
Jane Yao CC.
Publication year - 2013
Publication title -
journal of periodontal research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.31
H-Index - 83
eISSN - 1600-0765
pISSN - 0022-3484
DOI - 10.1111/jre.12028
Subject(s) - lysyl oxidase , periodontal fiber , matrix metalloproteinase , chemistry , metalloproteinase , biochemistry , biophysics , enzyme , dentistry , medicine , biology
Background and Objective Mechanical stretching modulates extracellular matrix ( ECM ) protein synthesis by periodontal ligament ( PDL ) cells. However, the mechanoregulation of lysyl oxidase ( LOX ), a key enzyme for collagen cross‐linking, is not fully understood. In the present study, we hypothesized that low‐level and high‐level mechanical stretching differentially regulates collagen deposition and the expression of LOX and the enzymes responsible for ECM degradation, such as MMP ‐2 in PDL cells. Material and Methods Human PDL cells were cultured on flexible‐bottom culture plates and subjected to cyclic mechanical stretching (3% and 10% elongation at 0.1 Hz) for 24 and 48 h in a Flexercell FX‐4000 strain unit. The levels of expression of type I collagen alpha 1 ( COL1A1 ), type III collagen alpha 1 ( COL3A1 ), lysyl oxidase ( LOX) , MMP2 and TIMP2 mRNAs were analyzed using an RT‐PCR technique. The cell layer and the culture medium were separately collected and processed for detection of the following ECM‐related molecules: (i) total collagen content using a Sircol dye‐binding method; (ii) LOX protein expression by western blotting; (iii) LOX activity using a fluorometric assay; and (iv) MMP‐2 enzyme activity by gelatin zymography. Results Low‐level (3%) mechanical stretching of PDL cells upregulated the expression of COL1A1 , COL3A1 and LOX mRNA s, enhanced the production of collagen and increased the LOX activity but did not change the level of expression of MMP2 or TIMP2 mRNA . The collagen content and LOX activity showed obvious elevation in the medium, but not in the cell layer. High‐level (10%) mechanical stretching downregulated COL1A1 mRNA but upregulated COL3A1 mRNA ; however, the effect on COL3A1 was smaller, and occurred earlier, compared with the effect on the COL1A1 gene. High‐level mechanical stretching upregulated the expression of MMP2 and TIMP2 mRNA s but did not change collagen production or LOX activity. Moreover, high‐level mechanical stretching increased the level of pro‐MMP‐2, especially in the cell layer. Conclusions This study substantiates the mechanoregulation of the expression of ECM‐related molecules in PDL cells. High‐level mechanical stretching upregulated the expression of MMP2 and TIMP2 mRNA s, but did not affect collagen production or LOX activity. In addition to increasing the transcription of COL1A1 , COL3A1 and LOX genes, low‐level mechanical stretching enhanced total collagen production and LOX activity, which should favor ECM stabilization. As an effective regulator of ECM remodeling, mechanical stretching can be exploited in periodontal regeneration and ligament tissue engineering via application of appropriate mechanical stimulation.