Increased cell proliferation and differential protein expression induced by low-level Er:YAG laser irradiation in human gingival fibroblasts: proteomic analysis
- 592 Downloads
Erbium-doped yttrium aluminum garnet (Er:YAG) laser treatment has demonstrated favorable wound healing effect after periodontal therapy. One of the reasons may be the positive biological effect of the low-level laser on the irradiated tissues, although the mechanism remains unclear. The aim of this study was to investigate the effect of low-level Er:YAG laser irradiation on cell proliferation and laser-induced differential expression of proteins in human gingival fibroblasts (HGFs) by proteomic analysis. In the first experiment, HGFs were exposed to low-level Er:YAG laser irradiation and the laser-induced cell proliferation and damage were evaluated on day 3. In the second experiment, proteomic analysis was performed on day 1 after irradiation. The peptides prepared from HGFs were analyzed by a hybrid ion trap-Fourier transform mass spectrometer, Mascot search engine, and UniProtKB database. A significant increase in cell proliferation without cell damage after irradiation was observed. Among the total identified 377 proteins, 59 proteins, including galectin-7, which was associated with the process of wound healing, were upregulated and 15 proteins were downregulated in laser-treated HGFs. In the third experiment, the increase in messenger RNA (mRNA) and protein expression of galectin-7 in the irradiated HGFs was validated by various analytical techniques. In addition, the effect of recombinant human galectin-7 on the modulation of HGFs proliferation was confirmed. The results indicate that low-level Er:YAG laser irradiation can promote HGF proliferation and induce a significant change in protein expression and the upregulation of galectin-7 expression may partly contribute to the increase in cell proliferation.
KeywordsEr:YAG laser Galectin-7 Gingival fibroblasts Low-level laser therapy Proteomics
This study was partially supported by the grant of the Global Center of Excellence Program of the International Research Center for Molecular Science in Tooth and Bone Diseases at TMDU funded by the Ministry of Education, Culture, Sports, Science and Technology of Japan and the grant of Grants-in-Aid for Scientific Research (C) (no. 22392308 and no. 23463212 for A.A.) from the Japan Society for the Promotion of Science. We would like to acknowledge Dr. Kengo Iwasaki and Dr. Yoichi Taniguchi at TMDU for their kind advices and supports.
- 16.Tsuchida S, Satoh M, Umemura H, Sogawa K, Kawashima Y, Kado S, Sawai S, Nishimura M, Kodera Y, Matsushita K, Nomura F (2012) Proteomic analysis of gingival crevicular fluid for discovery of novel periodontal disease markers. Proteomics 12(13):2190–2202. doi: 10.1002/pmic.201100655 CrossRefPubMedGoogle Scholar
- 17.Tsuchida S, Satoh M, Kawashima Y, Sogawa K, Kado S, Sawai S, Nishimura M, Ogita M, Takeuchi Y, Kobyashi H, Aoki A, Kodera Y, Matsushita K, Izumi Y, Nomura F (2013) Application of quantitative proteomic analysis using tandem mass tags for discovery and identification of novel biomarkers in periodontal disease. Proteomics 13(15):2339–2350. doi: 10.1002/pmic.201200510 CrossRefPubMedGoogle Scholar
- 23.Rondanino C, Poland PA, Kinlough CL, Li H, Rbaibi Y, Myerburg MM, Al-bataineh MM, Kashlan OB, Pastor-Soler NM, Hallows KR, Weisz OA, Apodaca G, Hughey RP (2011) Galectin-7 modulates the length of the primary cilia and wound repair in polarized kidney epithelial cells. Am J Physiol Renal Physiol 301(3):F622–633. doi: 10.1152/ajprenal.00134.2011 PubMedCentralCrossRefPubMedGoogle Scholar
- 33.Abe D, Kubota T, Morozumi T, Shimizu T, Nakasone N, Itagaki M, Yoshie H (2011) Altered gene expression in leukocyte transendothelial migration and cell communication pathways in periodontitis-affected gingival tissues. J Periodontal Res 46(3):345–353. doi: 10.1111/j.1600-0765.2011.01349.x CrossRefPubMedGoogle Scholar
- 34.Chidgey M, Brakebusch C, Gustafsson E, Cruchley A, Hail C, Kirk S, Merritt A, North A, Tselepis C, Hewitt J, Byrne C, Fassler R, Garrod D (2001) Mice lacking desmocollin 1 show epidermal fragility accompanied by barrier defects and abnormal differentiation. J Cell Biol 155(5):821–832. doi: 10.1083/jcb.200105009 PubMedCentralCrossRefPubMedGoogle Scholar
- 41.Berbee JF, Coomans CP, Westerterp M, Romijn JA, Havekes LM, Rensen PC (2010) Apolipoprotein CI enhances the biological response to LPS via the CD14/TLR4 pathway by LPS-binding elements in both its N- and C-terminal helix. J Lipid Res 51(7):1943–1952. doi: 10.1194/jlr.M006809 PubMedCentralCrossRefPubMedGoogle Scholar
- 43.Shefer G, Oron U, Irintchev A, Wernig A, Halevy O (2001) Skeletal muscle cell activation by low-energy laser irradiation: a role for the MAPK/ERK pathway. J Cell Physiol 187(1):73–80. doi: 10.1002/1097-4652(2001)9999:9999<::aid-jcp1053>3.0.co;2-9 CrossRefPubMedGoogle Scholar
- 45.Fischer A, Baljuls A, Reinders J, Nekhoroshkova E, Sibilski C, Metz R, Albert S, Rajalingam K, Hekman M, Rapp UR (2009) Regulation of RAF activity by 14-3-3 proteins: RAF kinases associate functionally with both homo- and heterodimeric forms of 14-3-3 proteins. J Biol Chem 284(5):3183–3194. doi: 10.1074/jbc.M804795200 CrossRefPubMedGoogle Scholar