PMID- 29214666 OWN - NLM STAT- MEDLINE DCOM- 20190529 LR - 20190529 IS - 1096-9101 (Electronic) IS - 0196-8092 (Linking) VI - 50 IP - 4 DP - 2018 Apr TI - Low-level ultrahigh-frequency and ultrashort-pulse blue laser irradiation enhances osteoblast extracellular calcification by upregulating proliferation and differentiation via transient receptor potential vanilloid 1. PG - 340-352 LID - 10.1002/lsm.22775 [doi] AB - BACKGROUND AND OBJECTIVE: Low-level laser irradiation (LLLI) exerts various biostimulative effects, including promotion of wound healing and bone formation; however, few studies have examined biostimulation using blue lasers. The purpose of this study was to investigate the effects of low-level ultrahigh-frequency (UHF) and ultrashort-pulse (USP) blue laser irradiation on osteoblasts. STUDY DESIGN/ MATERIALS AND METHODS: The MC3T3-E1 osteoblast cell line was used in this study. Following LLLI with a 405 nm newly developed UHF-USP blue laser (80 MHz, 100 fs), osteoblast proliferation, and alkaline phosphatase (ALP) activity were assessed. In addition, mRNA levels of the osteoblast differentiation markers, runt-related transcription factor 2 (Runx2), osterix (Osx), alkaline phosphatase (Alp), and osteopontin (Opn) was evaluated, and extracellular calcification was quantified. To clarify the involvement of transient receptor potential (TRP) channels in LLLI-induced biostimulation, cells were treated prior to LLLI with capsazepine (CPZ), a selective inhibitor of TRP vanilloid 1 (TRPV1), and subsequent proliferation and ALP activity were measured. RESULTS: LLLI with the 405 nm UHF-USP blue laser significantly enhanced cell proliferation and ALP activity, compared with the non-irradiated control and LLLI using continuous-wave mode, without significant temperature elevation. LLLI promoted osteoblast proliferation in a dose-dependent manner up to 9.4 J/cm(2) and significantly accelerated cell proliferation in in vitro wound healing assay. ALP activity was significantly enhanced at doses up to 5.6 J/cm(2) , and expression of Osx and Alp mRNAs was significantly increased compared to that of the control on days 3 and 7 following LLLI at 5.6 J/cm(2) . The extent of extracellular calcification was also significantly higher as a result of LLLI 3 weeks after the treatment. Measurement of TRPV1 protein expression on 0, 3, and 7 days post-irradiation revealed no differences between the LLLI and control groups; however, promotion of cell proliferation and ALP activity by LLLI was significantly inhibited by CPZ. CONCLUSION: LLLI with a 405 nm UHF-USP blue laser enhances extracellular calcification of osteoblasts by upregulating proliferation and differentiation via TRPV1. Lasers Surg. Med. 50:340-352, 2018. (c) 2017 Wiley Periodicals, Inc. CI - (c) 2017 Wiley Periodicals, Inc. FAU - Mikami, Risako AU - Mikami R AD - Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo, Japan. FAU - Mizutani, Koji AU - Mizutani K AD - Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo, Japan. FAU - Aoki, Akira AU - Aoki A AD - Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo, Japan. FAU - Tamura, Yukihiko AU - Tamura Y AD - Department of Bio-Matrix (Pharmacology), Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo, Japan. FAU - Aoki, Kazuhiro AU - Aoki K AD - Department of Basic Oral Health Engineering, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo, Japan. FAU - Izumi, Yuichi AU - Izumi Y AD - Department of Periodontology, Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University (TMDU), Tokyo, Japan. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20171207 PL - United States TA - Lasers Surg Med JT - Lasers in surgery and medicine JID - 8007168 RN - 0 (TRPV Cation Channels) RN - 0 (TRPV1 protein, mouse) RN - 0 (TRPV1 receptor) RN - 106441-73-0 (Osteopontin) RN - EC 3.1.3.1 (Alkaline Phosphatase) SB - IM MH - Alkaline Phosphatase/metabolism MH - Animals MH - Calcinosis/physiopathology MH - Cell Line/radiation effects MH - Cell Proliferation/radiation effects MH - Lasers, Dye/therapeutic use MH - Low-Level Light Therapy/*methods MH - Mice MH - Osteoblasts/*physiology/*radiation effects MH - Osteopontin/metabolism/radiation effects MH - Real-Time Polymerase Chain Reaction/methods MH - Sensitivity and Specificity MH - TRPV Cation Channels/*genetics/*radiation effects MH - Up-Regulation OTO - NOTNLM OT - TRPV1 OT - blue laser OT - calcification OT - cell differentiation OT - low-level laser irradiation OT - osteoblasts OT - proliferation EDAT- 2017/12/08 06:00 MHDA- 2019/05/30 06:00 CRDT- 2017/12/08 06:00 PHST- 2017/11/24 00:00 [accepted] PHST- 2017/12/08 06:00 [pubmed] PHST- 2019/05/30 06:00 [medline] PHST- 2017/12/08 06:00 [entrez] AID - 10.1002/lsm.22775 [doi] PST - ppublish SO - Lasers Surg Med. 2018 Apr;50(4):340-352. doi: 10.1002/lsm.22775. Epub 2017 Dec 7.