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Low-level laser irradiation modulates the proliferation and the osteogenic differentiation of bone marrow mesenchymal stem cells under healthy and inflammatory condition

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Abstract

The aim of this in vitro study was to evaluate the effects of low-level laser therapy (LLLT) at different energy intensities on proliferation and osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) under healthy and inflammatory microenvironments. Human BMSCs and BMSCs from inflammatory conditions (i-BMSCs, BMSCs treated with tumor necrosis factor α; TNF-α) were subject to LLLT (Nd:YAG;1064 nm) at different intensities. We designed one control group (without irradiation) and four testing groups (irradiation at 2, 4, 8, and 16 J/cm2) for both BMSCs and i-BMSCs. Cell proliferation was measured using colony-forming unit fibroblast assay and 3-(4,5-dimethylthiazol-2yl)-2,5-diphenyltetrazolium bromide assay. Osteogenic capacity of cells was determined by alkaline phosphatase (ALP) staining, ALP activity assay, Alizarin Red S staining and the mRNA transcript levels of genes runt-related transcription factor 2 (Runx2), ALP, and osteocalcin. Moreover, the effects of LLLT on secretion of TNF-α in BMSCs and i-BMSCs were measured by enzyme-linked immunosorbent assay. Our results demonstrated LLLT could significantly promote BMSC proliferation and osteogenesis at densities of 2 and 4 J/cm2. LLLT at density of 8 J/cm2 could promote the proliferation and osteogenesis of i-BMSCs. However, LLLT at 16 J/cm2 significantly suppressed the proliferation and osteogenesis of BMSCs both in healthy and in inflammatory microenvironment. Moreover, we also found that the expression of TNF-α was obviously inhibited by LLLT at 4, 8, and 16 J/cm2, in an inflammatory microenvironment. Considering these findings, LLLT could improve current in vitro methods of differentiating BMSCs under healthy and inflammatory microenvironments prior to transplantation.

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Funding

This research was financially supported by grants from the National Natural Science Foundation of China (No. 81701029), the Natural Science Foundation of Gansu Province (No. 1606RJZA185 and 17JR5RA335), and the Autonomous Research Project of State Key Laboratory of Military Stomatology (No. 2016KB02).

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Author notes

  1. Liying Wang and Fan Wu contributed equally to this work.

Authors and Affiliations

  1. Department of Stomatology, Lanzhou General Hospital, Lanzhou Command of PLA, 333 South Binhe Road, Qili River District, Lanzhou, 730050, Gansu, People’s Republic of China

    Liying Wang, Jiawen Guo, Yanwei Yang & Yinong Qiu

  2. Key Laboratory of Stem Cells and Gene Drugs of Gansu Province, Lanzhou General Hospital, Lanzhou Command of PLA, Lanzhou, Gansu, China

    Liying Wang

  3. Department of Laparoscope Surgery, The 451st Hospital of People’s Liberation Army, Xi’an, Shaanxi, China

    Fan Wu

  4. Department of General Dentistry, Stomatological Hospital of Xi’an Jiao Tong University, Xi’an, Shaanxi, China

    Chen Liu

  5. Department of Stomatology, The 323rd Hospital of People’s Liberation Army, Xi’an, Shaanxi, China

    Yang Song

Authors
  1. Liying Wang
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  2. Fan Wu
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  3. Chen Liu
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  4. Yang Song
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Corresponding author

Correspondence to Yinong Qiu.

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Ethics statement

This article does not contain any studies with human participants performed by any of the authors. Human cancellous bone fragments and blood were collected based on the treatment needs unrelated to this study. Each donor provided written informed consent. All procedures performed in studies were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

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The authors declare that they have no conflict of interest.

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Wang, L., Wu, F., Liu, C. et al. Low-level laser irradiation modulates the proliferation and the osteogenic differentiation of bone marrow mesenchymal stem cells under healthy and inflammatory condition. Lasers Med Sci 34, 169–178 (2019). https://doi.org/10.1007/s10103-018-2673-8

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  • DOI: https://doi.org/10.1007/s10103-018-2673-8

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