Effects of low-level laser irradiation on proliferation and osteoblastic differentiation of human mesenchymal stem cells seeded on a three-dimensional biomatrix: in vitro pilot study

Abstract

Mesenchymal stem cells (MSCs) from bone marrow are a recent source for tissue engineering. Several studies have shown that low-level laser irradiation has numerous biostimulating effects. The purpose of this trial was to evaluate the effects of Nd:Yag laser irradiation on proliferation and differentiation of MSCs induced into the osteoblastic lineage. MSCs were collected from adult human bone marrow, isolated, and cultured in complete medium (α-MEM). Subsequently, they were treated with osteogenic medium, seeded in three-dimensional collagen scaffolds, and incubated. We used six scaffolds, equally divided into three groups: two of these were irradiated with Nd:Yag laser at different power levels (15 Hz, 100 mJ, 1.5 W, and one with a power level of 15 Hz, 150 mJ, 2.25 W), and one was left untreated (control group). Evaluations with specific staining were performed at 7 and 14 days. After 7 days, proliferation was significantly increased in scaffolds treated with laser, compared with the control scaffold. After 14 days, however, laser irradiation did not appear to have any further effect on cell proliferation. As concerns differentiation, an exponential increase was observed after 14 days of laser irradiation, with respect to the control group. However, this was a pilot study with very limited sample size, we conclude, that low-level laser irradiation might lead to a reduction in healing times and potentially reduces risks of failure.

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References

  1. 1.

    Armitage GC (2000) Clinical evaluation of periodontal disease. Periodontol 1995(7):39–53

    Google Scholar 

  2. 2.

    Armitage GC (2000) Periodontal diagnosis and classification of period- ontal disease. Periodontol 2004(34):22–33

    Google Scholar 

  3. 3.

    McAllister BS, Haghighat K (2007) Bone augmentation techniques. J Periodontol 78:377–396

    PubMed  Article  Google Scholar 

  4. 4.

    Li J, Wang HL (2008) Common implant-related advanced bone grafting complications: classification, etiology, and management. Implant Dent 17(4):389–401

    PubMed  Article  Google Scholar 

  5. 5.

    Zippel N, Schulze M, Tobiasch E (2010) Biomaterials and mesenchymal stem cells for regenerative medicine. Recent Pat Biotechnol 4(1):1–22

    PubMed  Article  CAS  Google Scholar 

  6. 6.

    Caplan AI (2005) Review: mesenchymal stem cells: cell-based reconstructive therapy in orthopedics. Tissue Eng 11(7–8):1198–1211

    PubMed  Article  CAS  Google Scholar 

  7. 7.

    Gehron Robey P (2000) Stem cells near the century mark. J Clin Invest 105:1489–1491

    Article  Google Scholar 

  8. 8.

    Fibbe WE (2002) Mesenchymal stem cells. A potential source for skeletal repair. Ann Rheum Dis 61(suppl II):ii29–ii31

    PubMed  Google Scholar 

  9. 9.

    Kawaguchi H, Hirachi A, Hasegawa N, Iwata T, Hamaguchi H, Shiba H et al (2004) Enhancement of periodontal tissue regeneration by transplantation of bone marrow mesenchymal stem cells. J Periodontol 75:1281–1287

    PubMed  Article  Google Scholar 

  10. 10.

    Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD et al (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147

    PubMed  Article  CAS  Google Scholar 

  11. 11.

    Jaiswal N, Haynesworth SE, Caplan AI, Bruder SP (1997) Osteogenic differentiation of purified, culture-expanded human mesenchymal stem cells in vitro. J Cell Biochem 64:295–312

    PubMed  Article  CAS  Google Scholar 

  12. 12.

    Yang S, Leong K-F, Du Z, Chua C-K (2001) The design of scaffolds for use in tissue engineering. Part I. Traditional factors. Tissue Eng 7:679–689

    PubMed  Article  CAS  Google Scholar 

  13. 13.

    Hutmacher DW (2000) Scaffolds in tissue engineering bone and cartilage. Biomaterials 21:2529–2543

    PubMed  Article  CAS  Google Scholar 

  14. 14.

    El-Amin SF, Lu HH, Burems J, Mitchell J, Tuan RS, Laurencin CT (2003) Extracellular matrix production by human osteoblasts cultured on biodegradable polymers applicable for tissue engineering. Biomaterials 4:1213–1221

    Article  Google Scholar 

  15. 15.

    Gunatillake PA, Adhikari R (2003) Biodegradable synthetic polymers for tissue engineering. Eur Cell Mater 5:1–16

    PubMed  CAS  Google Scholar 

  16. 16.

    Meinel L, Karageorgiou V, Fajardo R, Snyder B, Shinde-Patil V, Zichner L et al (2004) Bone tissue engineering using human mesenchymal stem cells: effects of scaffold material and medium flow. Ann Biomed Eng 32:112–122

    PubMed  Article  Google Scholar 

  17. 17.

    Pittenger MF, Martin BJ (2004) Mesenchymal stem cells and their potential as cardiac therapeutics. Circ Res 95(1):9–20

    PubMed  Article  CAS  Google Scholar 

  18. 18.

    Mester E, Nagylucskay S, Tisza S, Mester A (1978) Stimulation of wound healing by means of laser rays. Acta Chir Acad Sci Hung 19:163–170

    PubMed  CAS  Google Scholar 

  19. 19.

    Mester E, Mester AF, Mester A (1985) The biomedical effects of laser application. Lasers Surg Med 5:31–39

    PubMed  Article  CAS  Google Scholar 

  20. 20.

    Ozawa Y, Shimizu N, Kariya G, Abiko Y (1998) Low-energy laser irradiation stimulates bone nodule formation at early stages of cell culture in rat calvarial cells. Bone 22:347–354

    PubMed  Article  CAS  Google Scholar 

  21. 21.

    Yamamoto M, Tamura K, Hiratsuka K, Abiko Y (2001) Stimulation of MCM3 gene expression in osteoblast by low level laser irradiation. Laser Med Sci 16:213–217

    Article  CAS  Google Scholar 

  22. 22.

    Hu WP, Wang JJ, Yu CL, Lan CC, Chen GS, Yu HS (2007) Helium-neon laser irradiation stimulates cell proliferation through photostimulatory effects in mitochondria. J Invest Dermatol 127:2048–2057

    PubMed  Article  CAS  Google Scholar 

  23. 23.

    Moriyama Y, Nguyen J, Akens M, Moriyama EH, Lilge L (2009) In vivo effects of low level laser therapy on inducible nitric oxide synthase. Lasers Surg Med 41:227–231

    PubMed  Article  Google Scholar 

  24. 24.

    Shiba H, Tsuda H, Kajiya M, Fujita T, Takeda K, Hino T, Kawaguchi H, Kurihara H (2009) Neodymium-doped yttrium-aluminium-garnet laser irradiation abolishes the increase in interleukin-6 levels caused by peptidoglycan through the p38 mitogen-activated protein kinase pathway in human pulp cells. J Endod 35:373–376

    PubMed  Article  Google Scholar 

  25. 25.

    Safavi SM, Kazemi B, Esmaeili M, Fallah A, Modarresi A, Mir M (2008) Effects of low-level He-Ne laser irradiation on the gene expression of IL-1beta, TNF-alpha, IFN-gamma, TGF-beta, bFGF, and PDGF in rat’s gingiva. Lasers Med Sci 23:331–335

    PubMed  Article  Google Scholar 

  26. 26.

    Karu T (1989) Photobiology of low-power laser effects. Health Phys 56:691–704

    PubMed  Article  CAS  Google Scholar 

  27. 27.

    Karu TI (2008) Mitochondrial signaling in mammalian cells activated by red and near-IR radiation. Photochem Photobiol 84:1091–1099

    PubMed  Article  CAS  Google Scholar 

  28. 28.

    Zhang L, Xing D, Zhu D, Chen Q (2008) Low-power laser irradiation inhibiting Abeta25-35-induced PC12 cell apoptosis via PKC activation. Cell Physiol Biochem 22:215–222

    PubMed  Article  CAS  Google Scholar 

  29. 29.

    Aimbire F, Santos FV, Albertini R, Castro-Faria-Neto HC, Mittmann J, Pacheco-Soares C (2008) Low-level laser therapy decreases levels of lung neutrophils anti-apoptotic factors by a NF-kappaB dependent mechanism. Int Immunopharmacol 8:603–605

    PubMed  Article  CAS  Google Scholar 

  30. 30.

    Lipovsky A, Nitzan Y, Lubart R (2008) A possible mechanism for visible lightinduced wound healing. Lasers Surg Med 40:509–514

    PubMed  Article  Google Scholar 

  31. 31.

    Ignatieva N, Zakharkina O, Andreeva I, Sobol E, Kamensky V, Lunin V (2008) Effects of laser irradiation on collagen organization in chemically induced degenerative annulus fibrosus of lumbar intervertebral disc. Lasers Surg Med 40:422–432

    PubMed  Article  Google Scholar 

  32. 32.

    Silveira LB, Prates RA, Novelli MD, Marigo HA, Garrocho AA, Amorim JC, Sousa GR, Pinotti M, Ribeiro MS (2008) Investigation of mast cells in human gingiva following low-intensity laser irradiation. Photomed Laser Surg 26:315–321

    PubMed  Article  CAS  Google Scholar 

  33. 33.

    Coombe AR, Ho CT, Darendeliler MA, Hunter N, Philips JR, Chapple CC, Yum LW (2001) The effects of low level laser irradiation on osteoblastic cells. Clin Orthod Res 4:3–14

    PubMed  Article  Google Scholar 

  34. 34.

    Bouvet-Gerbettaz S, Merigo E, Rocca J-P, Carle GF, Rochet N (2009) Effects of low-level laser therapy on proliferation and differentiation of murine bone marrow cells into osteoblasts and osteoclasts. Lasers Surg Med 41:291–297

    PubMed  Article  Google Scholar 

  35. 35.

    Hou JF, Zhang H, Yuan X, Li J, Wei YJ, Hu SS (2008) In vitro effects of low-level laser irradiation for bone marrow mesenchymal stem cells: proliferation, growth factors secretion and myogenic differentiation. Lasers Surg Med 40(10):726–733

    PubMed  Article  Google Scholar 

  36. 36.

    Tuby H, Maltz L, Oron U (2007) Low-level laser irradiation (LLLI) promotes proliferation of mesenchymal and cardiac stem cells in culture. Lasers Surg Med 39(4):373–378

    PubMed  Article  Google Scholar 

  37. 37.

    Bartold PM, McCulloch CA, Narayanan AS, Pitaru S (2000) Tissue engineering: a new paradigm for periodontal regeneration based on molecular and cell biology. Periodontol 2000(24):253–269

    Article  Google Scholar 

  38. 38.

    Slavkin HC, Bartold PM (2000) Challenges and potential in tissue engineering. Periodontol 2006(41):9–15

    Google Scholar 

  39. 39.

    Langer R, Vacanti JP (1993) Tissue engineering. Science 260:920–926

    PubMed  Article  CAS  Google Scholar 

  40. 40.

    Vacanti JP, Langer R (1999) Tissue engineering: the design and fabrication of living replacement devices for surgical reconstruction and transplantation. Lancet 354(Suppl 1):SI32–SI34

    PubMed  Google Scholar 

  41. 41.

    Li Y, Jin F, Du Y, Ma Z, Li F, Wu G, Shi J, Zhu X, Yu J, Jin Y (2008) Cementum and periodontal ligament-like tissue formation induced using bioengineered dentin. Tissue Eng Part A 14:1731–1742

    PubMed  Article  CAS  Google Scholar 

  42. 42.

    Tobita M, Uysal AC, Ogawa R, Hyakusoku H, Mizuno H (2008) Periodontal tissue regeneration with adipose-derived stem cells. Tissue Eng Part A 14:945–953

    PubMed  Article  CAS  Google Scholar 

Download references

Acknowledgments

This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors. All authors declare that there are no conflicting interests.

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Correspondence to A. Leonida.

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Leonida, A., Paiusco, A., Rossi, G. et al. Effects of low-level laser irradiation on proliferation and osteoblastic differentiation of human mesenchymal stem cells seeded on a three-dimensional biomatrix: in vitro pilot study. Lasers Med Sci 28, 125–132 (2013). https://doi.org/10.1007/s10103-012-1067-6

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Keywords

  • Mesenchymal stem cells
  • Tissue engineering
  • Low-level laser irradiation
  • Biomaterials