Lasers in Medical Science

, Volume 28, Issue 1, pp 125–132 | Cite as

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

  • A. LeonidaEmail author
  • A. Paiusco
  • G. Rossi
  • F. Carini
  • M. Baldoni
  • G. Caccianiga
Original Article


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.


Mesenchymal stem cells Tissue engineering Low-level laser irradiation Biomaterials 



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.


  1. 1.
    Armitage GC (2000) Clinical evaluation of periodontal disease. Periodontol 1995(7):39–53Google Scholar
  2. 2.
    Armitage GC (2000) Periodontal diagnosis and classification of period- ontal disease. Periodontol 2004(34):22–33Google Scholar
  3. 3.
    McAllister BS, Haghighat K (2007) Bone augmentation techniques. J Periodontol 78:377–396PubMedCrossRefGoogle Scholar
  4. 4.
    Li J, Wang HL (2008) Common implant-related advanced bone grafting complications: classification, etiology, and management. Implant Dent 17(4):389–401PubMedCrossRefGoogle Scholar
  5. 5.
    Zippel N, Schulze M, Tobiasch E (2010) Biomaterials and mesenchymal stem cells for regenerative medicine. Recent Pat Biotechnol 4(1):1–22PubMedCrossRefGoogle Scholar
  6. 6.
    Caplan AI (2005) Review: mesenchymal stem cells: cell-based reconstructive therapy in orthopedics. Tissue Eng 11(7–8):1198–1211PubMedCrossRefGoogle Scholar
  7. 7.
    Gehron Robey P (2000) Stem cells near the century mark. J Clin Invest 105:1489–1491CrossRefGoogle Scholar
  8. 8.
    Fibbe WE (2002) Mesenchymal stem cells. A potential source for skeletal repair. Ann Rheum Dis 61(suppl II):ii29–ii31PubMedGoogle 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 75:1281–1287PubMedCrossRefGoogle 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 284:143–147PubMedCrossRefGoogle 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 64:295–312PubMedCrossRefGoogle 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 7:679–689PubMedCrossRefGoogle Scholar
  13. 13.
    Hutmacher DW (2000) Scaffolds in tissue engineering bone and cartilage. Biomaterials 21:2529–2543PubMedCrossRefGoogle 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 4:1213–1221CrossRefGoogle Scholar
  15. 15.
    Gunatillake PA, Adhikari R (2003) Biodegradable synthetic polymers for tissue engineering. Eur Cell Mater 5:1–16PubMedGoogle 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 32:112–122PubMedCrossRefGoogle Scholar
  17. 17.
    Pittenger MF, Martin BJ (2004) Mesenchymal stem cells and their potential as cardiac therapeutics. Circ Res 95(1):9–20PubMedCrossRefGoogle 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 19:163–170PubMedGoogle Scholar
  19. 19.
    Mester E, Mester AF, Mester A (1985) The biomedical effects of laser application. Lasers Surg Med 5:31–39PubMedCrossRefGoogle 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 22:347–354PubMedCrossRefGoogle 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 16:213–217CrossRefGoogle 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 127:2048–2057PubMedCrossRefGoogle 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 41:227–231PubMedCrossRefGoogle 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 35:373–376PubMedCrossRefGoogle 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 23:331–335PubMedCrossRefGoogle Scholar
  26. 26.
    Karu T (1989) Photobiology of low-power laser effects. Health Phys 56:691–704PubMedCrossRefGoogle Scholar
  27. 27.
    Karu TI (2008) Mitochondrial signaling in mammalian cells activated by red and near-IR radiation. Photochem Photobiol 84:1091–1099PubMedCrossRefGoogle 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 22:215–222PubMedCrossRefGoogle 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 8:603–605PubMedCrossRefGoogle Scholar
  30. 30.
    Lipovsky A, Nitzan Y, Lubart R (2008) A possible mechanism for visible lightinduced wound healing. Lasers Surg Med 40:509–514PubMedCrossRefGoogle 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 40:422–432PubMedCrossRefGoogle 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 26:315–321PubMedCrossRefGoogle 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 4:3–14PubMedCrossRefGoogle 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 41:291–297PubMedCrossRefGoogle 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):726–733PubMedCrossRefGoogle 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):373–378PubMedCrossRefGoogle 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):253–269CrossRefGoogle Scholar
  38. 38.
    Slavkin HC, Bartold PM (2000) Challenges and potential in tissue engineering. Periodontol 2006(41):9–15Google Scholar
  39. 39.
    Langer R, Vacanti JP (1993) Tissue engineering. Science 260:920–926PubMedCrossRefGoogle 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–SI34PubMedGoogle 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 14:1731–1742PubMedCrossRefGoogle 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 14:945–953PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag London Ltd 2012

Authors and Affiliations

  • A. Leonida
    • 1
    Email author
  • A. Paiusco
    • 1
  • G. Rossi
    • 1
  • F. Carini
    • 2
  • M. Baldoni
    • 1
    • 3
  • G. Caccianiga
    • 4
  1. 1.Department of PeriodontologyUniversity of Milano-BicoccaMilanItaly
  2. 2.Department of Oral SurgeryUniversity of Milano-BicoccaMilanItaly
  3. 3.University of Milano-BicoccaMilanItaly
  4. 4.Department of OrthodonticsUniversity of Milano-BicoccaMilanItaly

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