Skip to main content

Advertisement

SpringerLink
Log in

Low level laser irradiation stimulates osteogenic phenotype of mesenchymal stem cells seeded on a three-dimensional biomatrix

  • Original Article
  • Published:
Lasers in Medical Science Aims and scope Submit manuscript

Abstract

Mesenchymal stem cells (MSCs) seeded on three-dimensional (3D) coralline (Porites lutea) biomatrices were irradiated with low-level laser irradiation (LLLI). The consequent phenotype modulation and development of MSCs towards ossified tissue was studied in this combined 3D biomatrix/LLLI system and in a control group, which was similarly grown, but was not treated by LLLI. The irradiated and non irradiated MSC were tested at 1–7, 10, 14, 21, 28 days of culturing via analysis of cellular distribution on matrices (trypan blue), calcium incorporation to newly formed tissue (alizarin red), bone nodule formation (von Kossa), fat aggregates formation (oil red O), alkaline phosphatase (ALP) activity, scanning electron microscopy (SEM) and electron dispersive spectrometry (EDS). The results obtained from the irradiated samples showed enhanced tissue formation, appearance of phosphorous peaks and calcium and phosphate incorporation to newly formed tissue. Moreover, in irradiated samples ALP activity was significantly enhanced in early stages and notably reduced in late stages of culturing. These findings of cell and tissue parameters up to 28 days of culture revealed higher ossification levels in irradiated samples compared with the control group. We suggest that both the surface properties of the 3D crystalline biomatrices and the LLLI have biostimulatory effects on the conversion of MSCs into bone-forming cells and on the induction of ex-vivo ossification.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1.
Fig. 2.
Fig. 3.
Fig. 4.
Fig. 5.
Fig. 6.
Fig. 7.

Similar content being viewed by others

References

  1. Prockop DJ (1997) Marrow stromal cells as stem cells for nonhematopoietic tissues. Science 276:71–74

    Article  PubMed  Google Scholar 

  2. Pittenger MF, Mackay AM, Beck SC, Jaiswal RK, Douglas R, Mosca JD, Moorman MA, Simonetti DW, Craig S, Marshak DR (1999) Multilineage potential of adult human mesenchymal stem cells. Science 284:143–147

    Article  PubMed  Google Scholar 

  3. Long MW (2001) Osteogenesis and bone-marrow-derived cells. Blood Cell Mol Dis. 27:677–690

    Article  Google Scholar 

  4. Caplan AI, Bruder S (2001) Mesenchymal stem cells: building blocks for molecular medicine in the 21st Century. Trends Mol Med. 7:259–264

    Article  PubMed  Google Scholar 

  5. Sukhikh GT, Malaitsev VV, Bogdanova IM, Dubrovina IV (2002) Mesenchymal stem cells. Bull Exp Biol Med.133:103–9

    Article  Google Scholar 

  6. Sikavitsas VI, Temenoff JS, Mikos AG (2001) Biomaterial and bone mechanotransduction. Biomaterials. 22:2581–2593

    Article  PubMed  Google Scholar 

  7. Roden GA, Harada S (1997) The missing bone. Cell. 89:677–680

    Article  PubMed  Google Scholar 

  8. Kuznetsov SA, Krebsbach PH, Satomura K, Kerr J, Riminucci M, Benayahu D, Robey PG (1997) Single-colony derived strains of human marrow stromal fibroblasts form bone after transplantation in vivo. J Biomed Mater Res 12:1335–1347

    Google Scholar 

  9. Dennis JE, Caplan AI (2004) Bone marrow mesenchymal stem cells. In: Sell S (eds) Stem cells handbook. Humana Press Inc, Totowa, NJ, pp 107–117

    Google Scholar 

  10. Dennis JE, Charbord P (2002) Origin and differentiation of human and murine stroma. Stem Cells 20:205–214

    Article  PubMed  Google Scholar 

  11. Cheng SL, Zhang SF, Nelson TL, Warlow PM, Civitelli R (1994) Stimulation of human osteoblast differentiation and function by ipriflavone and its metabolites. Calcif Tissue Int 55:356–362

    Article  PubMed  Google Scholar 

  12. Ter Brugge PJ, Jansen JA (2002) In vitro osteogenic differentiation of rat bone marrow cells subcultured with and without dexamethasone. Tissue Eng 8:321–331

    Article  PubMed  Google Scholar 

  13. 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  Google Scholar 

  14. Kana JS, Hutschenreiter G, Haina D, Waidelich W (1981) Effect of low-power density laser radiation on healing of open skin wound in rats. Arch Surg 116:293–296

    PubMed  Google Scholar 

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

    PubMed  Google Scholar 

  16. Boulton M, Marshall J (1986) He-Ne laser stimulation of human fibroblast proliferation and attachment in vitro. Lasers Life Sci 1:123–134

    Google Scholar 

  17. Van Breugel H, Bar PRD (1992) Power density and exposure time of He-Ne laser irradiation are more important than total energy dose in photo-biomodulation of human fibroblasts in vitro. Lasers Surg Med 12:528–537

    PubMed  Google Scholar 

  18. Balboni GC, Brandi ML, Zonefrati R, Repice F (1986) Effects of He-Ne/I.R. lasers irradiation on two lines of normal human fibroblasts in vitro. Arch Ital Embriol 91:179–188

    Google Scholar 

  19. Schultz RJ, Krishnamurthy S, Thelmo W, Rodriguez JE, Harvey G (1985) Effects of varying intensities of laser energy on articular cartilage. Lasers Surg Med 5:557–588

    Google Scholar 

  20. Abergel RP, Meeker CA, Lam TS, Dwyer RM, Lesavoy MA, Uitto J (1984) Control of connective tissue metabolism by laser: recent developments and future prospects. J Am Acad Dermatol 11:1142–1150

    PubMed  Google Scholar 

  21. Bosarta M, Jucci A, Olliaro P, Quacci D, Sacchi S (1984) In vitro fibroblast and dermis fibroblast activation by laser irradiation at low energy. Dermatologica 168:157–162

    PubMed  Google Scholar 

  22. Lam TS, Abergel RP, Meeker CA, Castel JC, Dwyer RM, Uitto J (1986) Laser stimulation of collagen synthesis in human skin fibroblast cultures. Lasers Life Sci 1:61–77

    Google Scholar 

  23. Honmura A, Yanase M, Obata J, Haruki E (1992) Therapeutic effect of Ga-Al-As diode laser irradiation on experimentally induced inflammation in rats. Lasers Surg Med 12:441–449

    PubMed  Google Scholar 

  24. Anders JJ, Borke RC, Woolery SK, Van de Merwe WP (1993) Low power laser irradiation alters the rate of regeneration of the rat facial nerve. Lasers Surg Med 13:72–82

    PubMed  Google Scholar 

  25. Ozawa Y, Shimizu N, Mishima H, Kariya G, Yamaguchi M, Takiguchi H, Iwasawa T, Abiko Y (1995) Stimulatory effects of low-power laser irradiation on bone formation in vitro. In: Altshuler GB, Blankenau RJ, Wigdor HA (eds) Advanced Laser Dentistry. Proc SPIE 1984. International Society for Optical Engineering, Washington, DC, pp 281–288

    Google Scholar 

  26. 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

    Article  PubMed  Google Scholar 

  27. Yamada K (1991) Biological effects of low power laser irradiation on clonal osteoblastic cells (MC3T3-E1). J Jap Orthop Assoc 65:787–799

    Google Scholar 

  28. Dortbudak O, Haas R, Mailath-Pokorny G (2000) Biostimulation of bone marrow cells with a diode soft laser. Clin Oral Impl Res 11:540–545

    Article  Google Scholar 

  29. 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

    Google Scholar 

  30. Trelles MA, Mayayo E (1987) Bone fracture consolidates faster with low power laser. Lasers Surg Med 7:36–45

    PubMed  Google Scholar 

  31. Lunger EJ, Rochkind S, Wollman Y, Kogan G, Dekel S (1998) Effect of low power laser irradiation on the mechanical properties of bone fracture healing in rats. Lasers Surg Med 22:97–102

    Article  PubMed  Google Scholar 

  32. Barushka O, Yaakobi T, Oron U (1995) Effect of low-energy laser (He-Ne) irradiation on the process of bone repair in the rat tibia. Bone 16:47–55

    Article  PubMed  Google Scholar 

  33. Kawasaki K, Shimizu N (2000) Effects of low-energy laser irradiation on bone remodeling during experimental tooth movement in rats. Lasers Surg Med 26:282–291

    Article  PubMed  Google Scholar 

  34. Saito S, Shimizu N (1997) Stimulatory effect of low-power laser irradiation on bone regeneration in mid-palatal suture during expansion in the rat. Am J Orthod Dentfac Orthop 111:525–532

    Google Scholar 

  35. Abbott A (2003) Biology’s new dimension. Nature 424:870–872

    Article  PubMed  Google Scholar 

  36. Yukna RA, Mayer ET, Brite DU (1984) Longitudinal evaluation of durapatite ceramic as an alloplastic implant in periodontal osseous defects after 3 years. J Periodontol 55:633–663

    PubMed  Google Scholar 

  37. Kattigan BD (1986) Bone regeneration with bone substitutes: an animal study. Springer Verlag, New York, pp 66–75

    Google Scholar 

  38. Daculsi G, Le Geros R, Heughebaert M, Barbieux I (1990) Formation of carbonate-apatite crystals after implantation of calcium-phosphate ceramics. Calcif Tissue Int 46: 20–27

    PubMed  Google Scholar 

  39. Ducheyne P, Oiu Q (1999) Bioactive ceramics: the effect of surface reactivity on bone formation and bone cell function. Biomaterials 20:2287–2303

    Article  PubMed  Google Scholar 

  40. Schmidt C, Ignatius AA, Claes LE (2001) Proliferation and differentiation parameters of human osteoblasts on titanium and steel surfaces. J Biomed Mater Res 54:209–215

    Article  PubMed  Google Scholar 

  41. Shea DE, Wang D, Franceschi RT, Moony DJ (2000) Engineered bone development from a pre-osteoblast cell line on three-dimensional scaffolds. Tissue Eng 6:605–617

    Article  PubMed  Google Scholar 

  42. Orban JM, Marra KG, Hollinger JO (2002) Composition options for tissue-engineered bone. Tissue Eng 8:529–539

    Article  PubMed  Google Scholar 

  43. Rizzi SC, Heath DJ, Coombes AGA, Bock N, Textor M, Downes S (2001) Biodegradable polymer/hydroxyapatite composites: Surface analysis and initial attachment of human osteoblasts. J Biomed Mater Res 55:475–486

    Article  PubMed  Google Scholar 

  44. Cancedda R, Dozin B, Giannoni P, Quarto R (2003) Tissue engineering and cell therapy of cartilage and bone. Mat Biol 22:81–91

    Article  Google Scholar 

  45. Demers C, Hamdy R, Corsi K, Chellat F, Tabrizian M, Yahia L (2002) Natural coral as a bone graft substitute: a review. Bio-Med Mater Eng 12:15–35

    Google Scholar 

  46. Green D, Walsh D, Mann S, Oreffo ROC (2002) The potential of biomimesis in bone tissue engineering: lessons from the design and synthesis of invertebrates skeletons. Bone 30:810–815

    Article  PubMed  Google Scholar 

  47. Abramovitch-Gottlib L, Katoshevski D, Vago R (2002) A computerized tank system for studying the effect of temperature on calcification of reef organisms. J Biochem Bioph Meth 50:245–252

    Article  Google Scholar 

  48. Roudier M, Bouchon C, Rouvillain JL, Amedee J, Bareille R, Rouais F, Fricain JC, Dupuy B, Kien P, Jeandot R, Bassecathalinat B (1995) The resorption of bone-implanted corals varies with porosity but also with the host-reaction. J Biomed Mater Res.29:909–915

    Article  Google Scholar 

  49. Guillemin G, Patat JL, Fournie J, Chetail M (1987) The use of coral as a bone-graft substitute. J Biomed Mater Res 21:557–567

    Article  PubMed  Google Scholar 

  50. Bou-Abboud NN, Ouhayoum JP (1998) Bone formation with discs or particles of natural coral skeleton plus polyglactin910 mesh: histologic evaluation in rat calvaria. Int J Oral Maxillofac Implants 13:115–120

    PubMed  Google Scholar 

  51. Vuola J, Böhling T, Kinnunen J, Hirvensalo E, Asko-Seljavaara S (2000) Natural coral as bone-defect-filling material. J Biomed Mater Res 51:117–122

    Article  PubMed  Google Scholar 

  52. Petite H, Viateau V, Bensaid W, Meunier A, de Pollak C, Bourguignon M, Oudina K, Sedel L, Guillemin G (2000) Tissue-engineered bone regeneration. Nature Biotech 18:959–963

    Google Scholar 

  53. Vago R, Plotquin D, Bunin A, Sinelnikov I, Atar D, Itzhak D (2002) Hard tissue remodeling using biofabricated coralline biomaterials. J Biochem Biophys Meth 50:253–259

    Article  PubMed  Google Scholar 

  54. Barnes DJ, Chalker BE (1990) Calcification and photosynthesis in reef-building corals and algae. In: Dubinsky Z, editor. Ecosystems of the World 25. Amsterdam: Elsevier, pp 109–131

  55. Dahan D, Vago R, Golan Y (2003) Skeletal architecture and microstructure of the calcifying coral Fungia simplex. Mater Sci Eng C 23:473–477

    Google Scholar 

  56. Weiner S, Addadi L (1997) Design strategies in mineralized biological materials. J Mater Chem 7:689–702

    Article  Google Scholar 

  57. Hu J, Fraser R, Russell JJ, Ben-Nissan B, Vago R (2000) Australian coral as a biomaterial: Characteristics. J Mater Sci Technol 16:591–595

    Google Scholar 

  58. Doherty MJ, Schlag G, Schwarz N, Mollan RAS, Nolam PS, Wilson DJ (1994) Biocompatibility of xenogeneic bone, commercially available coral, a bioceramic and tissue sealant for human osteoblasts. Biomaterials 15:601–608

    Article  PubMed  Google Scholar 

  59. Udea Y, Shimizu N (2003) Effects of pulse frequency of low-level laser therapy (LLLT) on bone nodule formation in rat calvarial cells. J Clin Laser Med Surg 21:271–277

    Article  PubMed  Google Scholar 

  60. Fricain JC, Bareille R, Ulyss F, Dupuy B, Amedee J (1998) Evaluation of proliferation and protein expression of human bone marrow cells cultured on coral crystallized in the aragonite or calcite form. J Biomed Mater Res 42: 96–102

    Article  PubMed  Google Scholar 

  61. Sauteir JM, Nefussi JR, Boulekbache H, Forest N (1990) In vitro bone formation on coral granules. In vitro. Cell Dev Biol 26:1079–1085

    Google Scholar 

  62. Grigoriadis AE, Heersche JNM, Aubin JE (1988) Differentiation of muscle, fat, cartilage, and bone from progenitor cells present in a bone-derived clonal cell-population - effect of dexamethasone. J Cell Biol 106:2139–2151

    Article  PubMed  Google Scholar 

  63. Asahina I, Sampath TK, Hauschka PV (1996) Human osteogenic protein-1 induces chondroblastic, osteoblastic, and/or adipocytic differentiation of clonal murine target cells. Exp Cell Res 222: 38–47

    Article  PubMed  Google Scholar 

  64. Mie M, Ohgushi H, Yanagida Y, Haruyama T, Kobatake E, Aizawa M (2000)Osteogenesis coordinated in C3H10T1/2 cells by adipogenesis-dependent BMP-2 expression system. Tissue Eng 6:9–18

    Article  PubMed  Google Scholar 

  65. Dennis JE, Merriam A, Awadallah A, Yoo JU, Johnstone B, Caplan AI (1999)A quadripotential mesenchymal progenitor cell isolated from the marrow of an adult mouse. J Bone Miner Res 14:700–709

    PubMed  Google Scholar 

  66. Chen D, Ji X, Harris MA, Feng JQ, Karsenty G, Celeste AJ, Rosen V, Mundy GR, Harris SE (1998) Differential roles for bone morphogenetic protein (BMP) receptor type IB and IA in differentiation and specification of mesenchymal precursor cells to osteoblast and adipocyte lineages. J Cell Biol 142:295–305

    Article  PubMed  Google Scholar 

  67. Thompson DL, Lum KD, Nygaard SC, Kuestner RE, Kelly KA, Gimble JM, Moore EE (1998) The derivation and characterization of stromal cell lines from the bone marrow of p53(-/-) mice: New insights into osteoblast and adipocyte differentiation. J Bone Miner Res 13:195–204

    PubMed  Google Scholar 

  68. Conget PA, Minguell JJ (1999) Phenotypical and functional properties of human bone marrow mesenchymal progenitor cells. J Cell Physiol 181:67–73

    Article  PubMed  Google Scholar 

  69. Harris SE, Harris MA, Mahy P, Wozney J, Feng JQ, Mundy GR (1994) Expression of bone morphogenetic protein messenger-RNA by normal rat and human prostate and prostate-cancer cells. Prostate 24:204–211

    PubMed  Google Scholar 

  70. Birnbaum RS, Bowsher RR, Wiren KM (1995) Changes in IGF-I and IGF-II expression and secretion during the proliferation and differentiation of normal rat osteoblasts. J Endocrinol 144:251–259

    PubMed  Google Scholar 

  71. Rubinov AN (2003) Physical grounds for biological effect of laser radiation. J Phys D: Appl Phys 36:2317–2330

    Google Scholar 

  72. Klebanov GI, Kreinina MV, Poltanov EA, Khristoforova TV, Vladimirov YA (2001) Mehanism of therapeutic effect of low-intensity infrared laser radiation. Bull Exp Biol Med 131:239–241

    Article  PubMed  Google Scholar 

Download references

Acknowledgments

The research was partially supported by the Binational Science Foundation (BSF) # 2001045 and by the James Franck Binational German-Israeli Program in Laser-Matter Interaction. The authors are grateful to Mrs. Aviva Kiryati for expert assistance in SEM and EDS analyses. The authors wish to thank Mrs. Inez Murinek for the fruitful discussions and comments on the manuscript.

Author information

Authors and Affiliations

  1. Department of Biotechnology Engineering, Ben-Gurion University of the Negev, Beer-Sheva, 84105, Israe

    Liat Abramovitch-Gottlib, Talia Gross, Shimona Geresh & Razi Vago

  2. The National Institute of Biotechnology, Ben-Gurion University of the Negev, Beer-Sheva, 84105, Israel

    Liat Abramovitch-Gottlib, Talia Gross & Razi Vago

  3. Department of Physics, Ben-Gurion University of the Negev, Beer-Sheva, 84105, Israel

    Doron Naveh, Salman Rosenwaks & Ilana Bar

  4. The Institutes for Applied Research, Ben-Gurion University of the Negev, Beer-Sheva, 84105, Israel

    Ilana Bar

Authors
  1. Liat Abramovitch-Gottlib
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Talia Gross
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Doron Naveh
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shimona Geresh
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Salman Rosenwaks
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ilana Bar
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Razi Vago
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Liat Abramovitch-Gottlib.

Additional information

This publication is dedicated to the memory of Professor Shimona Geresh, a colleague and friend, who passed away while this study was under way.

Salman Rosenwaks holds the Helen and Sanford Diller Family Chair in Chemical Physics

Rights and permissions

Reprints and permissions

About this article

Cite this article

Abramovitch-Gottlib, L., Gross, T., Naveh, D. et al. Low level laser irradiation stimulates osteogenic phenotype of mesenchymal stem cells seeded on a three-dimensional biomatrix. Lasers Med Sci 20, 138–146 (2005). https://doi.org/10.1007/s10103-005-0355-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10103-005-0355-9

Keywords

Search

Navigation