Skip to main content
SpringerLink
Log in

Initial effects of low-level laser therapy on growth and differentiation of human osteoblast-like cells

Initiale Effekte von Low Level Laser Therapie auf Wachstum und Differenzierung von humanen osteoblastären Zellen

  • Original Article
  • Published:
Wiener klinische Wochenschrift Aims and scope Submit manuscript

Zusammenfassung

Der Einsatz von Soft-Lasern im Rahmen einer Low level-Laser Therapie ist mittlerweile eine klinisch etablierte Behandlungsmethode. In vitro Studien haben gezeigt, dass Soft-Laser auch eine biostimulatorische Wirkung auf unterschiedlichste Zelltypen haben. Das Ziel dieser Untersuchung war die Effekte von Low level-Laser Therapie auf das initiale Wachstums- und Differenzierungsverhalten von in vitro kultivierten humanen osteoblastären Zellen zu untersuchen. SaOS-2 Zellen wurden mit Laser-Dosen von 1 J/cm2 und 2 J/cm2 mit einem Dioden Laser mit 670 nm Wellenlänge und einer Ausgangsleistung von 400 mW bestrahlt. Unbehandelte Zellen dienten als Kontrollgruppe. 24 h, 48 h und 72 h nach der Bestrahlung wurden die Zellen geerntet und ihre Vitalität bestimmt. Zusätzlich wurde die Aktivität der Alkalischen Phosphatase ermittelt und die Expression von Osteopontin und Collagen Typ I mittels semiquantitativer RT-PCR untersucht. Zellen, die mit 1 J/cm2 bestrahlt worden waren wiesen, sowohl eine höhere Vitalität als auch eine höhere Aktivität der Alkalischen Phosphatase gegenüber den Kontrollen auf. Auch die Expression von Osteopontin und Collagen Typ I mRNA war gegenüber der Kontrollgruppe erhöht. Hingegen führte eine Verdopplung der Laserleistung zu einer Abnahme der Zellviabilität in den ersten 48 h und zu einer konstant niedrigeren Alkalischen Phosphataseaktivität. Während die Expression von Collagen Typ I und Osteopontin mRNA in unbehandelten und mit 1 J/cm2 bestrahlten Zellen im Verlauf des Experiments leicht abnahm, konnte eine Zunahme ihrer Expression nach Bestrahlung mit 2 J/cm2 beobachtet werden. Unsere Beobachtungen deuten darauf hin, dass Low level-Laser Therapie eine biostimulatorische Wirkung auf SaOS-2 Zellen bereits in der inititalen Kulturphase hat. Diese Ergebnisse können dazu beitragen, neue Therapie-Konzepte in der Regeneration von Knochendefekten zu entwickeln. Weitere Untersuchungen über einen verlängerten Zeitraum wären hilfreich, dieses Potential genauer zu beurteilen.

Summary

Low-level laser therapy is a clinically well established tool for enhancement of wound healing. In vitro studies have also shown that low level laser therapy has a biostimulatory effect on cells of different origin. The aim of this in vitro study was to investigate the initial effect of low-level laser therapy on growth and differentiation of human osteoblast-like cells. SaOS-2 cells were irradiated with laser doses of 1 J/cm2 and 2 J/cm2 using a diode laser with 670 nm wave length and an output power of 400 mW. Untreated cells were used as controls. At 24 h, 48 h and 72 h post irradiation, cells were collected and assayed for viability of attached cells and alkaline phosphatase specific activity. In addition, mRNA expression levels of osteopontin and collagen type I were assessed using semi-quantitative RT-PCR. Over the observation period, cell viability, alkaline phosphatase activity and the expression of osteopontin and collagen type I mRNA were slightly enhanced in cells irradiated with 1 J/cm2 compared with untreated control cells. Increasing the laser dose to 2 J/cm2 reduced cell viability during the first 48 h and resulted in persistently lower alkaline phosphatase activity compared with the other two groups. The expression of osteopontin and collagen type I mRNA slightly decreased with time in untreated controls and cells irradiated with 1 J/cm2, but their expression was increased by treatment with 2 J/cm2 after 72 h. These results indicate that low-level laser therapy has a biostimulatory effect on human osteoblast-like cells during the first 72 h after irradiation. Further studies are needed to determine the potential of low-level laser therapy as new treatment concept in bone regeneration.

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

References

  • Rochkind S, Rousso M, Nissan M, Villarreal M, Barr-Nea L, Rees DG (1989) Systemic effects of low-power laser irradiation on the peripheral and central nervous system, cutaneous wounds and burns. Lasers Surg Med 9: 174–182

    Article  PubMed  CAS  Google Scholar 

  • Walsh LJ (1997) The current status of low level laser therapy in dentistry. Part I. Soft tissue applications. Aust Dent J 42: 247–254

    PubMed  CAS  Google Scholar 

  • Kemmotsu O, Sato K, Furomido H, Harada K, Takigawa C, Kaseno S (1991) Efficacy of low reactive-level laser therapy for pain attenuation of postherpetic neuralgia. Laser Therapy 3: 1–75

    Google Scholar 

  • Lizarelli RFZ, Lamano-Carvalho TL, Brentegani LG (1999) Histometrical evaluation of the healing of the dental alveolus in rats after irradiation with a low-powered GaAlAs laser. SPIE 3593: 49–55

    Article  Google Scholar 

  • Silva Junior AN, Pinheiro ALB, Oliveira MG, Weismann R, Ramalho LMP, Nicolau RA (2002) Computerized morphometric assessment of the effect of low-level laser therapy on bone repair: an experimental animal study. J Clin Laser Med Surg 20: 83–87

    Article  PubMed  Google Scholar 

  • Nissan J, Assif D, Gross MD, Yaffe A, Binderman I (2006) Effect of low intensity laser irradiation on surgically created bony defects in rats. J Oral Rehabil 33: 619–624

    Article  PubMed  CAS  Google Scholar 

  • Almeida-Lopes L, Rigau J, Zangaro RA, Guidugli-Neto J, Jaeger MM (2001) Comparison of the low level laser therapy effects on cultured human gingival fibroblasts proliferation using different irradiance and same fluence. Lasers Surg Med 29: 179–184

    Article  PubMed  CAS  Google Scholar 

  • Gaida K, Koller R, Isler C, Aytekin O, Al-Awami M, Meissl G, Frey M (2004) Low level laser therapy – a conservative approach to the burn scar? Burns 30: 362–367

    Article  PubMed  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Kamali F, Bayat M, Torkaman G, Ebrahimi E, Salavati M (2007) The therapeutic effect of low-level laser on repair of osteochondral defects in rabbit knee. J Photochem Photobiol B 88: 11–15

    Article  PubMed  CAS  Google Scholar 

  • Khadra M, Lyngstadaas SP, Haanaes HR, Mustafa K (2005) Effect of laser therapy on attachment, proliferation and differentiation of human osteoblast-like cells cultured on titanium implant material. Biomaterials 26: 3503–3509

    Article  PubMed  CAS  Google Scholar 

  • 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: 373–378

    Article  PubMed  Google Scholar 

  • Saperia D, Glassberg E, Lyons AF, Abergel RP, Dwyer RM, Uitto J (1986) Demonstration of elevated type I and type III procollagen mRNA levels in cutaneous wounds treated with Helium-Neon laser. Proposed mechanism for enhanced wound healing. Biochem Biophys Res Commun 138: 1123–1128

    Article  PubMed  CAS  Google Scholar 

  • Mester E, Jaszsagi-Nagy E (1973) The effect of laser radiation on wound healing and collagen synthesis. Studia Biophysica 35: 227–230

    CAS  Google Scholar 

  • Turhani D, Scheriau M, Kapral D, Benesch T, Jonke E, Bantleon HP (2006) Pain relief by single low-level laser irradiation in orthodontic patients undergoing fixed appliance therapy. Am J Orthod Dentofacial Orthop 30: 371–377

    Article  Google Scholar 

  • Kon K (1992) In vivo study of low power laser irradiation on new bone formation of rat calvaria defect. J Jpn Soc Laser Dent 3: 35–44

    Google Scholar 

  • Takeda Y (1988) Irradiation effect of low-energy laser on alveolar bone after tooth extraction: experimental study in rats. International Int J Oral Maxillofac Surg 17: 388–391

    Article  CAS  Google Scholar 

  • 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  CAS  Google Scholar 

  • Abramovitch-Gottlib L, Gross T, Naveh D, Geresh S, Rosenwaks S, Bar I, Vago R (2005) Low level laser irradiation stimulates osteogenic phenotype of mesenchymal stem cells seeded on a three-dimensional biomatrix. Lasers Med Sci 20: 138–146

    Article  PubMed  Google Scholar 

  • Khadra M, Ronold HJ, Lyngstadaas SP, Ellingsen JE, Haanes HR (2004) Low-level laser therapy stimulates bone-implant interaction: an experimental study in rabbits. Clin Oral Impl Res 15: 325–332

    Article  Google Scholar 

  • Shupnik MA, Tashjian AH Jr (1982) Epidermal growth factor and phorbol ester actions on human osteosarcoma cells. Characterization of response and nonresponsive cell lines. J Biol Chem 257: 12161–12164

    PubMed  CAS  Google Scholar 

  • 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  CAS  Google Scholar 

  • 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 

  • 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

    Article  Google Scholar 

  • 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 

  • 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

    Article  PubMed  CAS  Google Scholar 

  • 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

    Article  PubMed  CAS  Google Scholar 

  • Dörtbudak 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 

  • 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 Orthop Res 1: 3–14

    Article  Google Scholar 

  • 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  CAS  Google Scholar 

  • Jakse N, Payer M, Tangl S, Berghold A, Kirmeier R, Lorenzoni M (2007) Influence of low-level laser treatment on bone regeneration and osseointegration of dental implants following sinus augmentation: an experimental study on sheep. Clin Oral Impl Res 18: 517–524

    Article  Google Scholar 

  • Van Breughel HH, Bar PR (1992) Power density and exposure time of HeNe laser irradiation are more important than total energy dose in photo-biomodulation of human fibroblasts in vitro. Lasers Surg Med 12: 528–537

    Article  Google Scholar 

  • Ueda 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 

  • Soundry M, Franquin JC, Pourreau-Schreider N, Martin PM (1988) Action d'un laser helium-neon sur la croissance cellulaire: etude in vitro sur fibroblastes gingivaux humains. J Biol Buccale 16: 129–135

    Google Scholar 

  • Turhani D, Weißenböck M, Watzinger E, Cvikl B, Yerit K, Thurnher D, Ewers R (2005) Three dimensional composites manufactured with human mesenchymal cambial layer precursor cells as an alternative for sinus floor augmentation – an in vitro study. Clin Oral Impl Res 16: 417–424

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Cranio-Maxillofacial and Oral Surgery, Medical University of Vienna, Austria

    Elisabeth Stein, Jadranka Koehn, Walter Sutter, Felix Wanschitz & Dritan Turhani

  2. Department of Internal Medicine II, Department of Angiology, Medical University of Vienna, Austria

    Gabriele Wendtlandt & Mehrdad Baghestanian

  3. Department of Otorhinolaryngology, Head and Neck Surgery, Medical University of Vienna, Austria

    Dietmar Thurnher

Authors
  1. Elisabeth Stein
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jadranka Koehn
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Walter Sutter
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gabriele Wendtlandt
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Felix Wanschitz
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Dietmar Thurnher
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mehrdad Baghestanian
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Dritan Turhani
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Dritan Turhani.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Stein, E., Koehn, J., Sutter, W. et al. Initial effects of low-level laser therapy on growth and differentiation of human osteoblast-like cells. Wien Klin Wochenschr 120, 112–117 (2008). https://doi.org/10.1007/s00508-008-0932-6

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00508-008-0932-6

Keywords

Search

Navigation