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