Abstract Orthodontic tooth movement is a biological response to a mechanical force. One of the challenges in orthodontics is obtaining desired tooth movement during treatment. Accelerating tooth movement and decreasing demands on anchorage can reduce treatment times and overall satisfaction for patient and doctor. Low-level laser therapy (LLLT) is emerging as a technology that may decrease orthodontic treatment time. Many in vitro and in vivo studies have reported the effects of low level lasers at random time points and energy densities. None of the studies have optimized the dose required for osteoblast proliferation and differentiation. The purpose of this study was to find the optimum stimulatory dose of low level laser irradiation (LLLI) on human osteoblast cell proliferation and differentiation and to analyze our findings with reference to the Arndt-Shultz Law of applied energy. In this in vitro study a GaAlAs laser at 830nm, 20 mW with continuous exposure at various doses were used on a human osteoblast cell line. According to the Arndt-Shulz Law weak stimuli initiate vital activity, moderate stimuli enhance the cellular activity with subsequent peak stimulation and greater stimuli (beyond a threshold value) may not have any influence or inhibit the vital activity. The implications of LLLI on human osteoblasts and influencing tooth movement in orthodontics were discussed. Human osteoblasts were cultured in minimum essential medium (MEM) complete medium consisting 10% fetal bovine serum and 1% antibiotics. Cells grown in complete medium were plated onto 96 well plate, allowed to adhere for 4-5 hours and were exposed to GaAlAs lasers at 6 , 12, 18, 24, 30, 36, 45, 60, 75, and 90 seconds. The cells treated with xiii LLLI were assessed for cell proliferation at 24, 48 and 72 hour intervals. A calorimetric cell proliferation assay (WST-1) assay was performed according to manufacture's instructions. The results indicated that at 24 hours the 6 and 12 seconds doses significantly inhibited proliferation compared to the control. At 48 hours the 30 seconds exposure significantly increased proliferation. At 72 hours time interval, cell proliferation was observed in a dose dependent pattern with a minimum at 6 seconds with peak proliferation at 18 seconds. A gradual decrease in cell viability was observed in the cells treated beyond this dose with a maximum inhibition seen at 60 seconds. At 75 and 90 seconds no difference was observed between the control and experimental group. To establish efficient acquisition of adequate quantities of alkaline phosphatase, cells were grown in 12 well plates in complete medium or osteogenic medium. These cells were exposed to LLLI for 18, 48, and 60 seconds. The activity of early osteogenic differentiation marker alkaline phosphatase (ALP) was investigated 10 days post exposure. Our results demonstrated that alkaline phosphatase activity at 2.4 - 7.3 J/cm2 with 48 - 60 seconds of exposure, and an incident power ranging from 85-269mw significantly increased. The findings suggest that these irradiated cells obeyed the Arndt Shulz Law governing cellular response to applied energy. Further this research indicates the possible role of LLLT to accelerate tooth movement in orthodontics. Complete disclosure of low level laser parameters is essential in order to accurately compare findings of researchers.

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