Anabolic effects of low-intensity pulsed ultrasound on human gingival fibroblasts

ObjectiveLow-intensity pulsed ultrasound (LIPUS) demonstrated anabolic effects on cementoblasts, odontoblasts, and periodontal ligament cells. However, LIPUS effect on human gingival fibroblasts (HGF) remains to be investigated. Therefore, we evaluated the in vitro effects of LIPUS on HGF proliferation and differentiation to test its feasibility for periodontal therapy.DesignLIPUS treatment (1.5 MHz, 30 mW/cm2) was applied to HGF in the experimental groups after 24-h of culture (5 or 10 min/day for 28 days) and omitted in the control. Changes in HGF activities were evaluated in response to LIPUS treatment in dose-dependent (5 and 10 min) and time-dependent (weeks 1–4) manner. The effects of LIPUS on HGF cell viability (MTT), proliferation (total DNA content and growth pattern), alkaline phosphatase (ALP) activity, and gene expression by reverse-transcriptase polymerase chain reaction (RT-PCR) were determined.ResultsCell viability remained unchanged after LIPUS treatment during the 4 weeks of treatment as compared to the untreated control group which ensured a safe biological response. Both LIPUS treatments (5–10 min/day) did not yield any significant changes in the proliferation, and expression of proliferating cell nuclear antigen (PCNA) and collagen-I (COL-I). Conversely, LIPUS treatment enhanced osteogenic differentiation potential of HGF as determined by significant up-regulation of specific ALP activity and osteopontin (OPN) expression, with optimum effect following 3 weeks of 5 min/day LIPUS treatment.ConclusionLIPUS treatment at 30 mW/cm2 selectively enhanced HGF differentiation but not proliferation. The ability of LIPUS to enhance HGF differentiation is promising for its application in cell-based periodontal therapy.