Ultrasound-Mediated Microbubble Destruction Enhances GFP Gene Expression In Human Gingival Fibroblast Cells In Vitro

Objective:To investigate whether ultrasound-mediated microbubble destruction can effectively deliver pEGFP-N1 plasmid to human gingival fibroblasts.Methods: The primary cultured human gingival fibroblasts were divided into five groups. pEGFP-N1 plasmid was transfected to HGFs as a gene marker with ultrasound-mediated microbubble destruction. Experimental groups included naked plasmid, microbubble+plasmid, ultrasound+plasmid, and ultrasound+ microbubble+plasmid.Based on different conditions of transfection, the last experimental group was further divided into 24 groups.After 48 hours, phase-contrast fluorescent microscopy was employed to evaluate the expression of green fluorescent protein(GFP).The cell vitality was measured by the MTT assay.Results: The transfection efficiency of the fourth group was higher than the other experimental groups,where the optimal gene expression occurred with ultrasound parameter at 1.5 w/cm2 for 60s, microbubble at the concentration of 10% and plasmid at the concentration of 6.67 ug/ml. Conclusions: Under specific conditions, ultrasound mediated microbubble destruction can enhance the reporter gene transfection and expression.