The Study Of Ultrasound And Microbubble Destruction Delivering PEGFP-N1 To Human Periodontal Ligament Fibroblasts

Periodontitis is an infectious disease of the periodontal tissues, which is one of the main causes of periodontal pockets, progressive attachment loss and frontal resorption, finally leading to loosening of tooth and loss of the tooth. Since the regeneration capacity of periodontal tissues is finite,at present, the main treatment of periodontitis is'control the development of inflammation', which cannot completely recover the periodontium's physiological structure. Periodontal ligament cells play an important role in periodontal tissue regeneration. Periodontal ligament cells consist of periodontal ligament fibroblasts, osteoblasts, cementoblasts, et,al. When periodontal tissues are traumatized and inflamed, the quantity and biological function of the regenerative cells degrade, which hampers the complete regeneration.'Gene therapy'provides us with a new way to implement'periodontal tissue regeneration'. Using gene transfer technique, exogenous gene was transferred to target cells. Therefore, there was high expression of remedial protein in definite areas, which induced direct differentiation of cells;which in turn promoted periodontal tissue regeneration.Objective:To investigate whether ultrasound-mediated microbubble destruction can safely and effectively deliver pEGFP-N1 plasmid to HPDLFs.Methods:HPDLFs were gotten by the way of tissue pieces cultivation ,EGFP gene as the report gene, ultrasound-mediated microbubble destruction deliver pEGFP-N1 plasmid to HPDLFs.According to factors which influence ultrasound-mediated microbubble delivering,HPDLFs were divided into 24 groups, bolting groups of higher transfection efficiency. Group which was highest transfection efficiency as experiment group, respectively compared to plasmid group,ultrasound + plasmid group, microbubble + plasmid group and liposome + plasmid group in transfection efficiency. MTT was adopted to measure the cell vitality of every group.Results:The group in which ultrasound intensity was 0.5 W/cm2,bombardment time 60 s and microvesicle concentration 15% got higher transfection efficiency and less cell damage. Meanwhile,ultrasound-mediated microbubble and liposome's transfection efficiency were similar,but the cell vitality of ultrasound + microbubble + plasmid group was much higher than that of liposome+plasmid group.Conclusions: Based on ultrasound-mediated microbubble destruction delivering pEGFP-N1 plasmid to HPDLFs,comparing to liposome transfection,study the influence of it to HPDLFs vitality and screening suitable transfection condition.Confirm under specific conditions ultrasound mediated microbubble destruction can enhance the reporter gene transfection and expression in HPDLFs. It is a potential way of gene therapy with safe and effective quality in periodontal tissue regeneration.