pEGFP-N1 transfection of rat dental follicle cells under ultrasound-mediated lipid microbubble: transfected cells have a relatively stable biological property

doi:10.3969/j.issn.2095-4344.2014.50.020

Chinese Journal of Tissue Engineering Research ›› 2014, Vol. 18 ›› Issue (50): 8151-8155.doi: 10.3969/j.issn.2095-4344.2014.50.020

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pEGFP-N1 transfection of rat dental follicle cells under ultrasound-mediated lipid microbubble: transfected cells have a relatively stable biological property

Ran Ling¹, Li Xiao-qian², Jiang Xin-yi³, Deng Feng¹, Song Jin-lin¹, Cao Li¹

1Affiliated Hospital of Stomatology, Chongqing Medical University, Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, Chongqing 401147, China; 2the Stomatological Hospital of Chongqing Three Gorges Central Hospital, Chongqing 404199, China; 3Department of Stomatology, Mianyang Central Hospital, Mianyang 621000, Sichuan Province, China

Received:2014-11-18 Online:2014-12-03 Published:2014-12-03
Contact: Song Jin-lin, M.D., Professor, Chief physician, Master’s supervisor, Affiliated Hospital of Stomatology, Chongqing Medical University, Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, Chongqing 401147, China
About author:Ran Ling, Studying for master’s degree, Affiliated Hospital of Stomatology, Chongqing Medical University, Chongqing Key Laboratory for Oral Diseases and Biomedical Sciences, Chongqing 401147, China
Supported by:
the National Natural Science Foundation of China, No. 30870754; the Second Batch of Outstanding Technical Personnel of Chongqing Municipal Education Commission, No. 2010[72]

Abstract

Abstract:

BACKGROUND: With the interaction of ultrasound and microbubbles, cavitation and mechanical effects undermine the integrity of the cell membrane, resulting in temporary and reversible holes, increasing the permeability of cell membranes, enhancing gene transfer and improving gene transfection efficiency.
OBJECTIVE: To investigate the efficiency and safety of rat dental follicle cells transfected with pEGFP-N1 plasmid mediated by microbubble under ultrasonic irradiation.
METHODS: The primary dental follicle cells from newborn rats were cultured in vitro and passaged to the 4th generation. Under different conditions, pEGFP-N1 was used to transfect rat dental follicle cells. By combining the ultrasonic intensity (0.5, 1 W/cm2) with the irradiation time (15, 30, 45, 60 seconds), we got the best conditions of ultrasonic irradiation for the next experiment. There were five groups: plasmid, microbubble+plasmid, ultrasound+plasmid, ultrasound+microbubble+plasmid, and liposomes+plasmid groups. The expression of pEGFP was observed by inverted fluorescence microscope 48 hours after transfection, and meanwhile, the proliferation inhibition rate of rat dental follicle cells was determined by MTT method.
RESULTS AND CONCLUSION: Under the 0.5 W/cm2 ultrasound for 30 seconds, the transfection efficiency was obviously higher than that under the other combinations. Under the above-mentioned condition, the transfection efficiency of rat dental follicle cells with pEGFP-N1 plasmid was higher than that mediated by the traditional liposome, and the cell viability had no obvious changes. Under suitable conditions, ultrasound microbubble technology can safely and effecitively mediate the transfection of rat dental follicle cells with pEGFP-N1 plasmid, and transfected cells also have a stable biological property as normal dental follicle cells. Therefore, ultrasound microbubble technology can provide an ideal method of gene transfection in periodontal tissue engineering.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: dental sac, high-energy shock waves, genes, transfection

CLC Number:

R394.2

Ran Ling, Li Xiao-qian, Jiang Xin-yi, Deng Feng, Song Jin-lin, Cao Li. pEGFP-N1 transfection of rat dental follicle cells under ultrasound-mediated lipid microbubble: transfected cells have a relatively stable biological property[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(50): 8151-8155.

Figures/Tables 5

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pEGFP-N1 transfection of rat dental follicle cells under ultrasound-mediated lipid microbubble: transfected cells have a relatively stable biological property

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