Preparation of VP3 gene-loaded ultrasound microbubbles decorated with TATp and SDF-1 and targeting study in vitro / 重庆医学

ABSTRACT

Objective To prepare a innovative VP3 gene‐loaded ultrasound microbubble decorated with TATp and SDF‐1 , having the extracellular accumulation and intracellular permeation function ,and characterize their property .Methods VP3 gene‐loaded ultrasound microbubbles were prepared with the method of W/O/W double emulsion .SDF‐1 and TATp were covalently con‐gjugated to the surface of poly‐lactic/acid‐glycolic acid(PLGA) microbubble though thioether bonds to prepare gene‐loaded targeted ultrasound mirobubbles .Their particle size ,distribution and surface potential were determined by Malvern measuring instrument . The conjugation status of TATp and SDF‐1 were evaluated by flow cytometry and confocal microscopy .Their DNA protection were identified by digestion reaction test .The vitro targeting capacity was preliminarily assessed by light microscopy and flow cytometry , and the vitro ultrasound imaging was investigated under high frequency imaging condition .Results The gene‐loaded targeted ultra‐sound mirobubbles showed regularly sphericity .The diameter was (536 .00 ± 16 .55)nm ,and showed a narrow distribution .The zeta potential was(-0 .08 ± 0 .08)mV .The average gene loading was 0 .62% ,with the average rate of 36 .13% gene encapsulation effi‐ciency .The DNA protective effect sustained 60 min without damage .Connection rate of TATp and SDF‐1 coupled with PLGA mi‐crobubbles surface was 69 .84% .The vitro targeting study showed that more targeted microbubbles stably clustered together in the tongue SCC‐15 cell membranes with the connection rate of 91 .44% ,while non‐targeted microbubbles combination rate was 12 .96% .Moreover ,the vitro ultrasound imaging was tiny dot ,even high echo .Conclusion TATp‐SDF‐1‐VP3‐PEG‐PLGA micro‐bubbles were prepared successfully ,which can efficiently target to tongue SCC‐15 cells ,and pass through the cell membranes at a short time in company with outstanding ultrasound imagings in vitro .