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Microencapsulation of recombinant adenovirus within poly-DL-lactide-poly(ethylene glycol) microspheres for enhanced gene transfection efficiency and inhibitory effects on hepatocellular carcinoma cells in vitro

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When gene therapy is performed for the treatment of malignant tumors, gene transfer efficiency and selectivity are highly important. Polymer vehicle microspheres are a novel type of therapy, which have been developed rapidly in recent years and are able to control drug release, prolong the biological halflife of drugs, decrease side effects and achieve targeted delivery. The present study was designed to construct a polymer microsphere-encapsulated recombinant adenovirus with human tissue inhibitors of the matrix metalloproteinase1 (TIMP1) gene, and to discuss its characterization for the purpose of liver cancer gene therapy. The microsphere was prepared from biodegradable polyDLlactidepoly(ethylene glycol) (PELA) encapsulating rAdTIMP1, the recombinant adenovirus carrying TIMP1, by a modified doubleemulsion method. The particle morphology, diameter, virus encapsulation, loading rate and release kinetics of the rAdmicrospheres were determined in vitro. Hepatocellular carcinoma (HCC) HepG2 cells were transfected with the rAdmicrosphere and the efficiency of transfection was assessed by fluorescent microscopy. The production and expression of TIMP1 was identified by gelatin zymography and western blot analysis, and the invasiveness was detected by a matrigel matrix invasion assay. The microsphere encapsulating rAdTIMP1 was successfully constructed with a diameter of 1.965 µm, encapsulation efficiency of 60.0%, a viral load of 10.5x108/mg, a virus release of ~60% within 120 h and a total release time of >240 h. The resultant rAdmicrospheres were able to efficiently transfect HepG2 cells with the transfection efficiency enhanced by ~90%. As a result, the transfected HepG2 cells had significantly increased TIMP1 enzyme activity and the expression of TIMP1 was detected by western blot analysis. In addition, the proliferation and invasion ability of the HCC cells was markedly inhibited by the rAdmicrospheres. The resultant rAdmicrospheres, PELA-encapsulated recombinant TIMP1 adenovirus, had enhanced transfection efficiency and were able to markedly inhibit the in vitro biological behavior of HepG2 cells. This provides an experimental basis for this polymer application and may pave the way for prospective in vivo clinical trials and further comprehensive therapy for liver cancer.
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Document Type: Research Article

Affiliations: 1: Department of Gastrointestinal Surgery, Affiliated Hospital of Luzhou Medical College, Luzhou, Sichuan 646000, P.R. China 2: Department of Gastroenterology, Affiliated Hospital of Luzhou Medical College, Luzhou, Sichuan 646000, P.R. China

Publication date: August 1, 2015

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  • Molecular Medicine Reports is a monthly, peer-reviewed journal available in print and online, that includes studies devoted to molecular medicine, underscoring aspects including pharmacology, pathology, genetics, neurosciences, infectious diseases, molecular cardiology and molecular surgery. In vitro and in vivo studies of experimental model systems pertaining to the mechanisms of a variety of diseases offer researchers the necessary tools and knowledge with which to aid the diagnosis and treatment of human diseases.
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