Development of adenovirus immobilization strategies for in situ gene therapy

Wei Wen Hu, Michael W. Lang, Paul H. Krebsbach

研究成果: 雜誌貢獻期刊論文同行評審

22 引文 斯高帕斯(Scopus)

摘要

Background: Regenerative gene therapy using viral vectors enables transduced cells to express bioactive factors in vivo. Viral delivery with spatial control can enhance transduction efficiency and may limit systemic infection. Consequently, we tethered biotinylated adenovirus via interactions with avidin on chitosan surfaces to gain robust control for in situ transduction. Methods: Avidin was either directly conjugated to chitosan (virus-biotin- avidin-material; VBAM) or indirectly docked on biotinylated chitosan surfaces (virus-biotin-avidin-biotin-material; VBABM) to tether biotinylated adenovirus. Enzyme-linked immunosorbent assay (ELISA) and spectroscopic analysis were performed to demonstrate the binding profiles. Biotin-alkaline phosphatase and biotinylated adenovirus were used as different sized particles to evaluate binding efficiencies and were compared by the Sips isotherm adsorption method. Scanning electron microscopy (SEM) examination illustrated virus distribution, and the transduction efficiency was determined by in vitro cell transduction. Results: ELISA and spectroscopic analysis both demonstrated that the VBAM system led to multilayer avidin formation on biomaterial surfaces, whereas VBABM formed a monolayer of avidin. Sips isotherm adsorption indicated that the VBAM method increased heterogeneity and steric hindrance of binding sites. By contrast, the VBABM method docked avidin on chitosan surfaces and orientated the binding sites to facilitate ligand binding. In addition, SEM images illustrated that the VBABM method led to more even viral distribution. In vitro cell infection experiments also demonstrated that the VBABM system enhanced virus immobilization and thus improved cell transduction efficiency over the VBAM system. Conclusions The VBABM strategy is a superior method for in situ transduction from biomaterials. This strategy could be adapted for use with a variety of biomaterials as well as viral vectors, and thus may be an alternative method for in vivo regenerative gene therapy.

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頁(從 - 到)1102-1112
頁數11
期刊Journal of Gene Medicine
10
發行號10
DOIs
出版狀態已出版 - 2008

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