RESUMO
OBJECTIVE: To study the feasibility of delivering viral gene vector from a collagen-coated polyurethane (PU) film through a mechanism involving monoclonal antiviral antibody tethering. METHODS: Anti-adenoviral monoclonal antibodies were covalently bound to the collagen-coated PU surface. These antibodies enabled tethering of replication defective adenoviruses through highly specific antigen-antibody affinity. The PU film-based gene delivery using antibody-tethered adenovirus encoding green fluorescent protein (GEP) was tested in rat arterial smooth muscle cell (A10 cell) culture in vitro. The virus binding stability was studied by incubating the collagen-coated PU film in PBS solution at 37 degrees C for 20 days, followed with A10 cell cultures with the incubated films and the corresponding buffer solution. RESULTS: PU films with antibody-tethered adenovirus encoding GFP demonstrated efficient and highly localized gene delivery to A10 cells. Virus binding was stable for at least 10 days at physiological conditions, more than 77% of the originally bound virus remained in the film after 15 day's incubation. CONCLUSION: Gene delivery using PU film-based anti-viral antibody tethering of vectors exhibited potentials of applications in a wide array of single or multiple therapeutic gene strategies, and in further stent-based gene delivery therapeutic strategies.
Assuntos
Anticorpos Monoclonais/imunologia , Terapia Genética , Vetores Genéticos , Poliuretanos/química , Adenoviridae/genética , Especificidade de Anticorpos/imunologia , Técnicas de Transferência de Genes , Humanos , Ligação ProteicaRESUMO
In this paper, we studied a new preparation method of microcapsules for entrapment of genetically engineered cells. Polyvinyl alcohol microcapsules having well defined shape, high mechanical strength, good biochemical and permeability properties were prepared by using low temperature physical cross-linking method. Comparing with currently used alginate-polylysine-alginate microcapsules, polyvinyl alcohol microcapsules have much higher mechanical strength. The low temperature physical crosslinking procedure of polyvinyl alcohol is nontoxic to the genetically engineered E. coli DH5alpha cell, which attained high activity in decomposing and metabolizing urea in vitro studies.
