title: Release of bioactive adeno-associated virus from fibrin scaffolds: effects of fibrin glue concentrations.
abstract: Fibrin glue (FG) is used in a variety of clinical applications and in the laboratory for localized and sustained release of factors potentially important for tissue engineering. However, the effect of different fibrinogen concentrations on FG scaffold delivery of bioactive adeno-associated viruses (AAVs) has not been established. This study was performed to test the hypothesis that FG concentration alters AAV release profiles, which affect AAV bioavailability. Gene transfer efficiency of AAV-GFP released from FG was measured using HEK-293 cells. Bioactivity of AAV transforming growth factor-beta1 (TGF-β(1)) released from FG was assessed using the mink lung cell assay, and by measuring induction of cartilage-specific gene expression in human mesenchymal stem cells (hMSCs). Nondiluted FG had longer clotting times, smaller pore sizes, thicker fibers, and slower dissolution rate, resulting in reduced release of AAV. AAV release and gene transfer efficiency was higher with 25% and 50% FG than with the 75% and 100% FG. AAV-TGF-β(1) released from dilute-FG transduced hMSCs, resulting in higher concentrations of bioactive TGF-β(1) and greater upregulation of cartilage-specific gene expression compared with hMSC from undiluted FG. This study, showing improved release, transduction efficiency, and chondrogenic effect on hMSC of bioactive AAV-TGF-β(1) released from diluted FG, provides information important to optimization of this clinically available scaffold for therapeutic gene delivery, both in cartilage regeneration and for other tissue engineering applications.
authors: Cartilage Restoration Center, Department of Orthopaedic Surgery, University of Pittsburgh, Pittsburgh, Pennsylvania 15213, USA.
authors: Hannah H
authors: HH
authors: Lee
authors: Amgad M
authors: AM
authors: Haleem
authors: Veronica
authors: V
authors: Yao
authors: Juan
authors: J
authors: Li
authors: Xiao
authors: X
authors: Xiao
authors: Constance R
authors: CR
authors: Chu
mesh_terms: D001672:Biocompatible Materials / Q000494:pharmacology*; D001777:Blood Coagulation; D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D000229:Dependovirus / Q000187:drug effects / Q000378:metabolism*; D015718:Fibrin Tissue Adhesive / Q000032:analysis* / Q000737:chemistry*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism; D008855:Microscopy, Electron, Scanning; D013997:Time Factors; D054457:Tissue Scaffolds / Q000737:chemistry*; D014161:Transduction, Genetic; D053773:Transforming Growth Factor beta1 / Q000378:metabolism
chemical_list: D001672:Biocompatible Materials; D015718:Fibrin Tissue Adhesive; D053773:Transforming Growth Factor beta1; D049452:Green Fluorescent Proteins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate