title: Effective genetic modification and differentiation of hMSCs upon controlled release of rAAV vectors using alginate/poloxamer composite systems.
abstract: Viral vectors are common tools in gene therapy to deliver foreign therapeutic sequences in a specific target population via their natural cellular entry mechanisms. Incorporating such vectors in implantable systems may provide strong alternatives to conventional gene transfer procedures. The goal of the present study was to generate different hydrogel structures based on alginate (AlgPH155) and poloxamer PF127 as new systems to encapsulate and release recombinant adeno-associated viral (rAAV) vectors. Inclusion of rAAV in such polymeric capsules revealed an influence of the hydrogel composition and crosslinking temperature upon the vector release profiles, with alginate (AlgPH155) structures showing the fastest release profiles early on while over time vector release was more effective from AlgPH155+PF127 [H] capsules crosslinked at a high temperature (50°C). Systems prepared at room temperature (AlgPH155+PF127 [C]) allowed instead to achieve a more controlled release profile. When tested for their ability to target human mesenchymal stem cells, the different systems led to high transduction efficiencies over time and to gene expression levels in the range of those achieved upon direct vector application, especially when using AlgPH155+PF127 [H]. No detrimental effects were reported on either cell viability or on the potential for chondrogenic differentiation. Inclusion of PF127 in the capsules was also capable of delaying undesirable hypertrophic cell differentiation. These findings are of promising value for the further development of viral vector controlled release strategies.
authors.affiliation: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany; Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Farmacia, Universidad de Santiago de Compostela, Spain.
authors.forename: P
authors.initials: P
authors.lastname: Díaz-Rodríguez
authors.affiliation: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Rey-Rico
authors.affiliation: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany; Department of Orthopaedic Surgery, Saarland University Medical Center, Homburg/Saar, Germany.
authors.forename: H
authors.initials: H
authors.lastname: Madry
authors.affiliation: Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Farmacia, Universidad de Santiago de Compostela, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Landin
authors.affiliation: Center of Experimental Orthopaedics, Saarland University Medical Center, Homburg/Saar, Germany. Electronic address: mmcucchiarini@hotmail.com.
authors.forename: M
authors.initials: M
authors.lastname: Cucchiarini
mesh_terms: D000464:Alginates / Q000008:administration & dosage / Q000737:chemistry*; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D003692:Delayed-Action Preparations; D000229:Dependovirus / Q000235:genetics*; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage / Q000737:chemistry*; D020723:Glucuronic Acid / Q000008:administration & dosage / Q000737:chemistry; D006603:Hexuronic Acids / Q000008:administration & dosage / Q000737:chemistry; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000187:drug effects* / Q000502:physiology; D020442:Poloxamer / Q000008:administration & dosage / Q000737:chemistry*
chemical_list: D000464:Alginates; D003692:Delayed-Action Preparations; D006603:Hexuronic Acids; D020442:Poloxamer; D020723:Glucuronic Acid
keywords: Alginate/poloxamer systems; Differentiation potential; Human MSCs; Hydrogel structure; Release profile; rAAV gene transfer