Abstract
Recent advances in gene delivery into cells allow improved therapeutic effects in gene therapy trials. To increase the bioavailability of applied cells, it is of great interest that transfected cells remain at the application site and systemic spread is minimized. In this study, we tested clinically used biodegradable poly(lactic acid-co-glycolic acid) (PLGA) scaffolds (Vicryl & Ethisorb) as transient carriers for genetically modified cells. To this aim, we used human fibroblasts and examined attachment and proliferation of untransfected cells on the scaffolds in vitro, as well as the mechanical properties of the scaffolds at four time points (1, 3, 6 and 9 days) of cultivation. Furthermore, the adherence of cells transfected with green fluorescent protein (GFP) and vascular endothelial growth factor (VEGF165) and also VEGF165 protein secretion were investigated. Our results show that human fibroblasts adhere on both types of PLGA scaffolds. However, proliferation and transgene expression capacity were higher on Ethisorb scaffolds most probably due to a different architecture of the scaffold. Additionally, cultivation of the cells on the scaffolds did not alter their biomechanical properties. The results of this investigation could be potentially exploited in therapeutic regiments with areal delivery of transiently transfected cells and may open the way for a variety of applications of cell-based gene therapy, tissue engineering and regenerative medicine.
Highlights
New generation non-viral gene delivery systems, such as nanoparticles, novel cationic lipids and polymers, chemically coupling the nucleic acid to peptides and polymers [1,2,3,4,5,6,7,8,9] have shown to overcome some of the limitations of viral gene transfer and offer advantages in use of recombinant proteins [10, 11]
The culture medium was supplemented with 10% fetal calf serum (FCS: heat inactivated FCS-Gold, PAA, Coelbe, Germany) and antibiotic/antimycotic solution (AB/AM; PAA, Coelbe, Germany)
Hs27 cell proliferation could be observed on images of cell-seeded Vicryl and Ethisorb scaffolds captured with the light microscope in the course of 9 days (Fig 1)
Summary
New generation non-viral gene delivery systems, such as nanoparticles, novel cationic lipids and polymers, chemically coupling the nucleic acid to peptides and polymers [1,2,3,4,5,6,7,8,9] have shown to overcome some of the limitations of viral gene transfer and offer advantages in use of recombinant proteins [10, 11]. Biodegradable scaffolds are routinely used as reinforcement material for various surgical procedures, including hernia repair [13], tendon reconstruction [14], cranio-maxillo-facial surgery or neurosurgery [15,16,17,18]. Synthetic polymers such as polylactic acid (PLA), polyglycolic acid (PGA) and their copolymer poly(lactic acid-co-glycolic acid) (PLGA) are highly useful for the temporary management of pathologically altered tissue architectures including ligaments, skin, vascular tissues and skeletal muscle [19]. By utilizing easy manufactured off-the-shelf synthetic absorbable polymers, complications associated with antigenicity and disease transmission can be eliminated
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
