Delivery of Bioactive Gene Particles via Gelatin-Collagen-PEG-Based Electrospun Matrices.

Eleni K Tsekoura,Teo Dick,Hasan Uludağ,Yaman Boluk,Porntipa Pankongadisak,Daniel Graf

doi:10.3390/ph14070666

Abstract

The fabrication of fiber mats via electrospinning has been adopted in the last decades to produce high quality scaffolds for tissue engineering. However, an effective combination of electrospinning methods with gene delivery therapies remains a challenge. In this study, we describe how the delivery of gene complexes via electrospun mats that contain different volumes of gelatin (Gel), collagen (Col), and polyethylene glycol (PEG) can affect gene expression by transfected cells. Non-viral complexes were formulated by using lipid modified polyethylenimine (PEI) polymer and plasmid DNAs (pDNA) like the reporter Green Fluorescent Protein (GFP) and the therapeutically relevant Bone Morphogenetic Protein-2 (BMP-2) and electrospuned after being mixed with different volumes of Gel-Col-PEG mats and delivered to human myoblast (C2C12) and mouse osteoblast cells (MC3T3). The entrapment of GFP complexes via different homogeneous electrospun fiber mats revealed that a high fraction of collagen in the mats affected the quality of the fibers and led to reduced transfection efficiency on target cells. On the other hand, the fabrication of double-layered mats that contained collagen without complexes as a first layer and gelatin-collagen-PEG with complexes as a second layer successfully induced GFP expression and ALP activity in C2C12 cells. We conclude that this study has established the advantage of formulating multilayered bioactive collagen-based mats for gene delivery applications.

Highlights

IntroductionGene therapy has gained increasing attention in the fields of bone regeneration and tissue engineering [1]
Over the last decade, gene therapy has gained increasing attention in the fields of bone regeneration and tissue engineering [1]
Similar quality of fibers was observed from the Gelatin-Collagen-polyethylene glycol (PEG) (75-25-100) mats (Figure 1C,D); the presence of collagen in the electrospun mixture increased the fiber diameter to 140 ± 10 nm and bead’s size complexes (Figure 1H), good quality fibers were obtained, but the fiber size was increased to 177 ± 31 nm and the size of the observed beads was 770 ± 99 nm

Summary

Introduction

Gene therapy has gained increasing attention in the fields of bone regeneration and tissue engineering [1]. The delivery of gene-based therapeutic agents over protein delivery has been a superior method for the expression or deactivation of a desired protein in target cells or tissues [2,3]. A variety of viral and non-viral vectors has been developed and used for the safe delivery of plasmid DNA (pDNA) based expression systems to the cells. Gene activated matrices (GAMs) have been developed for administration of pDNA in an anatomical area of interest while simultaneously offering a structural support for new tissue and matrix deposition [4,5]. The selection of the most appropriate natural or synthetic scaffold along with the method of processing should be chosen in consideration of the type of interactions between the complexes and the scaffold. The stability of the gene complexes during scaffold fabrication as well as Pharmaceuticals 2021, 14, 666.

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