Comparative Efficiency of Gene-Activated Matrices Based on Chitosan Hydrogel and PRP Impregnated with BMP2 Polyplexes for Bone Regeneration

Abstract

Gene therapy is one of the most promising approaches in regenerative medicine. Gene-activated matrices provide stable gene expression and the production of osteogenic proteins in situ to stimulate osteogenesis and bone repair. In this study, we developed new gene-activated matrices based on polylactide granules (PLA) impregnated with BMP2 polyplexes and included in chitosan hydrogel or PRP-based fibrin hydrogel. The matrices showed high biocompatibility both in vitro with mesenchymal stem cells and in vivo when implanted intramuscularly in rats. The use of porous PLA granules allowed the inclusion of a high concentration of polyplexes, and the introduction of the granules into hydrogel provided the gradual release of the plasmid constructs. All gene-activated matrices showed transfecting ability and ensured long-term gene expression and the production of target proteins in vitro. At the same time, the achieved concentration of BMP-2 was sufficient to induce osteogenic differentiation of MSCs. When implanted into critical-size calvarial defects in rats, all matrices with BMP2 polyplexes led to new bone formation. The most significant effect on osteoinduction was observed for the PLA/PRP matrices. Thus, the developed gene-activated matrices were shown to be safe and effective osteoplastic materials. PLA granules and PRP-based fibrin hydrogel containing BMP2 polyplexes were shown to be the most promising for future applications in bone regeneration.

Osteogenesis Enhancement with 3D Printed Gene-Activated Sodium Alginate Scaffolds

“…It should also be highlighted that 3D printing technology ensured the simplification of the GAS fabrication process, eliminating the loss of DNA during its incorporation into the scaffold, and provided a high integration of porous scaffold with the tissue in the implantation zone. That resulted in the more distinct osteoinduction properties of the 3D printed SA-based scaffolds compared to the traditional hydrogel scaffolds [ 55 ] and even to the artificially modified (physically and chemically) matrices [ 56 , 57 ]. Thus, the developed platform of the gene-activated scaffold formation for bone regeneration is a promising methodology that is expected to be widely applied in bone defect restoration and bone disease treatment.…”

Adenovirus-Based Gene Therapy for Bone Regeneration: A Comparative Analysis of In Vivo and Ex Vivo BMP2 Gene Delivery

“…Platelet-rich plasma (PRP) was obtained according to the previously described method [ 11 ]. To obtain PLA/PRP matrices, PLA were mixed with PRP in a volume ratio of 1:1.…”

“…The treatment of bone defects by gene therapy requires the use of carrier matrices to ensure the targeted delivery of genetic constructs or transduced cells to the damaged area in order to achieve a high therapeutic concentration of osteoinductors and minimize the risk of heterotopic osteogenesis outside the regeneration site [ 10 ]. Highly porous polylactide particles (PLA), proven to be a biocompatible material with a high capacity for filling with gene constructs and proteins [ 11 , 12 , 13 , 14 ], can serve as a promising material for depositing genetic vectors. The efficiency of bone regeneration largely depends on the rate of vascularization [ 15 , 16 ].…”

Women’s contribution to stem cell research for osteoarthritis: an opinion paper

“…Different systems (micelles, hydrogels, solid scaffolds) have been used to generate GAMs capable of delivering a variety of gene transfer vectors. Nonviral vectors were incorporated in GAMs by the groups of A. L. Bertone, of W. Richter, and of M. Murphy* and also by Curtin et al , Raftery et al , and Tierney et al to carry PTH(1–34) (collagen) ( Backstrom et al, 2004 ), VEGF (collagen) ( Geiger et al, 2005 ), BMP-2 (collagen, nano-hydroxyapatite - nHA, alginate, chondroitin sulfate) ( Curtin et al, 2012 ; Loozen et al, 2013 ; Nedorubova et al, 2022 ; Husteden et al, 2023 ), BMP-2/VEGF (collagen, nHA, chitosan) ( Curtin et al, 2015 ; Raftery et al, 2017 ; Raftery et al, 2019 ; Walsh et al, 2021 ), BMP-2/BMP-7 (collagen, nHA, chitosan) ( Raftery et al, 2018 ), SOX9 (collagen, alginate) ( Ledo et al, 2020 ), ephrinB2 (collagen, nHA) ( Tierney et al, 2013 ), and the stromal-derived factor 1 alpha (SDF-1α) (collagen, nHA) ( Power et al, 2022 ) to stimulate MSC chondro-/osteogenesis and bone healing. GAMs formulating rAAV vectors carrying TGF-β (pluronics, carbon dots - CDs, poly (ε-caprolactone) - PCL) ( Rey-Rico et al, 2017b ; Meng et al, 2020 ; Venkatesan et al, 2021 ), IGF-I (alginate) ( Maihöfer et al, 2021 ), and SOX9 (pluronics, PCL, CDs) ( Rey-Rico et al, 2018 ; Madry et al, 2020b ; Urich et al, 2020 ; Venkatesan et al, 2020 ) were also used by the group of M. Cucchiarini* to stimulate MSC chondrogenesis, cartilage repair, and prevent OA.…”