Healing and Angiogenic Properties of Collagen/Chitosan Scaffolds Enriched with Hyperstable FGF2-STAB(R) Protein: In Vitro, Ex Ovo and In Vivo Comprehensive Evaluation

Healing and Angiogenic Properties of Collagen/Chitosan Scaffolds Enriched with Hyperstable FGF2-STAB(R) Protein: In Vitro, Ex Ovo and In Vivo Comprehensive Evaluation

WoS Article

Wound healing is a process regulated by a complex interaction of multiple growth factors including fibroblast growth factor 2 (FGF2). Although FGF2 appears in several tissue engineered studies, its applications are limited due to its low stability both in vitro and in vivo. Here, this shortcoming is overcome by a unique nine-point mutant of the low molecular weight isoform FGF2 retaining full biological activity even after twenty days at 37 degrees C. Crosslinked freeze-dried 3D porous collagen/chitosan scaffolds enriched with this hyper stable recombinant human protein named FGF2-STAB(R) were tested for in vitro biocompatibility and cytotoxicity using murine 3T3-A31 fibroblasts, for angiogenic potential using an ex ovo chick chorioallantoic membrane assay and for wound healing in vivo with 3-month old white New Zealand rabbits. Metabolic activity assays indicated the positive effect of FGF2-STAB(R) already at very low concentrations (0.01 mu g/mL). The angiogenic properties examined ex ovo showed enhanced vascularization of the tested scaffolds. Histological evaluation and gene expression analysis by RT-qPCR proved newly formed granulation tissue at the place of a previous skin defect without significant inflammation infiltration in vivo. This work highlights the safety and biocompatibility of newly developed crosslinked collagen/chitosan scaffolds involving FGF2-STAB(R) protein. Moreover, these sponges could be used as scaffolds for growing cells for dermis replacement, where neovascularization is a crucial parameter for successful skin regeneration.

Wound healing is a process regulated by a complex interaction of multiple growth factors including fibroblast growth factor 2 (FGF2). Although FGF2 appears in several tissue engineered studies, its applications are limited due to its low stability both in vitro and in vivo. Here, this shortcoming is overcome by a unique nine-point mutant of the low molecular weight isoform FGF2 retaining full biological activity even after twenty days at 37 degrees C. Crosslinked freeze-dried 3D porous collagen/chitosan scaffolds enriched with this hyper stable recombinant human protein named FGF2-STAB(R) were tested for in vitro biocompatibility and cytotoxicity using murine 3T3-A31 fibroblasts, for angiogenic potential using an ex ovo chick chorioallantoic membrane assay and for wound healing in vivo with 3-month old white New Zealand rabbits. Metabolic activity assays indicated the positive effect of FGF2-STAB(R) already at very low concentrations (0.01 mu g/mL). The angiogenic properties examined ex ovo showed enhanced vascularization of the tested scaffolds. Histological evaluation and gene expression analysis by RT-qPCR proved newly formed granulation tissue at the place of a previous skin defect without significant inflammation infiltration in vivo. This work highlights the safety and biocompatibility of newly developed crosslinked collagen/chitosan scaffolds involving FGF2-STAB(R) protein. Moreover, these sponges could be used as scaffolds for growing cells for dermis replacement, where neovascularization is a crucial parameter for successful skin regeneration.

collagen; chitosan; scaffold; FGF2; skin regeneration; tissue engineering

collagen; chitosan; scaffold; FGF2; skin regeneration; tissue engineering

VOJTOVÁ, L.; PAVLIŇÁKOVÁ, V.; MUCHOVÁ, J.; KACVINSKÁ, K.; BRTNÍKOVÁ, J.; KNOZ, M.; LIPOVÝ, B.; FALDYNA, M.; GÖPFERT, E.; HOLOUBEK, J.; PAVLOVSKÝ, Z.; VÍCENOVÁ, M.; BLAHNOVÁ, V.; HEARNDEN, V.; FILOVA, E.

2022

01.06.2021

MDPI

BASEL

2227-9059

Biomedicines

9

6

Swiss Confederation

1

27

27

https://www.mdpi.com/2227-9059/9/6/590

http://hdl.handle.net/11012/200493

Vojtova_Biomedicines_10-3390-biomedicines9060590