The Mechanical and Biological Properties of Chitosan Scaffolds for Tissue Regeneration Templates Are Significantly Enhanced by Chitosan from Gongronella butleri

Abstract

Chitosan with a molecular weight (MW) of 10⁴ Da and 13% degree of acetylation (DA) was extracted from the mycelia of the fungus Gongronella butleri USDB 0201 grown in solid substrate fermentation and used to prepare scaffolds by the freeze-drying method. The mechanical and biological properties of the fungal chitosan scaffolds were evaluated and compared with those of scaffolds prepared using chitosans obtained from shrimp and crab shells and squid bone plates (MW 10⁵-10⁶ Da and DA 10-20%). Under scanning electron microscopy, it was observed that all scaffolds had average pore sizes of approximately 60-90 mm in diameter. Elongated pores were observed in shrimp chitosan scaffolds and polygonal pores were found in crab, squid and fungal chitosan scaffolds. The physico-chemical properties of the chitosans had an effect on the formation of pores in the scaffolds, that consequently influenced the mechanical and biological properties of the scaffolds. Fungal chitosan scaffolds showed excellent mechanical, water absorption and lysozyme degradation properties, whereas shrimp chitosan scaffolds (MW 10⁶Da and DA 12%) exhibited the lowest water absorption properties and lysozyme degradation rate. In the evaluation of biocompatibility of chitosan scaffolds, the ability of fibroblast NIH/3T3 cells to attach on all chitosan scaffolds was similar, but the proliferation of cells with polygonal morphology was faster on crab, squid and fungal chitosan scaffolds than on shrimp chitosan scaffolds. Therefore fungal chitosan scaffold, which has excellent mechanical and biological properties, is the most suitable scaffold to use as a template for tissue regeneration.