Biomimetic Scaffold of Chitosan from Litopenaeus vannamei Shrimp Shells Incorporated with Collagen and Hydroxyapatite for Bone Tissue Regeneration.

Abstract

Introduction: The goal of this study was to create a biocompatible bone tissue engineering scaffold from Litopenaeus vannamei shrimp shell chitosan with hydroxyapatite (HAP) and collagen type I (COL I), and to investigate its physicochemical and biological properties relative to commercial chitosan scaffolds.

Methods: The chitosan was extracted by following steps: deproteination, demineralization, and deacetylation processes, further characterizing it through composite FTIR spectroscopy. The scaffolds were fabricated using lyophilization, followed by the evaluation of their morphology, porosity, swelling ratio, degradation rate, and compressive strength. The scaffold morphology and porosity were observed using Scanning Electron Microscopy (SEM) and the solvent displacement method. Swelling and degradation ratio were investigated in phosphate buffer saline, while compressive strength was investigated with the Universal Testing Method. For the assessment of cytocompatibility, MG-63 osteoblast-like cells were subjected to the MTT assay, and cell morphology on the scaffold was observed under SEM.

Results: As shown in the FTIR results, the derived chitosan exhibited comparable functional groups to those of commercial chitosan, confirming its successful extraction. Both scaffolds exhibited interconnected pores and suitable diameters and porosity for bone scaffold tissue engineering. Chitosan from shrimp shells showed a reduction in swelling (278.55±36.49% at 24h) and a slower degradation rate (41.87±7.27% at 4 weeks) compared to commercial scaffolds, possibly due to higher residual minerals and a lower degree of deacetylation. However, compressive strength 0.991±0.01 MPa and attachment and proliferation of MG-63 cells were similar, suggesting good osteoconductivity of the biomaterials.

Conclusion: Chitosan-derived shrimp shells are a sustainable biomaterial candidate for bone tissue engineering. The scaffold based on shrimp shells exhibited relatively lower degradation and moderate swelling, adequate mechanical stability, and bioactivity to support osteoblast-like cell adhesion and viability, suggesting it is adequate for bone tissue engineering applications.