In vitro and in vivo response of composites based on chitosan, hydroxyapatite and collagen

Maurício Bordini Do Amaral,Rommel Bezerra Viana,Katúcia Bezerra Viana,Ana Maria De Guzzi Plepis,Aline Bassi Denis,Cristina Aparecida Diagone

doi:10.4025/actascitechnol.v42i1.41102

Abstract

The goal of this study was to evaluate the in vitro cellular toxicity and biological behavior of new bone graft composites after subcutaneous implantation during remodeling and wound-healing processes. We developed composites based on hydroxyapatite (obtained by deproteinizing bovine bone), collagen (obtained from bovine tendon) and chitosan (obtained from gladii of the squid species Loligo), that were characterized by different techniques (X-ray, FT-IR, Thermogravimetry, DSC and SEM). Three biomaterials were evaluated here: B1 (collagen/chitosan/hydroxyapatite), B2 (collagen/hydroxyapatite) B3 (collagen/hydroxyapatite). For in vitro cytotoxicity tests, two cell lines were used: HEp human larynx tumor cells (ATCC-CCL-23) and VERO cells from African green monkey (Cercopithecus aethiops). These toxicity tests demonstrated that the evaluated composites are not toxic. In biocompatibility tests, the results of a histological analysis showed that all three biomaterials present a low inflammatory tissue reaction. The tissue response was most favorable for sample B3, followed by B2 and B1, in that order. Based on these results, we conclude that all three biomaterials show good biocompatibility and no evidence of cytotoxicity; thus, these materials represent good candidates for tissue and graft engineering for use in bone regeneration.

Highlights

IntroductionBiomaterials for bone tissue are used, for example, in the filling of extensive bone defects (Bongio, van den Beucken, Leeuwenburgh, & Jansen, 2015) and in the development of implants that will be subjected to mechanical forces with applications in orthopedics (Ong, Yun, & White, 2015), odontology (Li, Chow, & Matinlinna, 2014), and oral and maxillofacial surgery (Keranen et al, 2011)
The purpose of this study was to characterize and study the biological behavior of composites based on polyionic collagen, natural hydroxyapatite and chitosan that are intended for use in bone tissue reconstruction
In the 200-400°C range, 0.48% of the organic matter remaining in the sample was degraded, and in the 400-700°C range, 0.45% of the material was carbonized, yielding a residue of 98.6%; these results indicate the purity of the hydroxyapatite samples used

Summary

Introduction

Biomaterials for bone tissue are used, for example, in the filling of extensive bone defects (Bongio, van den Beucken, Leeuwenburgh, & Jansen, 2015) and in the development of implants that will be subjected to mechanical forces with applications in orthopedics (Ong, Yun, & White, 2015), odontology (Li, Chow, & Matinlinna, 2014), and oral and maxillofacial surgery (Keranen et al, 2011). In this context, it is important to emphasize that the advancement of the dentistry science was accompanied by the development of biomaterials, from their manufacturing to understanding the obtained cellular responses (Shin, 2007). The stabilizing effect in hydroxyapatite/collagen scaffolds by incorporating carbodiimide (Kozlowska, Sionkowska, Osyczka, & Dubiel, 2017), graphene oxide (Wang et al, 2017), carbon nanotube (Jing et al, 2017), calcium phosphate (Zhao et al, 2017), PLGA and alginate microparticles (Quinlan et al, 2015) were performed to improve the mechanical and biological properties

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