Osteoblast responses to injectable bone substitutes of kappa-carrageenan and nano hydroxyapatite

Abstract

The combination of kappa-carrageenan (κ-CG) and hydroxyapatite (HA) to generate a bone substitute material has been underexplored to date. Carrageenans (CGs) have remarkable characteristics such as biocompatibility, hydrophilicity, and structural similarities with natural glycosaminoglycans (GAGs), and they have demonstrated the ability to stimulate cellular adhesion and proliferation. Hydroxyapatite nanoparticles have been one of the most investigated materials for bone regeneration due to their excellent biocompatibility, bioactivity and osteoconductivity. In particular, this study presents an approach for the preparation of new bioactive composites of κ-CG/nHA for numerous bone regeneration applications. We performed a set of in vitro experiments to evaluate the influence of the bone substitutes on human osteoblasts. Cell culture studies indicated that all samples tested were cytocompatible. Relative to control substrates, cellular attachment and proliferation were better on all the scaffold surfaces that were tested. The S2 and S3 samples, those permeated by 1.5 and 2.5 wt% of CG, respectively, exhibited an enhancement in cell spreading capacity compared to the S1 test materials which were comprised of 1 wt% of CG. Excellent osteoblast viability and adhesion were observed for each of the tested materials. Additionally, the bone substitutes developed for this study presented a distinct osteoconductive environment. Data supporting this claim were derived from alkaline phosphatase (ALP) and calcium deposition analyses, which indicated that, compared to the control species, ALP expression and calcium deposition were both improved on test κ-CG/nHA surfaces. In summary, the injectable bone substitute developed here demonstrated great potential for numerous bone regeneration applications, and thus, should be studied further. STATEMENT OF SIGNIFICANCE: The novelty of this work lies in the determination of the in vitro cytocompatibility behavior of carrageenan and hydroxyapatite composite materials used as injectable bone substitutes. This injectable biomaterial can fill in geometric complex defects, and it displays bioactivity as well as high bone regeneration capacity. In this study, we evaluated the behaviors of osteoblast cells in contact with the scaffolds, including cellular adhesion and proliferation, cellular metabolism, and mineralization on the fabricated injectable bone substitutes. The results show than the carrageenan and hydroxyapatite substitutes provided a biomaterial with a great capacity for promoting cellular growth, adhesion, and proliferation, as well as contributing an osteoinductive environment for osteoblast differentiation and osteogenesis.

Keywords: Biocompatibility; Carrageenan; Injectable bone substitute; Nano hydroxyapatite; Osteoblasts.