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

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.

Evaluation of cytotoxicity and antimicrobial activity of an injectable bone substitute of carrageenan and nano hydroxyapatite.

A successful post-surgical implant is associated with accelerated recovery periods, involving the efficient regeneration of lost or non-viable tissue and a reduction in microbial growth. Alternatively, the long-term success of an implant is guided by the selection of an engineered biomimetic material that is biocompatible, non-biodegradable, and stable at the site of implantation, without invoking any non-essential or undesirable biological responses. The potential for developing an injectable bone substitute (IBS) was investigated here. In particular, carrageenan (CG) and nano-hydroxyapatite (nHA) injectable composites were fabricated by chemical cross-linking, and the in vitro behavior of mammalian cells and bacteria on the IBS surface structures were evaluated. Formulations consisting of 1%, 1.5%, and 2.5% CG and 60% nHA by weight were then evaluated for their interactions with human osteoblasts (or bone forming cells). MTS viability testing indicated that osteoblast adhesion and viability on the IBS were excellent and uniform among various formulation types. Bacteria assays were also performed to assess antimicrobial functions on the CG/nHA composite against both Gram-negative and Gram-positive strains. A higher CG content, as found in some samples, correlated with improved Pseudomonas aeruginosa growth inhibition, although other bacteria strains appeared unaffected by the IBS. In summary, this study highlights CG/nHA composites as innovative biomaterials that should be further studied for reduced bacteria activity and promoted osteoblast responses which was achieved without using pharmaceutical drugs. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2984-2993, 2018.

Porous bodies of hydroxyapatite produced by a combination of the gel-casting and polymer sponge methods.

A combination of gel-casting and polymeric foam infiltration methods is used in this study to prepare porous bodies of hydroxyapatite (HA), to provide a better control over the microstructures of samples. These scaffolds were prepared by impregnating a body of porous polyurethane foam with slurry containing HA powder, and using a percentage of solids between 40% and 50% w/v, and three different types of monomers to provide a better performance. X-Ray Diffraction (XRD), and Fourier Transformed Infrared (FTIR) and Scanning Electron Microscopy (SEM) were employed to evaluate both the powder hydroxyapatite and the scaffolds obtained. In addition, porosity and interconnectivity measurements were taken in accordance with the international norm. Bioactivity was checked using immersion tests in Simulated Body Fluids (SBF). After the sintering process of the porous bodies, the XRD results showed peaks characteristic of a pure and crystalline HA (JCPDS 9-432) as a single phase. SEM images indicate open and interconnected pores inside the material, with pore sizes between 50 and 600 µm. Also, SEM images demonstrate the relatively good bioactivity of the HA scaffolds after immersion in SBF. All results for the porous HA bodies suggest that these materials have great potential for use in tissue engineering.