The aim of this study was to evaluate biocompatibility of hydroxyapatite (HAP) from fish waste using in vitro and in vivo assays. Fish samples (whitemouth croaker - Micropogonias furnieri) from the biowaste was used as HAP source. Pre-osteoblastic MC3T3-E1 cells were used in vitro study. In addition, bone defects were artificially created in rat calvaria and filled with HAP in vivo. The results demonstrated that HAP reduced cytotoxicity in pre-osteoblast cells after 3 and 6 days following HAP exposure. DNA concentration was lower in the HAP group after 6 days. Quantitative RT-PCR did not show any significant differences (p > 0.05) between groups. In vivo study revealed that bone defects filled with HAP pointed out moderate chronic inflammatory cells with slight proliferation of blood vessels after 7 and 15 days. Chronic inflammatory infiltrate was absent after 30 days of HAP exposure. There was also a decrease in the amount of biomaterial, being followed by newly formed bone tissue. All experimental groups also demonstrated strong RUNX-2 immoexpression in the granulation tissue as well as in cells in close contact with biomaterial. The number of osteoblasts inside the defect area was lower in the HAP group when compared to control group after 7 days post-implantation. Similarly, the osteoblast surface as well as the percentage of bone surface was higher in control group when compared with HAP group after 7 days post-implantation. Taken together, HAP from fish waste is a promising possibility that should be explored more carefully by tissue-engineering or biotechnology.
Research on biomaterials of natural origin has gained prominence in the literature. Above all, marine sponges, due to their architecture and structural components, present a promising potential for the engineering of bone tissue. In vitro studies demonstrate that a biosilica of marine sponges has osteogenic potential. However, in vivo works are needed to elucidate the interaction of biosilica (BS) and bone tissue. The objective of the study was to evaluate the morphological and chemical characteristics of BS compared to Bioglass (BG) by scanning electron microscopy (SEM) and X-ray dispersive energy (EDX) spectroscopy. In addition, to evaluate the biological effects of BS, through an experimental model of tibial bone defect using histopathological, histomorphometric, immunohistochemical (IHC) and mechanical tests. SEM and EDX demonstrated the successful extraction of BS. Histopathological analysis demonstrated that Control Group (GC) had greater formation of newly formed bone tissue compared to BG and BS, yet BG bone neoformation was greater than BS. However, BS showed material degradation and granulation tissue formation, with HIGHLIGHTS BS was isolated from specimens of the marine sponge Tedania ignis collected in the north coast of São Paulo, Brazil. BS derived from marine sponges has been used as a promising raw material for bone grafts substitutes and tissue engineering proposes. The present work successfully obtained crystalline BS from marine sponges, able of properly interacting with bone tissue. The results are very inspiring toward further in vivo investigations. Cruz, M. A..; et al.
One promising skin substitutes in the wound healing are the bacterial cellulose membranes (BCM). These biomaterials present nanostructures composed of microfibrils capable of forming three-dimensional pores that allow cell. In association with these biopolymers, several treatments are used, such as enrichment by growth factors and/or the application of photobiomodulation (PBM). Therefore, the aim of this study was to investigate the viability, proliferation and cytotoxicity of a BCM (culturing of Komagataeibacter xylinus), with or without FGF-2 in association with PBM therapy. In the characterization of BCM we saw that the membrane does not show great variations in pH and with the scanning electron microscopy it was possible to observe that the BCM has a denser and a porous side that allows the adhesion of fibroblasts, confirmed by histological staining and DAPI/Phalloidin. In vitro evaluation showed that the immunofluorescence (CaAM/EthD-1) for live and dead cells presented, in the groups with combined treatment at long-term of PBM and FGF-2, a greater quantity of live cells than with these isolated treatments and/or at short-term. However, in the short-term of combined treatment PBM and FGF-2 supplementation, fibroblasts and macrophages were more viable by Alamar Blue, in direct and indirect contact respectively. The comet assay did not show cytotoxicity for DNA damage in fibroblasts indirect contact with membrane extract. The results highlight the potential of association of FGF-2 supplementation with the application of PBM for use with BCM, due to its promoted increased cell density at long-term and improved viability in fibroblasts and macrophages at short-term.
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