BIOCOMPATIBILIDADE EM FILMES DE HIDROXIAPATITA REFORÇADA COM NANOTUBOS DE CARBONO

As características químicas e estruturais individuais de nanotubos de carbono (NTC) e hidroxiapatita (HAp), principal constituinte da fase inorgânica do osso, possibilitam seu uso na área médica como materiais biocompatíveis em implantes e próteses. O objetivo deste trabalho é estudar a biocompatibilidade do compósito HAp/NTC eletrodepositadas comitantemente em aço 316. Para a produção dos compósitos os NTC foram dispersos na solução eletrolítica em concentrações de 1% e 3%. Os resultados obtidos in vitro, utilizando o ensaio da Lactato Desidrogenase (LDH), demostraram que nenhuma amostra causou citotoxicidade nas células osteoblásticas. Neste estudo, também foram avaliados genes relacionados à osteogênese, tais como: fosfatase alcalina (ALP), osteopontina (OPN), osteocalcina (OC) por RT-qPCR (Transcrição Reversa – Reação em Cadeia da Polimerase quantitativa), após 14 dias. Os osteoblastos em contato com a HAp/NTC apresentaram aumento da expressão de genes importantes no processo de mineralização e maturação óssea. Os resultados demonstraram um grande potencial de aplicação clínica deste compósito

Electrospun nanofibrous poly (Lactic Acid)/Titanium dioxide nanocomposite membranes for cutaneous scar minimization

The animal study was reviewed and approved by 10/2015-CEUA/ICT/CJSC-UNESP.Poly (lactic acid) (PLA) has been increasingly used in cutaneous tissue engineering due to its low cost, ease of handling, biodegradability, and biocompatibility, as well as its ability to form composites. However, these polymers possess a structure with nanoporous that mimic the cellular environment. In this study, nanocomposites are prepared using PLA and titanium dioxide (TiO2) (10 and 35%—w/w) nanoparticles that also function as an active anti-scarring agent. The nanocomposites were prepared using an electrospinning technique. Three different solutions were prepared as follows: PLA, 10% PLA/TiO2, and 35% PLA/TiO2 (w/w%). Electrospun PLA and PLA/TiO2 nanocomposites were characterized morphologically, structurally, and chemically using electron scanning microscopy, transmission electron microscopy, goniometry, and X-ray diffraction. L929 fibroblast cells were used for in vitro tests. The cytotoxic effect was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays. Versicam (VCAN), biglicam (BIG), interleukin-6 (IL6), interleukin-10 (IL-10), and type-1 collagen (COL1A1) genes were evaluated by RT-qPCR. In vivo tests using Wistar rats were conducted for up to 15 days. Nanofibrous fibers were obtained for all groups that did not contain residual solvents. No cytotoxic effects were observed for up to 168 h. The genes expressed showed the highest values of versican and collagen-1 (p < 0.05) for PLA/TiO2 nanocomposite scaffolds when compared to the control group (cells). Histological images showed that PLA at 10 and 35% w/w led to a discrete inflammatory infiltration and expression of many newly formed vessels, indicating increased metabolic activity of this tissue. To summarize, this study supported the potential of PLA/TiO2 nanocomposites ability to reduce cutaneous scarring in scaffolds.This work was supported by the National Council for Scientificand Technological Development (CNPq, #303752/2017-3 and#404683/2018-5 to AL and #304133/2017-5 and #424163/2016-0 to FM). AZ acknowledges financial support of the FCT throughUID/CTM/00264/2019 and Investigator FCT Research contract(IF/00071/2015) and the project PTDC/CTM-TEX/28295/2017 financed by FCT, FEDER, and POCI

In Vitro Osteogenesis Stimulation via Nano-Hydroxyapatite/Carbon Nanotube Thin Films on Biomedical Stainless Steel

We evaluated the electrophoretic deposition of nanohydroxyapatite/superhydrop hilic multiwalled carbon nanotube composites (nHAp/MWCNT) onto stainless steel biomedical alloys for applications in bone tissue engineering. First, nHAp/MWCNT composites were dispersed into 0.042 mol&middot;L&minus;1 of Ca(NO3)2&middot;4H2O + 0.025 mol&middot;L&minus;1 NH4H2PO4 electrolytes (pH = 4.8) at two different concentrations. Next, a voltage of &minus;2 V was applied using 316L stainless steel as a working electrode (0.27 cm2), a high-purity platinum coil wire was used as the auxiliary electrode, and an Ag/AgCl (3 M) electrode was used as the reference electrode. The nHAp/MWCNT composites were characterized by transmission electron microscopy. The deposited nHAp and nHAp/MWCNT films were characterized by profilometry, scanning electron microscopy, X-ray diffractometry and Raman spectroscopy. Human osteoblast cells were cultivated with the different materials and in vitro cytotoxicity was evaluated using lactate dehydrogenase (LDH) assay. The osteogenesis process was evaluated by mRNA levels of the three genes that are directly related to bone repair: Alkaline Phosphatase, Osteopontin and Osteocalcin. We showed that rough, crystalline apatite thin films containing phases of nHAp were successfully deposited onto 316L stainless steel alloys. Also, we noticed that nHAp/MWCNT thin films deposited onto 316L stainless steel alloys upregulated the expression of important genes related to bone mineralization and maturation. Our results strongly support the possibility of this new alternative to modify the surface of metallic biomedical alloys to promote bone tissue regeneration

Glial cells modulate heparan sulfate proteoglycan (HSPG) expression by neuronal precursors during early postnatal cerebellar development

Cerebellum controls motor coordination, balance, eye movement, and has been implicated in memory and addiction. As in other parts of the CNS, correct embryonic and postnatal development of the cerebellum is crucial for adequate performance in the adult. Cellular and molecular defects during cerebellar development can lead to severe phenotypes, such as ataxias and tumors. Knowing how the correct development occurs can shed light into the mechanisms of disease. Heparan sulfate proteoglycans are complex molecules present in every higher eukaryotic cells and changes in their level of expression as well as in their structure lead to drastic functional alterations. This work aimed to investigate changes in heparan sulfate proteoglycans expression during cerebellar development that could unveil control mechanisms. Using real time RT-PCR we evaluated the expression of syndecans, glypicans and modifying enzymes by isolated cerebellar granule cell precursors, and studied the influence of soluble glial factors on the expression of those genes. We evaluated the possible involvement of Runx transcription factors in the response of granule cell precursors to glial factors. Our data show for the first time that cerebellar granule cell precursors express members of the Runx family and that the expression of those genes can also be controlled by glial factors. Our results also show that the expression of all genes studied vary during postnatal development and treatment of precursors with glial factors indicate that the expression of heparan sulfate proteoglycan genes as well as genes encoding heparan sulfate modifying enzymes can be modulated by the microenvironment, reflecting the intricate relations between neuron and glia. (C) 2010 ISDN. Published by Elsevier B.V. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Investiga Institutos de PesquisaUniversidade Federal de São Paulo, Dept Bioquim, BR-04044020 São Paulo, BrazilInvestiga Inst Pesquisa, BR-13084791 Campinas, SP, BrazilUniversidade Federal de São Paulo, Dept Bioquim, BR-04044020 São Paulo, BrazilWeb of Scienc

Nanohydroxyapatite/Graphene Nanoribbons Nanocomposites Induce in Vitro Osteogenesis and Promote in Vivo Bone Neoformation

Nanomaterials based on graphene oxide nanoribbons (GNR) and nanohydroxyapatite (nHAp) serve as attractive materials for bone tissue engineering. Herein, we evaluated the potential of nHAp/GNR toward in vitro analysis of specific genes related to osteogenesis and in vivo bone regeneration using animal model. Three different concentrations of nHAp/GNR composites were analyzed in vitro using a cytotoxicity assay, and osteogenic potential was determined by <i>ALP</i>, <i>OPN</i>, <i>OCN</i>, <i>COL1</i>, and <i>RUNX2</i> genes and alkaline phosphatase assays. In vivo bone neoformation using a well-established in vivo rat tibia defect model was used to confirm the efficiency of the optimized composite. The scaffolds were nontoxic, and the osteogenesis process was dose-dependent (at 200 μg mL^–1 of nHAp/GNR) compared to controls. The in vivo results showed higher bone neoformation after 15 days of nHAp/GNR implantation compared to all groups. After 21 days, both nHAp/GNR composites showed better lamellar bone formation compared to control. We attributed this enhanced bone neoformation to the high bioactivity and surface area presented by nHAp/GNR composites, which was systematically evaluated in previous studies. These new in vivo results suggest that nHAp/GNR composites can be exploited for a range of strategies for the improved development of novel dental and orthopedic bone grafts to accelerate bone regeneration