RESUMO
Titanium with nanotopography (Ti Nano) favors osteoblast differentiation and attenuates the osteoclast inhibitory effects on osteoblasts. Because the interactions between nanotopography and osteoclasts are underexplored, the aims of this study were to evaluate the effects of Ti Nano on osteoclast differentiation and activity, and the influence of osteoblasts on osteoclast-Ti Nano interaction. The discs were conditioned with a mixture of 10 N H2SO4 and 30% aqueous H2O2 to create Ti Nano and non-conditioned Ti discs were used as control (Ti Control). Osteoclasts were cultured on Ti Control and Ti Nano in the presence of osteoblasts in an indirect co-culture system. Also, osteoclasts were cultured on polystyrene and calcium phosphate plates in conditioned media by osteoblasts grown on Ti Control and Ti Nano. While Ti Control exhibited an irregular and smooth surface, Ti Nano presented nanopores distributed throughout the whole surface. Additionally, anisotropy was higher on Ti Nano than Ti Control. Nanotopography favored the gene expression of osteoclast markers but inhibited osteoclast differentiation and activity, and the presence of osteoblasts enhanced the effects of Ti Nano on osteoclasts. Such findings were mimicked by conditioned medium of osteoblasts cultured on Ti Nano, which reduced the osteoclast differentiation and activity. In conclusion, our results indicated that nanotopography regulates osteoblast-osteoclast crosstalk and further investigations should focus the impact of these bone cell interactions on Ti osseointegration.
Assuntos
Osteoclastos , Titânio , Titânio/farmacologia , Titânio/metabolismo , Peróxido de Hidrogênio/farmacologia , Osteoblastos , Diferenciação CelularRESUMO
BACKGROUND: The biogenic synthesis of metallic nanoparticles is a green alternative that reduces the toxicity of this nanomaterials and may enable a synergy between the metallic core and the biomolecules employed in the process enhancing biological activity. The aim of this study was to synthesize biogenic titanium nanoparticles using the filtrate of the fungus Trichoderma harzianum as a stabilizing agent, to obtain a potential biological activity against phytopathogens and mainly stimulate the growth of T. harzianum, enhancing its efficacy for biological control. RESULTS: The synthesis was successful and reproductive structures remained in the suspension, showing faster and larger mycelial growth compared to commercial T. harzianum and filtrate. The nanoparticles with residual T. harzianum growth showed inhibitory potential against Sclerotinia sclerotiorum mycelial growth and the formation of new resistant structures. A great chitinolytic activity of the nanoparticles was observed in comparison with T. harzianum. In regard to toxicity evaluation, an absence of cytotoxicity and a protective effect of the nanoparticles was observed through MTT and Trypan blue assay. No genotoxicity was observed on V79-4 and 3T3 cell lines while HaCat showed higher sensitivity. Microorganisms of agricultural importance were not affected by the exposure to the nanoparticles, however a decrease in the number of nitrogen cycling bacteria was observed. In regard to phytotoxicity, the nanoparticles did not cause morphological and biochemical changes on soybean plants. CONCLUSION: The production of biogenic nanoparticles was an essential factor in stimulating or maintaining structures that are important for biological control, showing that this may be an essential strategy to stimulate the growth of biocontrol organisms to promote more sustainable agriculture.
Assuntos
Hypocreales , Nanopartículas Metálicas , Trichoderma , Trichoderma/química , Trichoderma/metabolismo , Titânio/farmacologia , Titânio/metabolismo , Nanopartículas Metálicas/toxicidadeRESUMO
Titanium dioxide nanoparticles (Np-TiO2) have become the common component of sunscreen cosmetic products. Np-TiO2 can affect especially aquatic ecosystems health, including aquatic organisms such as fish. It is therefore necessary to acquire a better understanding of the effect of Np-TiO2 on aquatic organisms. This study evaluated the biological effects of Np-TiO2 on Danio rerio, such as survival rate and weight change and, in particular, the Ti content or retention in the intestine and liver, as well as the activities of catalase and superoxide dismutase enzymes. In addition, the structure of the intestine, kidney, and liver was investigated through histological analysis. Ninety zebrafish were used, randomly divided into three treatment-groups: a control group (fed with food without adding Np-TiO2) and two groups of fish fed with food containing Np-TiO2 exposed for 7 and 14 days. The amount of Ti in the liver and intestine was measured using atomic absorption spectrophotometry coupled to a graphite furnace (GFAAS). Morphological analysis and enzyme catalase and superoxide dismutase assays were likewise performed. Ti was detected in all fish even in control group; probably Ti must have been introduced during production by the fish food industry. Structural changes were detected in fish fed with Np-TiO2 as vacuolization and disruption of the apical cytoplasm of epithelial cells that covered the intestinal villi. Although kidney morphology appeared intact, the lumen of the proximal tubule was enlarged, and the cells of the distal tubule were vacuolated. No morphological changes in the liver were detected; however, superoxide dismutase activity decreased, suggesting that liver changes occurred at the molecular level. Thus, Np-TiO2 causes morphological changes in the intestine, kidney, and liver of zebrafish and biochemical changes in the liver exposed for 7 and 14 days. Although not highly lethal, Np-TiO2 in the food chain can interfere with the morphophysiology of aquatic organisms. Neither mortalities nor body weight losses were recorded among fish in all groups over the duration of the experiment.
Assuntos
Mucosa Intestinal/efeitos dos fármacos , Rim/efeitos dos fármacos , Fígado/efeitos dos fármacos , Nanopartículas/toxicidade , Titânio/toxicidade , Poluentes Químicos da Água/toxicidade , Peixe-Zebra , Animais , Bioacumulação , Catalase , Relação Dose-Resposta a Droga , Ecossistema , Cadeia Alimentar , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patologia , Rim/metabolismo , Rim/patologia , Fígado/metabolismo , Fígado/patologia , Nanopartículas/metabolismo , Distribuição Aleatória , Protetores Solares/química , Titânio/metabolismo , Poluentes Químicos da Água/metabolismoRESUMO
The Organisation for Economic Co-operation and Development has listed thirteen engineered nanomaterials (ENM) in order to investigate their toxicity on human health. Silicon dioxide (SiO2) and titanium dioxide (TiO2) are included on that list and we added indium tin oxide (ITO) nanoparticles (NPs) to our study, which is not listed on OECD suggested ENM to be investigated, however ITO NPs has a high potential of industrial production. We evaluate the physicochemical properties of SiO2 NPs (10-20 nm), TiO2 nanofibers (NFs; 3 µm length) and ITO NPs (<50 nm) and the impact of protein-corona formation on cell internalization. Then, we evaluated the toxicity of uncoated ENM on human lung epithelial cells exposed to 10 and 50 µg/cm2 for 24 h. TiO2 NFs showed the highest capability to adsorb proteins onto the particle surface followed by SiO2 NPs and ITO NPs after acellular incubation with fetal bovine serum. The protein adsorption had no impact on Alizarin Red S conjugation, intrinsic properties for reactive oxygen (ROS) formation or cell uptake for all types of ENM. Moreover, TiO2 NFs induced highest cell alterations in human lung epithelial cells exposed to 10 and 50 µg/cm2 while ITO NPs induced moderated cytotoxicity and SiO2 NPs caused even lower cytotoxicity under the same conditions. DNA, proteins and lipids were mainly affected by TiO2 NFs followed by SiO2 NPs with toxic effects in protein and lipids while limited variations were detected after exposure to ITO NPs on spectra analyzed by Fourier Transform Infrared Spectroscopy.
Assuntos
Nanoestruturas/química , Nanoestruturas/toxicidade , Coroa de Proteína/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Células A549 , Tamanho Celular , Grânulos Citoplasmáticos/metabolismo , Grânulos Citoplasmáticos/ultraestrutura , Células Epiteliais/metabolismo , Humanos , Metabolismo dos Lipídeos/efeitos dos fármacos , Mucosa Respiratória/citologia , Mucosa Respiratória/metabolismo , Dióxido de Silício/química , Dióxido de Silício/metabolismo , Dióxido de Silício/toxicidade , Propriedades de Superfície , Titânio/química , Titânio/metabolismo , Titânio/toxicidade , Cicatrização/efeitos dos fármacosRESUMO
Although, the excellent level of success of titanium surfaces is based on the literature, there are some biological challenges such as unfavorable metabolic conditions or regions of poor bone quality where greater surface bioactivity is desired. Seeking better performance, we hypothesized that silica-based coating via sol-gel route with immersion in potassium hydroxide basic solution induces acceleration of bone mineralization. This in vitro experimental study coated titanium surfaces with bioactive glass synthesized by route sol-gel via hydrolysis and condensation of chemical alkoxide precursor, tetraethylorthosilicate (TEOS) and/or deposition of chemical compound potassium hydroxide (KOH) to accelerate bone apposition. The generated surfaces titanium(T), titanium with potassium hydroxide deposition (T + KOH), titanium with bioactive glass deposition synthesized by sol-gel route via tetraethylorthosilicate hydrolysis (TEOS), titanium with bioactive glass deposition synthesized by sol-gel route via tetraethylorthosilicate hydrolysis with potassium hydroxide deposition (TEOS + KOH) were characterized by 3D optical profilometry, scanning electron microscopy (SEM), transmission electron microscopy (TEM), contact angle by the sessile drop method, x-ray excited photoelectron spectroscopy (XPS) and energy dispersive x-ray spectrometer (EDX). The addition of the KOH group on the pure titanium (T) or bioactive glass (TEOS) surfaces generated a tendency for better results for mineralization. Groups covered with bioactive glass (TEOS, TEOS + KOH) tended to outperform even groups with titanium substrate (T, T + KOH). The addition of both, bioactive glass and KOH, in a single pure titanium substrate yielded the best results for the mineralization process.
Assuntos
Materiais Revestidos Biocompatíveis/química , Géis/química , Hidróxidos/química , Compostos de Potássio/química , Dióxido de Silício/química , Titânio/química , Animais , Calcificação Fisiológica , Adesão Celular , Proliferação de Células , Materiais Revestidos Biocompatíveis/metabolismo , Implantação Dentária , Humanos , Camundongos , Osteogênese , Silanos/química , Propriedades de Superfície , Titânio/metabolismoRESUMO
Titanium (Ti), used in many dailyuse products, such as shampoos and sunscreen filters, in the form of TiO2 nanoparticles (NPs), may elicit adverse marine biota effects. Marine mammal Ti data is scarce, and subcellular distribution and detoxification information is non-existent. Ti concentrations and metalloprotein detoxification in Pontoporia blainvillei and Steno bredanensis dolphins from Southeastern Brazil were assessed. Metallothionein (MT) concentrations were determined spectrophotometrically, total and subcellular Ti, by ICP-MS and detoxification, by HPLC-ICP-MS. Ti detoxification occurred through MT complexation. Statistical Ti-MT associations were observed in S. bredanensis liver, indicating TiO2 NPs contamination, as Ti binds to MT only as NPs. MT-Ti correlations were observed for both the coastal (P. blainvillei) and offshore (S. bredanensis) dolphins, evidencing oceanic TiO2 diffusion. Ti detoxification through binding to reduced glutathione occurred in both species. Thermostable subcellular fractions are a valuable tool for cetacean Ti detoxification assessments and should be applied to conservation efforts.
Assuntos
Golfinhos/metabolismo , Titânio/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , Brasil , Inativação MetabólicaRESUMO
Magnetic Co3 O4 nanoparticles (NPs) have great potential for applications in biomedicine, as contrast enhancement agents for magnetic resonance imaging, or for drug delivery. Although these NPs are so attractive, their potential toxicity raises serious questions about decreasing cellular viability. In this context, Co3 O4 NPs were prepared via sol-gel method and encapsulated with a layer of TiO2 , a biocompatible oxide, and subjected to structural, magnetic and toxicity characterization. X-ray diffractograms of the samples demonstrate the successful synthesis of the spinel and Raman spectroscopy confirms the coating of the Co3 O4 spinel with TiO2 . The Co3 O4 cores showed a very intense superparamagnetic character; however, this behavior is strongly suppressed when the material is covered with TiO2 . According to the neutral red uptake assay, the coating of the cores with TiO2 significantly decreases the cytotoxic character of the Co3 O4 particles and, as it can be observed with the zeta (ξ) potential measurements, they form a stable colloidal dispersion at cytoplasmic pH. The effect of the thermal treatment enhances the biocompatibility even further, with no statistically significant effect on cell viability even at the highest analyzed concentration. The proposed pathway presents a successful sol-gel method for the preparation of Co3 O4 @TiO2 core-shell nanoparticles. This work opens up possibilities for future application of these materials not only for magnetic resonance imaging but also in catalysis and hyperthermia.
Assuntos
Materiais Revestidos Biocompatíveis/química , Cobalto/química , Nanopartículas Metálicas/química , Nanocompostos/química , Óxidos/química , Titânio/química , Animais , Linhagem Celular , Permeabilidade da Membrana Celular , Sobrevivência Celular/efeitos dos fármacos , Materiais Revestidos Biocompatíveis/metabolismo , Cricetulus , Fibroblastos/citologia , Humanos , Magnetismo , Propriedades de Superfície , Titânio/metabolismoRESUMO
Metals are subject to internal subcellular compartmentalization, altering their bioavailability. Thus, subcellular metal assessments are crucial in biomonitoring efforts. Metal distribution in three subcellular fractions (insoluble - ISF, thermolabile - TLF and thermostable - TSF) were determined by ICP-MS in Steno bredanensis specimens from Southeastern Brazil. Associations between metals, metallothionein (MT) and reduced glutathione (GSH) were also investigated. Differential metal-detoxification mechanisms were observed. MT detoxification was mostly noted for As, Cd, and Pb, while Cu, Cr, Hg, Ni, Se and Ti displayed lower MT-associations. Fe, Zn and Se, on the other hand, were poorly associated to MT, and mostly present in the ISF, indicating low bioavailability. This is the first report on subcellular Sn and Ti distribution in cetaceans and the first in this species in Brazil. Potential protective roles of essential metals against toxic elements are postulated. This study indicates that important biochemical detoxification information is obtained through subcellular fraction analyses in marine mammals.
Assuntos
Golfinhos/metabolismo , Monitoramento Ambiental/métodos , Metalotioneína/metabolismo , Metais/metabolismo , Frações Subcelulares/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , Brasil , Glutationa/metabolismo , Inativação Metabólica , Metais Pesados/análise , Estanho/metabolismo , Titânio/metabolismoRESUMO
The release of the prototypic DAMP High Mobility Group Box 1 (HMGB1) into extracellular environment and its binding to the Receptor for Advanced Glycation End Products (RAGE) has been described to trigger sterile inflammation and regulate healing outcome. However, their role on host response to Ti-based biomaterials and in the subsequent osseointegration remains unexplored. In this study, HMGB1 and RAGE inhibition in the Ti-mediated osseointegration were investigated in C57Bl/6 mice. C57Bl/6 mice received a Ti-device implantation (Ti-screw in the edentulous alveolar crest and a Ti-disc in the subcutaneous tissue) and were evaluated by microscopic (microCT [bone] and histology [bone and subcutaneous]) and molecular methods (ELISA, PCR array) during 3, 7, 14, and 21 days. Mice were divided into 4 groups: Control (no treatment); GZA (IP injection of Glycyrrhizic Acid for HMGB1 inhibition, 4 mg/Kg/day); RAP (IP injection of RAGE Antagonistic Peptide, 4 mg/Kg/day), and vehicle controls (1.5% DMSO solution for GZA and 0.9% saline solution for RAP); treatments were given at all experimental time points, starting 1 day before surgeries. HMGB1 was detected in the Ti-implantation sites, adsorbed to the screws/discs. In Control and vehicle groups, osseointegration was characterized by a slight inflammatory response at early time points, followed by a gradual bone apposition and matrix maturation at late time points. The inhibition of HMGB1 or RAGE impaired the osseointegration, affecting the dynamics of mineralized and organic bone matrix, and resulting in a foreign body reaction, with persistence of macrophages, necrotic bone, and foreign body giant cells until later time points. While Control samples were characterized by a balance between M1 and M2-type response in bone and subcutaneous sites of implantation, and also MSC markers, the inhibition of HMGB1 or RAGE caused a higher expression M1 markers and pro-inflammatory cytokines, as well chemokines and receptors for macrophage migration until later time points. In conclusion, HMGB1 and RAGE have a marked role in the osseointegration, evidenced by their influence on host inflammatory immune response, which includes macrophages migration and M1/M2 response, MSC markers expression, which collectively modulate bone matrix deposition and osseointegration outcome.
Assuntos
Antígenos de Neoplasias/metabolismo , Artroplastia/métodos , Materiais Biocompatíveis/metabolismo , Proteínas HMGB/metabolismo , Inflamação/imunologia , Macrófagos/imunologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Titânio/metabolismo , Animais , Materiais Biocompatíveis/química , Biomarcadores/metabolismo , Matriz Óssea/efeitos dos fármacos , Movimento Celular , Ácido Glicirrízico/administração & dosagem , Proteínas HMGB/antagonistas & inibidores , Humanos , Imunomodulação , Camundongos , Camundongos Endogâmicos C57BL , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Osseointegração , Peptídeos/administração & dosagem , Titânio/químicaRESUMO
The toxicity of titanium dioxide nanoparticles (TiO2 -NP) in the blood, liver, muscle, and brain of a Neotropical detritivorous fish, Prochilodus lineatus, was tested. Juvenile fish were exposed to 0, 1, 5, 10, and 50 mg L-1 of TiO2 -NP for 48 hours (acute exposure) or 14 days (subchronic exposure) to evaluate changes in hematology, red blood cell (RBC) genotoxicity/mutagenicity, liver function (reactive oxygen species (ROS) production, antioxidant responses, detoxification, and histopathology), acetylcholinesterase (AChE) activity in muscles and brain, and Ti bioaccumulation. TiO2 -NP did not cause genetic damage to RBC, but acutely decreased white blood cells (WBC) and increased monocytes. Subchronically, RBC decreased, mean cell volume and hemoglobin increased, and WBC and lymphocytes decreased. Therefore, NP has the potential to affect immune system and increase energy expenditure, reducing the fish's ability to avoid predator and to resist pathogens. In the liver, acute exposure decreased ROS and increased glutathione (GSH) content, while subchronic exposure decreased superoxide dismutase activity and increased glutathione-S-transferase (GST) activity and GSH content. GSH and GST seem to play an essential role in metabolizing NP and ROS, likely increasing hepatocytes' metabolic rate, which may be the cause of observed cell hypertrophy, disarrangement of hepatic cords and degenerative morphological alterations. Although most studies indicate that the kidney is responsible for metabolizing and/or eliminating TiO2 -NP, this study shows that the liver also has a main role in these processes. Nevertheless, Ti still accumulated in the liver, muscle, and brain and decreased muscular AChE activity after acute exposure, showing neurotoxic potential. More studies are needed to better understand the biochemical pathways TiO2 -NP are metabolized and how its bioaccumulation may affect fish homeostasis and survival in the environment.