RESUMO
Despite the well-known advantages of the titanium-based implant systems, they still lack an optimal balance between biofunctionality and mechanical strength, especially regarding the modulation of cellular response and a desired implant osseointegration. In this work, we fabricated a nanocomposite based on porous commercially pure grade 4 titanium (c.p. Ti) reinforced with carbon nanotubes (CNT) at 5% and 10% w/w, with the aim of obtaining a nanocomposite with lower stiffness compared to traditional titanium-based implants and with the mechanical strength and bioactivity owed by the CNT. Results obtained by scanning electron microscopy, X-ray photoelectron spectroscopy, and atomic force microscopy characterization showed that the CNT dispersed and incorporated into the porous c.p. Ti matrix. Interestingly, CNT were associated with a higher twining within neighbor Ti grains, which was indeed consistent with an increased in nano-hardness. Biological evaluation by MTT and Comet assay revealed that the nanocomposites did not induce genotoxicity and cytotoxicity on two different cells lines despite the presence of nickel at the surface. Accordingly, a purification step would be required before these CNT can be used for biomedical applications. Our results indicate that incorporation of CNT into porous c.p. Ti is a promising avenue to achieve an adequate balance between biofunctionality and mechanical strength in Ti-based scaffolds for tissue replacement. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 719-731, 2019.
Assuntos
Teste de Materiais , Nanotubos de Carbono/química , Titânio/química , Linhagem Celular Tumoral , Humanos , PorosidadeRESUMO
Surgical clipping and endovascular coiling are well recognized as conventional treatments of Penetrating Brain Injury aneurysms. These clinical approaches show partial success, but often result in thrombus formation and the rupture of aneurysm near arterial walls. The authors address these challenging brain traumas with a unique combination of a highly biocompatible biopolymer hydrogel rendered magnetic in a flexible and resilient membrane coating integrated to a scaffold stent platform at the aneurysm neck orifice, which enhances the revascularization modality. This work focuses on the in situ diagnosis of nano-mechanical behavior of bacterial nanocellulose (BNC) membranes in an aqueous environment used as tissue reconstruction substrates for cerebral aneurysmal neck defects. Nano-mechanical evaluation, performed using instrumented nano-indentation, shows with very low normal loads between 0.01 to 0.5 mN, in the presence of deionized water. Mechanical testing and characterization reveals that the nano-scale response of BNC behaves similar to blood vessel walls with a very low Young´s modulus, E (0.0025 to 0.04 GPa), and an evident creep effect (26.01 ± 3.85 nm s-1 ). These results confirm a novel multi-functional membrane using BNC and rendered magnetic with local adhesion of iron-oxide magnetic nanoparticles.
Assuntos
Revascularização Cerebral/métodos , Procedimentos Endovasculares/métodos , Hidrogéis/uso terapêutico , Aneurisma Intracraniano/cirurgia , Nanopartículas de Magnetita/uso terapêutico , Celulose/uso terapêutico , Procedimentos Endovasculares/efeitos adversos , Gluconacetobacter xylinus/metabolismo , Humanos , Aneurisma Intracraniano/fisiopatologia , Fenômenos Mecânicos , Instrumentos CirúrgicosRESUMO
A new generation of biomaterials are evolving from being biologically inert toward bioactive surfaces, which can further interact with biological components at the nanoscale. Here, we present directed irradiation synthesis (DIS) as a novel technology to selectively apply plasma ions to bombard any type of biomaterial and tailor the nanofeatures needed for in vitro growth stimulation. In this work, we demonstrate for the first time, the influence of physiochemical cues (e.g., self-organized topography at nanoscale) of medical grade Ti6Al4V results in control of cell shape, adhesion, and proliferation of human aortic smooth muscle stem cells. The control of surface nanostructures was found to be correlated to ion-beam incidence angle linked to a surface diffusive regime during irradiation synthesis with argon ions at energies below 1 keV and a fluence of 2.5 × 1017 cm-2. Cell viability and cytoskeleton morphology were evaluated at 24 h, observing an advance cell attachment state on post-DIS surfaces. These modified surfaces showed 84% of cell biocompatibility and an increase in cytoplasmatic protusions ensuring a higher cell adhesion state. Filopodia density was promoted by a 3-fold change for oblique incidence angle DIS treatment compared to controls (e.g., no patterning) and lamellipodia structures were increased more than a factor of 2, which are indicators of cell attachment stimulation due to DIS modification. In addition, the morphology of the nanofeatures were tailored, with high fidelity control of the main DIS parameters that control diffusive and erosive regimes of self-organization. We have correlated the morphology and the influence in cell behavior, where nanoripple formation is the most active morphology for cell stimulation.
RESUMO
Current treatments for brain aneurysms are invasive, traumatic, and not suitable in most patients with increased risks. A new alternative method is using scaffold stents to create a local and focal attraction force of cells for an in situ reconstruction of the tunica media. For this purpose, a nanostructured bioactive coating is designed to render an asymmetric region of the stent scaffold magnetic and biomimetic, which utilizes bacterial nanocellulose (BNC) as a platform for both magnetic and cell attraction as well as proliferation. The magnetization of the BNC is realized through the reaction of Fe III and II, precipitating superparamagnetic iron oxide nanoparticles (SPION). Subsequently, magnetic bacterial nanocellulose (MBNC) is coated with polyethylene glycol to improve its biocompatibility. Cytotoxicity and biocompatibility are evaluated using porcine aortic smooth muscle cells. Preliminary cellular migration assays demonstrate the behavior between MBNC and cells labeled with SPION. In this work, (1) synthesis of BNC impregnated with magnetic nanoparticles is successfully demonstrated; (2) a viable, resilient, and biocompatible hydrogel membrane is tested for neuroendovascular application using a stent scaffold; (3) cell viability and minimal cytotoxicity is achieved; (4) cell migration tests and examination of cellular magnetic attraction confirm the viability of MBNC as a multifunctional coating.
Assuntos
Celulose/química , Compostos Férricos/química , Aneurisma Intracraniano/terapia , Nanopartículas de Magnetita/química , Stents , Animais , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Celulose/farmacologia , Compostos Férricos/farmacologia , Humanos , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Hidrogel de Polietilenoglicol-Dimetacrilato/uso terapêutico , Aneurisma Intracraniano/fisiopatologia , Nanopartículas de Magnetita/uso terapêutico , Teste de Materiais , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/patologia , Nanoestruturas/química , Nanoestruturas/uso terapêutico , Polietilenoglicóis/química , SuínosRESUMO
Metallic nanoparticles (such as gold and silver) have been intensely studied for wound healing applications due to their ability to be easily functionalized, possess antibacterial properties, and their strong potential for targeted drug release. In this study, rod-shaped silver nanorods (AgNRs) and gold nanorods (AuNRs) were fabricated by electron beam physical vapor deposition (EBPVD), and their cytotoxicity toward human skin fibroblasts were assessed and compared to sphere-shaped silver nanospheres (AgNSs) and gold nanospheres (AuNSs). Results showed that the 39.94 nm AgNSs showed the greatest toxicity with fibroblast cells followed by the 61.06 nm AuNSs, â¼556 nm × 47 nm (11.8:1 aspect ratio) AgNRs, and the â¼534 nm × 65 nm (8.2:1 aspect ratio) AuNRs demonstrated the least amount of toxicity. The calculated IC50 (50% inhibitory concentration) value for the AgNRs exposed to fibroblasts was greater after 4 days of exposure (387.3 µg mL(-1)) compared to the AgNSs and AuNSs (4.3 and 23.4 µg mL(-1), respectively), indicating that these spherical metallic nanoparticles displayed a greater toxicity to fibroblast cells. The IC50 value could not be measured for the AuNRs due to an incomplete dose response curve. The reduced cell toxicity with the presently developed rod-shaped nanoparticles suggests that they may be promising materials for use in numerous biomedical applications.
Assuntos
Fibroblastos/citologia , Ouro/toxicidade , Nanosferas/toxicidade , Nanotubos/toxicidade , Prata/toxicidade , Adesão Celular , Proliferação de Células , Endocitose , Ouro/química , Ouro/metabolismo , Humanos , Nanosferas/química , Nanosferas/metabolismo , Nanosferas/ultraestrutura , Nanotubos/química , Nanotubos/ultraestrutura , Prata/química , Prata/metabolismoRESUMO
Gold nanoparticles are materials with unique optical properties that have made them very attractive for numerous biomedical applications. With the increasing discovery of techniques to synthesize novel nanoparticles such as star-shaped gold nanoparticles for biomedical applications, the safety and performance of these new nanomaterials must be systematically assessed before use. In this study, gold nanostars (AuNSTs) with multibranched surface structures were synthesized, and their influence on the cytotoxicity of human skin fibroblasts and rat fat pad endothelial cells (RFPECs) were assessed and compared with that of gold nanospheres (AuNSPs) with unbranched surfaces. Results showed that the AuNSPs with diameters of approximately 61.46 nm showed greater toxicity with fibroblast cells and RFPECs compared with the synthesized AuNSTs with diameters of approximately 33.69 nm. The AuNSPs were lethal at concentrations of 40 µg/mL for both cell lines, whereas the AuNSTs were less toxic at higher concentrations (400 µg/mL). The calculated IC50 (50% inhibitory concentration) values of the AuNSPs exposed to fibroblast cells were greater at 1 and 4 days of culture (26.4 and 27.7 µg/mL, respectively) compared with the RFPECs (13.6 and 13.8 µg/mL, respectively), indicating that the AuNSPs have a greater toxicity to endothelial cells. It was proposed that possible factors that could be promoting the reduced toxicity effects of the AuNSTs to fibroblast cells and RFPECs, compared with the AuNSPs may be size, surface chemistry, and shape of the gold nanoparticles. The reduced cell toxicity observed with the AuNSTs suggests that AuNSTs may be a promising material for use in biomedical applications.
Assuntos
Ouro/química , Nanopartículas Metálicas/química , Nanosferas/química , Tecido Adiposo/citologia , Animais , Morte Celular , Proliferação de Células , Forma Celular , Células Cultivadas , Células Endoteliais/citologia , Humanos , Hidrodinâmica , Nanopartículas Metálicas/ultraestrutura , Nanosferas/ultraestrutura , Fenômenos Ópticos , Tamanho da Partícula , Ratos , Propriedades de SuperfícieRESUMO
Microarrays allow researchers to measure the expression of thousands of genes in a single experiment. Before statistical comparisons can be made, the data must be assessed for quality and normalisation procedures must be applied, of which many have been proposed. Methods of comparing the normalised data are also abundant, and no clear consensus has yet been reached. The purpose of this paper was to compare those methods used by the EADGENE network on a very noisy simulated data set. With the a priori knowledge of which genes are differentially expressed, it is possible to compare the success of each approach quantitatively. Use of an intensity-dependent normalisation procedure was common, as was correction for multiple testing. Most variety in performance resulted from differing approaches to data quality and the use of different statistical tests. Very few of the methods used any kind of background correction. A number of approaches achieved a success rate of 95% or above, with relatively small numbers of false positives and negatives. Applying stringent spot selection criteria and elimination of data did not improve the false positive rate and greatly increased the false negative rate. However, most approaches performed well, and it is encouraging that widely available techniques can achieve such good results on a very noisy data set.