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1.
J Funct Biomater ; 13(4)2022 Nov 17.
Artigo em Inglês | MEDLINE | ID: mdl-36412893

RESUMO

Three-dimensional bioprinting is a powerful technique for manufacturing improved engineered tissues. Three-dimensional bioprinted hydrogels have significantly advanced the medical field to repair cartilage tissue, allowing for such constructs to be loaded with different components, such as cells, nanoparticles, and/or drugs. Cartilage, as an avascular tissue, presents extreme difficulty in self-repair when it has been damaged. In this way, hydrogels with optimal chemical and physical properties have been researched to respond to external stimuli and release various bioactive agents to further promote a desired tissue response. For instance, methacryloyl gelatin (GelMA) is a type of modified hydrogel that allows for the encapsulation of cells, as well as oxygen-releasing nanoparticles that, in the presence of an aqueous medium and through controlled porosity and swelling, allow for internal and external environmental exchanges. This review explores the 3D bioprinting of hydrogels, with a particular focus on GelMA hydrogels, to repair cartilage tissue. Recent advances and future perspectives are described.

2.
J Funct Biomater ; 12(2)2021 Jun 07.
Artigo em Inglês | MEDLINE | ID: mdl-34200191

RESUMO

BACKGROUND: Several studies proved that anodic oxidation improves osseointegration. This study aimed to optimize osseointegration through anodization in dental implants, obtaining anatase phase and controlled nanotopography. METHODS: The division of the groups with 60 titanium implants was: control (CG); sandblasted (SG); anodized (AG): anodized pulsed current (duty cycle 30%, 30 V, 0.2 A and 1000 Hz). Before surgery, surface characterization was performed using Atomic Force Microscopy (AFM), Scanning Electron Microscopy (SEM), X-ray Dispersive Energy Spectroscopy (EDS) and Raman Spectroscopy. For in vivo tests, 10 New Zealand white rabbits received an implant from each group. The sacrifice period was 2 and 6 weeks (n = 5) and the specimens were subjected to computed microtomography (µCT) and reverse torque test. RESULTS: AFM and SEM demonstrated a particular nanotopography on the surface in AG; the anatase phase was proved by Raman spectroscopy. In the µCT and in the reverse torque test, the AG group presented better results than the other groups. CONCLUSION: The chemical composition and structure of the TiO2 film were positively affected by the anodizing technique, intensifying the biological characteristics in osseointegration.

3.
J Funct Biomater ; 12(2)2021 May 02.
Artigo em Inglês | MEDLINE | ID: mdl-34063270

RESUMO

Different strategies have been employed to provide adequate nutrients for engineered living tissues. These have mainly revolved around providing oxygen to alleviate the effects of chronic hypoxia or anoxia that result in necrosis or weak neovascularization, leading to failure of artificial tissue implants and hence poor clinical outcome. While different biomaterials have been used as oxygen generators for in vitro as well as in vivo applications, certain problems have hampered their wide application. Among these are the generation and the rate at which oxygen is produced together with the production of the reaction intermediates in the form of reactive oxygen species (ROS). Both these factors can be detrimental for cell survival and can severely affect the outcome of such studies. Here we present calcium peroxide (CPO) encapsulated in polycaprolactone as oxygen releasing microparticles (OMPs). While CPO releases oxygen upon hydrolysis, PCL encapsulation ensures that hydrolysis takes place slowly, thereby sustaining prolonged release of oxygen without the stress the bulk release can endow on the encapsulated cells. We used gelatin methacryloyl (GelMA) hydrogels containing these OMPs to stimulate survival and proliferation of encapsulated skeletal myoblasts and optimized the OMP concentration for sustained oxygen delivery over more than a week. The oxygen releasing and delivery platform described in this study opens up opportunities for cell-based therapeutic approaches to treat diseases resulting from ischemic conditions and enhance survival of implants under severe hypoxic conditions for successful clinical translation.

4.
Colloids Surf B Biointerfaces ; 205: 111850, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-34015729

RESUMO

Currently, oxygen supply for in vitro cell culture is one of the major challenges in tissue engineering, especially in three-dimensional (3D) structures, such as polymeric hydrogels, because oxygen is an essential element for cells survival. In this context, oxygen levels must be maintained in articular cartilage to promote the differentiation, viability, and proliferation of chondrocytes due to the low level of oxygen presence in this region. Although some technologies employ oxygen-generating materials to add sufficient oxygen levels, the limitations and challenges of current technologies include the lack of controlled, sustained, and prolonged release of the oxygen. Moreover, the fabrication methods may leave some impurities or residues resulting in toxicity to the cells. "Click" chemistry is a facile, versatile, and compatible chemical strategy to engineer hydrogels for tissue engineering applications. Herein, we disclose the engineering of oxygen-generating microparticles in chondrocytes-laden hydrogels through a versatile catalyst-free tetrazine and norbornene inverse electron demand Diels‒Alder (iEDDA) click reaction. The hydrogels combine chondroitin sulfate (CS) and poly(ethylene glycol) (PEG) crosslinked in situ, displaying tunable rheological and mechanical properties, for sustained and prolonged oxygen-release. Gene expression analysis of the chondrocytes by real-time reverse transcription polymerase chain reaction (RT-PCR) demonstrated promising cell response within the engineered hydrogel.


Assuntos
Condrócitos , Hidrogéis , Química Click , Oxigênio , Engenharia Tecidual
5.
Int J Nanomedicine ; 16: 667-682, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33531806

RESUMO

BACKGROUND: Nanostructured surface modifications of Ti-based biomaterials are moving up from a highly-promising to a successfully-implemented approach to developing safe and reliable implants. METHODS: The study's main objective is to help consolidate the knowledge and identify the more suitable experimental strategies related to TiO2 nanotubes-modified surfaces. In this sense, it proposes the thorough investigation of two optimized nanotubes morphologies in terms of their biological activity (cell cytotoxicity, alkaline phosphatase activity, alizarin red mineralization test, and cellular adhesion) and their electrochemical behavior in simulated body fluid (SBF) electrolyte. Layers of small-short and large-long nanotubes were prepared and investigated in their amorphous and crystallized states and compared to non-anodized samples. RESULTS: Results show that much more than the surface area development associated with the nanotubes' growth; it is the heat treatment-induced change from amorphous to crystalline anatase-rutile structures that ensure enhanced biological activity coupled to high corrosion resistance. CONCLUSION: Compared to both non-anodized and amorphous nanotubes layers, the crystallized nano-structures' outstanding bioactivity was related to the remarkable increase in their hydrophilic behavior, while the enhanced electrochemical stability was ascribed to the thickening of the dense rutile barrier layer at the Ti surface beneath the nanotubes.


Assuntos
Nanotubos/química , Próteses e Implantes , Titânio/química , Fosfatase Alcalina/metabolismo , Adesão Celular , Linhagem Celular Tumoral , Sobrevivência Celular , Corrosão , Eletroquímica , Eletrólitos/química , Humanos , Nanotubos/ultraestrutura , Molhabilidade
6.
Mater Sci Eng C Mater Biol Appl ; 120: 111776, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-33545906

RESUMO

A biomineralization processes is disclosed for engineering nanomaterials that support bone repair. The material was fabricated through a hot press process using electrospun poly(lactic acid) (PLA) matrix covered with hybrid composites of carbon nanotubes/graphene nanoribbons (GNR) and nanohydroxyapatite (nHA). Various scaffolds were devised [nHA/PLA, PLA/GNR, and PLA/nHA/GNR (1 and 3%)] and their structure and morphology characterized through Scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDS), and Atomic force microscope (AFM). Moreover, thorough biocompatibility and toxicity studies were performed. Here, in vivo studies on toxicity and cytotoxicity were conducted in aqueous dispersions of the biomaterials at concentrations of 30, 60, and 120 µg/mL using the Allium cepa test. Further toxicity studies were performed through hemolysis toxicity tests and genotoxicity tests evaluating the damage index and damage frequencies of DNAs through comet assays with samples of the animals' peripheral blood, marrow, and liver. Additionally, the regenerative activity of the scaffolds was analyzed by measuring the cortical tibiae of rats oophorectomized implanted with the biomaterials. Biochemical analyzes [glutamic pyruvic transaminase (GPT), glutamic oxaloacetic transaminase (GOT), urea, calcium, phosphorus, and alkaline phosphatase (ALP)] were also performed on blood samples. The results suggested a toxicity and cytotoxicity level for the GNR biomaterials at a concentration of 60 and 120 µg/mL, but non-toxicity and cytotoxicity for the 30 µg/mL concentration. The scaffolds obtained at a concentration of 0.3 mg/cm2 were not toxic in the hemolysis test and demonstrated no cytotoxicity, genotoxicity, and mutagenicity in the blood, marrow, and liver analyzes of the animals, corroborating data from the biochemical markers of GPT, GOT, and urea. Tissue regeneration was performed in all groups and was more pronounced in the group containing the combination of nHA/GNR (3%), which is consistent with the data obtained for the calcium, serum phosphorus, and ALP concentrations. Consequently, the study indicates that the engineered nanobiomaterial is a promising candidate for bone tissue repair and regenerative applications. STATEMENT OF SIGNIFICANCE: The scientific contribution of this study is the engineering of a synthetic hybrid biomaterial, in nanoscale by a pressing and heating process. A biodegradable polymeric matrix was covered on both sides with a carbonated hybrid bioceramic/graphene nanoribbons (GNR), which has hydrophilic characteristics, with chemical elements stoichiometrically similar to bone mineral composition. The nanomaterial displayed promising bone regeneration ability, which is the first example to be used in an osteoporotic animal model. Moreover, detailed biocompatibility and toxicity studies were performed on the nanomaterials and their compositions, which is of great interest for the scientific community.


Assuntos
Durapatita , Nanotubos de Carbono , Animais , Biomineralização , Regeneração Óssea , Ratos , Engenharia Tecidual , Alicerces Teciduais
7.
Commun Biol ; 4(1): 233, 2021 02 19.
Artigo em Inglês | MEDLINE | ID: mdl-33608611

RESUMO

The engineering of multifunctional surgical bactericidal nanofibers with inherent suitable mechanical and biological properties, through facile and cheap fabrication technology, is a great challenge. Moreover, hernia, which is when organ is pushed through an opening in the muscle or adjacent tissue due to damage of tissue structure or function, is a dire clinical challenge that currently needs surgery for recovery. Nevertheless, post-surgical hernia complications, like infection, fibrosis, tissue adhesions, scaffold rejection, inflammation, and recurrence still remain important clinical problems. Herein, through an integrated electrospinning, plasma treatment and direct surface modification strategy, multifunctional bactericidal nanofibers were engineered showing optimal properties for hernia repair. The nanofibers displayed good bactericidal activity, low inflammatory response, good biodegradation, as well as optimal collagen-, stress fiber- and blood vessel formation and associated tissue ingrowth in vivo. The disclosed engineering strategy serves as a prominent platform for the design of other multifunctional materials for various biomedical challenges.


Assuntos
Antibacterianos/farmacologia , Materiais Biocompatíveis , Gelatina/farmacologia , Hérnia Abdominal/cirurgia , Herniorrafia/instrumentação , Metacrilatos/farmacologia , Nanofibras , Poliésteres/farmacologia , Infecção da Ferida Cirúrgica/prevenção & controle , Alicerces Teciduais , Animais , Antibacterianos/química , Modelos Animais de Doenças , Gelatina/química , Hérnia Abdominal/patologia , Metacrilatos/química , Camundongos , Células NIH 3T3 , Nanomedicina , Poliésteres/química , Ratos , Infecção da Ferida Cirúrgica/microbiologia , Cicatrização/efeitos dos fármacos
8.
J Funct Biomater ; 12(1)2021 Feb 05.
Artigo em Inglês | MEDLINE | ID: mdl-33562592

RESUMO

Electrospun ultrathin fibrous scaffold filed with synthetic nanohydroxyapatite (nHAp) and graphene nanoribbons (GNR) has bioactive and osteoconductive properties and is a plausible strategy to improve bone regeneration. Poly(butylene-adipate-co-terephthalate) (PBAT) has been studied as fibrous scaffolds due to its low crystallinity, faster biodegradability, and good mechanical properties; however, its potential for in vivo applications remains underexplored. We proposed the application of electrospun PBAT with high contents of incorporated nHAp and nHAp/GNR nanoparticles as bone grafts. Ultrathin PBAT, PBAT/nHAp, and PBAT/nHAp/GNR fibers were produced using an electrospinning apparatus. The produced fibers were characterized morphologically and structurally using scanning electron (SEM) and high-resolution transmission electron (TEM) microscopies, respectively. Mechanical properties were analyzed using a texturometer. All scaffolds were implanted into critical tibia defects in rats and analyzed after two weeks using radiography, microcomputed tomography, histological, histomorphometric, and biomechanical analyses. The results showed through SEM and high-resolution TEM characterized the average diameters of the fibers (ranged from 0.208 µm ± 0.035 to 0.388 µm ± 0.087) and nHAp (crystallite around 0.28, 0.34, and 0.69 nm) and nHAp/GNR (200-300 nm) nanoparticles distribution into PBAT matrices. Ultrathin fibers were obtained, and the incorporated nHAp and nHAp/GNR nanoparticles were well distributed into PBAT matrices. The addition of nHAp and nHAp/GNR nanoparticles improved the elastic modulus of the ultrathin fibers compared to neat PBAT. High loads of nHAp/GNR (PBATnH5G group) improved the in vivo lamellar bone formation promoting greater radiographic density, trabecular number and stiffness in the defect area 2 weeks after implantation than control and PBAT groups.

9.
Colloids Surf B Biointerfaces ; 194: 111192, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32599503

RESUMO

Cartilage is one of our body's tissues which are not repaired automatically by itself. Problems associated with cartilage are very common worldwide and are considered the leading cause of pain and disability. Smart biomaterial or "Four dimensional" (4D) biomaterials has started emerging as a suitable candidate, which are principally three dimensional (3D) materials that change their morphology or generate a response measured at space and time to physiologic stimuli. In this context, the release of oxygen through hydrogels in contact with water is considered as 4D biomaterials. The objective of this study is to develop strategies to release oxygen in a sustainable and prolonged manner through hydrogels systems to promote chondrocytes survival in oxygen-free environment. The 4D biomaterials are engineered from gelatin methacryloyl (GelMA) loaded with calcium peroxide (CPO), which have the ability to generate oxygen in a controlled and sustained manner for up to 6 days. The incorporation of CPO into the hydrogel system provided materials with enhanced mechanical and porosity properties. Furthermore, the hydrogels promoted chondrocyte survival and reduced cell death under oxygen-free conditions.


Assuntos
Condrócitos , Hidrogéis , Gelatina , Oxigênio , Engenharia Tecidual
10.
J Funct Biomater ; 11(1)2020 Feb 22.
Artigo em Inglês | MEDLINE | ID: mdl-32098431

RESUMO

Stereolithography technology associated with the employment of photocrosslinkable, biocompatible, and bioactive hydrogels have been widely used. This method enables 3D microfabrication from images created by computer programs and allows researchers to design various complex models for tissue engineering applications. This study presents a simple and fast home-made stereolithography system developed to print layer-by-layer structures. Polyethylene glycol diacrylate (PEGDA) and gelatin methacryloyl (GelMA) hydrogels were employed as the photocrosslinkable polymers in various concentrations. Three-dimensional (3D) constructions were obtained by using the stereolithography technique assembled from a commercial projector, which emphasizes the low cost and efficiency of the technique. Lithium phenyl-2,4,6-trimethylbenzoyl phosphonate (LAP) was used as a photoinitiator, and a 404 nm laser source was used to promote the crosslinking. Three-dimensional and vascularized structures with more than 5 layers and resolutions between 42 and 83 µm were printed. The 3D printed complex structures highlight the potential of this low-cost stereolithography technique as a great tool in tissue engineering studies, as an alternative to bioprint miniaturized models, simulate vital and pathological functions, and even for analyzing the actions of drugs in the human body.

11.
Spectrochim Acta A Mol Biomol Spectrosc ; 228: 117818, 2020 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-31780307

RESUMO

Raman spectroscopy and scanning electron microscopy (SEM) were used to investigate the effect of coating materials and acidulated phosphate fluoride gel (APF) treatment on dentin before and after erosion-abrasion cycles. A multi-walled carbon nanotube/graphene oxide hybrid carbon-based material (MWCNTO-GO), nanohydroxyapatite (nHAp), or a combined composite (nHAp/MWCNTO-GO) were used as a coating. Seventy root dentin fragments obtained from 40 bovine teeth were prepared and divided into groups (n = 10): negative control, artificial saliva - C, positive control - APF; nHAp; MWCNTO-GO; APF_nHAp; APF_MWCNTO-GO and APF_nHAp/MWCNTO-GO. All samples were subjected to cycles of demineralization (orange juice, pH ~3.7, room temperature, 1 min) followed by remineralization (saliva, 37 °C, 1 h). The remineralization procedures were followed by tooth brushing (150 strokes). The above cycle was repeated 3×/day for 5 days. The previous APF treatment of dentin allowed a better affinity of nHAp and MWCNTO-GO with the inorganic and organic portion of dentin, respectively. This interaction indicates the formation of a protective layer for the dentin surface and for the collagen giving possible protection against erosion. SEM micrographs illustrated the formation of a protective layer after application of the biomaterials and that it was partially or totally removed after the erosion and abrasion. Raman spectroscopy combined with multivariate analysis could distinguish samples with respect to treatment efficacy. The APF_nHAP/MWCNT-GO composite has shown to be a promising material since it has binding characteristics both to the inorganic and organic portion of the dentin and reduced solubility. Mineral-to-matrix ratio (MMR) parameter analysis confirmed the binding capability of MWCNTO-GO-based materials to dentin.


Assuntos
Carbono/química , Esmalte Dentário/efeitos dos fármacos , Dentina/química , Fluoretos/química , Nanoestruturas/química , Nanotubos de Carbono/química , Análise Espectral Raman/métodos , Animais , Materiais Biocompatíveis/química , Bovinos , Durapatita/química , Grafite/química , Microscopia Eletrônica de Varredura , Análise Multivariada , Saliva/metabolismo , Solubilidade , Temperatura
12.
Mater Sci Eng C Mater Biol Appl ; 103: 109778, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31349506

RESUMO

Herein, we electrospun ultrathin core-shell fibers based on polycaprolactone (PCL), polyethylene glycol (PEG), gelatin and osteogenic growth peptide (OGP), and evaluated their potential to upregulate human osteoblast cells (hFOB) and to reduce Gram-positive and Gram-negative bacteria. We also evaluated the fiber morphology, chemical structure and peptide delivery efficacy. The employment of core-shell fibers compared to fibers without a core-shell showed improved mechanical strength, comparable to the strength of pure PCL, as well as improved hydrophilicity and wettability. The careful selection of polymer combination and core-shell strategy promoted a controlled and sustained release of OGP. Moreover, increased calcium deposition (CD) (1.3-fold) and alkaline phosphate (ALP) activity was observed when hFOBs were cultivated onto core-shell fibers loaded with OGP after 21 days of culture. Our developed scaffolds were also able to reduce the amount of Pseudomonas aeruginosa (ATCC 25668) bacteria by a factor of two compared to raw PCL without the use of any antibiotics. All of these results demonstrate a promising potential of the developed core-shell electrospun scaffolds based on PCL:PEG:Gelatin:OGP for numerous bone tissue applications.


Assuntos
Diferenciação Celular/efeitos dos fármacos , Histonas , Peptídeos e Proteínas de Sinalização Intercelular , Staphylococcus aureus Resistente à Meticilina/crescimento & desenvolvimento , Osteoblastos/metabolismo , Pseudomonas aeruginosa/crescimento & desenvolvimento , Engenharia Tecidual , Alicerces Teciduais/química , Linhagem Celular , Histonas/química , Histonas/farmacologia , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/química , Peptídeos e Proteínas de Sinalização Intercelular/farmacologia
13.
J Mater Sci Mater Med ; 30(7): 81, 2019 Jun 28.
Artigo em Inglês | MEDLINE | ID: mdl-31254104

RESUMO

Carbon nanotubes combine high bend and mechanical strength, which is advantageous for many structural and biomedical purposes. Recently, some biomaterials, based on carbon nanostructures and nanohydroxyapatite (nHAp), have been investigated as bone substitutes in order to improve regeneration. The aim of this study was to access the expression of some RNA transcripts (involved in the process of osteoblast differentiation) by mesenchymal stem cells cultured over different nanocomposite surfaces. A multi-walled carbon nanotube (MWCNT) was firstly grown using chemical vapor deposition and then exfoliated using chemical and oxygen plasma treatments to obtain graphene nanoribbons (GNR). The hybrid composites nHAp/GNR were prepared using the wet method assisted by ultrasound irradiation with different amounts of GNR (1.0, 2.0 and 3.0 wt %). Five groups were tested in cell cultures. Group 1: synthesized nHAp; Group 2: synthesized GNR; Group 3: nHAp and 1.0% of GNR; Group 4: nHAp and 2.0% of GNR and group 5: nHAp and 3.0% of GNR. Real time reverse transcription polymerase chain reactions were performed, and all data was submitted to Kruskal Wallis and Dunn tests, at a significance level of 5%. As a result, three nanocomposites with different proportions of GNR were successfully produced. After cell culture, the expression of osteogenic genes demonstrated no significant differences among the groups and periods. However, bone morphogenetic protein II (BMP II), integrin binding sialoprotein (IBSP), and Osterix highest expressions were observed in the group containing 3.0% of GNR. In conclusion, our hybrid composites may be useful in bone interventions requiring mesenchymal stem cell differentiation into osteoblasts for healing.


Assuntos
Durapatita/química , Grafite/química , Nanotubos de Carbono/química , Osteogênese , Engenharia Tecidual/métodos , Bioensaio , Proteína Morfogenética Óssea 2/metabolismo , Células Cultivadas , Humanos , Sialoproteína de Ligação à Integrina/metabolismo , Células-Tronco Mesenquimais/citologia , Nanocompostos/química , Osteoblastos/citologia , Oxigênio/química , RNA/análise , Fator de Transcrição Sp7/metabolismo , Estresse Mecânico , Alicerces Teciduais/química
14.
Materials (Basel) ; 12(8)2019 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-31022828

RESUMO

Here, butylene adipate-co-terephthalate/polypyrrole with nanohydroxyapatite (PBAT/PPy/nHAp) scaffolds were fabricated and characterized. The electrospinning process was carried out using 12 kV, a needle of 23 G, an infusion pump set at 0.3 mL/h, and 10 cm of distance. Afterwards, nHAp was directly electrodeposited onto PBAT/PPy scaffolds using a classical three-electrode apparatus. For in vivo assays (comet assay, acute and chronic micronucleus), 60 male albino Wistar rats with 4 groups were used in each test (n = 5): PBAT/PPy; PBAT/PPy/nHAp; positive control (cyclophosphamide); and the negative control (distilled water). Peripheral blood samples were collected from the animals to perform the comet test after 4 h (for damage) and 24 h (for repair). In the comet test, it was shown that the scaffolds did not induce damage to the % DNA tail and neither for tail length. After the end of 48 h (for acute micronucleus) and 72 h (for chronic micronucleus), bone marrow was collected from each rat to perform the micronucleus test. All of the produced scaffolds did not present genotoxic effects, providing strong evidence for the biological application of PBAT/PPy/nHAp scaffolds.

15.
Int J Nanomedicine ; 14: 1725-1736, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30880976

RESUMO

BACKGROUND: Nanophase surface properties of titanium alloys must be obtained for a suitable biological performance, particularly to facilitate cell adhesion and bone tissue formation. Obtaining a bulk nanostructured material using severe plastic deformation is an ideal processing route to improve the mechanical performance of titanium alloys. By decreasing the grain size of a metallic material, a superior strength improvement can be obtained, while surface modification of a nanostructured surface can produce an attractive topography able to induce biological responses in osteoblastic cells. METHODS: Aiming to achieve such an excellent synergetic performance, a processing route, which included equal channel angular pressing (ECAP), hot and cold extrusion, and heat treatments, was used to produce a nanometric and ultrafine-grained (UFG) microstructure in the Ti-6Al-7Nb alloy (around of 200 nm). Additionally, UFG samples were surface-modified with acid etching (UFG-A) to produce a uniform micron and submicron porosity on the surface. Subsequently, alkaline treatment (UFG-AA) produced a sponge-like nanotopographic substrate able to modulate cellular interactions. RESULTS: After several kinds of biological tests for both treatment conditions (UFG-A and UFG-AA), the main results have shown that there was no cytotoxicity, expressed alkaline phosphatase activity and total protein amounts without statistical differences compared to control. However, the UFG-AA samples presented an attractive effect on the cell membranes, and cell adhesions were preferentially induced as compared with UFG-A. Both conditions demonstrated cell projections, but for UFG-AA, cells were more widely dispersed, and more quantities of filopodia formation could be observed. CONCLUSION: Herein, the reasons for such behaviors are discussed, and further results are presented in addition to those mentioned above.


Assuntos
Tecnologia Biomédica/métodos , Tamanho da Partícula , Titânio/farmacologia , Líquidos Corporais/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Humanos , Teste de Materiais , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Molhabilidade
16.
Materials (Basel) ; 12(6)2019 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-30871217

RESUMO

Treatment of articular cartilage lesions remains an important challenge. Frequently the bone located below the cartilage is also damaged, resulting in defects known as osteochondral lesions. Tissue engineering has emerged as a potential approach to treat cartilage and osteochondral defects. The principal challenge of osteochondral tissue engineering is to create a scaffold with potential to regenerate both cartilage and the subchondral bone involved, considering the intrinsic properties of each tissue. Recent nanocomposites based on the incorporation of nanoscale fillers into polymer matrix have shown promising results for the treatment of osteochondral defects. In this present study, it was performed using the recently developed methodologies (electrodeposition and immersion in simulated body fluid) to obtain porous superhydrophilic poly(d,l-lactic acid)/vertically aligned carbon nanotubes/nanohydroxyapatite (PDLLA/VACNT-O:nHAp) nanocomposite scaffolds, to analyze cell behavior and gene expression of chondrocytes, and then assess the applicability of this nanobiomaterial for osteochondral regenerative medicine. The results demonstrate that PDLLA/VACNT-O:nHAp nanocomposite supports chondrocytes adhesion and decreases type I Collagen mRNA expression. Therefore, these findings suggest the possibility of novel nanobiomaterial as a scaffold for osteochondral tissue engineering applications.

17.
Int J Nanomedicine ; 14: 865-874, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30774339

RESUMO

BACKGROUND: It has been difficult to find bioactive compounds that can optimize bone repair therapy and adequate osseointegration for people with osteoporosis. The nano-hydroxyapatite (nHAp)/carbon nanotubes with graphene oxides, termed graphene nanoribbons (GNR) composites have emerged as promising materials/scaffolds for bone regeneration due to their bioactivity and osseointegration properties. Herein, we evaluated the action of nHAp/GNR composites (nHAp/GNR) to promote bone regeneration using an osteoporotic model. MATERIALS AND METHODS: First, three different nHAp/GNR (1, 2, and 3 wt% of GNR) were produced and characterized. For in vivo analyses, 36 Wistar rats (var. albinus, weighing 250-300 g, Comissão de Ética no Uso de Animais [CEUA] n.002/17) were used. Prior to implantation, osteoporosis was induced by oophorectomy in female rats. After 45 days, a tibial fracture was inflicted using a 3.0-mm Quest trephine drill. Then, the animals were separated into six sample groups at two different time periods of 21 and 45 days. The lesions were filled with 3 mg of one of the above samples using a curette. After 21 or 45 days of implantation, the animals were euthanized for analysis. Histological, biochemical, and radiographic analyses (DIGORA method) were performed. The data were evaluated through ANOVA, Tukey test, and Kolmogorov-Smirnov test with statistical significance at P<0.05. RESULTS: Both nHAp and GNR exhibited osteoconductive activity. However, the nHAp/GNR exhibited regenerative activity proportional to their concentration, following the order of 3% >2% >1% wt. CONCLUSION: Therefore, it can be inferred that all analyzed nanoparticles promoted bone regeneration in osteoporotic rats independent of analyzed time.


Assuntos
Regeneração Óssea , Durapatita/química , Grafite/química , Nanotubos de Carbono/química , Osteoporose/patologia , Fosfatase Alcalina/metabolismo , Animais , Regeneração Óssea/efeitos dos fármacos , Modelos Animais de Doenças , Feminino , Nanotubos de Carbono/ultraestrutura , Osteoporose/fisiopatologia , Ovariectomia , Ratos Wistar , Tíbia/diagnóstico por imagem , Alicerces Teciduais/química
18.
Int J Nanomedicine ; 13: 7891-7903, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30538466

RESUMO

BACKGROUND: Here, electrospun fibers based on a blend of polycaprolactone (PCL), poly(ethylene glycol) (PEG), and gelatin methacryloyl (GelMA) were developed. The careful choice of this polymer combination allowed for the preparation of a biomaterial that preserved the mechanical strength of PCL, while at the same time improving the hydrophilicity of the blended material and human osteoblast maturation. METHODS: The morphology, chemical structure, wettability, and mechanical properties before and after UV photocrosslinking were evaluated. Furthermore, human osteoblasts (hFOB) were cultivated for up to 21 days on the scaffolds, and their potential to upregulate cell proliferation, alkaline phosphatase (ALP) activity, and calcium deposition were investigated. RESULTS: Contact angle measurement results showed that the developed scaffolds presented hydrophilic properties after PEG and GelMA incorporation before (25°) and after UV photocross-linking (69°) compared to pure PCL (149°). PCL:PEG:GelMA-UV displayed a slight increase in mechanical strength (elastic modulus ~37 MPa) over PCL alone (~33 MPa). Normally, an increase in strength of fibers leads to a decrease in elongation at break, due to the material becoming less deformable and stiffer, thus leading to breaks at low strain. This behavior was observed by comparing PCL (elongation at break ~106%) and PCL:PEG:GelMA-UV (~50%). Moreover, increases in ALP activity (10-fold at day 14) and calcium deposition (1.3-fold at day 21) by hFOBs were detected after PEG and GelMA incorporation after UV photocross-linking compared to pure PCL. Ultrathin and hydrophilic fibers were obtained after PEG and GelMA incorporation after UV photocrosslinking, but the strength of PCL was maintained. Interestingly, those ultrathin fiber characteristics improved hFOB functions. CONCLUSION: These findings appear promising for the use of these electrospun scaffolds, based on the combination of polymers used here for numerous orthopedic applications.


Assuntos
Teste de Materiais , Nanofibras/química , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Fosfatase Alcalina/metabolismo , Animais , Módulo de Elasticidade , Gelatina/química , Humanos , Metacrilatos/química , Nanofibras/ultraestrutura , Osteoblastos/citologia , Osteogênese/fisiologia , Poliésteres/química , Polietilenoglicóis/química , Espectroscopia de Infravermelho com Transformada de Fourier , Tensão Superficial , Sus scrofa
19.
PLoS One ; 13(12): e0209386, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30571704

RESUMO

Herein, we report the design of electrospun ultrathin fibers based on the combination of three different polymers polycaprolactone (PCL), polyethylene glycol (PEG), and gelatin methacryloyl (GelMA), and their potential bactericidal activity against three different bacteria Staphylococcus aureus (S. aureus), Pseudomonas aeruginosa (P. aeruginosa), and Methicillin-resistant Staphylococcus aureus (MRSA). We evaluated the morphology, chemical structure and wettability before and after UV photocrosslinking of the produced scaffolds. Results showed that the developed scaffolds presented hydrophilic properties after PEG and GelMA incorporation. Moreover, they were able to significantly reduce gram-positive, negative, and MRSA bacteria mainly after UV photocrosslinking (PCL:PEG:GelMa-UV). Furthermore, we performed a series of study for gaining a better mechanistic understanding of the scaffolds bactericidal activity through protein adsorption study and analysis of the reactive oxygen species (ROS) levels. Furthermore, the in vivo subcutaneous implantation performed in rats confirmed the biocompatibility of our designed scaffolds.


Assuntos
Materiais Biocompatíveis/farmacologia , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Nanofibras/química , Pseudomonas aeruginosa/efeitos dos fármacos , Alicerces Teciduais/química , Animais , Materiais Biocompatíveis/química , Linhagem Celular , Gelatina/química , Humanos , Masculino , Teste de Materiais/métodos , Metacrilatos/química , Testes de Sensibilidade Microbiana , Modelos Animais , Poliésteres/química , Polietilenoglicóis/química , Ratos , Ratos Wistar , Engenharia Tecidual/métodos , Infecção dos Ferimentos/microbiologia , Infecção dos Ferimentos/prevenção & controle
20.
Microsc Res Tech ; 81(12): 1456-1466, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30295365

RESUMO

This study evaluated the antierosive effect applying a modified acidulated phosphate fluoride (APF) gel containing nano-hydroxyapatite (nHAp) on the enamel surface before erosion. After polishing, the exposed flat enamel surfaces (n = 7/group) from bovine incisors were treated with artificial saliva (S - negative control), orange juice (ERO), APF gel (positive control) and APF_nHAp gel. All samples were subjected to six cycles of demineralization (orange juice, pH ~ 3.5, 10 min) followed by remineralization (saliva, 37°C, 1 hr). The enamel surface morphology, topography, and inorganic composition were analyzed using scanning electron microscopy, roughness testing, and micro energy-dispersive X-ray fluorescence spectrometry, respectively. The mean (standard deviation) roughness values (Ra, µm) were S, 0.13 (0.05); ERO, 0.25 (0.07); APF, 0.22 (0.08); and APF_nHAp, 0.17 (0.04). Ra values were significantly higher after ERO (p < .01) and APF (p < .05) treatments than after S. The enamel surface morphology was altered by the treatments, except for the S specimens. The mineral content of the enamel showed a clear trend with Ca and P reduction in the order of APF < S < APF_nHAp < ERO and APF < S < ERO < APF_nHAp, respectively. We can conclude that APF gel increased mineral concentration on the enamel. Moreover, the APF_nHAp material modified the composition and morphology of the enamel surface.


Assuntos
Fluoreto de Fosfato Acidulado/administração & dosagem , Esmalte Dentário/efeitos dos fármacos , Durapatita/administração & dosagem , Erosão Dentária/prevenção & controle , Fluoreto de Fosfato Acidulado/química , Animais , Bovinos , Esmalte Dentário/metabolismo , Esmalte Dentário/ultraestrutura , Durapatita/química , Géis/administração & dosagem , Géis/química , Humanos , Microscopia Eletrônica de Varredura , Minerais/metabolismo , Desmineralização do Dente/metabolismo , Desmineralização do Dente/prevenção & controle , Erosão Dentária/metabolismo
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