Development and characterization of ceramic-polymeric hybrid scaffolds for bone regeneration: incorporating of bioactive glass BG-58S into PDLLA matrix.

In recent years, there has been a notable surge of interest in hybrid materials within the biomedical field, particularly for applications in bone repair and regeneration. Ceramic-polymeric hybrid scaffolds have shown promising outcomes. This study aimed to synthesize bioactive glass (BG-58S) for integration into a bioresorbable polymeric matrix based on PDLLA, aiming to create a bioactive scaffold featuring stable pH levels. The synthesis involved a thermally induced phase separation process followed by lyophilization to ensure an appropriate porous structure. BG-58S characterization revealed vitreous, bioactive, and mesoporous structural properties. The scaffolds were analyzed for morphology, interconnectivity, chemical groups, porosity and pore size distribution, zeta potential, pH, in vitro degradation, as well as cell viability tests, total protein content and mineralization nodule production. The PDLLA scaffold displayed a homogeneous morphology with interconnected macropores, while the hybrid scaffold exhibited a heterogeneous morphology with smaller diameter pores due to BG-58S filling. The hybrid scaffold also demonstrated a pH buffering effect on the polymer surface. In addition to structural characteristics, degradation tests indicated that by incorporating BG-58S modified the acidic degradation of the polymer, allowing for increased total protein production and the formation of mineralization nodules, indicating a positive influence on cell culture.

Development, characterization, and biological study of bioglass coatings 45S5 and BioK on zirconia implant surfaces.

Zirconia implants are gaining attention as a viable alternative to titanium implants due to their comparable osseointegration development, improved soft tissue adaptation, and enhanced aesthetics. An encouraging avenue for improving zirconia implant properties involves the potential application of bioactive coatings to their surfaces. These coatings have shown potential for inducing hydroxyapatite formation, crucial for bone proliferation, and improving implant mechanical properties. This study aimed to evaluate the effect of coating zirconia implants with two bioactive glasses, 45S5 and BioK, on osteogenesis in vitro and osseointegration in vivo. Zirconia samples and implants were prepared using Zpex zirconia powder and blocks, respectively. The samples were divided into three groups: polished zirconia (ZRC), zirconia coated with 45S5 bioglass (Z + 45S5), and zirconia coated with BioK glass (Z + BK). Coatings were applied using a brush and sintered at 1200°C. Chemical analysis of the coatings was carried out using x-ray diffraction and Fourier Transform Infrared Spectroscopy. Surface topography and roughness were characterized using scanning electron microscopy and a roughness meter. In vitro experiments used mesenchymal cells from Wistar rat femurs, and the coated zirconia implants were found to promote cell viability, protein synthesis, alkaline phosphatase activity, and mineralization, indicating enhanced osteogenesis. In vivo experiments with 18 rats showed positive results for bone formation and osseointegration through histological and histomorphometric analysis and a push-out test. The findings indicate that bioactive glass coatings have the potential to improve cell differentiation, bone formation, and osseointegration in zirconia implants.

Chlorinated-based bioceramics incorporated in polycaprolactone membranes.

The development of bioactive membranes with bone repair properties is great interest in the field of tissue engineering. In this study, we aimed to fabricate and characterize a composite membrane composed of sol-gel synthesized bioceramics and electrospun polycaprolactone (PCL) fibers for bone tissue regeneration applications. The bioceramics were prepared using the sol-gel method with nitrate (N) and chloride (CL) as precursors. PCL and bioceramic solutions were electrospun to obtain ultrafine fiber mats. Raman spectroscopy, x-ray diffraction (XRD), Fourier Transform Infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and scanning electron microscopy (SEM) were used to characterize the materials. The results showed that both chlorinated and non-chlorinated bioceramics contained NBOs (non-bridge bonds) and crystallized the α-wollastonite phase, with the chlorinated version doing so at lower temperatures. In vitro tests were performed to evaluate cytotoxicity, cell adhesion, and mineralized matrix formation on the membranes. The composite membranes showed improved cell viability and promoted mineralization nodules formation. This study presents a promising approach for the development of bioactive membranes for bone tissue engineering, with potential applications in bone regeneration therapies.

Carbon fiber: Characterization and evaluation of the inflammatory response and toxicity in rats.

This study aimed to evaluate the Carbon Fiber obtained from PAN textile and cotton fiber in their different forms of presentation: non-activated carbon fiber felt (NACFF), activated carbon fiber felt (ACFF), silver activated carbon fiber felt (Ag-ACFF), and activated carbon fiber tissue (ACFT), to obtain scaffolds as a potential material with properties related to the synthetic bone graft. Characterization tests performed: surface wettability, traction, swelling, and in vivo tests: evaluation of the inflammatory response by implanting the materials in the subcutaneous tissue of 14 Wistar rats, evaluation of collagen fibers by picrosirius red staining and assessment of toxicity in the following organs: heart, spleen, liver, and kidney. In the wettability test, NACFF and ACFT were hydrophobic (θ124° and 114°), ACFF and Ag-ACFF were hydrophilic. For maximum stress, ACFF was more resistant (2.983 ± 1.059) p < .05. In the swelling test, the Ag-ACFF and ACFF groups showed the highest absorption percentage for the PBS solution and distilled water (p < .001). The organs showed no signs of acute systemic toxicity. The implant regions showed mild to moderate inflammatory infiltrate at 7 and 21 days. Only the ACFT group did not show the maturation of type I collagen fibers in 21 days. Through the conducted analyses, the ACFT shows little potential to be indicated as a possible scaffold. Therefore NACFF, ACFF, and Ag-ACFF have the potential to be considered scaffolds due to the following characteristics presented: good absorption rate, hydrophilicity, and non-toxic.

The Local Release of Teriparatide Incorporated in 45S5 Bioglass Promotes a Beneficial Effect on Osteogenic Cells and Bone Repair in Calvarial Defects in Ovariectomized Rats.

With the increase in the population's life expectancy, there has also been an increase in the rate of osteoporosis, which has expanded the search for strategies to regenerate bone tissue. The ultrasonic sonochemical technique was chosen for the functionalization of the 45S5 bioglass. The samples after the sonochemical process were divided into (a) functionalized bioglass (BG) and (b) functionalized bioglass with 10% teriparatide (BGT). Isolated mesenchymal cells (hMSC) from femurs of ovariectomized rats were differentiated into osteoblasts and submitted to in vitro tests. Bilateral ovariectomy (OVX) and sham ovariectomy (Sham) surgeries were performed in fifty-five female Wistar rats. After a period of 60 days, critical bone defects of 5.0 mm were created in the calvaria of these animals. For biomechanical evaluation, critical bone defects of 3.0 mm were performed in the tibias of some of these rats. The groups were divided into the clot (control) group, the BG group, and the BGT group. After the sonochemical process, the samples showed modified chemical topographic and morphological characteristics, indicating that the surface was chemically altered by the functionalization of the particles. The cell environment was conducive to cell adhesion and differentiation, and the BG and BGT groups did not show cytotoxicity. In addition, the experimental groups exhibited characteristics of new bone formation with the presence of bone tissue in both periods, with the BGT group and the OVX group statistically differing from the other groups (p < 0.05) in both periods. Local treatment with the drug teriparatide in ovariectomized animals promoted positive effects on bone tissue, and longitudinal studies should be carried out to provide additional information on the biological performance of the mutual action between the bioglass and the release of the drug teriparatide.

Polycaprolactone/chlorinated bioglass scaffolds doped with Mg and Li ions: Morphological, physicochemical, and biological analysis.

The objective was to synthesize and characterize fine polycaprolactone (PCL) fibers associated with a new 58S bioglass obtained by the precipitated sol-gel route, produced by the electrospinning process in order to incorporate therapeutic ions (Mg and Li). In PCL/acetone solutions were added 7% pure bioglass, bioglass doped with Mg(NO3 )2 and Li2 CO3 and were subjected to electrospinning process. The fibers obtained were characterized morphologically, chemically and biologically. The results showed the presence of fine fibers at the nanometric scale and with diameters ranging from 0.67 to 1.92 µm among groups. Groups containing bioglass showed particles both inside and on the surface of the fibers. The components of the polymer, bioglass and therapeutic ions were present in the fibers produced. The produced fibers showed cell viability and induced the formation of mineralization nodules. It was observed the applicability of that methodology in making an improved biomaterial, which adds the osteoinductive properties of the bioglass to PCL and to those of therapeutic ions, applicable to guided bone regeneration.

Electrospun polylactic acid scaffolds with strontium- and cobalt-doped bioglass for potential use in bone tissue engineering applications.

The development of nanoscale biomaterials associated with polymers has been growing over the years, due to their important structural characteristics for applications in biological systems. The present study aimed to produce and test polymeric scaffolds composed of polylactic acid (PLA) fibers associated with a 58S bioglass doped with therapeutic ions for use in tissue engineering. Three 58S Bioglass was obtained by the sol-gel route, pure and doped with 5% strontium and cobalt ions. Solutions of 7% PLA was used as control and added the three different bioglass, 4% of 58S bioglass (PLA-BG), 4% bioglass-doped strontium (PLA-BGSr) and 4% bioglass-doped cobalt (PLA-BGCo). Scaffolds were produced through electrospinning process, and was characterized chemical and morphologically. The in vitro tests were performed using mesenchymal cells cultures from femurs of nine rats, grown in osteogenic supplemented total culture medium. After osteoblastic differentiation induction cell viability, alkaline phosphatase activity, total protein content quantification, and visualization of mineralization nodule tests were performed. Analysis of normal distribution used the Shapiro-Wilk test (nanofibers diameter and biological assay). Data were compared using the Kruskal-Wallis nonparametric test (p = 0.05). The bioglasses produced proved to be free of nitrate, chlorinated and nano-sized, with effective incorporation of therapeutic ions in their structure. All materials showed cell viability (>70%), total protein production, and alkaline phosphatase activity. It was possible to develop polylactic acid scaffolds associated with 58S bioglass doped with therapeutic ions without cytotoxicity. Scaffolds characteristics appear to sustain its application in bone tissue engineering.

Characterization of Optimized TiO2 Nanotubes Morphology for Medical Implants: Biological Activity and Corrosion Resistance.

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.

Evaluation of pulsed electromagnetic field protocols in implant osseointegration: in vivo and in vitro study.

OBJECTIVES: The present study aims to evaluate two protocols of pulsed electromagnetic field (PEMF) on osseointegration and establish one that addresses ideal parameters for its use in dentistry, especially in the optimization of the implants osseointegration process. MATERIALS AND METHODS: Sixty male rats (Wistar) were allocated into three experimental groups: control (GC), test A (GTA, 3 h exposed), and test B (GTB, 1 h exposed). All animals received titanium implants in both tibias, and PEMF application (15 Hz, ± 1 mT, 5 days/week) occurred only in the test groups. They were euthanized at 03, 07, 21, and 45 days after PEMF therapy. Removal torque, histomorphometric measurements, three-dimensional radiographic evaluation, and in vitro biological assay analyses were performed. RESULTS: GTB showed better results compared with GTA in removal torque tests, in bone volume and bone mineral density, cell viability, total protein content, and mineralization nodules (p < 0.05). GTA showed better performance in trabecular bone thickness and cell proliferation compared with GTB (p < 0.05), especially at osseointegration early periods. In the histomorphometric analysis and number of trabeculae, there were no differences in the test groups. CONCLUSION: PEMF as a biostimulator was effective in optimizing the events in bone tissue that lead to osseointegration, especially when applied for a shorter time and in the initial periods of bone healing. CLINICAL RELEVANCE: The PEMF therapy is an effective alternative method for optimizing bone healing.

Biological and microbiological interactions of Ti-35Nb-7Zr alloy and its basic elements on bone marrow stromal cells: good prospects for bone tissue engineering.

BACKGROUND: An effective biomaterial for bone replacement should have properties to avoid bacterial contamination and promote bone formation while inducing rapid cell differentiation simultaneously. Bone marrow stem cells are currently being investigated because of their known potential for differentiation in osteoblast lineage. This makes these cells a good option for stem cell-based therapy. We have aimed to analyze, in vitro, the potential of pure titanium (Ti), Ti-35Nb-7Zr alloy (A), niobium (Nb), and zirconia (Zr) to avoid the microorganisms S. aureus (S.a) and P. aeruginosa (P.a). Furthermore, our objective was to evaluate if the basic elements of Ti-35Nb-7Zr alloy have any influence on bone marrow stromal cells, the source of stem cells, and observe if these metals have properties to induce cell differentiation into osteoblasts. METHODS: Bone marrow stromal cells (BMSC) were obtained from mice femurs and cultured in osteogenic media without dexamethasone as an external source of cell differentiation. The samples were divided into Ti-35Nb-7Zr alloy (A), pure titanium (Ti), Nb (niobium), and Zr (zirconia) and were characterized by scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDS). After predetermined periods, cell interaction, cytotoxicity, proliferation, and cell differentiation tests were performed. For monotypic biofilm formation, standardized suspensions (106 cells/ml) with the microorganisms S. aureus (S.a) and P. aeruginosa (P.a) were cultured for 24 h on the samples and submitted to an MTT test. RESULTS: All samples presented cell proliferation, growth, and spreading. All groups presented cell viability above 70%, but the alloy (A) showed better results, with statistical differences from Nb and Zr samples. Zr expressed higher ALP activity and was statistically different from the other groups (p < 0.05). In contrast, no statistical difference was observed between the samples as regards mineralization nodules. Lower biofilm formation of S.a and P.a. was observed on the Nb samples, with statistical differences from the other samples. CONCLUSION: Our results suggest that the basic elements present in the alloy have osteoinductive characteristics, and Zr has a good influence on bone marrow stromal cell differentiation. We also believe that Nb has the best potential for reducing the formation of microbial biofilms.

Synergistic effect of adding bioglass and carbon nanotubes on poly (lactic acid) porous membranes for guided bone regeneration.

Polymer membranes have been widely used in guided bone regeneration (GBR), especially when it comes to their use in dentistry. Poly (lactic acid) (PLA) have good mechanical properties such as flexibility, which allows the material to be moldable and also has biocompatibility and biodegradation. Besides that, bioglass (BG) incorporated into the polymer matrix can promote osteoinduction properties and osteoconduction properties to the polymer-ceramic biocomposite. The membranes are also required to exhibit antimicrobial activity to prevent or control the proliferation of pathogenic microorganisms, and the addition of carbon nanotubes (CNT) can assist in this property. The porous membranes of PLA with the addition of different contents of BG and CNT were obtained by solvent casting in controlled humidity method, and the synergistic effect of the addition of both fillers were investigated. The membranes showed pores (3-11 µm) on their surface. The addition of 5 wt% BG causes an increase in the surface porosity and bioactivity properties of the PLA. The agar diffusion test showed antimicrobial activity in the membranes with addition of CNT. In vitro results showed that the porous membranes were not cytotoxic and allowed cell activity and differentiation. Thus, BG collaborated to increase biological activity while CNT contributed to microbial activity, creating a synergistic effect on PLA porous membranes, being this effect more evident for PLA/5BG/1.0CNT. These results indicated a promising use of this new biomaterial for the production of porous membranes for GBR.

Scaffolds of PCL combined to bioglass: synthesis, characterization and biological performance.

Biomaterials may be useful in filling lost bone portions in order to restore balance and improve bone regeneration. The objective of this study was to produce polycaprolactone (PCL) membranes combined with two types of bioglass (Sol-Gel and melt-quenched) and determine their physical and biological properties. Membranes were produced through electrospinning. This study presented three experimental groups: pure PCL membranes, PCL-Melt-Bioglass and PCL-Sol-gel-Bioglass. Membranes were characterized using Scanning Electron Microscopy, Fourier Transform Infrared Spectrophotometry (FTIR), Energy-Dispersive Spectroscopy and Zeta Potential. The following in vitro tests were performed: MTT assay, alkaline phosphatase activity, total protein content and mineralization nodules. Twenty-four male rats were used to observe biological performance through radiographic, fracture energy, histological and histomorphometric analyses. The physical and chemical analysis results showed success in manufacturing bioactive membranes which significantly enhanced cell viability and osteoblast differentiation. The new formed bone from the in vivo experiment was similar to that observed in the control group. In conclusion, the electrospinning enabled preparing PCL membranes with bioglass incorporated into the structure and onto the surface of PCL fibers. The microstructure of the PCL membranes was influenced by the bioglass production method. Both bioglasses seem to be promising biomaterials to improve bone tissue regeneration when incorporated into PCL.

Systemic and local effects of radiotherapy: an experimental study on implants placed in rats.

OBJECTIVES: Evaluate the modulating effect of ionizing radiation, blood cytokine levels, and bone remodeling of the interface around the implant to understand the radiation mechanisms which can impair the implants receptor site. MATERIAL AND METHODS: Sixty rats were submitted to grade V titanium implants in the femurs and were divided into the following groups: no-irradiation (N-Ir): control group with implant only; early-irradiation (E-Ir): implant + irradiation after 24 h; late-irradiation (L-Ir): implant + irradiation after 4 weeks; and previous-irradiation (P-Ir): irradiation + implant after 4 weeks. The animals in the E-Ir, L-Ir, and P-Ir groups were irradiated in two fractional stages of 15 Gy. At 3 days, 2 weeks, and 7 weeks after the final procedure, five animals were randomly euthanized per group. Serum levels of TNF-ɑ, IL-1ß, TGF-ß, IL-6, M-CSF, and IL-10 were measured from blood collected prior to euthanasia using the ELISA test. The pieces containing the implants were subjected to immunohistochemical labeling using the tartrate acid resistant to phosphatase, osteocalcin, and caspase-3 markers and mCT. The ANOVA test was used for statistical analysis, and the Tukey multiple comparison test (p < 0.05) was applied. RESULTS: The results indicated that ionizing radiation modifies the production of pro- and anti-inflammatory serum cytokines, the expression of proteins involved in bone remodeling and cellular apoptosis, as well as changes in bone formation. CONCLUSIONS: The results suggests that a longer period between radiotherapy and implant placement surgery when irradiation occurs prior to implant installation would allow the recovery and renewal of bone cells and avoid future failures in osseointegration. CLINICAL RELEVANCE: The search for modifications caused by ionizing irradiation in bone tissue can indicate the ideal period for implant placement without affecting the osseointegration process.

Biological and microbiological behavior of calcium aluminate cement-based blend for filling of bone defects.

Calcium aluminate cement (CAC) as a biomaterial has been evaluated for its physical, mechanical and biocompatibility properties. Furthermore, the application of CAC for bone repair is due to its composition and coefficient of thermal expansion, which is similar to that of human bone. Thus, the aim of this study was to evaluate compositions of CAC-based blends as substitutes for bone defects. Five compositions of blends (alumina, zirconia, hydroxyapatite, tricalcium phosphate, chitosan), in addition to the base cement consisting of homogeneous CAC were evaluated as a substitute for bone repair. Additionally, the monotypic biofilm formation was assessed. Creation of a monocortical bone defect was performed on the femurs of rats, which were randomly filled with the different materials. The polymethylmethacrylate (PMMA) group was used as a control. All the animals were euthanized 04 weeks after the surgery procedure. Subsequently, computerized microtomography, histological and histomorphometric analyses were performed to verify the bone repair. To evaluate the formation of biofilms, reference strains of Staphylococcus aureus, Streptococcus mutans and Pseudomonas aeruginosa were cultured on the samples, and the biofilm formed was quantified by the MTT method. In the microtomography and histomorphometry results, it was observed that the blends exhibited better results than the control group, with statistically significant differences (p < 0.05) for alumina and zirconia blends. In the biofilm formation, a statistical difference (p < 0.05) in general was observed between the alumina blends and the control group (p < 0.05). It was concluded that CAC-based blends with alumina and zirconia are promising for use in fillings for bone repair.

Titanium alloys: in vitro biological analyzes on biofilm formation, biocompatibility, cell differentiation to induce bone formation, and immunological response.

Biological effects of titanium (Ti) alloys were analyzed on biofilms of Candida albicans, Enterococcus faecalis, Pseudomonas aeruginosa, Staphylococcus aureus, Streptococcus mutans, and Streptococcus sanguinis, as well as on osteoblast-like cells (MG63) and murine macrophages (RAW 264.7). Standard samples composed of aluminum and vanadium (Ti-6Al-4V), and sample containing niobium (Ti-35Nb) and zirconium (Ti-13Nb-13Zr) were analyzed. Monomicrobial biofilms were formed on the Ti alloys. MG63 cells were grown with the alloys and the biocompatibility (MTT), total protein (TP) level, alkaline phosphatase (ALP) activity, and mineralization nodules (MN) formation were verified. Levels of interleukins (IL-1ß and IL-17), tumor necrosis factor alpha (TNF-α), and oxide nitric (NO) were checked, from RAW 264.7 cells supernatants. Data were statically analyzed by one-way analysis of variance (ANOVA) and Tukey's test, or T-test (P ≤ 0.05). Concerning the biofilm formation, Ti-13Nb-13Zr alloy showed the best inhibitory effect on E. faecalis, P. aeruginosa, and S. aureus. And, it also acted similarly to the Ti-6Al-4V alloy on C. albicans and Streptococcus spp. Both alloys were biocompatible and similar to the Ti-6Al-4V alloy. Additionally, Ti-13Nb-13Zr alloy was more effective for cell differentiation, as observed in the assays of ALP and MN. Regarding the stimulation for release of IL-1ß and TNF-α, Ti-35Nb and Ti-13Nb-13Zr alloys inhibited similarly the synthesis of these molecules. However, both alloys stimulated the production of IL-17. Additionally, all Ti alloys showed the same effect for NO generation. Thus, Ti-13Nb-13Zr alloy was the most effective for inhibition of biofilm formation, cell differentiation, and stimulation for release of immune mediators.

In vitro osteogenesis process induced by hybrid nanohydroxyapatite/graphene nanoribbons composites.

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.

Diamond-like carbon films over reconstructive TMJ prosthetic materials: Effects in the cytotoxicity, chemical and mechanical properties.

Increasingly more young patients have been submitted to reconstruction of the Temporomandibular Joint (TMJ), so, the prostheses must to present more functional longevity. OBJECTIVE: To evaluate the effect of diamond-like carbon film (DLC) over titanium alloy (Ti6Al4V) and polyethylene (UHWPE) samples, their mechanical and chemical properties and cellular cytotoxicity. METHODS: Titanium and UHWPE specimens, with 2.5 cm in diameter and 2 mm thickness were coated through plasma enhanced chemical vapor deposition (PECVD) with DLC or DLC doped with silver (DLC-Ag). Scanning electron microscopy (SEM) morphological analysis, Energy-dispersive spectroscopy (EDS) chemical analysis, scratching test, mechanical fatigue test, surface roughness analysis, and cellular cytotoxicity were performed. Data were statistically analyzed using one-way ANOVA (p < 0.05) or two-way ANOVA and multiple comparison Tukey test. RESULTS: In the SEM analysis, morphological differences were observed on substrates after DLC deposition. The film chemically modified the substrate surfaces, according to the EDS analysis. The initial critical load failure occurred at 6.1 N for DLC and 9.7 N for the DLC-Ag film. The DLC film deposition over the polyethylene promoted a decrease in the polymer's damaged area after mechanical fatigue cycling. The cytotoxicity analysis demonstrated less biocompatibility in experimental groups, when compared to control, however, increased biocompatibility was observed, at 10 days, in all groups. CONCLUSION: The diamond-like carbon coating enhanced the chemical and mechanical properties from substrates, however modified biological interaction course of the titanium alloy (Ti6Al4V) and polyethylene (UHWPE) samples. Parameters for film deposition remain to be improved in order to obtain best biocompatibility.

Influence of Chronic Alcohol Use on Osteoblastic Differentiation of Bone Marrow Cells, Bone Properties, and Hepatic and Renal Morphology of Rats.

Chronic alcohol exposure can affect the osteoblastic activity and the proliferation and differentiation of cells due to its toxic effect, which can affect negatively bone repair and bone microarchitecture. The aim of this study was to evaluate the effects of chronic use of 20% alcohol on rats regarding osteoblastic differentiation, extrinsic and intrinsic properties of the tibia, and hepatic and renal morphology. Wistar rats were divided into three groups (n = 9) in accordance with a 24-week diet. After euthanasia, kidneys, liver, and tibias were removed for analysis and femurs mesenchymal cells were collected. The results showed that chronic use of 20% alcohol influenced neither the alkaline phosphatase production nor total protein (p > 0.05) in rats, with similar formation of nodules in all groups (p > 0.05). However, significant changes in the liver and kidneys and adverse effects on the mechanical properties of the tibia were observed. According to the results, it can be concluded that the chronic use of alcohol for 24 weeks had no negative influence on the activity and differentiation of osteoblasts, but the mechanical properties of the tibia were impaired and the organs responsible for metabolism and excretion were also affected due to the consumption of alcohol.

In vitro and in vivo biological performance of porous Ti alloys prepared by powder metallurgy.

Titanium (Ti) and Ti-6 Aluminium-4 Vanadium alloys are the most common materials in implants composition but ß type alloys are promising biomaterials because they present better mechanical properties. Besides the composition of biomaterial, many factors influence the performance of the biomaterial. For example, porous surface may modify the functional cellular response and accelerate osseointegration. This paper presents in vitro and in vivo evaluations of powder metallurgy-processed porous samples composed by different titanium alloys and pure Ti, aiming to show their potential for biomedical applications. The porous surfaces samples were produced with different designs to in vitro and in vivo tests. Samples were characterized with scanning electron microscopy (SEM), energy dispersive spectroscopy (EDS) and elastic modulus analyses. Osteogenic cells from newborn rat calvaria were plated on discs of different materials: G1-commercially pure Ti group (CpTi); G2-Ti-6Al-4V alloy; G3-Ti-13 Niobium-13 Zirconium alloy; G4-Ti-35 Niobium alloy; G5-Ti-35 Niobium-7 Zirconium-5 Tantalum alloy. Cell adhesion and viability, total protein content, alkaline phosphatase activity, mineralization nodules and gene expression (alkaline phosphatase, Runx-2, osteocalcin and osteopontin) were assessed. After 2 and 4 weeks of implantation in rabbit tibia, bone ingrowth was analyzed using micro-computed tomography (µCT). EDS analysis confirmed the material production of each group. Metallographic and SEM analysis revealed interconnected pores, with mean pore size of 99,5µm and mean porosity of 42%, without significant difference among the groups (p>0.05). The elastic modulus values did not exhibit difference among the groups (p>0.05). Experimental alloys demonstrated better results than CpTi and Ti-6Al-4V, in gene expression and cytokines analysis, especially in early experimental periods. In conclusion, our data suggests that the experimental alloys can be used for biomedical application since they contributed to excellent cellular behavior and osseointegration besides presenting lower elastic modulus.