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The present paper investigates the impact behaviour of both pristine carbon-fibre-reinforced-plastic (CFRP) composite laminates and repaired CFRP laminates. For the patch-repaired CFRP specimen, the pristine CFRP panel specimen has been damaged by cutting out a central disc of the CFRP material and then repaired using an adhesively bonded patch of CFRP to cover the hole. Drop-weight, impact tests are performed on these two types of specimens and a numerical elastic-plastic, three-dimensional damage model is developed and employed to simulate the impact behaviour of both types of specimen. This numerical model is meso-scale in nature and assumes that cracks initiate in the CFRP at a nano-scale, in the matrix around fibres, and trigger sub-micrometre intralaminar matrix cracks during the impact event. These localized regions of intralaminar cracking then lead to interlaminar, i.e. delamination, cracking between the neighbouring plies which possess different fibre orientations. These meso-scale, intralaminar and interlaminar, damage processes are modelled using the numerical finite-element analysis model with each individual ply treated as a continuum. Good agreement is found between the results from the experimental studies and the predictions from the numerical simulations. This article is part of the theme issue 'Nanocracks in nature and industry'.
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Plasma electrolytic oxidation (PEO) has been demonstrated to be an effective surface treatment for enhancing the osteoconduction and osseointegration of commercially pure α-Ti (CP α-Ti) dental implant materials for clinical application. To explore the feasibility of extending the application of PEO to low-modulus ß-type titanium alloys for load-bearing orthopaedic implants, a thorough understanding of the effect of substrate material on the biological performance of the PEO-treated surface is required. A 10 kW 50 Hz KeroniteTM processing unit was used to modify the surface of low-modulus near ß-Ti13Nb13Zr and ß-Ti45Nb substrates. CP α-Ti and (α + ß)-Ti6Al4V were also used in parallel as reference materials. In vitro culture of foetal human osteoblast (fHOb) cells on PEO-treated low-modulus near ß-Ti13Nb13Zr and ß-Ti45Nb alloys revealed comparable behaviour to that seen with CP α-Ti and (α + ß)-Ti6Al4V with respect to metabolic activity, collagen production, matrix formation and matrix mineralisation. No difference was observed in TNF-α and IL-10 cytokine release from CD14+ monocytes as markers of inflammatory response across samples. Cell interdigitation into the porous structure of the PEO coatings was demonstrated and cell processes remained adherent to the porous structure despite rigorous sonication. This study shows that PEO technology can be used to modify the surface of low-modulus ß-type titanium alloys with porous structure facilitating osseointegration, without impeding osteoblast activity or introducing an untoward inflammatory response.
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Eletrólise , Osteoblastos/citologia , Gases em Plasma/química , Titânio/farmacologia , Ligas , Calcificação Fisiológica/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Forma Celular/efeitos dos fármacos , Células Cultivadas , Colágeno Tipo I/biossíntese , Citocinas/metabolismo , Matriz Extracelular/metabolismo , Humanos , Osteoblastos/efeitos dos fármacos , Osteoblastos/metabolismo , Osteoblastos/ultraestrutura , Osteogênese/efeitos dos fármacos , Oxirredução , Propriedades de Superfície , Fatores de TempoRESUMO
Scaffolds for tissue engineering applications should be highly permeable to support mass transfer requirements while providing a 3-D template for the encapsulated biological cells. High porosity and cell interconnectivity result in highly compliant scaffolds. Overstraining occurs easily with such compliant materials and can produce misleading results. In this paper, the cell structure of freeze-dried collagen scaffolds, in both dry and hydrated states, was characterised using X-ray tomography and 2-photon confocal microscopy respectively. Measurements have been made of the scaffold's Young's modulus using conventional mechanical testing and a customised see-saw testing configuration. Specific permeability was measured under constant pressure gradient and compared with predictions. The collagen scaffolds investigated here have a coarse cell size (â¼100-150 µm) and extensive connectivity between adjacent cells (â¼10-30 µm) in both dry and hydrated states. The Young's modulus is very low, of the order of 10 kPa when dry and 1 kPa when hydrated. There is only a single previous study concerning the specific permeability of (hydrated) collagen scaffolds, despite its importance in nutrient diffusion, waste removal and cell migration. The experimentally measured value reported here (5 × 10(-)(10)m(2)) is in good agreement with predictions based on Computational Fluid Dynamics simulation and broadly consistent with the Carman-Kozeny empirical estimate. It is however about three orders of magnitude higher than the single previously-reported value and this discrepancy is attributed at least partly to the high pressure gradient imposed in the previous study. STATEMENT OF SIGNIFICANCE: The high porosity and interconnectivity of tissue engineering scaffolds result in highly compliant structures (ie large deflections under low applied loads). Characterisation is essential if these scaffolds are to be systematically optimised. Scaffold overstraining during characterisation can lead to misleading results. In this study, the stiffness (in dry and hydrated states) and specific permeability of freeze-dried collagen scaffolds have been measured using techniques customised for low stiffness structures. The scaffold cell structure is investigated using X-ray computed tomography, which has been applied previously to visualise such materials, without extracting any structural parameters or simulating fluid flow. These are carried out in this work. 2-photon confocal microscopy is used for the first time to study the structure in hydrated state.
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Colágeno/química , Liofilização , Engenharia Tecidual/métodos , Alicerces Teciduais/química , Água/química , Fenômenos Biomecânicos , Colágeno/ultraestrutura , Força Compressiva , Módulo de Elasticidade , Glicosaminoglicanos/metabolismo , Glicosaminoglicanos/ultraestrutura , Permeabilidade , TomografiaRESUMO
The requirement for revision surgery of total joint replacements is increasing and modular joint replacement implants have been developed to provide adjustable prosthetic revision systems with improved intra-operative flexibility. An electrochemical study of the corrosion resistance of the interface between the distal and proximal modules of a modular prosthesis was performed in combination with a cyclic fatigue test. The complexity resides in the existence of interfaces between the distal part, the proximal part, and the dynamometric screw. A new technique for evaluating the resistance to cyclic dynamic corrosion with crevice stimulation was used and the method is presented. In addition, two components of the proximal module of explanted Ti6Al4V and Ti6Al7Nb prostheses were investigated by optical and electron microscopy. Our results reveal that: The electrolyte penetrates into the interface between the distal and proximal modules during cyclic dynamic fatigue tests, the distal module undergoes cracking and corrosion was generated at the interface between the two models; The comparison of the explanted proximal parts with the similar prostheses evaluated following cyclic dynamic crevice corrosion testing showed that there were significant similarities indicating that this method is suitable for evaluating materials used in the fabrication of modular prostheses.
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Teste de Materiais , Próteses e Implantes , Estresse Mecânico , Ligas , Cátions , Corrosão , Eletricidade , Técnicas Eletroquímicas , Humanos , Titânio/químicaRESUMO
The use of a porous coating on prosthetic components to encourage bone ingrowth is an important way of improving uncemented implant fixation. Enhanced fixation may be achieved by the use of porous magneto-active layers on the surface of prosthetic implants, which would deform elastically on application of a magnetic field, generating internal stresses within the in-growing bone. This approach requires a ferromagnetic material able to support osteoblast attachment, proliferation, differentiation, and mineralization. In this study, the human osteoblast responses to ferromagnetic 444 stainless steel networks were considered alongside those to nonmagnetic 316L (medical grade) stainless steel networks. While both networks had similar porosities, 444 networks were made from coarser fibers, resulting in larger inter-fiber spaces. The networks were analyzed for cell morphology, distribution, proliferation, and differentiation, extracellular matrix production and the formation of mineralized nodules. Cell culture was performed in both the presence of osteogenic supplements, to encourage cell differentiation, and in their absence. It was found that fiber size affected osteoblast morphology, cytoskeleton organization and proliferation at the early stages of culture. The larger inter-fiber spaces in the 444 networks resulted in better spatial distribution of the extracellular matrix. The addition of osteogenic supplements enhanced cell differentiation and reduced cell proliferation thereby preventing the differences in proliferation observed in the absence of osteogenic supplements. The results demonstrated that 444 networks elicited favorable responses from human osteoblasts, and thus show potential for use as magnetically active porous coatings for advanced bone implant applications.
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Fenômenos Magnéticos , Imãs/química , Osteoblastos/citologia , Próteses e Implantes , Aço Inoxidável/farmacologia , Adsorção , Humanos , Microscopia de Fluorescência , Osteoblastos/efeitos dos fármacos , Osteoblastos/ultraestrutura , Fenótipo , Cloreto de Tolônio/metabolismoRESUMO
Collagen fibres are ubiquitous macromolecular assemblies in nature, providing the structures that support tensile mechanical loads within the human body. Aligned type I collagen fibres are the primary structural motif for tendon and ligament, and therefore biomaterials based on these structures are considered promising candidates for mediating regeneration of these tissues. However, despite considerable investigation, there remains no collagen-fibre-based biomaterial that has undergone clinical evaluation for this application. Recent research in this area has significantly enhanced our understanding of these complex and challenging biomaterials, and is reinvigorating interest in the development of such structures to recapitulate mechanical function. In this review we describe the progress to date towards a ligament or tendon regeneration template based on collagen fibre scaffolds. We highlight reports of particular relevance to the development of the underlying biomaterials science in this area. In addition, the potential for tailoring and manipulating the interactions between collagen fibres and biological systems, as hybrid biomaterial-biological ensembles, is discussed in the context of developing novel tissue engineering strategies for tendon and ligament.
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Materiais Biocompatíveis/química , Colágeno/química , Ligamentos/fisiologia , Tendões/fisiologia , Engenharia Tecidual/métodos , Alicerces Teciduais , Humanos , Modelos Biológicos , RegeneraçãoRESUMO
This study examined HIV testing behaviours, perceived vulnerability to HIV and correlates of sexual risk behaviours of young adult Latino and African American male gang members in Los Angeles, California. Data were collected from 249 gang members aged 18-26 years. The majority (59%) of gang members reported unprotected vaginal intercourse (UVI) in the past 12 months. Only one-third (33.2%) of gang members had ever been tested for HIV. In our multivariate analysis, gang members who reported UVI were more likely to have engaged in the following behaviours: had sex with someone they just met (adjusted odds ratio [AOR] = 4.51), had sex with someone they think or know had a sexually transmitted infection (STI; AOR = 4.67) or had sex while incarcerated (AOR = 8.92). In addition, gang members with a higher perceived vulnerability to HIV were less likely to report UVI in the previous 12 months (AOR = 0.75). These findings offer implications for development of an HIV prevention intervention for young Latino and African American male gang members.
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Infecções por HIV/epidemiologia , Infecções por HIV/transmissão , Aceitação pelo Paciente de Cuidados de Saúde/estatística & dados numéricos , Assunção de Riscos , Adolescente , Adulto , Negro ou Afro-Americano , Infecções por HIV/diagnóstico , Hispânico ou Latino , Humanos , Los Angeles , Masculino , Adulto JovemRESUMO
OBJECTIVE: The profile of women with gynecologic malignancies treated with pelvic exenteration has changed since the initial description of this procedure. We sought to evaluate our experience with pelvic exenteration over the last 20 years. METHODS: Patients who underwent anterior, posterior, or total pelvic exenteration for vulvar, vaginal, and cervical cancer at Barnes-Jewish Hospital between January 1, 1990 and August 1, 2009 were identified through hospital databases. Patient characteristics, the indications for the procedure, procedural modifications, and patient outcomes were retrospectively assessed. Categorical variables were analyzed with chi-square method, and survival data was analyzed using the Kaplan-Meier method and log rank test. RESULTS: Fifty-four patients were identified who had pelvic exenteration for cervical, vaginal, or vulvar cancer. Recurrent cervical cancer was the most common procedural indication. One year overall survival from pelvic exenteration for the entire cohort was 64%, with 44% of patients still living at 2 years and 34% at 50 months. Younger age was associated with improved overall survival after exenteration (p = 0.01). Negative margin status was associated with a longer disease-free survival (p=0.014). Nodal status at the time of exenteration was not associated with time to recurrence or progression, site of recurrence, type of post-operative treatment, early or late complications, or survival. CONCLUSIONS: Despite advances in imaging and increased radical techniques, outcomes and complications after total pelvic exenteration in this cohort are similar to those described historically. Pelvic exenteration results in sustained survival in select patients, especially those that are young with recurrent disease and pathologically negative margins.
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Neoplasias dos Genitais Femininos/cirurgia , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Pessoa de Meia-Idade , Exenteração Pélvica , Estudos Retrospectivos , Taxa de Sobrevida , Adulto JovemRESUMO
A new deposition method is presented, based on electrospraying, that can build bioceramic structures with desirable surface properties. This technology allows nanoapatite crystals, including hydroxyapatite (nHA), carbonate-substituted HA (nCHA) and silicon-substituted HA (nSiHA), to be electrosprayed on glass substrates. Human osteoblast cells cultured on nSiHA showed enhanced cell attachment, proliferation and protein expression, namely alkaline phosphatase, type 1 collagen and osteocalcin, as compared to nHA and nCHA. The modification of nanoapatite by the addition of silicon into the HA lattice structure renders the electrosprayed surface more hydrophilic and electronegatively charged.
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Substitutos Ósseos/química , Galvanoplastia/métodos , Hidroxiapatitas/química , Nanopartículas/química , Osteoblastos/citologia , Osteoblastos/fisiologia , Engenharia Tecidual/métodos , Adesão Celular , Técnicas de Cultura de Células/métodos , Proliferação de Células , Sobrevivência Celular , Células Cultivadas , Cristalização/métodos , Humanos , Interações Hidrofóbicas e Hidrofílicas , Teste de Materiais , Nanopartículas/ultraestrutura , Tamanho da Partícula , Eletricidade Estática , MolhabilidadeRESUMO
Silicon has been shown to have important effects on skeletal development and repair, and soluble silicate ions have been found to stimulate the expression of type-I collagen in osteoblast-like cell cultures. Furthermore, silicon has been incorporated into the hydroxyapatite lattice and enhanced metabolic activity of human osteosarcoma cells was observed when cells were cultured on this material. In vivo assessments have demonstrated enhanced bioactivity of silicon-substituted hydroxyapatite (Si-HA) over pure HA. However, detailed mechanisms for the stimulative effects of Si-HA have not been described. In this study, we found that silicon substitution into hydroxyapatite affects the adhesion of human osteoblast-like cells (HOBs) in culture, with 0.8 wt % silicon substitution being optimal. In addition, metabolic activity and proliferation of HOBs were increased by supplementation of the growth medium with 30 microM silicon. It was determined that this response may depend on the proportion of cells at different stages of differentiation within the cultures.
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Adesão Celular/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Osteoblastos/efeitos dos fármacos , Silicatos/farmacologia , Linhagem Celular Tumoral , Durapatita , Citometria de Fluxo , Humanos , Osteoblastos/citologia , Difração de Raios XRESUMO
Polyimides have been widely used for biosensor encapsulation and more recently as substrates for neural implants. They have excellent thermal stability, high chemical resistance, and can be prepared as thin, flexible films. Photosensitive polyimides present similar physical properties to polyimides, and have the advantage that they can be photo-lithographically patterned. However, to date little data on their biocompatibility has been reported. Two commercially available polyimides (PI) and one photo-sensitive polyimide (PSPI) were evaluated in vitro using the ISO 10993 standard on biocompatibility. The materials were Dupont Kapton foil HN, HD Microsystem PI2611, and Fujifilm Durimide 7020 (PSPI). PI2611 and Durimide 7020 were spin-coated on silicon wafers, cured at temperatures ranging from 150 to 450 degrees C, and sterilized by autoclave. All materials were evaluated using a scanning electron microscope pre- and postcell culture. Cell viability was determined by an MTS assay. Their mechanical properties and stability during cell culture as a function of time and environment were investigated by nanoindentation. The MTS results show that PSPI is noncytotoxic compared with the negative control of polyethylene and the conventional PIs tested. Fibroblast adhesion, morphology, and spreading were good and better on the PSPI substrate than on the PI2611. Schwann cell appearance was similar on each of the PIs and the PSPI tested. The results suggest that PSPIs may have potential use for biological microsystem and neuroprosthetic applications.
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Materiais Biocompatíveis/química , Imidas , Teste de Materiais , Polímeros , Próteses e Implantes , Animais , Adesão Celular , Forma Celular , Sobrevivência Celular , Células Cultivadas , Desenho de Equipamento , Fibroblastos/citologia , Fenômenos Mecânicos , Camundongos , Células de Schwann/citologiaRESUMO
Electrohydrodynamic spraying is a well established process used to deposit, coat, analyse and synthesise materials within the biomedical remit. Recently, electrohydrodynamic printing has been developed to afford structures for potential applications in the biomedical and medical engineering fields. Both of these processes rely on the formation of an electrically-induced jet, however the resulting products can be made strikingly different and offer potential in broader applications. Here we show how spraying and printing are linked by elucidating the ease of transition between the processes. Changes in the deposition distance can result in either spray (>10 mm) or print formation (<3 mm), with an overlap of the two in between this range. For the optimal printing distance of 0.5 mm, gradual changes in the applied voltage (0-4.5 kV) encounters transitional printing modes (dripping, micro-dripping, rapid micro-dripping, unstable and stable jetting) which can be utilised for patterning. The results indicate the robustness of the electrohydrodynamic route in the nano-materials processing arena, with emphasis on biomedical materials.
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Materiais Revestidos Biocompatíveis/química , Durapatita/química , Nanopartículas/química , Nanotecnologia/métodos , Substitutos Ósseos , Eletroquímica/métodos , Etanol/química , Vidro , Teste de Materiais , Metais/química , Microscopia Confocal , Microscopia Eletrônica de Transmissão/métodos , Propriedades de Superfície , Titânio/química , Difração de Raios XRESUMO
Nanometer scale carbonate-substituted hydroxyapatite (nanoCHA) particles were prepared and examined using transmission electron microscopy, which revealed their polycrystalline nature with a rod-like morphology (20-30 nm in width and 50-80 nm in length). In vitro cytotoxicity study showed that there was some evidence of lactate dehydrogenase (LDH) release when macrophages were in contact with high concentrations of nanoCHA particles. The levels of LDH release decreased significantly with a reduction in nanoCHA concentration. A similar trend was observed for the inflammatory cytokine TNF-alpha. nanoCHA particles with high carbonate content induced a high level of TNF-alpha release. Biological testing using a human osteoblast (HOB) cell model found that HOB cells were able to grow and proliferate on a nanoCHA deposited surface. Well organized actin fibers were observed for HOB cells in contact with nanoCHA particles with low carbonate content and the cell proliferation rate was higher on these particles in comparison with those of high carbonate nanoCHA particles. Therefore, low carbonate nanoCHA particles were incorporated into poly-(2-hydroxyethylmethacrylate) matrix to make a nanocomposite. It was found that the nanoCHA composite was hydrophilic and became rubber-like after hydration. Both 20 wt % and 40 wt % composites were able to induce the formation of bone-like apatite after immersion in simulated body fluid. A high bioactivity of the composite was obtained with high loading of the nanoCHA filler. These results demonstrate the potential of formulating nanocomposites for biomedical applications.
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Materiais Biocompatíveis/química , Carbonatos/química , Durapatita/química , Nanocompostos/química , Poli-Hidroxietil Metacrilato/química , Células Cultivadas , Humanos , Hidrogéis/química , Lactato Desidrogenases/metabolismo , Macrófagos/metabolismo , Nanopartículas/química , Osteoblastos/metabolismo , Solubilidade , Fator de Necrose Tumoral alfa/metabolismoRESUMO
Electrohydrodynamic spraying has been used to produce patterns of line width up to 100 microm in size on glass discs, using nanohydroxyapatite (nHA). A human osteoblast (HOB)-like cell model was then used to study the interaction between the HOB cells and nHA patterns in vitro. Growth of the cells was significantly increased (p < 0.05) on the nHA surfaces. In addition, HOBs attached and spread well, secreting extracellular matrix. It was found that a confluent, aligned cell layer was achieved on nHA patterns by day 9. Immunofluorescent staining indicated that these cells showed elongated nuclei, enhanced adhesion (vinculin adhesion plaques) and a well-aligned cytoskeleton (actin stress fibres). This work suggests that this type of spraying may provide a route for the production of nanoscale features on implants for biomedical applications.
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Materiais Revestidos Biocompatíveis , Durapatita , Nanoestruturas , Osteoblastos/metabolismo , Células Cultivadas , Durapatita/síntese química , HumanosRESUMO
INTRODUCTION: We performed a questionnaire study to establish the frequency and consequences of the detection of orthopaedic implants by airport security and to help us advise patients correctly. All published literature on this subject is based on experimental studies and no 'real-life' data are available. PATIENTS AND METHODS: A total of 200 patients with a variety of implants were identified. All patients were sent a postal questionnaire enquiring about their experience with airport security since their surgery. RESULTS: Of the cohort, 154 (77%) patients responded. About half of the implants (47%) were detected, but the majority of patients (72%) were not significantly inconvenienced. When detected, only 9% of patients were asked for documentary evidence of their implant. We also found that patients with a total knee replacement (TKR) had a greater chance of detection as compared to those with a total hip replacement (THR; 71% versus 31%; P = 0.03). CONCLUSIONS: All patients, and in particular those with a TKR, can be re-assured that, although they have a fair chance of detection by airport security, a major disruption to their journey is unlikely. We advise that documentation to prove the presence of an orthopaedic implant should be offered to those who are concerned about the potential for inconvenience, but such documentation is not required routinely.
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Aviação , Metais , Próteses e Implantes , Medidas de Segurança , Adolescente , Adulto , Idoso , Humanos , Pessoa de Meia-IdadeRESUMO
Latinos are under-represented in HIV/AIDS medical research in the US. Although they are disproportionately impacted by HIV/AIDS, Latinos may be reluctant to participate in HIV vaccine trials. Three focus groups were conducted with 32 Spanish-speaking Latinos recruited from two community-based healthcare organizations in Los Angeles, California. A qualitative focus group interview guide was developed to explore concerns, motivators and intentions in regard to participation in HIV vaccine trials. Mistrust and fear of government emerged as important themes related to reluctance to participate in an HIV vaccine trial. Specific concerns regarding trial participation included: (1) fear of vaccine-induced HIV infection, (2) physical side effects, (3) stigma and (4) false-induced HIV-positive test results and their social repercussions. Motivators for enrolling in an HIV vaccine trial included: (1) incentives, (2) convenience of participating in a study, (3) sufficient and appropriate study information, (4) personal benefits and (5) altruism. Interventions to facilitate participation by Latinos in HIV vaccine trials should address mistrust and fear of government-sponsored HIV/AIDS medical research, increase access to and convenience of clinical trials, address fear of vaccine-induced infection, combat HIV/AIDS stigma and raise awareness of the relevance of HIV/AIDS to Latino communities.
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Vacinas contra a AIDS/uso terapêutico , Infecções por HIV/prevenção & controle , Conhecimentos, Atitudes e Prática em Saúde , Hispânico ou Latino/psicologia , Recusa de Participação/psicologia , Adolescente , Adulto , Ensaios Clínicos como Assunto/psicologia , Medo/psicologia , Feminino , Grupos Focais/métodos , Governo , Infecções por HIV/psicologia , Humanos , Los Angeles/epidemiologia , Masculino , Pessoa de Meia-Idade , Confiança/psicologiaRESUMO
Human osteoblasts were cultured on hydroxyapatite (HA), 0.8 wt % silicon substituted hydroxyapatite (Si-HA) and 1.5 wt % Si-HA discs. The influence of these substrates on cell behaviour in vitro was assessed by measuring total protein in the cell lysate and the production of several phenotypic markers: collagen type I (COL I), alkaline phosphatase (ALP), osteocalcin (OC), and the formation of bone mineral. After 7 days, beta-glycerophosphate and physiological levels of hydrocortisone were added to the culture medium to stimulate cell differentiation and mineral production. There was a significantly higher production of ALP on 1.5 wt % Si-HA at day 7 following which, the addition of hydrocortisone promoted the differentiation of cells on the other two substrates. Hydrocortisone addition also decreased the production of OC. During the period, when hydrocortisone was present, no significant difference in behavior was seen between cells on Si-HA and HA; however, following removal of hydrocortisone, cells responded to 0.8 wt % Si-HA with a significant increase in protein production. Using fluorescence microscopy, nodular structures labeled with tetracycline were observed on the surface of all substrates after 21 days. These structures were deposited on areas of high cell density but were not related to the presence or level of silicon in the substrate. These results indicate that human osteoblasts are affected by the presence of silicon in the HA substrate and that the timing of these effects may be dependent upon the level of silicon substitution.
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Durapatita/química , Durapatita/farmacologia , Osteoblastos/efeitos dos fármacos , Silício/química , Células Cultivadas , Colágeno Tipo I/metabolismo , Humanos , Minerais/metabolismo , Osteoblastos/metabolismo , Osteocalcina/metabolismo , Diester Fosfórico Hidrolases/metabolismo , Pirofosfatases/metabolismoRESUMO
In healthy bone, resorption and synthesis are in perfect coordination. In previous studies we demonstrated that the incorporation of silicon into the hydroxyapatite (HA) lattice enhances the proliferation and differentiation of human osteoblasts. Therefore, the aim of this study was to demonstrate the effect of silicon-substituted HA (0.8 and 1.5 wt % Si-HA) on the differentiation of mononuclear cells into osteoclasts, using two different starting cultures, peripheral blood mononuclear cells (PBMC) and monocytes expressing the CD14 antigen (CD14+). Through this study, it was possible to demonstrate that Si-HA allows the differentiation of mononuclear cells into mature osteoclasts, independent of the starting culture, PBMC or CD14+. Most of the cells on the surface of the materials expressed osteoclastic markers: actin rings, several nuclei, positivity for tartrate-resistant acid phosphatase (TRAP), and vitronectin receptor. In the presence of osteoclasts, a higher release of calcium and phosphate into the medium from the 1.5 wt % Si-HA substrate was detected when compared to the HA substrate; therefore, these results indicate higher osteoclastic resorptive activity on the 1.5 wt % Si-HA surface. Si-HA can be resorbed by cellular mechanisms and have a stimulatory effect on osteoclasts, although the underlying mechanism is still poorly understood.
Assuntos
Diferenciação Celular , Durapatita/química , Osteoblastos/citologia , Silício/química , Receptores de Lipopolissacarídeos/imunologia , Microscopia de Fluorescência , Monócitos/citologia , Monócitos/imunologiaRESUMO
Suspensions containing nano-sized silicon substituted hydroxyaptite (nSiHA) particles were produced and processed for electrostatic spray deposition. No secondary phases were detected by X-ray diffraction, which indicated that the nSiHA was phase pure. Electrostatic spraying of nSiHA in cone-jet mode was achieved at flow rate of 10(-9) m3s(-1) with an applied voltage between the needle and the ring-shaped ground electrode set at 6 to 8 kV. Micrometer- and submicrometer-scaled islands of nSiHA have been deposited on glass and titanium substrates. The surface roughness of such nHA and nSiHA islands was in the range 60 to 80 nm, as measured from atomic force microscopy in tapping mode. The growth of primary human osteoblast (HOB) cells on the nSiHA deposited substrates increased with time during the 4 days of culture, and the increase was related with the Si content in substituted HA, indicating that nSiHA was able to promote and support the growth of HOB cells. Scanning electron microscopy (SEM) revealed that extracellular matrix (ECM) produced by the HOB cells on these nSiHA deposits was well organized. In addition, the presence of Ca and P containing nodules in the ECM were also confirmed by Energy Dispersive X-ray (EDX) analysis, indicating early signs of calcification fronts. The results showed that nSiHA produced by electrostatic spray deposition was able to promote the attachment and the growth of HOB cells. Therefore, electrostatic spray deposition offers great potential for the creation of bioactive surfaces to provide improved interfacial bonding with host tissues.
Assuntos
Durapatita , Silício , Adesão Celular , Divisão Celular , Células Cultivadas , Humanos , Microscopia Eletrônica de Varredura , Nanotecnologia , Osteoblastos/citologia , Eletricidade EstáticaRESUMO
Phase pure hydroxyapatite (HA) and two silicate-substituted hydroxyapatites (0.8 and 1.5 wt% Si, or 2.6 and 4.9 wt% SiO4) were prepared by aqueous precipitation methods. The filter-cakes of HA and silicate-substituted hydroxyapatite (SiHA) compositions were processed into granules 1.0-2.0 mm in diameter and sintered at 1200 degrees C for 2 h. The sintered granules underwent full structural characterisation, prior to assessment in an ovine defect model by implantation for a period of 6 and 12 weeks. The results indicate that HA and SiHA implants were well accepted by the host tissue, with no evidence of inflammation. New bone formation was observed directly on the surfaces and in the spaces between the granular implants. Quantitative histomorphometry as determined by the percentage of bone ingrowth and bone coverage for both SiHA implant compositions was significantly greater than that for phase pure HA. These findings indicate that the in vivo bioactivity of hydroxyapatite was significantly improved by the incorporation of silicate ions into the HA structure, making SiHA ceramics attractive alternatives to conventional HA materials for use as bone graft substitute ceramics.