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1.
J Funct Biomater ; 15(1)2024 Jan 05.
Artículo en Inglés | MEDLINE | ID: mdl-38248685

RESUMEN

Platelet-rich Fibrin (PRF), a second-generation blood concentrate, offers a versatile structure for bone regeneration due to its composition of fibrin, growth factors, and cytokines, with adaptations like denatured albumin-enriched with liquid PRF (Alb-PRF), showing potential for enhanced stability and growth factor dynamics. Researchers have also explored the combination of PRF with other biomaterials, aiming to create a three-dimensional framework for enhanced cell recruitment, proliferation, and differentiation in bone repair studies. This study aimed to evaluate a combination of Alb-PRF with nanostructured carbonated hydroxyapatite microspheres (Alb-ncHA-PRF), and how this association affects the release capacity of growth factors and immunomodulatory molecules, and its impact on the behavior of MG63 human osteoblast-like cells. Alb-PRF membranes were prepared and associated with nanocarboapatite (ncHA) microspheres during polymerization. MG63 cells were exposed to eluates of both membranes to assess cell viability, proliferation, mineralization, and alkaline phosphatase (ALP) activity. The ultrastructural analysis has shown that the spheres were shattered, and fragments were incorporated into both the fibrin mesh and the albumin gel of Alb-PRF. Alb-ncHA-PRF presented a reduced release of growth factors and cytokines when compared to Alb-PRF (p < 0.05). Alb-ncHA-PRF was able to stimulate osteoblast proliferation and ALP activity at lower levels than those observed by Alb-PRF and was unable to positively affect in vitro mineralization by MG63 cells. These findings indicate that the addition of ncHA spheres reduces the biological activity of Alb-PRF, impairing its initial effects on osteoblast behavior.

2.
Biomed Mater ; 18(6)2023 11 01.
Artículo en Inglés | MEDLINE | ID: mdl-37844570

RESUMEN

This study aimed to analyze the physicochemical and histological properties of nanostructured hydroxyapatite and alginate composites produced at different temperatures with and without sintering and implanted in rabbit tibiae. Hydroxyapatite-alginate (HA) microspheres (425-600 µm) produced at 90 and 5 °C without (HA90 and HA5) or with sintering at 1000 °C (HA90S and HA5S) were characterized and applied to evaluate thein vitrodegradation; also were implanted in bone defects on rabbit's tibiae (n= 12). The animals were randomly divided into five groups (blood clot, HA90S, HA5S, HA90, and HA5) and euthanized after 7 and 28 d. X-ray diffraction and Fourier-transform infrared analysis of the non-sintered biomaterials showed a lower crystallinity than sintered materials, being more degradablein vitroandin vivo. However, the sinterization of HA5 led to the apatite phase's decomposition into tricalcium phosphate. Histomorphometric analysis showed the highest (p< 0.01) bone density in the blood clot group, similar bone levels among HA90S, HA90, and HA5, and significantly less bone in the HA5S. HA90 and HA5 groups presented higher degradation and homogeneous distribution of the new bone formation onto the surface of biomaterial fragments, compared to HA90S, presenting bone only around intact microspheres (p< 0.01). The elemental distribution (scanning electron microscope and energy dispersive spectroscopy andµXRF-SR analysis) of Ca, P, and Zn in the newly formed bone is similar to the cortical bone, indicating bone maturity at 28 d. The synthesized biomaterials are biocompatible and osteoconductive. The heat treatment directly influenced the material's behavior, where non-sintered HA90 and HA5 showed higher degradation, allowing a better distribution of the new bone onto the surface of the biomaterial fragments compared to HA90S presenting the same level of new bone, but only on the surface of the intact microspheres, potentially reducing the bone-biomaterial interface.


Asunto(s)
Durapatita , Trombosis , Animales , Conejos , Durapatita/química , Sincrotrones , Materiales Biocompatibles/química , Cerámica , Alginatos/química
3.
Materials (Basel) ; 14(19)2021 Oct 07.
Artículo en Inglés | MEDLINE | ID: mdl-34640255

RESUMEN

Successful biomaterials for bone tissue therapy must present different biocompatible properties, such as the ability to stimulate the migration and proliferation of osteogenic cells on the implantable surface, to increase attachment and avoid the risks of implant movement after surgery. The present work investigates the applicability of a three-dimensional (3D) model of bone cells (osteospheres) in the evaluation of osteoconductive properties of different implant surfaces. Three different titanium surface treatments were tested: machined (MA), sandblasting and acid etching (BE), and Hydroxyapatite coating by plasma spray (PSHA). The surfaces were characterized by Scanning Electron Microscopy (SEM) and atomic force microscopy (AFM), confirming that they present very distinct roughness. After seeding the osteospheres, cell-surface interactions were studied in relation to cell proliferation, migration, and spreading. The results show that BE surfaces present higher densities of cells, leaving the aggregates towards than titanium surfaces, providing more evidence of migration. The PSHA surface presented the lowest performance in all analyses. The results indicate that the 3D model allows the focal analysis of an in vitro cell/surfaces interaction of cells and surfaces. Moreover, by demonstrating the agreement with the clinical data observed in the literature, they suggest a potential use as a predictive preclinical tool for investigating osteoconductive properties of novel biomaterials for bone therapy.

4.
J Biomed Mater Res B Appl Biomater ; 108(6): 2610-2620, 2020 08.
Artículo en Inglés | MEDLINE | ID: mdl-32096353

RESUMEN

Zinc is an important element for bone structure and metabolism. Its interaction with hydroxyapatite has been investigated for the improvement of bone repair. The objective of this study was to evaluate the in vitro and in vivo biological response to nanostructured calcium alginate-hydroxyapatite (HA) and zinc-containing HA (ZnHA). Cytocompatibility was evaluated by applying PrestoBlue reagent after exposing murine pre-osteoblast cells to extracts of each biomaterial microspheres. After physical and chemical characterization, the biomaterial microspheres were implanted in a critical size calvaria defect (8 mm) in Wistar rats (n = 30) that were randomly divided into the HA and ZnHA groups. Tissue samples were evaluated through histological and histomorphometric analyses after 1, 3, and 6 months (n = 5). The results showed cellular viability for both groups compared to the negative control, and no differences in metabolic activity were observed. The HA group presented a significant reduction of biomaterial compared with the ZnHA group in all experimental periods; however, a considerable amount of new bone formation was observed surrounding the ZnHA spheres at the 6-month time point compared with the HA group (p < .05). Both biomaterials were biocompatible, and the combination of zinc with hydroxyapatite was shown to improve bone repair.


Asunto(s)
Alginatos/farmacología , Regeneración Ósea/efectos de los fármacos , Durapatita/farmacología , Zinc/farmacología , Alginatos/administración & dosificación , Animales , Materiales Biocompatibles , Supervivencia Celular , Durapatita/administración & dosificación , Femenino , Masculino , Ensayo de Materiales , Microesferas , Osteoblastos , Ratas , Ratas Wistar , Zinc/administración & dosificación
5.
J Biomed Mater Res B Appl Biomater ; 108(4): 1351-1362, 2020 05.
Artículo en Inglés | MEDLINE | ID: mdl-31496111

RESUMEN

Drug delivery technology is a promising way to enhance the therapeutic efficacy of drugs. The purpose of this study is to evaluate the physical and chemical properties of hydroxyapatite ceramic microspheres loaded with doxycycline (HADOX), their effects on in vitro osteoblast viability, and their antimicrobial activity, and to determine the effects of DOX on the healing of rat sockets after tooth extraction. The internal microsphere porosity was sensitive to the treatment used to adsorb DOX onto microsphere surface; HA microspheres without DOX presented 26% of pores, whereas HADOX0.15 microspheres presented 52.0%. An initial drug release of 49.15 µg/ml was observed in the first 24 hr. The minimal inhibitory concentration (MIC) tested against Enterococcus faecalis demonstrated that bacterial growth was inhibited for up to 7 days. Results of cell viability and cell proliferation did not indicate statistical differences in the metabolic activity of HADOX samples relative to HA without DOX microspheres (p > .05). After 1 week, a discreet inflammation reaction was observed in the control group, and after 6 weeks, newly-formed bone was observed in the HADOX0.15 (p < .05). The HADOX did not interfere in the bone repair and controlled the early inflammatory response. HADOX could be a promising biomaterial to promote bone repair in infected sites.


Asunto(s)
Cerámica , Doxiciclina , Sistemas de Liberación de Medicamentos , Durapatita , Microesferas , Osteoblastos/metabolismo , Animales , Antibacterianos/química , Antibacterianos/farmacocinética , Antibacterianos/farmacología , Cerámica/química , Cerámica/farmacocinética , Cerámica/farmacología , Doxiciclina/química , Doxiciclina/farmacocinética , Doxiciclina/farmacología , Durapatita/química , Durapatita/farmacocinética , Durapatita/farmacología , Enterococcus faecalis/crecimiento & desarrollo , Femenino , Masculino , Ratones , Ratas , Ratas Wistar
6.
Int J Nanomedicine ; 14: 4559-4571, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31417258

RESUMEN

Background and objective: Tetracycline and its derivatives, combined with calcium phosphates, have been proposed as a delivery system to control inflammatory processes and chronic infections. The objective of this study was to evaluate the microspheres of alginate encapsulated minocycline-loaded nanocrystalline carbonated hydroxyapatite (CHAMINO) as a biomimetic device to carry out target-controlled drug delivery for alveolar bone repair. Methods: CHAMINO microspheres were implanted in a rat central incisor socket after 7 and 42 days. New bone was formed in both groups between 7 and 42 days of implantation. However, the bone growth was significantly higher for the CHAMINO microspheres. Results: The minocycline (MINO) loading capacity of the nanocrystaline carbonated hydroxyapatite (CHA) nanoparticles was 25.1±2.2 µg MINO/mg CHA for adsorption over 24 hrs. The alginate microspheres containing minocycline-loaded CHA were biologically active and inhibited the Enterococcus faecalis culture growth for up to seven days of the MINO release. An osteoblastic cell viability assay based on the resazurin reduction was conducted after the cells were exposed to the CHAMINO powder and CHAMINO microspheres. Thus, it was found that the alginate extracts encapsulated the minocycline-loaded CHA microspheres and did not affect the osteoblastic cell viability, while the minocycline-doped CHA powder reduced the cell viability by 90%. Conclusion: This study concluded that the alginate microspheres encapsulating the minocycline-loaded nanocrystalline carbonated hydroxyapatite exhibited combined antibacterial activity against Enterococcus faecalis with cytocompatibility and osteoconduction properties. The significant improvement in the new bone formation after 42 days of implantation suggests that the CHAMINO microsphere has potential in clinical applications of bone regeneration.


Asunto(s)
Alginatos/química , Regeneración Ósea/efectos de los fármacos , Carbonatos/química , Durapatita/química , Microesferas , Minociclina/farmacología , Nanopartículas/química , Animales , Antibacterianos/farmacología , Supervivencia Celular/efectos de los fármacos , Sistemas de Liberación de Medicamentos , Liberación de Fármacos , Enterococcus/efectos de los fármacos , Humanos , Masculino , Ratones , Pruebas de Sensibilidad Microbiana , Osteoblastos/efectos de los fármacos , Ratas Wistar , Difracción de Rayos X
7.
Int J Nanomedicine ; 14: 3471-3490, 2019.
Artículo en Inglés | MEDLINE | ID: mdl-31190805

RESUMEN

Background: Zinc-doped hydroxyapatite has been proposed as a graft biomaterial for bone regeneration. However, the effect of zinc on osteoconductivity is still controversial, since the release and resorption of calcium, phosphorus, and zinc in graft-implanted defects have rarely been studied. Methods: Microspheres containing alginate and either non-doped carbonated hydroxyapatite (cHA) or nanocrystalline 3.2 wt% zinc-doped cHA (Zn-cHA) were implanted in critical-sized calvarial defects in Wistar rats for 1, 3, and 6 months. Histological and histomorphometric analyses were performed to evaluate the volume density of newly formed bone, residual biomaterial, and connective tissue formation. Biomaterial degradation was characterized by transmission electron microscopy (TEM) and synchrotron radiation-based X-ray microfluorescence (SR-µXRF), which enabled the elemental mapping of calcium, phosphorus, and zinc on the microsphere-implanted defects at 6 months post-implantation. Results: The bone repair was limited to regions close to the preexistent bone, whereas connective tissue occupied the major part of the defect. Moreover, no significant difference in the amount of new bone formed was found between the two microsphere groups. TEM analysis revealed the degradation of the outer microsphere surface with detachment of the nanoparticle aggregates. According to SR-µXRF, both types of microspheres released high amounts of calcium, phosphorus, and zinc, distributed throughout the defective region. The cHA microsphere surface strongly adsorbed the zinc from organic constituents of the biological fluid, and phosphorus was resorbed more quickly than calcium. In the Zn-cHA group, zinc and calcium had similar release profiles, indicating a stoichiometric dissolution of these elements and non-preferential zinc resorption. Conclusions: The nanometric size of cHA and Zn-cHA was a decisive factor in accelerating the in vivo availability of calcium and zinc. The high calcium and zinc accumulation in the defect, which was not cleared by the biological medium, played a critical role in inhibiting osteoconduction and thus impairing bone repair.


Asunto(s)
Alginatos/química , Regeneración Ósea , Calcio/metabolismo , Durapatita/química , Microesferas , Nanopartículas/química , Zinc/química , Zinc/metabolismo , Animales , Materiales Biocompatibles/química , Disponibilidad Biológica , Regeneración Ósea/efectos de los fármacos , Carbonatos/química , Muerte Celular , Línea Celular , Supervivencia Celular , Femenino , Ratones , Nanopartículas/ultraestructura , Ratas Wistar , Cráneo/fisiología , Espectroscopía Infrarroja por Transformada de Fourier , Difracción de Rayos X
8.
J Biomed Mater Res A ; 107(9): 1965-1976, 2019 09.
Artículo en Inglés | MEDLINE | ID: mdl-31035306

RESUMEN

Nanostructured carbonated hydroxyapatite (nCHA) is a promising biomaterial for bone tissue engineering due to its chemical properties, similar to those of the bone mineral phase and its enhanced in vivo bioresorption. However, the biological effects of nCHA nanoparticles on cells and tissues are not sufficiently known. This study assessed the impact of exposing pre-osteoblasts to suspensions with high doses of nCHA nanoparticles with high or low crystallinity. MC3T3-E1 pre-osteoblasts were cultured for 1 or 7 days in a culture medium previously exposed to CHA nanoparticles for 1 day. Control groups were produced by centrifugation for removal of bigger nCHA aggregates before exposure. Interaction of nanoparticles with the culture medium drastically changed medium composition, promoting Ca, P, and protein adsorption. Transmission Electron microscopy revealed that exposed cells were able to internalize both materials, which seemed concentrated inside endosomes. No cytotoxicity was observed for both materials, regardless of centrifugation, and the exposure did not induce alterations in the release of pro-and anti-inflammatory cytokines. Morphological analysis revealed strong interactions of nCHA aggregates with cell surfaces, however without marked alterations in morphological features and cytoskeleton ultrastructure. The overall in vitro biocompatibility of nCHA materials, regardless of physicochemical characteristics such as crystallinity, encourages further studies on their clinical applications.


Asunto(s)
Citoesqueleto/metabolismo , Durapatita/química , Ensayo de Materiales , Nanopartículas/química , Osteoblastos/metabolismo , Animales , Línea Celular , Citoesqueleto/ultraestructura , Ratones , Osteoblastos/ultraestructura
9.
Mater Sci Eng C Mater Biol Appl ; 79: 802-811, 2017 Oct 01.
Artículo en Inglés | MEDLINE | ID: mdl-28629083

RESUMEN

Insulin-loaded calcium phosphate nanoparticles have been proposed as a potential drug delivery system for the oral treatment of diabetes and to stimulate bone cell proliferation and bone mineralization. The kinetics of insulin incorporation onto hydroxyapatite (HA) and Sr (SrHA)- and Zn (ZnHA)-substituted hydroxyapatite nanoparticles was investigated using X-ray photoelectron spectroscopy (XPS), Fourier transform infrared (FTIR) spectroscopy, zeta potential measurements and circular dichroism (CD) spectroscopy. The increase in insulin concentration on HA, SrHA and ZnHA was a typical physical adsorption process controlled by electrostatic forces and followed a Freundlich isotherm model. Zn substitution enhanced the capacity of the apatite surface to adsorb insulin, whereas Sr substitution inhibited insulin uptake. The surface stoichiometry and mesopore specific area induced by Zn and Sr substitution are proposed as the main causes of the difference in insulin adsorption. Despite the weak interaction between insulin and the apatite surface, the CD spectra revealed a decrease in the insulin ellipticity when the protein was adsorbed on the HA, SrHA and ZnHA nanoparticles. A reduction in alpha-helical structures and an increase in beta sheets were observed when insulin interacted with the HA surface. A less pronounced effect was found for ZnHA, for which a subtle decrease in alpha-helical structures was followed by an increase in turn structures. Interaction with the SrHA surface did not change the native insulin conformation. In vitro cell culture experiments lasting 24h using F-OST stromal cells showed that the insulin loaded on HA and ZnHA did not affect cell proliferation but the insulin loaded on SrHA improved cell proliferation. These results suggest that the stability of the native protein conformation is an important factor to consider when cells interact with insulin adsorbed on metal-substituted HA surfaces.


Asunto(s)
Durapatita/química , Adsorción , Insulina , Espectroscopía Infrarroja por Transformada de Fourier , Estroncio , Zinc
10.
J Biomed Mater Res B Appl Biomater ; 104(2): 274-82, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25716009

RESUMEN

Various synthetic bone substitutes have been developed to reconstruct bone defects. One of the most prevalent ceramics in bone treatment is hydroxyapatite (HA) that is a useful material as bone substitute, however, with a low rate of biodegradation. Its structure allows isomorphic cationic and anionic substitutions to be easily introduced, which can alter the crystallinity, morphology, biocompatibility, and osteoconductivity. The objective of this study was to investigate the in vitro and in vivo biological responses to strontium-containing nanostructured carbonated HA/sodium alginate (SrCHA) spheres (425<ϕ <600 µm) that were used for sinus lifts in rabbits using nanostructured carbonated HA/sodium alginate (CHA) as a reference. Cytocompatibility was determined using a multiparametric assay after exposing murine preosteoblasts to the extracts of these materials. Twelve male and female rabbits underwent bilateral sinus lift procedures and were divided into two groups (CHA or SrCHA) and in two experimental periods (4 and 12 weeks), for microscopic and histomorphometric analyses. The in vitro test revealed the overall viability of the cells exposed to the CHA and SrCHA extracts; thus, these extracts were considered cytocompatible, which was confirmed by three different parameters in the in vitro tests. The histological analysis showed chronic inflammation with a prevalence of macrophages around the CHA spheres after 4 weeks, and this inflammation decreased after 12 weeks. Bone formation was observed in both groups, and smaller quantities of SrCHA spheres were observed after 12 weeks, indicating greater bioresorption of SrCHA than CHA. SrCHA spheres are biocompatible and osteoconductive and undergo bioresorption earlier than CHA spheres.


Asunto(s)
Alginatos , Sustitutos de Huesos , Senos Craneales/cirugía , Durapatita , Nanoestructuras/química , Estroncio , Alginatos/química , Alginatos/farmacología , Animales , Regeneración Ósea/efectos de los fármacos , Sustitutos de Huesos/química , Sustitutos de Huesos/farmacología , Línea Celular , Senos Craneales/lesiones , Senos Craneales/metabolismo , Evaluación Preclínica de Medicamentos , Durapatita/química , Durapatita/farmacología , Femenino , Ácido Glucurónico/química , Ácido Glucurónico/farmacología , Ácidos Hexurónicos/química , Ácidos Hexurónicos/farmacología , Masculino , Ensayo de Materiales/métodos , Ratones , Conejos , Estroncio/química , Estroncio/farmacología
11.
J Mater Sci Mater Med ; 26(4): 166, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25791461

RESUMEN

Hydroxyapatite (HA) has been investigated as a delivery system for antimicrobial and antibacterial agents to simultaneously stimulate bone regeneration and prevent infection. Despite evidence supporting the bactericidal efficiency of these HA carriers, few studies have focused on the effect of this association on bone regeneration. In this work, we evaluated the physico-chemical properties of hydroxyapatite microspheres loaded with chlorhexidine (CHX) at two different concentrations, 0.9 and 9.1 µgCHX/cm2 HA, and characterized their effects on in vitro osteoblast viability and bone regeneration. Ultraviolet-visible spectroscopy, scanning and transmission electron microscopy associated with energy-dispersive X-ray spectroscopy and electron energy loss spectroscopy were used to characterize the association of CHX and HA nanoparticles. The high CHX loading dose induced formation of organic CHX plate-like aggregates on the HA surface, whereas a Langmuir film was formed at the low CHX surface concentration. Quantitative evaluation of murine osteoblast viability parameters, including adhesion, mitochondrial activity and membrane integrity of cells exposed to HA/CHX extracts, revealed a cytotoxic effect for both loading concentrations. Histomorphological analysis upon implantation into the dorsal connective tissues and calvaria of rats for 7 and 42 days showed that the high CHX concentration induced the infiltration of inflammatory cells, resulting in retarded bone growth. Despite a strong decrease in in vitro cell viability, the low CHX loading dose did not impair the biocompatibility and osteoconductivity of HA during bone repair. These results indicate that high antimicrobial doses may activate a strong local inflammatory response and disrupt the long-term osteoconductive properties of CHX-HA delivery systems.


Asunto(s)
Fenómenos Fisiológicos Bacterianos/efectos de los fármacos , Sustitutos de Huesos/administración & dosificación , Clorhexidina/administración & dosificación , Implantes de Medicamentos/administración & dosificación , Osteoblastos/fisiología , Osteogénesis/fisiología , Células 3T3 , Animales , Antibacterianos/administración & dosificación , Antibacterianos/química , Sustitutos de Huesos/síntesis química , Cápsulas/administración & dosificación , Cápsulas/síntesis química , Línea Celular , Proliferación Celular/efectos de los fármacos , Proliferación Celular/fisiología , Clorhexidina/química , Terapia Combinada , Difusión , Implantes de Medicamentos/química , Durapatita/administración & dosificación , Durapatita/química , Masculino , Ratones , Osteoblastos/citología , Osteogénesis/efectos de los fármacos , Ratas , Ratas Wistar
12.
Mater Sci Eng C Mater Biol Appl ; 41: 309-19, 2014 Aug 01.
Artículo en Inglés | MEDLINE | ID: mdl-24907765

RESUMEN

The effect of zinc-substituted calcium phosphate (CaP) on bone osteogenesis was evaluated using an in vivo normalized ISO 10993-6 protocol. Zinc-containing hydroxyapatite (ZnHA) powder with 0.3% by wt zinc (experimental group) and stoichiometric hydroxyapatite (control group) were shaped into cylindrical implants (2×6 mm) and were sintered at 1000 °C. Thermal treatment transformed the ZnHA cylinder into a biphasic implant that was composed of Zn-substituted HA and Zn-substituted ß-tricalcium phosphate (ZnHA/ßZnTCP); the hydroxyapatite cylinder was a highly crystalline and poorly soluble HA implant. In vivo tests were performed in New Zealand White rabbits by implanting two cylinders of ZnHA/ßZnTCP in the left tibia and two cylinders of HA in the right tibia for 7, 14 and 28 days. Incorporation of 0.3% by wt zinc into CaP increased the rate of Zn release to the biological medium. Microfluorescence analyses (µXRF-SR) using synchrotron radiation suggested that some of the Zn released from the biomaterial was incorporated into new bone near the implanted region. In contrast with previous studies, histomorphometric analysis did not show significant differences between the newly formed bone around ZnHA/ßZnTCP and HA due to the dissolution profile of Zn-doped CaP. Despite the great potential of Zn-containing CaP matrices for future use in bone regeneration, additional in vivo studies must be conducted to explain the mobility of zinc at the CaP surface and its interactions with a biological medium.


Asunto(s)
Materiales Biocompatibles/química , Fosfatos de Calcio/química , Zinc/química , Animales , Materiales Biocompatibles/metabolismo , Materiales Biocompatibles/uso terapéutico , Durapatita/química , Femenino , Fracturas Óseas/terapia , Masculino , Prótesis e Implantes , Conejos , Tibia/patología , Factores de Tiempo , Zinc/metabolismo
13.
J Mater Sci Mater Med ; 24(5): 1271-83, 2013 May.
Artículo en Inglés | MEDLINE | ID: mdl-23494616

RESUMEN

The incorporation of zinc into the hydroxyapatite structure (ZnHA) has been proposed to stimulate osteoblast proliferation and differentiation. Another approach to improve cell adhesion and hydroxyapatite (HA) performance is coating HA with adhesive proteins or peptides such as RGD (arginine-glycine-aspartic acid). The present study investigated the adhesion of murine osteoblastic cells to non-sintered zinc-substituted HA disks before and after the adsorption of RGD. The incorporation of zinc into the HA structure simultaneously changed the topography of disk's surface on the nanoscale and the disk's surface chemistry. Fluorescence microscopy analyses using RGD conjugated to a fluorescein derivative demonstrated that ZnHA adsorbed higher amounts of RGD than non-substituted HA. Zinc incorporation into HA promoted cell adhesion and spreading, but no differences in the cell density, adhesion and spreading were detected when RGD was adsorbed onto ZnHA. The pre-treatment of disks with fetal bovine serum (FBS) greatly increased the cell density and cell surface area for all RGD-free groups, overcoming the positive contribution of zinc to cell adhesion. The presence of RGD on the ZnHA surface impaired the effects of FBS pre-treatment possibly due to competition between FBS proteins and RGD for surface binding sites.


Asunto(s)
Materiales Biocompatibles Revestidos/farmacología , Durapatita/química , Oligopéptidos/farmacología , Osteoblastos/efectos de los fármacos , Zinc/química , Adsorción , Animales , Adhesión Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Células Cultivadas , Materiales Biocompatibles Revestidos/química , Durapatita/farmacología , Ensayo de Materiales , Ratones , Osteoblastos/citología , Osteoblastos/fisiología , Albúmina Sérica Bovina/química , Propiedades de Superficie/efectos de los fármacos , Andamios del Tejido/química , Zinc/farmacología
14.
Colloids Surf B Biointerfaces ; 83(1): 1-9, 2011 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-21109408

RESUMEN

Bovine serum albumin (BSA) may have an inhibitory or promoter effect on hydroxyapatite (HA) nucleation when apatite is precipitated in a medium containing the protein. In this study we evaluated the influence of BSA on the precipitation of calcium phosphate phases (CP) from simulated body fluid (SBF) when the protein was previously bounded to HA surface. The kinetics of BSA immobilization onto hydroxyapatite surface was performed in different buffers and protein concentrations in order to adjust experimental conditions in which BSA was tightly linked to HA surface for long periods in SBF solution. It was shown that for BSA concentration higher than 0.1mg/mL the adsorption to HA surface followed Langmuir-Freundlich mechanisms, which confirmed the existence of cooperative protein-protein interactions on HA surface. Fourier Transformed Infrared Attenuated Total Reflectance Microscopy (FTIRM-ATR) evidenced changes in BSA conformational state in favor of less-ordered structure. Analyses from high resolution grazing incident X-ray diffraction using synchrotron radiation (GIXRD), Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM) showed that a poorly crystalline calcium phosphate was precipitated on the surface of HA discs coated with BSA, after the immersion in SBF for 4 days. The new bioactive layer had morphological characteristics similar to the one formed on the HA surface without protein. It was identified as a carbonated apatite with preferential crystal growth along apatite 002 direction. The GIXRD results also revealed that BSA layer bound to the surface inhibited the HA dissolution leading to a reduction on the formation of new calcium phosphate phase.


Asunto(s)
Materiales Biocompatibles/metabolismo , Durapatita/metabolismo , Albúmina Sérica Bovina/metabolismo , Adsorción , Animales , Tampones (Química) , Calcio/análisis , Bovinos , Simulación por Computador , Humanos , Microscopía de Fuerza Atómica , Microscopía Electrónica de Rastreo , Fósforo/análisis , Polvos , Espectrofotometría Atómica , Espectroscopía Infrarroja por Transformada de Fourier , Propiedades de Superficie , Temperatura , Difracción de Rayos X
15.
Rio de Janeiro; s.n; 2003. [150] p. mapas, tab, graf.
Tesis en Portugués | LILACS | ID: lil-392473

RESUMEN

A concentração de metais poluentes, como o chumbo (Pb), tem aumentado em áreas populosas e desenvolvidas devido a poluição atmosférica e outras atividades humanas. Consequêntemente, o potencial para este elemento alcançar a cadeia alimentar também aumentou. O chumbo é muito tóxico para o ser humano, especialmente para crianças, e a exposição ao chumbo pode causar efeitos adversos à saúde humana. A hidroxiapatita (Hap), um composto de fosfato de cálcio, é o principal constituinte inorgânico dos ossos e dentes. Pesquisas concluiram que a hidroxiapatita sintética tem uma alta capacidade para remover metais pesados divalentes da água em condições ambientais. Existem várias tecnologias empregadas para tratamento de água e solos contaminados. Estes tratamentos além de serem caros, muitas vezes são destrutivos. Torna-se então necessário, desenvolver novos métodos para remover íons tóxicos de solos e esgotos pela redução da solubilidade do Pb e consequentemente sua bio-disponibilidade. Visto ser a hidroxiapatita um regulador da concentração de cálcio, fósforo e chumbo no meio ambiente, este material poderia ser usado com baixos custos, na remoção de chumbo em solos e águas contaminadas. O objetivo deste trabalho é elucidar os mecanismos envolvidos na remoção de chumbo de soluções aquosas, investigando-se a transformação do Pb para uma forma mais estável: a piromorfita


Asunto(s)
Cadmio , Cobre , Exposición a Riesgos Ambientales , Plomo , Fosfatos , Metales Pesados
16.
Environ Sci Technol ; 36(7): 1625-9, 2002 Apr 01.
Artículo en Inglés | MEDLINE | ID: mdl-11999075

RESUMEN

The sorption of lead by synthetic hydroxyapatite (HA) from solutions containing Pb2+ initial concentrations up to 1770 mg L(-1) was studied. X-ray diffractometry (XRD) associated with Rietveld methodology for refining the spectra pattern was used in order to characterize the mechanisms of lead uptake. It is shown that the dissolution of hydroxyapatite is followed by the formation of a solid solution, Pb(10-x)Ca(x)(PO4)6(OH)2, with Pb ions mostly occupying Ca(II) sites. The Ca/Pb molar ratio of this solid solution decreases continuously until it reaches the structure of a pure hydroxypyromorphite. The cell parameters and the crystallite mean size behavior of both mineral phases reinforce the hypothesis that hydroxypyromorphite, PbHA, formation is the end of a process in which Pb(10-x)Ca(x)(PO4)6(OH)2 crystallites are continuously dissolved and recrystallized producing crystals with lower calcium content. Combination of Inductively Coupled Plasma spectrometry (ICP), chemical analysis, and XRD results permitted the conclusion that lead ions are not completely immobilized by precipitating Pb(10-x)Ca(x)(PO4)6(OH)2. Additional surface mechanisms also contribute to Pb2+ uptake. During Pb2+ sorption process, pH variations of the solution phase showed a more complex pattern than previously reported. Contribution of surface mechanisms, in addition to the hydroxyapatite dissolution, could explain this behavior.


Asunto(s)
Materiales Biocompatibles/química , Durapatita/química , Plomo/química , Cristalización , Contaminación Ambiental/prevención & control , Solubilidad , Difracción de Rayos X
17.
Rio de Janeiro; s.n; 1999. 105 p. ilus, tab, graf.
Tesis en Portugués | LILACS | ID: lil-239235

RESUMEN

A concentraçäo de metais poluentes, como o chumbo (Pb), tem aumentado em áreas populosas e desenvolvidas devido a poluiçäo atmosférica e outras atividades humanas. Conseqüentemente, o potencial para este elemento alcançar a cadeia alimentar aumentou. O chumbo é muito tóxico para o ser humano, especialmente para crianças, e a exposiçäo ao chumbo pode causar efeitos adversos à saúde humana. A hidroxiapatita (Hap), um composto de fosfato de cálcio, é o principal constituinte inorgânico dos ossos e dentes. Pesquisas concluiram que a hidroxiapatita sintética tem uma alta capacidade para remover metais pesados divalentes da água em condiçöes ambientais. Existem várias tecnologias empregadas para tratamento de água e solos contaminados. Estes tratamentos além de serem caros, muitas vezes säo destrutivos. Torna-se entäo necessário, desenvolver novos métodos para remover íons tóxicos de solos e esgotos pela reduçäo da solubilidade do Pb e conseqüentemente sua bio-disponibilidade. Visto ser a hidroxiapatita um regulador da concentraçäo de cálcio, fósforo e chumbo no meio ambiente, este material poderia ser usado com baixos custos, na remoçäo de chumbo em solos e águas contaminadas. O objetivo deste trabalho é elucidar os mecanismos envolvidos na remoçäo de chumbo de soluçöes aquosas, investigando-se a transformaçäo do Pb para uma forma mais estável: a piromorfita.


Asunto(s)
Durapatita , Exposición a Riesgos Ambientales , Intoxicación por Plomo
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