RESUMO
Many biomaterials are used for Bone Morphogenetic Proteins (BMPs) delivery in bone tissue engineering. The BMP carrier system's primary function is to hold these growth factors at the wound's site for a prolonged time and provide initial support for cells to attach and elaborate the extracellular matrix for bone regeneration. This study aimed to evaluate the nanostructured carbonated hydroxyapatite microspheres (nCHA) as an rhBMP-2 carrier on rats calvaria. A total of fifteen male Wistar rats were randomly divided into three groups (n = 5): clot (control group), rhBMP-2 associated with collagen membrane (COL/rhBMP-2) or associated with the microspheres (nCHA/rhBMP-2). After 45 days, the calvaria defect samples were evaluated through histological, histomorphometric, and SR-µCT analyses to investigate new-formed bone and connective tissue volume densities. The descriptive histological analysis showed that nCHA/rhBMP-2 improved bone formation compared to other groups. These results were confirmed by histomorphometric and SR-µCT analysis that showed substantially defect area filling with a higher percentage of newly formed (36.24 ± 6.68) bone than those with the COL/rhBMP-2 (0.42 ± 0.40) and Clot (3.84 ± 4.57) (p < 0.05). The results showed that nCHA is an effective carrier for rhBMP-2 encouraging bone healing and an efficient alternative to collagen membrane for rhBMP-2 delivery.
RESUMO
To develop an orthopedic material for bone substitution, the substitute material must mimic living tissue from an anatomical and physiological point of view. The high wear and impact resistance besides the low friction coefficient, make ultra-high molecular weight polyethylene (UHMWPE) a suitable material to be used in orthopedic applications. However, UHMWPE is a bioinert material, not providing a proper interaction with the bone tissue surrounding to the implant. One way to mitigate this issue is improving UHMWPE bioactivity. This can be done by adding bioactive fillers in the polymeric matrix. In this work, UHMWPE composites were prepared by twin-screw extrusion. The fillers used were carbonated hydroxyapatite (CHA) and hybrids formed by precipitating CHA in collagens (hydrolyzed and type II). The results show that the fillers used caused a slight reduction in UHMWPE crystallinity degree, while both crystallization and melting temperatures remained almost unchanged. Dynamic-mechanical thermal analysis indicated a weak adhesion between filler and polymeric matrix, which is good from the biological point of view since the bioactive filler surface will be available to apatite deposition. The obtained materials exhibited good mechanical properties and in vitro bioactivity assay showed that all of the prepared materials are bioactive.
Assuntos
Durapatita , Polietilenos , Fricção , Teste de Materiais , Peso MolecularRESUMO
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.
Assuntos
Alginatos/química , Regeneração Óssea/efeitos dos fármacos , Carbonatos/química , Durapatita/química , Microesferas , Minociclina/farmacologia , Nanopartículas/química , Animais , Antibacterianos/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Enterococcus/efeitos dos fármacos , Humanos , Masculino , Camundongos , Testes de Sensibilidade Microbiana , Osteoblastos/efeitos dos fármacos , Ratos Wistar , Difração de Raios XRESUMO
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.
Assuntos
Alginatos/química , Regeneração Óssea , Cálcio/metabolismo , Durapatita/química , Microesferas , Nanopartículas/química , Zinco/química , Zinco/metabolismo , Animais , Materiais Biocompatíveis/química , Disponibilidade Biológica , Regeneração Óssea/efeitos dos fármacos , Carbonatos/química , Morte Celular , Linhagem Celular , Sobrevivência Celular , Feminino , Camundongos , Nanopartículas/ultraestrutura , Ratos Wistar , Crânio/fisiologia , Espectroscopia de Infravermelho com Transformada de Fourier , Difração de Raios XRESUMO
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<Ï <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.
Assuntos
Alginatos , Substitutos Ósseos , Cavidades Cranianas/cirurgia , Durapatita , Nanoestruturas/química , Estrôncio , Alginatos/química , Alginatos/farmacologia , Animais , Regeneração Óssea/efeitos dos fármacos , Substitutos Ósseos/química , Substitutos Ósseos/farmacologia , Linhagem Celular , Cavidades Cranianas/lesões , Cavidades Cranianas/metabolismo , Avaliação Pré-Clínica de Medicamentos , Durapatita/química , Durapatita/farmacologia , Feminino , Ácido Glucurônico/química , Ácido Glucurônico/farmacologia , Ácidos Hexurônicos/química , Ácidos Hexurônicos/farmacologia , Masculino , Teste de Materiais/métodos , Camundongos , Coelhos , Estrôncio/química , Estrôncio/farmacologiaRESUMO
A hidroxiapatita (HA) tem sido amplamente utilizada como um importante substituto ósseo. Quando em dimensão nanométrica, assemelha-se em tamanho e morfologia à apatita biológica, podendo ser considerada um biomaterial promissor para aplicação clínica. O estrôncio contribui por atuar na redução da reabsorção óssea e indução na atividade osteoblástica, enquanto os carbonatos favorecem a bioabsorção. Objetivos: caracterizar físico-quimicamente e analisar histologicamente, e de forma comparativa, a hidroxiapatita carbonatada contendo 5% de estrôncio com a hidroxiapatita estequiométrica. Material e métodos: foram utilizados 12 coelhos brancos Nova Zelândia, divididos em: hidroxiapatita carbonatada nanoestruturada contendo 5% de estrôncio (nSrcHA-experimental) e hidroxiapatita carbonatada nanoestruturada (ncHA-controle). Após a confecção dos sítios cirúrgicos, foram implantadas nas cavidades dos seios maxilares microesferas de ncSrHA e ncHA, nos lados esquerdo e direito, respectivamente. Os animais foram eutanasiados para análise histológica após quatro e 12 semanas. Resultados: após quatro semanas, o grupo ncHA apresentou osso neoformado e pavimentação osteoblástica próximo da parede do defeito. No grupo nSrcHA, o biomaterial apresentou-se de forma difusa com uma maior deposição de matriz osteogênica em torno do biomaterial, tecido ósseo neoformado próximo das paredes e no interior dos defeitos. No período de 12 semanas o grupo ncHA exibiu biomaterial no interior do defeito e osso neoformado, enquanto no grupo nSrcHA observou-se uma intensa formação óssea no interior do defeito com presença de osteócitos. Conclusão: ambos os materiais foram biocompatíveis e osteocondutores.
Hydroxyapatite (HA) has been widely used as an important bone substitute. Its nanometer scale is similar in size and morphology of biological apatite, which can be considered a promising biomaterial for clinical application. Strontium contributes to reduce bone resorption and induces osteoblast activity, whereas the carbonates favor bio-absorption. Objectives: to perform physico-chemical and histological characterization of nanostructured carbonated hydroxyapatite containing 5% strontium and the stoichiometric hydroxyapatite. Material and methods: twelve white New Zealand rabbits were used in this study and divided into nanostructured carbonated hydroxyapatite containing 5% strontium (ncSrHA-experimental) and nanostructured carbonated hydroxyapatite (ncHA-control). Two surgical defects were created in the maxillary sinus cavities and received microspheres of ncSrHA and ncHA in the left and right sides, respectively. After the experimental periods of 4 and 12 weeks, the animals were euthanized for histological analysis. Results: after four weeks, the ncHA group showed new bone formation and osteoblastic layer near the defect wall. For ncSrHA, a diffuse, increased osteogenic matrix deposition was seen around the biomaterial, with newly formed bone near the walls and inside the defects. At 12 weeks, the ncHA group exhibited biomaterial inside the defect and new bone formation, while in the ncSrHA group an intense bone formation within the defect with presence of osteocytes was observed. Conclusion: both materials are biocompatible and osteoconductive.