RESUMO
The aim of the study was to develop an orthopedic implant coating in the form of vancomycin-loaded collagen/hydroxyapatite layers (COLHA+V) that combine the ability to prevent bone infection with the ability to promote enhanced osseointegration. The ability to prevent bone infection was investigated employing a rat model that simulated the clinically relevant implant-related introduction of bacterial contamination to the bone during a surgical procedure using a clinical isolate of Staphylococcus epidermidis. The ability to enhance osseointegration was investigated employing a model of a minipig with terminated growth. Six weeks following implantation, the infected rat femurs treated with the implants without vancomycin (COLHA+S. epidermidis) exhibited the obvious destruction of cortical bone as evinced via a cortical bone porosity of up to 20% greater than that of the infected rat femurs treated with the implants containing vancomycin (COLHA+V+S. epidermidis) (3%) and the non-infected rat femurs (COLHA+V) (2%). The alteration of the bone structure of the infected COLHA+S. epidermidis group was further demonstrated by a 3% decrease in the average Ca/P molar ratio of the bone mineral. Finally, the determination of the concentration of vancomycin released into the blood stream indicated a negligible systemic load. Six months following implantation in the pigs, the quantified ratio of new bone indicated an improvement in osseointegration, with a two-fold bone ingrowth on the COLHA (47%) and COLHA+V (52%) compared to the control implants without a COLHA layer (27%). Therefore, it can be concluded that COLHA+V layers are able to significantly prevent the destruction of bone structure related to bacterial infection with a minimal systemic load and, simultaneously, enhance the rate of osseointegration.
RESUMO
A composite nanofibrous layer containing collagen and hydroxyapatite was deposited on selected surface areas of titanium acetabular cups. The layer was deposited on the irregular surface of these 3D objects using a specially developed electrospinning system designed to ensure the stability of the spinning process and to produce a layer approximately 100 micrometers thick with an adequate thickness uniformity. It was verified that the layer had the intended nanostructured morphology throughout its entire thickness and that the prepared layer sufficiently adhered to the smooth surface of the model titanium implants even after all the post-deposition sterilization and stabilization treatments were performed. The resulting layers had an average thickness of (110 ± 30) micrometers and an average fiber diameter of (170 ± 49) nanometers. They were produced using a relatively simple and cost-effective technology and yet they were verifiably biocompatible and structurally stable. Collagen- and hydroxyapatite-based composite nanostructured surface modifications represent promising surface treatment options for metal implants.
Assuntos
Nanoestruturas , Eletricidade Estática , Nanoestruturas/química , Nanoestruturas/ultraestrutura , Análise Espectral RamanRESUMO
Infections of the musculoskeletal system present a serious problem with regard to the field of orthopedic and trauma medicine. The aim of the experiment described in this study was to develop a resorbable nanostructured composite layer with the controlled elution of antibiotics. The layer is composed of collagen, hydroxyapatite nanoparticles, and vancomycin hydrochloride (10 wt%). The stability of the collagen was enhanced by means of cross-linking. Four cross-linking agents were studied, namely an ethanol solution, a phosphate buffer solution of N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride/N-hydroxysuccinimide, genipin, and nordihydroguaiaretic acid. High performance liquid chromatography was used so as to characterize the in vitro release rates of the vancomycin and its crystalline degradation antibiotically inactive products over a 21-day period. The maximum concentration of the released active form of vancomycin (approximately 265 mg/L) exceeded the minimum inhibitory concentration up to an order of 17 times without triggering the burst releasing effect. At the end of the experiment, the minimum inhibitory concentration was exceeded by up to 6 times (approximately 100 mg/L). It was determined that the modification of collagen with hydroxyapatite nanoparticles does not negatively influence the sustainable release of vancomycin. The balance of vancomycin and its degradation products was observed after 14 days of incubation.
Assuntos
Colágeno/química , Preparações de Ação Retardada/química , Sistemas de Liberação de Medicamentos/métodos , Nanoestruturas/química , Vancomicina/química , Carbodi-Imidas/química , Portadores de Fármacos/química , Durapatita , Metilaminas/química , Nanopartículas/químicaRESUMO
Nanocomposite scaffolds which aimed to imitate a bone extracellular matrix were prepared for bone surgery applications. The scaffolds consisted of polylactide electrospun nano/sub-micron fibres, a natural collagen matrix supplemented with sodium hyaluronate and natural calcium phosphate nano-particles (bioapatite). The mechanical properties of the scaffolds were improved by means of three different cross-linking agents: N-(3-dimethylamino propyl)-N'-ethylcarbodiimide hydrochloride and N-hydroxysuccinimide in an ethanol solution (EDC/NHS/EtOH), EDC/NHS in a phosphate buffer saline solution (EDC/NHS/PBS) and genipin. The effect of the various cross-linking conditions on the pore size, structure and mechanical properties of the scaffolds were subsequently studied. In addition, the mass loss, the swelling ratio and the pH of the scaffolds were determined following their immersion in a cell culture medium. Furthermore, the metabolic activity of human mesenchymal stem cells (hMSCs) cultivated in scaffold infusions for 2 and 7 days was assessed. Finally, studies were conducted of cell adhesion, proliferation and penetration into the scaffolds. With regard to the structural stability of the tested scaffolds, it was determined that EDC/NHS/PBS and genipin formed the most effectively cross-linked materials. Moreover, it was discovered that the genipin cross-linked scaffold also provided the best conditions for hMSC cultivation. In addition, the infusions from all the scaffolds were found to be non-cytotoxic. Thus, the genipin and EDC/NHS/PBS cross-linked scaffolds can be considered to be promising biomaterials for further in vivo testing and bone surgery applications.
Assuntos
Substitutos Ósseos/síntese química , Colágeno/química , Transplante de Células-Tronco Mesenquimais/instrumentação , Células-Tronco Mesenquimais/fisiologia , Nanocompostos/química , Alicerces Teciduais , Materiais Biocompatíveis/síntese química , Matriz Óssea/química , Adesão Celular/fisiologia , Proliferação de Células/fisiologia , Sobrevivência Celular/fisiologia , Reagentes de Ligações Cruzadas/química , Desenho de Equipamento , Análise de Falha de Equipamento , Humanos , Teste de Materiais , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/citologia , Nanocompostos/ultraestrutura , Tamanho da PartículaRESUMO
Hydroxyapatite (HA) is considered to be a bioactive material that favorably influences the adhesion, growth, and osteogenic differentiation of osteoblasts. To optimize the cell response on the hydroxyapatite composite, it is desirable to assess the optimum concentration and also the optimum particle size. The aim of our study was to prepare composite materials made of polydimethylsiloxane, polyamide, and nano-sized (N) or micro-sized (M) HA, with an HA content of 0%, 2%, 5%, 10%, 15%, 20%, 25% (v/v) (referred to as N0-N25 or M0-M25), and to evaluate them in vitro in cultures with human osteoblast-like MG-63 cells. For clinical applications, fast osseointegration of the implant into the bone is essential. We observed the greatest initial cell adhesion on composites M10 and N5. Nano-sized HA supported cell growth, especially during the first 3 days of culture. On composites with micro-size HA (2%-15%), MG-63 cells reached the highest densities on day 7. Samples M20 and M25, however, were toxic for MG-63 cells, although these composites supported the production of osteocalcin in these cells. On N2, a higher concentration of osteopontin was found in MG-63 cells. For biomedical applications, the concentration range of 5%-15% (v/v) nano-size or micro-size HA seems to be optimum.
Assuntos
Fenômenos Fisiológicos Celulares/efeitos dos fármacos , Durapatita/química , Durapatita/farmacologia , Nanopartículas/química , Tamanho da Partícula , Linhagem Celular Tumoral , Proteínas do Citoesqueleto/metabolismo , Humanos , Microesferas , Osteoblastos , Osteocalcina/metabolismo , Osteopontina/metabolismo , Solubilidade , Espectrometria por Raios XRESUMO
Composite materials based on a polyamide fabric (aramid) and a polydymethylsiloxane (PDMS) matrix were designed for application in bone surgery. In order to increase the bioactivity, 2, 5, 10, 15, 20, and 25 vol.% of nano/micro hydroxyapatite (HA) and tricalcium phosphate (TCP) were added. We studied the effect of the additives on the biocompatibility of the composite. It appears that nano additives have a more favorable effect on mechanical properties than microparticles. 15 vol.% of nano hydroxyapatite additive is an optimum amount for final application of the composites as substitutes for bone tissue: in this case both the mechanical properties and the biological properties are optimized without distinct changes in the inner structure of the composite.