RESUMO
Prior studies on two-dimensional cell cultures suggest that hyaluronic acid (HA) stimulates cell-mediated regeneration of extracellular matrix structures, specifically those containing elastin, though such biologic effects are dependent on HA fragment size. Towards being able to regenerate three-dimensional (3-D) elastic tissue constructs, the present paper studies photo-crosslinked hydrogels containing glycidyl methacrylate (GM)-derivatized bio-inert high molecular weight (HMW) HA (1 × 10(6)Da) and a bioactive HA oligomer mixture (HA-o: MW â¼0.75 kDa). The mechanical (rheology, degradation) and physical (apparent crosslinking density, swelling ratio) properties of the gels varied as a function of incorporated HA oligomer content; however, overall, the mechanics of these hydrogels were too weak for vascular applications as stand-alone materials. Upon in vivo subcutaneous implantation, only a few inflammatory cells were evident around GM-HA gels, however their number increased as HA-o content within the gels increased, and the collagen I distribution was uniform. Smooth muscle cells (SMC) were encapsulated into GM hydrogels, and calcein acetoxymethyl detection revealed that the cells were able to endure twofold the level of UV exposure used to crosslink the gels. After 21 days of culture, SMC elastin production, measured by immunofluorescence quantification, showed HA-o to increase cellular deposition of elastic matrix twofold relative to HA-o-free GM-HA gels. These results demonstrate that cell response to HA/HA-o is not altered by their methacrylation and photo-crosslinking into a hydrogel, and that HA-o incorporation into cell-encapsulating hydrogel scaffolds can be useful for enhancing their production of elastic matrix structures in a 3-D space, important for regenerating elastic tissues.
Assuntos
Reagentes de Ligações Cruzadas/farmacologia , Elastina/metabolismo , Compostos de Epóxi/farmacologia , Ácido Hialurônico/farmacologia , Hidrogel de Polietilenoglicol-Dimetacrilato/farmacologia , Metacrilatos/farmacologia , Alicerces Teciduais/química , Animais , Materiais Biocompatíveis/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Elasticidade/efeitos dos fármacos , Compostos de Epóxi/síntese química , Compostos de Epóxi/química , Imunofluorescência , Ácido Hialurônico/síntese química , Ácido Hialurônico/química , Inflamação/patologia , Metacrilatos/síntese química , Metacrilatos/química , Microscopia Eletrônica de Varredura , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/metabolismo , Tamanho da Partícula , Ratos , Ratos Sprague-Dawley , Espectroscopia de Infravermelho com Transformada de Fourier , Fatores de Tempo , Viscosidade/efeitos dos fármacosRESUMO
There are numerous reports in the literature using animal models of osteomyelitis for investigating pathogenesis, diagnosis, and treatment of bone infections. Rabbits, rats, and dogs are commonly used animals, and, less frequently, chickens, guinea pigs, miniature pigs, goats, and sheep. Commonly used bones for creating local osteomyelitis include tibia, femur, and radius, and, less frequently, mandible and spine. When designing a specific model, one should consider which animal and which bone will be used, which route for inoculation (either local injection or systemically through vascular injection), which bacterial species and how many bacteria should be applied, if and what sclerosing agent, foreign body or implant should be employed, and if local trauma is needed. Basic methods of evaluation include clinical observation, radiography, microbiology, and histology.
Assuntos
Modelos Animais de Doenças , Osteomielite/patologia , Animais , Osteomielite/microbiologiaRESUMO
Several species belonging to Staphylococcus genus, other than Staphylococcus aureus and Staphylococcus epidermidis (non Sau/ non Sep species), exhibit increasing abilities as opportunistic pathogens in the colonisation of periprosthetic tissues. Consequently, the availability of means for accurate identification is crucial to assess the pathogenic characteristics and to clarify clinical relevance of the individual species. Here, 146 clinical staphylococcal isolates belonging to non Sau/ non Sep species from prosthesis-associated orthopedic infections were analyzed by conventional enzymatic galleries and by automated ribotyping. Twelve different species were recognised: S. capitis, S. caprae, S. cohnii, S. equorum, S. haemolyticus, S. hominis, S. lugdunensis, S. pasteuri, S. sciuri, S. simulans, S. warneri, S. xylosus. Ribotype identifications were compared with the phenotypes obtained by the Api 20 Staph system and/or ID 32 Staph system. ID 32 Staph profiles were more consistent with ribotyping results than Api Staph profiles. Across the different staphylococcal species investigated, correct identifications with Api Staph were 45%, while with ID 32 Staph they were 59%. It has, however, to be mentioned that ID 32 Staph was mostly applied to discriminate unmatched ribotyping and Api Staph identifications, thus to a subpopulation of strains with ""atypical"" metabolic profile. Automated ribotyping provided a correct identification for 91% of the isolates. These results confirm automated ribotyping as a convenient rapid technique, still subject to improvements, which will accurately and rapidly recognise the newly emerging staphylococcal pathogens in implant-related orthopedic infections.
Assuntos
Automação , Equipamentos Ortopédicos/microbiologia , Ribotipagem , Infecções Estafilocócicas/microbiologia , Staphylococcus/isolamento & purificação , Humanos , Especificidade da Espécie , Infecções Estafilocócicas/diagnóstico , Staphylococcus/classificação , Staphylococcus/genéticaRESUMO
Supercritical phase CO2 is a promising method for sterilizing implantable devices and tissue grafts. The goal of this study is to evaluate the biocompatibility of titanium implants sterilized by supercritical phase CO2 in a rat subcutaneous implantation model. At 5 weeks post implantation titanium implants sterilized by supercritical phase CO2 produce a soft tissue reaction that is comparable to other methods of sterilization (steam autoclave, ultraviolet light radiation, ethylene oxide gas, and radio-frequency glow-discharge), as indicated by the thickness and density of the foreign body capsule, although there were some differences on the capillary density. Overall the soft tissue response to the implants was similar among all methods of sterilization, indicating supercritical phase CO2 treatment did not compromise the biocompatibility of the titanium implant.