Efectos biológicos del empleo de implantes ortopédicos de polipropileno en perros

Este trabajo pretende aportar conocimientos sobre el desarrollo de nuevos materiales para implantes ortopédicos en Medicina Veterinaria y Humana, de bajo costo y sin efectos biológicos indeseables. Para esto se pretende verificar los posibles efectos biológicos de los implantes de polipropileno, mediante una evaluación clínica, bioquímica e histopatológica en 12 caninos a los que se les implantará una placa ortopédica, por las técnicas quirúrgicas de rutina. Sobre el material de implante (polipropileno) se efectuará un estudio de homogeneidad química y de los posibles efectos del organismo sobre el mismo, mediante espectroscopía infrarroja por transformada de Fourier y espectroscopía Uv-visible. Objetivos generales Aportar conocimientos para: desarrollar nuevos materiales para implantes ortopédicos, de bajo costo, buena adaptabilidad mecánica y toxicidad mínima. Objetivos específicos 1. Verificar los posibles efectos biológicos de los implantes (placas ortopédicas) de polipropileno en perros, mediante una evaluación clínica, bioquímica e histopatológica de los animales implantados. 2. Determinar la homogeneidad química, la presencia y concentración de antioxidantes y los posibles estados superficiales del material utilizado para la confección de las placas, mediante espectroscopía infrarroja por transformada de Fourier. 3. Determinar el efecto del organismo sobre el polímero mediante espectroscopía UV-visible.

Propriedades mecânicas e de corrosão de dois aços inoxidáveis austeníticos utilizados na fabricação de implantes ortopédicos

ASTM F 138 austenitic stainless steel is extensively used as an orthopedic implant material. However, some aspects, such as low strength in the annealed condition and susceptibility to localized corrosion, limit wider use of this kind of steel. Recently, a high-nitrogen austenitic stainless steel, specified in the standard ISO 5832-9, has been indicated as an alternative to ASTM F 138 steel for more severe loading and permanent application inside the human body. In this work, microstructure, mechanical properties, corrosion resistance and fatigue behavior of both steels were determined and compared. ISO 5832-9 steel displayed better mechanical and corrosion behaviors than did ASTM F 138 steel The combination of these features lead ISO steel to enhanced fatigue performance in both neutral and aggressive environments. Analyzed were the role of nitrogen in solid solution, combined with niobium in the Z-phase, and the factors that led to superior ISO 5832-9 properties.

Avaliação microbiológica da superfície dos implantes ortopédicos metálicos usados para fixar fraturas ósseas

In the majority of cases of bone fracture requiring surgery, orthopedic implants (screw-plate and screw) are used for osteosynthesis and the infections associated with such implants are due to the growth of microorganisms in biofilms. The objective of this study was to identify microorganisms recovered from osteosynthesis implants used to fix bone fractures, to assess the viability of the cells and the ability of staphylococci to adhere to a substrate and to determine their sensitivity/resistance to antimicrobials. After surgical removal, the metal parts of austenitic stainless steel (ASTM F138/F139 or ISO NBR 5832-1/9) were transported to the Laboratory of Clinical Microbiology, washed in buffer and subjected to ultrasonic bath at 40±2 kHz for 5 minutes. The sonicated fluid was used to seed solid culture media and cell viability was assessed under the microscope by with the aid of a fluorescent marker. The production of extracellular polysaccharide by Staphylococcus spp. was investigated by means of adhesion to a polystyrene plate. The profile of susceptibility to antimicrobials was determined by the disk diffusion assay. The most frequently isolated bacteria included coagulase-negative Staphylococcus resistant to erythromycin, clindamycin and oxacillin. Less frequent were Pseudomonas aeruginosa resistant to trimethoprim/sulfamethoxazole and ampicillin, Acinetobacter baumannii resistant to ceftazidime, Enterobacter cloacae resistant to cephalothin, cefoxitin, cefazolin, levofloxacin and ciprofloxacin, Bacillus spp. and Candida tropicalis. The observation of slides by fluorescence microscope showed clusters of living cells embedded in a transparent matrix. The test for adherence of coagulase-negative Staphylococcus to a polystyrene plate showed that these microorganisms produce extracellular polysaccharide. In conclusion, the metal parts were colonized by bacteria related to orthopedic implant infection, which were resistant to multiple antibiotics.

Análisis de distribución de tensiones asociada a la utilización de implantes ortopédicos de materiales superelásticos en el tratamiento de deficiencias óseas.

Durante el desarrollo de un niño pueden ocurrir diversas anomalías y distorsiones en el crecimiento de los huesos que resultan en distintos problemas. Independientemente de las razones por las que ocurren, si son detectadas y tratadas con tiempo sus secuelas pueden ser minimizadas o eliminadas. Este trabajo continúa el proyecto integrador de ingeniería mecánica del ingeniero Matías Korten. En ese trabajo se realizaron ensayos termomecánicos a alambres de NiTi para caracterizar las fuerzas desarrolladas y la influencia de diferentes parámetros geométricos y físicos en las mismas. El Dr. J. Groiso propone la fabricación de un dispositivo pseudoelástico y biocompatible de NiTi para corregir deformaciones angulares en huesos. En este trabajo, se buscó caracterizar nuevamente el material NiTi, validar los resultados obtenidos en ese trabajo y diseñar un dispositivo que permita la caracterización del material de forma más confiable. Se desarrolló un modelo computacional de un hueso en 3D que permite obtener el campo de tensiones sobre la placa de crecimiento al aplicar una fuerza superficial sobre los tornillos. Se analizaron los resultados con distintas formas de aplicación de la fuerza, la zona afectada por la prótesis al cambiar la posición de los tornillos y se hizo un análisis de sensibilidad en el rango (según bibliografía) del módulo de elasticidad de la placa de crecimiento. Se concluye que la aplicación de un dispositivo de NiTi puede generar las tensiones necesarias para impulsar el crecimiento del hueso en la dirección correcta.

Análisis de distribución de tensiones asociada a la utilización de implantes ortopédicos de materiales superelásticos en el tratamiento de deficiencias óseas.

Durante el desarrollo de un niño pueden ocurrir diversas anomalías y distorsiones en el crecimiento de los huesos que resultan en distintos problemas. Independientemente de las razones por las que ocurren, si son detectadas y tratadas con tiempo sus secuelas pueden ser minimizadas o eliminadas. Este trabajo continúa el proyecto integrador de ingeniería mecánica del ingeniero Matías Korten. En ese trabajo se realizaron ensayos termomecánicos a alambres de NiTi para caracterizar las fuerzas desarrolladas y la influencia de diferentes parámetros geométricos y físicos en las mismas. El Dr. J. Groiso propone la fabricación de un dispositivo pseudoelástico y biocompatible de NiTi para corregir deformaciones angulares en huesos. En este trabajo, se buscó caracterizar nuevamente el material NiTi, validar los resultados obtenidos en ese trabajo y diseñar un dispositivo que permita la caracterización del material de forma más confiable. Se desarrolló un modelo computacional de un hueso en 3D que permite obtener el campo de tensiones sobre la placa de crecimiento al aplicar una fuerza superficial sobre los tornillos. Se analizaron los resultados con distintas formas de aplicación de la fuerza, la zona afectada por la prótesis al cambiar la posición de los tornillos y se hizo un análisis de sensibilidad en el rango (según bibliografía) del módulo de elasticidad de la placa de crecimiento. Se concluye que la aplicación de un dispositivo de NiTi puede generar las tensiones necesarias para impulsar el crecimiento del hueso en la dirección correcta.

Estudo mecânico comparativo da estabilização de fratura ilíaca com dois tipos de implantes (parafusos com placa de compressão dinâmica e parafusos associados ao polimetilmetacrilato) em hemipelves caninas sintéticas

Pós-graduação em Cirurgia Veterinária - FCAV

Estudo eletroquímico comparativo do aço ISO 5832-9 em diferentes meios de interesse biológico

Foi estudado o comportamento eletroquímico a 37°C do aço inoxidável ISO 5832-9, em meios de NaCl 0,9 %, de Ringer Lactato e meio mínimo de Eagle (MEM), por voltametria linear e análises da superfície por microscopia eletrônica de varredura (MEV) e por espectroscopia por dispersão de energia (EDS). Foram feitos ensaios mecânicos e testes de toxicidade. O aço ISO 5832-9 se encontra passivado no potencial de corrosão e não apresenta corrosão por pite nos três meios estudados em toda faixa de potencial investigada, desde o potencial de corrosão até 50 mV acima do potencial de transpassivação. Em meio de MEM, no entanto, as análises por MEV e EDS mostraram que o referido aço, nesse valor mais elevado de potencial, apresentou um comportamento diferente, com perda das inclusões de óxido de manganês. Os potenciais de corrosão, Ecorr (potencial de circuito aberto estacionário) bem como os valores de densidade de corrente de passivação, variaram na seguinte ordem: Ecorr, RL < Ecorr, NaCl < Ecorr, MEM. e jMEM << jRL ≅ jNaCl. No ensaio de citotoxicidade, o aço foi caracterizado como não-tóxico.

Biochemical and physical surface functionalization of polycaprolactone as a key mediator of osteogenic differentiation and vascularized tissue morphogenesis

Tese de Doutoramento em Engenharia de Tecidos, Medicina Regenerativa e Células Estaminais.

Antibacterial properties of multifunctional coatings Ag-ZrCN for biomedical devices: a novel approach

PhD in Sciences Specialty in Physics

Avaliação do estresse oxidativo induzido por superfícies de titânio

Commercially pure Titanium (cp Ti) is a material largely used in orthopedic and dental implants due to its biocompatibility properties. Changes in the surface of cp Ti can determine the functional response of the cells such as facilitating implant fixation and stabilization, and increased roughness of the surface has been shown to improve adhesion and cellular proliferation. Various surface modification methods have been developed to increase roughness, such as mechanical, chemical, electrochemical and plasma treatment. An argon plasma treatment generates a surface that has good mechanical proprieties without chemical composition modification. Besides the topography, biological responses to the implant contribute significantly to its success. Oxidative stress induced by the biomaterials is considered one of the major causes of implant failure. For this reason the oxidative potential of titanium surfaces subjected to plasma treatment was evaluated on this work. CHO-k1 cells were cultivated on smooth or roughed Ti disks, and after three days, the redox balance was investigated measuring reactive oxygen species (ROS) generation, total antioxidant capacity and biomarkers of ROS attack. The results showed cells grown on titanium surfaces are subjected to intracellular oxidative stress due to hydrogen peroxide generation. Titanium discs subjected to the plasma treatment induced less oxidative stress than the untreated ones, which resulted in improved cellular ability. Our data suggest that plasma treated titanium may be a more biocompatible biomaterial.

Análise da estabilidade genética de células-tronco mesenquimais humanas

Human multipotent mesenchymal stromal cells (MSCs), also known as mesenchymal stem cells, have become an important and attractive therapeutic tool since they are easily isolated and cultured, have in vitro expansion potential, substantial plasticity and secrete bioactive molecules that exert trophic effects. The human umbilical cord as a cell source for cell therapy will help to avoid several ethical, political, religious and technical issues. One of the main issues with SC lines from different sources, mainly those of embryonic origin, is the possibility of chromosomal alterations and genomic instability during in vitro expansion. Cells isolated from one umbilical cord exhibited a rare balanced paracentric inversion, likely a cytogenetic constitutional alteration, karyotype: 46,XY,inv(3)(p13p25~26). Important genes related to cancer predisposition and others involved in DNA repair are located in 3p25~26. Titanium is an excellent biomaterial for bone-implant integration; however, the use can result in the generation of particulate debris that can accumulate in the tissues adjacent to the prosthesis, in the local bone marrow, in the lymph nodes, liver and spleen. Subsequently may elicit important biological responses that aren´t well studied. In this work, we have studied the genetic stability of MSC isolated from the umbilical cord vein during in vitro expansion, after the cryopreservation, and under different concentrations and time of exposition to titanium microparticles. Cells were isolated, in vitro expanded, demonstrated capacity for osteogenic, adipogenic and chondrogenic differentiation and were evaluated using flow cytometry, so they met the minimum requirements for characterization as MSCs. The cells were expanded under different concentrations and time of exposition to titanium microparticles. The genetic stability of MSCs was assessed by cytogenetic analysis, fluorescence in situ hybridization (FISH) and analysis of micronucleus and other nuclear alterations (CBMN). The cells were able to internalize the titanium microparticles, but MSCs preserve their morphology, differentiation capacity and surface marker expression profiles. Furthermore, there was an increase in the genomic instability after long time of in vitro expansion, and this instability was greater when cells were exposed to high doses of titanium microparticles that induced oxidative stress. It is necessary always assess the risks/ benefits of using titanium in tissue therapy involving MSCs, considering the biosafety of the use of bone regeneration using titanium and MSCs. Even without using titanium, it is important that the therapeutic use of such cells is based on analyzes that ensure quality, security and cellular stability, with the standardization of quality control programs appropriate. In conclusion, it is suggested that cytogenetic analysis, FISH analysis and the micronucleus and other nuclear alterations are carried out in CTMH before implanting in a patient

Análise do potencial genotóxico da superfície de titânio modificada por plasma

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior

Efeito do oxigênio nas propriedades anelásticas da liga Ti-10Mo para aplicação biomédica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Caracterização fenotípica e genotípica de bactérias recuperadas de implante ortopédico de conjunto placa-parafuso e parafuso de aço inoxidável austenítico após remoção cirúrgica

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Efeito de elementos intersticiais nas propriedades físicas e biocompatibilidade da liga 'TI'-13'NB'-13'ZR'

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)