Made available in DSpace on 2015-04-14T13:58:54Z (GMT). No. of bitstreams: 1 443125.pdf: 1527521 bytes, checksum: 2c9a9ef7ac02480e7ed3b2734a8acc04 (MD5) Previous issue date: 2012-08-22 / Peripheral nerve injury accident is a common type of lesion that can cause loss of mobility and decrease the quality of life. Despite advances in the biomedical area, there are few cases where there is complete functional recovery of a damaged nerve. The application of biodegradable polymers such as poly(lactide-co-glycolide) (PLGA), shows a viable alternative of treatment in peripheral nerves injuries for presenting mechanical strength, biocompatibility and controlled degradation, being also an excellent material for drug delivery. A PLGA scaffold should be porous with pore sizes on the order of micro-and nanometer, which offers the possibility to insert growth factors, promoting a constant release of these biological structures. The aim of this research is to produce porous scaffolds of PLGA. This was achieved by dissolving the polymer in chloroform, adding a porogenic compound. A titanium rod with 2mm in diameter was deeped in the suspension obtained and dried in a hood, obtaining a layer of PLGA / porogen agent on the surface of the rod. In sequence, the rod was immersed in a solution of PLGA with chloroform. After dried for 2 hours in a hood, the titanium rods were submerged in deionized water for 48 hours, during which the water was changed every 8 hours in order to remove the porogenic agent. After this process, the tubular structures formed were easily removed from the rods. Finally, the prepared tubes were divided into groups and treated with different concentrations of sodium hydroxide, to create pores in the order of nanometers, and subsequently vacuum dried for 48 hours. The structures obtained were characterized by XRD, FTIR, SEM and the in vitro degradation was monitored by GPC. / Danos a nervos perif?ricos s?o formas comuns de les?es que podem causar perda de mobilidade e diminuir a qualidade de vida. Apesar dos avan?os no campo biom?dico, existem poucos casos em que h? recupera??o funcional completa de um nervo danificado. A aplica??o de pol?meros biodegrad?veis tais como PLGA, apresenta uma alternativa vi?vel no tratamento de les?es de nervos perif?ricos por possuir resist?ncia mec?nica, biocompatibilidade e degrada??o controlada. Uma matriz de PLGA deve apresentar poros na ordem de micr?metros e nan?metros, oferecendo a possibilidade de inclus?o de fatores de crescimento e possivelmente promovendo uma liberta??o constante destas estruturas biol?gicas. O objetivo desta pesquisa ? produzir matrizes porosas de PLGA. Isto foi realizado atrav?s da dissolu??o do pol?mero em clorof?rmio, seguida da adi??o de um composto porog?nico. Uma haste de tit?nio com 2mm de di?metro foi imersa na suspens?o obtida e quando retirada, foi seca em capela, obtendo uma camada de PLGA/agente porog?nico na superf?cie da haste. Na sequ?ncia, a haste foi imersa em uma solu??o de PLGA com clorof?rmio. Ap?s seca durante 2h em capela, as hastes de tit?nio foram submersas em ?gua deionizada durante 48h, durante as quais, a ?gua foi trocada a cada 8h, para remo??o do agente porog?nico. Ap?s este processo, as estruturas tubulares formadas s?o facilmente removidas das hastes. Finalmente, os tubos confeccionados foram divididos em grupos, e tratados com diferentes concentra??es de hidr?xido de s?dio, para cria??o de poros na ordem de nan?metros, sendo, subsequentemente, secos em forno a v?cuo durante 48h. As estruturas obtidas foram caracterizadas por XRD, FTIR, MEV e a degrada??o in vitro foi acompanhada por GPC.

Submitted by PPG Medicina e Ci?ncias da Sa?de (medicina-pg@pucrs.br) on 2018-07-12T13:10:54Z No. of bitstreams: 1 TATIANA_ZOTTI_PIT?GORAS.pdf: 16837328 bytes, checksum: 65a299720852db7830bef151dfdba46a (MD5) / Approved for entry into archive by Sheila Dias (sheila.dias@pucrs.br) on 2018-07-17T14:10:24Z (GMT) No. of bitstreams: 1 TATIANA_ZOTTI_PIT?GORAS.pdf: 16837328 bytes, checksum: 65a299720852db7830bef151dfdba46a (MD5) / Made available in DSpace on 2018-07-17T14:31:09Z (GMT). No. of bitstreams: 1 TATIANA_ZOTTI_PIT?GORAS.pdf: 16837328 bytes, checksum: 65a299720852db7830bef151dfdba46a (MD5) Previous issue date: 2018-03-28 / Introduction: Peripheral nerve transection injuries are common and occur frequently in clinical practice and are responsible for serious problems such as pain, loss of sensibility and motor, and permanent sequelae. Great advances were obtained in the treatment of peripheral nerve lesions due to greater knowledge about their physiology and the development of techniques such as microsurgery. The results of electrical stimulation are encouraging and suggest its use as an alternative treatment. Objective: To evaluate, in vivo, the use of electrostimulation (EE) associated with the tubulization technique on the repair of sciatic nerve injury in Wistar rats. Materials and Methods: Ninety male Wistar isogenic rats weighing between 200 and 300 grams were divided into three groups of 30 rats (electrostimulation associated with the tubing technique (EE), tubing suture (ST) and suture(SUT), which were subdivided into three other groups with ten animals according to the evaluation time, with three, six and nine weeks postoperative. Regeneration was assessed by ?walking track? analysis and histological analysis in which determination of the area and number of Schwann cells in the nerve samples. Results: In the functional evaluation, the group that received only one EE with 20 Hz, 3 V for one hour presented a superior result, statistically significant when compared to those stimulated more than once, however when comparing the three different types of treatment at the end of the nine weeks there was statistically significant superiority in the treated with standard suture. Regarding the number of cells, no statistically significant result was found in either group. The findings related to the nerve area, in those submitted to EE at three weeks, when compared to the other groups in the same period, were significant, and in the comparison within the same group, those with suture and tube at six weeks obtained a regenerated nerve with area in a statistically significant way. / Introdu??o: Traumatismos por transec??o de nervo perif?rico s?o comuns e ocorrem, frequentemente, na pr?tica cl?nica, sendo respons?veis por problemas graves como dor, perda de sensibilidade e motricidade, al?m de sequelas permanentes. Grandes avan?os foram obtidos no tratamento das les?es dos nervos perif?ricos, devido ao maior conhecimento sobre sua fisiologia e ao desenvolvimento de t?cnicas como a microcirurgia. Os resultados da estimula??o el?trica s?o encorajadores e sugerem seu uso como alternativa de tratamento. Objetivo: Avaliar, in vivo, o uso da eletroestimula??o (EE) associada ? t?cnica de tubuliza??o sobre o reparo de les?o de nervo ci?tico, em ratos Wistar. Materiais e m?todos: Noventa ratos isog?nicos da ra?a Wistar, machos, adultos, pesando entre 200 e 300 gramas de massa corporal, foram divididos em tr?s grupos de 30 ratos (eletroestimula??o associada ? t?cnica de tubuliza??o (EE), sutura com tubuliza??o (ST) e sutura padr?o (SUT). Os tr?s grupos foram ainda subdivididos em tr?s grupos com 10 animais, de acordo com o tempo de avalia??o (tr?s, seis e nove semanas de p?s-operat?rio). A regenera??o foi avaliada atrav?s da an?lise da marcha e an?lise histol?gica, em que houve a determina??o da ?rea e do n?mero de c?lulas de Schwann nas amostras de nervo. Resultados: Na avalia??o funcional, o grupo que recebeu somente uma EE com 20 Hz e 3V por uma hora apresentou um resultado superior se comparado com os estimulados mais de uma vez. Entretanto, ao se comparar os tr?s diferentes tipos de tratamento ao final das nove semanas, os ratos com sutura padr?o apresentaram resultados melhores. Com rela??o ao n?mero de c?lulas, n?o foi encontrado resultado estatisticamente significativo em nenhum dos grupos. Em rela??o aos achados relacionados ? ?rea do nervo, naqueles submetidos ? EE com tr?s semanas, quando comparados aos outros grupos no mesmo per?odo, houve signific?ncia, sendo que na compara??o dentro do mesmo grupo, aqueles com sutura e tubo com seis semanas, obtiveram um nervo regenerado com ?rea maior.

Submitted by Setor de Tratamento da Informa??o - BC/PUCRS (tede2@pucrs.br) on 2016-03-17T12:57:37Z No. of bitstreams: 1 DIS_GIOVANNA_RAMOS_GARCEZ_COMPLETO.pdf: 5976123 bytes, checksum: 9c877744ccee252f8823d2473ec77be8 (MD5) / Made available in DSpace on 2016-03-17T12:57:37Z (GMT). No. of bitstreams: 1 DIS_GIOVANNA_RAMOS_GARCEZ_COMPLETO.pdf: 5976123 bytes, checksum: 9c877744ccee252f8823d2473ec77be8 (MD5) Previous issue date: 2015-08-31 / Peripheral nerve injuries are a common clinical problem and the development of new approaches to assist in its repair has been shown to be a growth area. The use of microstructured channels has been investigated, in order to promote cell growth guided by assisting in the damaged tissue recovery. Biocompatible and biodegradable materials can be applied for the construction of these devices by eliminating the need for a second procedure to remove the implant. This work proposes the preparation of microstructured film along channels aligned with polydimethylsiloxane (PDMS) and blends of polycaprolactone (PCL) and poly (lacticco- glycolic acid) (PLGA) for potential application in peripheral nerve regeneration. PDMS films were prepared by the method of mixing the solution with oligomers based curing agent. Blends of PCL (Mn = 80.00) and PLGA (85:15) with different ratios of PCL / PLGA (100/0, 90/10, 80/20 and 70/30) were prepared by the solvent evaporation method. The mold used to produce the films consisted of channels with different widths (5, 10, 15 and 20 ?m) and constant height (25 ?m). The materials were characterized by Scanning Electron Microscopy (SEM), Infrared Spectroscopy Fourier Transform (ATR / FT-IR), thermal gravimetric analysis (TGA), Atomic Force Microscopy (AFM). To evaluate the degradation time of the films was carried out in vitro degradation test of PCL blends and PCL / PLGA observing weight loss 0.4 to 9.9% until 90 days testing. It was found that the addition of the PLGA PCL is an effective way to control the degradation rate of the blend and there is no significant difference in the degradation of flat and microstructured films. / Les?es no nervo perif?rico s?o um problema cl?nico comum e a constru??o de novos dispositivos para auxiliar no seu reparo tem se mostrado uma ?rea em crescimento. A utiliza??o de condutos microestruturados vem sendo investigada, com o objetivo de promover um crescimento celular guiado, auxiliando na recupera??o do tecido lesado. Materiais biocompat?veis e biodegrad?veis podem ser aplicados para a constru??o destes dispositivos eliminando a necessidade de um segundo procedimento para remo??o do implante. Este trabalho prop?e a prepara??o de filmes microestruturados com canais longitudinalmente alinhados de polidimetilsiloxano (PDMS) e blendas de policaprolactona (PCL) e poli(?cido l?cticoco- glic?lico) (PLGA) para potencial aplica??o em regenera??o de nervo perif?rico. Filmes de PDMS foram preparados pelo m?todo de mistura da solu??o base de olig?meros com agente de cura. Blendas de PCL (Mn = 80,00) e PLGA (85:15) com diferentes raz?es de PCL/PLGA (100/0, 90/10, 80/20 e 70/30) foram preparadas pelo m?todo de evapora??o do solvente. O molde utilizado para produzir os filmes era constitu?do de canais com diferentes larguras (5, 10, 15 e 20 ?m) e altura constante (25 ?m). Os materiais foram caracterizados por Microscopia Eletr?nica de Varredura (MEV), Espectroscopia no Infravermelho por Transformada de Fourier (ATR/FT-IR), An?lise termogravim?trica (TGA), Microscopia de For?a At?mica (AFM). Para verificar o tempo de degrada??o dos filmes foi realizado ensaio de degrada??o in vitro das blendas de PCL e PCL/PLGA, observando-se perda de massa de 0,4-9,9 % at? o per?odo de 90 dias de ensaio. Constatou-se que a adi??o de PLGA ao PCL ? uma forma efetiva de controlar a taxa de degrada??o das blendas e que n?o h? diferen?a significativa entre a degrada??o dos filmes lisos e microestruturados.

Made available in DSpace on 2015-04-14T13:35:35Z (GMT). No. of bitstreams: 1 443023.pdf: 2231703 bytes, checksum: 8fa714244d8e44469edc5dc83e9b3038 (MD5) Previous issue date: 2012-08-29 / Injuries to the peripheral nervous system present a challenge for regenerative medicine. The study investigated the effects of platelet-rich plasma (PRP) and erythropoietin (EPO), alone or in combination, in the regeneration of peripheral nerve in rats. The work was developed in three phases using 70 rats. The first phase, evaluated the platelet concentration and growth factors production/release (TGF-β, PDGF, VEGF and αβ) in three protocols for obtaining PRP (A, B and C) obtained by aortic or cardiac puncture. Cardiac puncture in association with A protocol showed the best results. In the second stage, using best results methods at the first stage, was produced two PRP concentrations (1,000,000 and 2,000,000 platelets.μl-1; PRP1 and PRP2, respectively) for applying in the rats sciatic nerve. Comparison of concentrations with autologous nerve graft (ANG) and saline solution (SS) groups showed benefits in the use of PRP2. In the third phase (same experimental model) were added PRP2 + EPO and EPO groups. The animals of phases 1 and 2 were evaluated by walking track test and histomorphometric analysis (toluidine blue) and showed benefits in autologous nerve graft (ANG) with respect to tubulization (independent of regenerative factor). For tubularized animals, the study showed advantages for PRP and EPO simultaneous use followed by EPO group. Finally, it was observed that PRP 2,000,000 platelets.μl-1 has beneficial effects when compared to PRP 1,000,000 platelets.μl-1 for peripheral neuroregeneration rats. / Les?es ao sistema nervoso perif?rico representam um desafio para a medicina regenerativa. O estudo objetivou conhecer os efeitos do plasma rico em plaquetas (PRP) e da eritropoietina (EPO), isolados ou combinados, na regenera??o do nervo perif?rico de ratos. O trabalho desenvolveu-se em tr?s etapas utilizando 70 ratos. A primeira fase avaliou a concentra??o plaquet?ria e produ??o/libera??o de fatores de crescimento (TGF-β, PDGF-αβ e VEGF) em tr?s protocolos de obten??o do PRP (A, B e C) obtidos por pun??o a?rtica ou card?aca. A pun??o card?aca em associa??o com protocolo A gerou os melhores resultados. Na segunda fase, usando os m?todos com melhores resultados na primeira etapa, produziram-se duas concentra??es de PRP (1.000.000 e 2.000.000 plaquetas. μl-1; PRP1 e PRP2, respectivamente) para aplica??o no nervo ci?tico tubulizado de ratos. Compara??o das concentra??es entre si e com grupos auto enxerto (AE) e solu??o fisiol?gica (SF) evidenciaram benef?cios no uso de PRP2. Na terceira fase (mesmo modelo experimental), foram adicionados os grupos EPO e EPO+PRP2. Os animais das fases 1 e 2 foram avaliados por teste de marcha e histomorfometria (azul de toluidina) e demonstraram que o auto enxerto apresentou benef?cios sobre a tubuliza??o (independente do fator regenerativo utilizado). Para os animais tubulizados, o estudo demostrou melhores resultados para o uso simult?neo de PRP e EPO, seguido do grupo EPO. Finalizando, observou-se que a aplica??o de PRP na concentra??o de 2.000.000 plaquetas.μl-1 em compara??o com 1.000.000 platelets.μl- 1 apresentou vantagens para neuroregenera??o perif?rica de ratos.

Made available in DSpace on 2015-04-14T13:36:01Z (GMT). No. of bitstreams: 1 464259.pdf: 3964531 bytes, checksum: 04739dfcb6adbc45be2c0bbc0dde167b (MD5) Previous issue date: 2014-08-29 / Introduction : Injuries in peripheral nerves are common in clinical practice being responsible for serious problems and often permanent as loss of motor, sensitive functions, acute and chronic pain. Although surgical, physiotherapy and diagnostic techniques have evolved in recent decades, the clinical outcomes of nerve recovery still unsatisfactory, which encourages many researchers to seek drugs and devices that may help regeneration (NOBLE et al, 1998; OLIVEIRA et al, 2004). Various trophic factors, also known as growth factors (GF) are used and tested in vitro and in vivo in an attempt to assist the regeneration of peripheral nerves. These proteins act directly on the proliferation and differentiation of various cell types, being able to promote tissue repair and functional recovery (BOYD AND GORDON, 2003). Among them, we highlight the FK506 (Tacrolimus), a ligand for immunophilins with strong immunosuppressive effect that has potential neurotrophic and neuroprotective actions. Studies have shown that FK506 increases the rate of axonal regeneration and also the level of necessary reinnervation after peripheral nerve injury, besides enhancing recovery from nerve damage. Objective : Evaluating in vivo the availability and effect of FK 506 on the repair of injury of rat sciatic nerve. Material and Methods : Thirty-six male adult Wistar rats were randomized into three groups (I - control, II - Systemic FK506 III - FK506 site;). To evaluate the effect of different treatments, the sciatic nerve of animals was transected, creating a defect of 10 mm and then tubulized with the silicone chamber. In the control group, no additional treatment was performed. In systemic group, FK506 (dose 0.1 mg/kg/day) was administered systemically via osmotic minipump ALZET while in the local group the administration also occurred through the ALZET osmotic minipump, but adapted to release into the silicone chamber the same dose. The performed treatment lasted 90 days. Results : Data from the present study suggest that the immunosuppressive drug FK506 has a positive effect on the nervous functional improvement and the use of osmotic minipumps is effective in continuous dispensing of the drug. The most important finding of this research is that the degree of functional recovery, measured by the rate of sciatic function after injury with loss of substance and repair through tubing, it is better when FK506 is administered locally whether compared to systemic administration in animal models. No other studies have used local application of the drug with constant release rate for comparison were found. / Introdu??o : Les?es em nervos perif?ricos s?o frequentes na pr?tica cl?nica sendo respons?veis por problemas graves e muitas vezes permanentes como perda de fun??es motoras, sensitivas, dor aguda e cr?nica. Embora as t?cnicas cir?rgicas, fisioter?picas e de diagn?stico tenham evolu?do nas ?ltimas d?cadas, os desfechos cl?nicos de recupera??o nervosa continuam insatisfat?rios, o que estimula diversos pesquisadores a buscarem drogas e dispositivos que possam auxiliar na regenera??o (NOBLE et al, 1998; OLIVEIRA et al, 2004). Diversos fatores tr?ficos, tamb?m conhecidos como fatores de crescimento (FC), s?o utilizados e testados in vitro e in vivo como uma tentativa de auxiliar a regenera??o de nervos perif?ricos.Essas prote?nas atuam diretamente na prolifera??o e diferencia??o de diversos tipos celulares, sendo capazes de promover reparo tecidual e recupera??o funcional (BOYD E GORDON, 2003). Dentre eles, destaca-se o FK506 (Tacrolimus), um ligante de imunofilinas com forte efeito imunossupressor que apresenta potenciais a??es neurotr?ficas e neuroprotetoras. Estudos t?m mostrado que o FK506 aumenta a taxa de regenera??o axonal e tamb?m o n?vel de reinerva??o necess?ria ap?s les?o do nervo perif?rico, al?m de melhorar a recupera??o ap?s les?es no nervo. Objetivo : Avaliar in vivo a viabilidade e efeito do FK 506 sobre o reparo de les?o de nervo ci?tico de ratos Material e M?todos : Trinta e seis ratos Wistar, machos, adultos foram randomicamente distribu?dos em tr?s grupos (I controle; II FK506 sist?mico; III FK506 local;). Para avaliar o efeito dos diferentes tratamentos, o nervo ci?tico dos animais foi transeccionado criando um defeito de 10mm e, em seguida, tubulizado com c?mara de silicone. No grupo controle, nenhum tratamento adicional foi realizado. No grupo sist?mico, o FK506 (dose 0,1mg/kg/dia) foi administrado sistemicamente via minibomba osm?tica ALZET enquanto que no grupo local a administra??o tamb?m ocorreu atrav?s da minibomba osm?tica ALZET, mas adaptada para libera??o dentro da c?mara de silicone na mesma dose. O tratamento empregado teve dura??o de 90 dias. Resultados : Os dados do presente estudo sugerem que a droga imunossupressora FK506 possui efeito positivo na melhora funcional nervosa e o uso de minibombas osm?ticas ? eficaz na dispensa??o cont?nua da droga. O dado mais importante dessa pesquisa ? que o grau de recupera??o funcional, medido atrav?s do ?ndice de fun??o ci?tica ap?s les?o com perda de subst?ncia e reparo atrav?s de tubuliza??o, ? melhor quando o FK506 ? administrado localmente se comparado ? administra??o sist?mica em modelos animais. N?o foram encontrados outros trabalhos que tenham usado aplica??o local da droga com taxa de libera??o constante para efeitos de compara??o.