Submitted by Luciana Ferreira (lucgeral@gmail.com) on 2017-04-03T11:39:27Z No. of bitstreams: 2 Dissertação - Kamilla Dias Ferreira - 2017.pdf: 4419008 bytes, checksum: 3c26845aebdde46ec46944acf9ca0b04 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-04-03T11:40:34Z (GMT) No. of bitstreams: 2 Dissertação - Kamilla Dias Ferreira - 2017.pdf: 4419008 bytes, checksum: 3c26845aebdde46ec46944acf9ca0b04 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-04-03T11:40:34Z (GMT). No. of bitstreams: 2 Dissertação - Kamilla Dias Ferreira - 2017.pdf: 4419008 bytes, checksum: 3c26845aebdde46ec46944acf9ca0b04 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-02-06 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / This study aimed to evaluate the viability of the decellularization process in bovine cartilaginous tissue by histological analysis. For this purpose, 20 bovine elastic cartilages were used, immersed in alkaline solution for 72 hours, followed by washing, freezing and lyophilization of the material. Histological processing was then performed to perform the analyzes. In the histological analyzes the decellularization and removal of the amorphous substance from the tissue was observed, but the structure of elastic fibers and collagen remained. It was concluded that the alkaline treatment was efficient to obtain biomaterial from elastic cartilaginous tissue. This study also evaluated, through thermographic examination, the feasibility of using implants with biomaterials obtained from elastic cartilages treated in alkaline solution. The intensity of the inflammatory process was observed through the variation of cutaneous temperature comparing before and after the implantation of the biomaterials in rabbits (Oryctolagus cuniculus). A total of 24 rabbits were evaluated, divided into two groups, of 12 animals each, according to the implanted biomaterial. Evaluations were performed in the preoperative period and after the implantation of biomaterials. In the (GI) group, polypropylene mesh and elastic cartilage (GII) were treated with alkaline solution, evaluated in the periods of seven and 35 days. From the results obtained, after performing statistical tests, the group (GI) polypropylene mesh, obtained upper abdominal skin temperatures, compared to the cartilage group, providing subsidies that the use of elastic cartilage treated in alkaline solution, promoted lower inflammatory intensity due to the processing to which it was submitted. It is concluded that treated elastic cartilage has been shown to be a superior biomaterial relative to the mesh polypropylene. / Este estudo objetivou avaliar a viabilidade do processamento de descelularização em tecido cartilaginoso bovino por meio de análises histológicas. Pra tanto, foram utilizadas 20 cartilagens elásticas de bovinos, submetidas à imersão em solução alcalina por 72 horas, seguidos de lavagens, congelamento e liofilização do material. Em seguida foi realizado o processamento histológico para serem feitas as analises. Nas analises histológicas foi observado a descelularização e retirada da substancia amorfa do tecido, porém manteve a estrutura das fibras elásticas e de colágeno. Concluiu-se que o tratamento alcalino foi eficiente para obtenção de biomaterial a partir de tecido cartilaginoso elástico. Este estudo avaliou também, através do exame termográfico, a viabilidade no uso de implantes com biomateriais obtidos de cartilagens elásticas tratadas em solução alcalina. Foram observados através da variação da temperatura cutânea, a intensidade do processo inflamatório comparando antes e após a implantação dos biomateriais em coelhos (Oryctolagus cuniculus). Para tanto foram avaliados um total de 24 coelhos, divididos em dois grupos, de 12 animais, de acordo com o biomaterial implantado. Foram realizadas avaliações no período pré-operatório e após o implante dos biomateriais. No grupo (GI) foi implantada a tela de polipropileno e (GII) a cartilagem elástica tratada em solução alcalina, avaliados nos períodos de sete e 35 dias. Dos resultados obtidos, após realização de testes estatísticos, o grupo (GI), obteve temperaturas cutâneas abdominais superiores, comparadas ao grupo cartilagem, fornecendo, subsídios de que a utilização da cartilagem elástica tratada em solução alcalina, promoveu menor intensidade inflamatória devido ao processamento ao qual foi submetida. Conclui-se que a cartilagem elástica tratada demonstrou ser um biomaterial superior em relação à tela de polipropileno.

Hintergrund: Elastischer Ohrknorpel ist eine potentielle Quelle fuer autologe Zellen um die luminalen Oberflaechen von links ventrikularen assist systemen (LVAD''s) zu besiedeln zu dem Zweck die Biokompatibilitaet dieser Systeme zu verbessern. Wir prueften dieses Potential der Ohrknorpelzellen in vitro und in vivo in einem Tiermodel (Kalb). Methoden: In vitro, Elastischer Knorpel wurde von dem Ohr eines Kalbes unter Lokalanaesthesie gewonnen, isoliert, und in Gewebekultur gezuechtet. Chondrocytes wurden immunocytochemisch analysiert, transferriert in Zellkulturmedium, zweimal umgesetzt, und auf die blutkontaktierenden, luminalen Oberflaechen von vier LVAD''s (HeartMate(R); Thermo Cardiosystems, Inc., Woburn, MA) gesiedelt. Die beschichteten Zelloberflaechen wurden preconditioniert unter Flussbedingungen in vitro, um die Zellhaftung an den luminalen Oberflaechen der LVAD''s zu beguenstigen. Die Effiezienz der Zellbesiedelung und der kumulative Zellverlust unter Flussbedingungen in vitro wurden quantifiziert. In vivo, eine der vier chondrocyte-beschichteten und preconditionierten LVAD''s wurde in das Kalb implantiert, welches urspruenglich Ohrknorpelspender diente. Das LVAD wurde fuer sieben Tage betrieben, dann explantiert und pathologisch und histologisch evaluiert unter der Benutzung eines Scanningelektronenmikroskopes und eines Transmissionelektronenmikroskopes. Resultate: Die luminalen Oberflaechen von vier LVAD''s wurden mit autologen Ohrknorpelzellen besiedelt. Die Effiezienz der Zellbesiedelung betrug 95.11± 4.23% (n = 4). Der kumulative Zellverlust unter Flussbedingungen in vitro war geringer als 12% (n = 4). Nach sieben Tagen der in vivo Implantation zeigten die luminalen Oberflaechen des implantierten LVAD eine intakte, fest haftende Zellbesiedelung. Es wurden keine Thromboembolien waehrend der Nekropsie festgestellt. Schlussfolgerungen: Ohrknorpel stellt eine einfach und schnell erreichbare Quelle fuer autologe Ohrknorpelzellen dar, dessen Collagen Typ II und andere wichtige Bestandteile der extrazellulaeren Matrix sie dazu befaehigt, fest an den luminalen Oberflaechen der LVAD''s zu haften. Die hier beschriebene einfache Methode der Knorpelgewinnung und der Zellisolierung kann benutzt werden, um eine autologe Zellbeschichtung von LVAD''s und anderen implantierbaren kardiovaskulaeren Systemen vorzunehmen mit dem Ziel die Biokompatibilitaet dieser zu verbessern. Unsere erfolgreiche Feasibilitystudie in einem Kalbmodel fordert weitere in vivo Studien. / Background: Auricular elastic cartilage is a potential source of autologous cells for lining the luminal surfaces of left ventricular assist devices (LVAD''s) to improve long-term biocompatibility. We evaluated this potential in vitro and in vivo in a calf model. Methods: In vitro, auricular cartilage was harvested from the anesthetized ear of a calf, isolated, and cultured on tissue culture dishes. Primary chondrocytes were typed by immunocytochemistry, transferred into culture media, passaged twice, and seeded onto the blood-contacting luminal surfaces of four LVAD''s (HeartMate(R); Thermo Cardiosystems, Inc., Woburn, MA). The seeded cell linings were preconditioned under flow conditions in vitro to promote cell adhesion to the luminal surfaces. Seeding efficiency and cumulative cell loss under flow conditions in vitro were quantitated. In vivo, one of the four preconditioned, autologous chondrocyte-lined LVAD''s was implanted into the tissue-donor calf; run for 7 days; explanted; and finally evaluated grossly, by scanning electron microscopy, and by transmission electron microscopy. Results: Autologous chondrocytes were seeded onto the luminal surfaces of the four LVAD''s. The seeding efficiency was 95.11± 4.23% (n = 4). Cumulative cell loss during preconditioning under flow conditions in vitro did not exceed 12% (n = 4). After 7 days of in vivo implantation, the luminal surfaces of the implanted LVAD demonstrated an intact, strongly adherent cellular lining. There was no evidence of thromboembolic events at necropsy. Conclusions: Auricular elastic cartilage is a ready and easily accessible source of chondrocytes whose ability to produce collagen II and other important extracellular matrix constituents allows them to adhere strongly to the luminal surfaces of LVAD''s. The simple method of isolating and expanding auricular chondrocytes presented here could be used to provide autologous cell linings for LVAD''s and other cardiovascular devices to improve their long-term biocompatibility. Our successful short-term feasibility study in a calf model warrants further study in vivo.

1 Abstract Introduction Autologous transplants of the cartilage tissue from the pinna is commonly used in reconstructive surgery of the nasal skeleton. The present study used animal models to elucidate responses of the auricular cartilage to its damage or transplantation to ectopic sites. Histomorphological analysis of changes observed in auricular cartilage including immunohistochemical study of different isoforms of actin and S-100 proteins was performed. Human articular cartilage prepared by in vitro cultivation using artificial scaffolds was also studied after its transplantation. Aims of the study The aim was to study histological changes and expression of chondrocytic markers (α- SMA and S-100 proteins) in intact, artificially traumatised, or in a human auricular cartilage cultivated in culture medium. An attempt to grow human auricular cartilage chondrocytes implanted in vitro into various types of three dimensional scaffolds aimed at testing chondrocyte survival and phenotype both in the culture and after transplantation to immunodeficient mice. A human auricular cartilage transplanted into the nasal skeleton of patients during a reconstruction surgery should be submitted to a histomorphological examination. Research assumed also comparison of the auricular cartilage responses to a damage,...

1 Abstract Introduction Autologous transplants of the cartilage tissue from the pinna is commonly used in reconstructive surgery of the nasal skeleton. The present study used animal models to elucidate responses of the auricular cartilage to its damage or transplantation to ectopic sites. Histomorphological analysis of changes observed in auricular cartilage including immunohistochemical study of different isoforms of actin and S-100 proteins was performed. Human articular cartilage prepared by in vitro cultivation using artificial scaffolds was also studied after its transplantation. Aims of the study The aim was to study histological changes and expression of chondrocytic markers (α- SMA and S-100 proteins) in intact, artificially traumatised, or in a human auricular cartilage cultivated in culture medium. An attempt to grow human auricular cartilage chondrocytes implanted in vitro into various types of three dimensional scaffolds aimed at testing chondrocyte survival and phenotype both in the culture and after transplantation to immunodeficient mice. A human auricular cartilage transplanted into the nasal skeleton of patients during a reconstruction surgery should be submitted to a histomorphological examination. Research assumed also comparison of the auricular cartilage responses to a damage,...