Desenvolvimento, caracterização e aplicação de membranas à base de celulose bacteriana e do nanocompósito celulose bacteriana/policaprolactona na ceratoplastia lamelar experimental em coelhos

Detalhes bibliográficos
Ano de defesa: 2012
Autor(a) principal: Queiroz, Paulo Victor da Silva
Orientador(a): Borges, Andrea Pacheco Batista lattes
Banca de defesa: Benjamin, Laércio dos Anjos lattes, Barud, Hernane da Silva lattes
Tipo de documento: Dissertação
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Universidade Federal de Viçosa
Programa de Pós-Graduação: Mestrado em Medicina Veterinária
Departamento: Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de
País: BR
Palavras-chave em Português:
Palavras-chave em Inglês:
Área do conhecimento CNPq:
Link de acesso: http://locus.ufv.br/handle/123456789/5122
Resumo: Corneal conditions are among the main causes of blindness in both humans and animals. Many surgical techniques are described to recover the cornea s physical structure and refractive state; among them is the use of different types of biomaterials. Due to the cornea s transparency, viscoelasticity and lack of vessels, biomaterials to be applied on them must undergo a careful selection. Bacterial cellulose (BC) and polycaprolactone (PCL) are two well known biomaterials, studied extensively in separate. The purpose of the present study was to produce and characterize membranes made of BC and PCL to be tested as corneal tissue substitutes in rabbit s cornea. Membranes of BC were produced through static culture of the bacteria Gluconacetobacter xylinus. Membranes were then washed, dried and swelled in acetone for 48 hours, followed by immersion in 25ml of 3% PCL solution for 72 hours. Next, membranes of the BC/PCL composite underwent several washes in acetone and were then dried to be characterized by scanning electron microscopy (SEM), X-ray diffraction, thermal and mechanical trials and luminous transmittance. In vivo study was conducted with the BC/PCL composite and with pure BC to evaluate their interactions with corneal tissue. At first, membranes were cut in 8mm diameter to be then applied in corneal tisse. Thirty six female rabbits were separated in three groups of 12 animals each. An ulcer of 5mm in diameter and 0.2mm in depth was created on the right cornea of all rabbits. From the base of the ulcer, the stroma was separated creating an interlamellar pocket, and each group of 12 animals received 8mm diameter BC/PCL composite membrane and the other group received BC membranes on the same size, both adjusted in the pocket. The last group did not receive any membrane, used as the control. Animals were followed for 45 days, where clinical evaluations for signs of pain and inflammation were conducted. Three animals of each group were euthanized on days 3, 7, 21 and 45 after surgery to collect samples for histology (HE and Picrosirius) and SEM. BC/PCL presented 84% of light transmission while pure BC presented 45%. Lower tension resistance and Young s module was observed on the composite compared to pure BC, while the composite presented greater elongation at rupture what demonstrates a greater viscoelasticity of this material. Eyes that received membranes presented moderate inflammation until 30 days after surgery, decreasing progressively until 45 days. Histology revealed the presence of lymphohistiocytic inflammatory infiltrate at 21 and 45 days in implanted groups, at a moderate intensity, as well as giant cells, suggesting chronic inflammatory and foreign body reactions. Fibroplasia was observed around the membranes, a positive sign of biocompatibility. Epithelization was complete only in control group, what shows that membranes surfaces did not allow epithelium growth and maintenance. Picrosirius staining showed a greater proportion of type III collagen at the end of 45 days in both implanted groups, suggesting that the repair process around the membranes was incomplete. SEM showed extracellular matrix and cells adhered to implant surfaces, suggesting integration at 45 days. Both membranes made of BC/PCL composite and BC alone stimulated chronic inflammatory and foreign body reactions and did not allow complete epithelization on their surfaces. Based on these conclusions, we believe that implants made of BC/PCL and BC using the here presented process are not indicated as a substitute for corneal tissue in rabbits.
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spelling Queiroz, Paulo Victor da Silvahttp://lattes.cnpq.br/0612690489308163Pontes, Kelly Cristine de Sousahttp://lattes.cnpq.br/2423435012302575Carlo, Ricardo Junqueira Delhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788143A9Borges, Andrea Pacheco Batistahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4767373D8Benjamin, Laércio dos Anjoshttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797917E7Barud, Hernane da Silvahttp://lattes.cnpq.br/70204672926901122015-03-26T13:47:10Z2013-07-042015-03-26T13:47:10Z2012-09-04QUEIROZ, Paulo Victor da Silva. Development, characterization and application of membranes made of bacterial cellulose and of the nanocomposite bacterial cellulose/polycaprolactone on experimental lamellar keratoplasty in rabbits. 2012. 83 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.http://locus.ufv.br/handle/123456789/5122Corneal conditions are among the main causes of blindness in both humans and animals. Many surgical techniques are described to recover the cornea s physical structure and refractive state; among them is the use of different types of biomaterials. Due to the cornea s transparency, viscoelasticity and lack of vessels, biomaterials to be applied on them must undergo a careful selection. Bacterial cellulose (BC) and polycaprolactone (PCL) are two well known biomaterials, studied extensively in separate. The purpose of the present study was to produce and characterize membranes made of BC and PCL to be tested as corneal tissue substitutes in rabbit s cornea. Membranes of BC were produced through static culture of the bacteria Gluconacetobacter xylinus. Membranes were then washed, dried and swelled in acetone for 48 hours, followed by immersion in 25ml of 3% PCL solution for 72 hours. Next, membranes of the BC/PCL composite underwent several washes in acetone and were then dried to be characterized by scanning electron microscopy (SEM), X-ray diffraction, thermal and mechanical trials and luminous transmittance. In vivo study was conducted with the BC/PCL composite and with pure BC to evaluate their interactions with corneal tissue. At first, membranes were cut in 8mm diameter to be then applied in corneal tisse. Thirty six female rabbits were separated in three groups of 12 animals each. An ulcer of 5mm in diameter and 0.2mm in depth was created on the right cornea of all rabbits. From the base of the ulcer, the stroma was separated creating an interlamellar pocket, and each group of 12 animals received 8mm diameter BC/PCL composite membrane and the other group received BC membranes on the same size, both adjusted in the pocket. The last group did not receive any membrane, used as the control. Animals were followed for 45 days, where clinical evaluations for signs of pain and inflammation were conducted. Three animals of each group were euthanized on days 3, 7, 21 and 45 after surgery to collect samples for histology (HE and Picrosirius) and SEM. BC/PCL presented 84% of light transmission while pure BC presented 45%. Lower tension resistance and Young s module was observed on the composite compared to pure BC, while the composite presented greater elongation at rupture what demonstrates a greater viscoelasticity of this material. Eyes that received membranes presented moderate inflammation until 30 days after surgery, decreasing progressively until 45 days. Histology revealed the presence of lymphohistiocytic inflammatory infiltrate at 21 and 45 days in implanted groups, at a moderate intensity, as well as giant cells, suggesting chronic inflammatory and foreign body reactions. Fibroplasia was observed around the membranes, a positive sign of biocompatibility. Epithelization was complete only in control group, what shows that membranes surfaces did not allow epithelium growth and maintenance. Picrosirius staining showed a greater proportion of type III collagen at the end of 45 days in both implanted groups, suggesting that the repair process around the membranes was incomplete. SEM showed extracellular matrix and cells adhered to implant surfaces, suggesting integration at 45 days. Both membranes made of BC/PCL composite and BC alone stimulated chronic inflammatory and foreign body reactions and did not allow complete epithelization on their surfaces. Based on these conclusions, we believe that implants made of BC/PCL and BC using the here presented process are not indicated as a substitute for corneal tissue in rabbits.As afecções corneais representam uma das principais causas de cegueira tanto em humanos como em animais. São descritas várias técnicas cirúrgicas para recuperar a estrutura física e o estado refrativo da córnea, dentre essas, encontra-se a aplicação de diferentes tipos de biomateriais. Devido à transparência, à falta de vasos e à viscoelasticidade que a córnea apresenta, os biomateriais para nela serem aplicados devem passar por criteriosa seleção. A celulose bacteriana (CB) e a policaprolactona (PCL) são dois consagrados biomateriais, bastante estudados isoladamente. Foi proposto, neste estudo, a produção e a caracterização de membranas à base de CB e de PCL para serem testadas como substituto tecidual em córneas de coelhos. Membranas de CB foram produzidas por meio do cultivo estático da bactéria Gluconacetobacter xylinus. Após serem lavadas e secas, as membranas foram intumescidas em acetona por 48 horas e então imersas em 25 ml de solução a 3% de PCL diluída em acetona durante 72 horas. Na sequência, as membranas do compósito CB/PCL passaram por repetidas lavagens em acetona, e foram secas para serem caracterizadas por microscopia eletrônica de varredura (MEV), difração de raio-X, ensaios térmico e mecânico e transmissão óptica. Foi realizado o estudo in vivo com membranas do compósito CB/PCL e da CB pura para avaliar suas interações com o tecido corneal. Inicialmente elas foram cortadas em 8 mm de diâmetro para então serem implantadas nas córneas dos animais. Foram utilizadas 36 coelhas divididas em três grupos de 12 animais. Foi criada uma úlcera de 5 mm de diâmetro e 0,2 mm de profundidade na córnea direita de todos os animais. A partir da base da úlcera o estroma foi separado, criando-se um bolso interlamelar, sendo que um grupo recebeu membranas do compósito CB/PCL e outro grupo recebeu membranas da CB, que foram ajustadas dentro do bolso. O último grupo não recebeu tratamento sendo utilizado como controle. Os animais foram acompanhados por até 45 dias, sendo feitas avaliações clínicas para os sinais de dor e inflamação. Três animais de cada grupo foram eutanasiados aos três, sete, 21 e 45 dias após a cirurgia, para coleta de amostras para análises histológicas (HE e Picrosirius) e MEV. O compósito CB/PCL apresentou 84% de transmissão de luz, enquanto que a CB pura apresentou 65%. Houve redução da resistência à tração e do módulo de Young do compósito CB/PCL em relação à CB pura. Em contrapartida, o compósito apresentou aumento na ruptura ao alongamento, demonstrando incremento na elasticidade do material. Os olhos que receberam implantes apresentaram inflamação moderada até os 30 dias de pós-operatório, reduzindo progressivamente até os 45 dias da cirurgia. A análise histológica demonstrou, aos 21 e aos 45 dias, presença de infiltrado inflamatório linfo-histiocitário em intensidade moderada e células gigantes nos animais implantados, sugerindo resposta inflamatória persistente e do tipo corpo estranho. Também foi observada fibroplasia em torno das membranas, sendo este um sinal positivo de biocompatibilidade. A epitelização foi completa somente no grupo controle, demonstrando que as superfícies das membranas não permitiram o crescimento e manutenção do epitélio. Pela coloração de Picrosirius, evidenciou-se maior proporção do colágeno tipo III ao final dos 45 dias nos grupos com implantes, sugerindo que o processo de reparação em torno das membranas estava incompleto. A MEV demonstrou matriz extracelular e células aderidas às superfícies dos implantes, sinalizando integração no período de 45 dias. As membranas tanto do compósito CB/PCL como da CB incitaram resposta inflamatória crônica e de corpo estranho, e não permitiram a completa epitelização sobre suas superfícies. Baseado nisso, acredita-se que os implantes CB/PCL e CB, da maneira como foram desenvolvidos, não devem ser indicados para substituir o tecido corneal de coelhos.application/pdfporUniversidade Federal de ViçosaMestrado em Medicina VeterináriaUFVBRBiotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. deÚlcera de córneaBiomaterialSubstituto tecidualCorneal ulcerBiomaterialTissue substituteCNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::CLINICA E CIRURGIA ANIMAL::CLINICA CIRURGICA ANIMALDesenvolvimento, caracterização e aplicação de membranas à base de celulose bacteriana e do nanocompósito celulose bacteriana/policaprolactona na ceratoplastia lamelar experimental em coelhosDevelopment, characterization and application of membranes made of bacterial cellulose and of the nanocomposite bacterial cellulose/polycaprolactone on experimental lamellar keratoplasty in rabbitsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINALtexto completo.pdfapplication/pdf2556452https://locus.ufv.br//bitstream/123456789/5122/1/texto%20completo.pdfd5563353a41b374f1b598abd2f574cf3MD51TEXTtexto completo.pdf.txttexto completo.pdf.txtExtracted texttext/plain171581https://locus.ufv.br//bitstream/123456789/5122/2/texto%20completo.pdf.txta10b04f407b857ec4e6494ef4c2b7c05MD52THUMBNAILtexto completo.pdf.jpgtexto completo.pdf.jpgIM Thumbnailimage/jpeg3625https://locus.ufv.br//bitstream/123456789/5122/3/texto%20completo.pdf.jpg8939fe6db2d811910ed169892e65b4f7MD53123456789/51222016-04-11 23:09:28.3oai:locus.ufv.br:123456789/5122Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-12T02:09:28LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false
dc.title.por.fl_str_mv Desenvolvimento, caracterização e aplicação de membranas à base de celulose bacteriana e do nanocompósito celulose bacteriana/policaprolactona na ceratoplastia lamelar experimental em coelhos
dc.title.alternative.eng.fl_str_mv Development, characterization and application of membranes made of bacterial cellulose and of the nanocomposite bacterial cellulose/polycaprolactone on experimental lamellar keratoplasty in rabbits
title Desenvolvimento, caracterização e aplicação de membranas à base de celulose bacteriana e do nanocompósito celulose bacteriana/policaprolactona na ceratoplastia lamelar experimental em coelhos
spellingShingle Desenvolvimento, caracterização e aplicação de membranas à base de celulose bacteriana e do nanocompósito celulose bacteriana/policaprolactona na ceratoplastia lamelar experimental em coelhos
Queiroz, Paulo Victor da Silva
Úlcera de córnea
Biomaterial
Substituto tecidual
Corneal ulcer
Biomaterial
Tissue substitute
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::CLINICA E CIRURGIA ANIMAL::CLINICA CIRURGICA ANIMAL
title_short Desenvolvimento, caracterização e aplicação de membranas à base de celulose bacteriana e do nanocompósito celulose bacteriana/policaprolactona na ceratoplastia lamelar experimental em coelhos
title_full Desenvolvimento, caracterização e aplicação de membranas à base de celulose bacteriana e do nanocompósito celulose bacteriana/policaprolactona na ceratoplastia lamelar experimental em coelhos
title_fullStr Desenvolvimento, caracterização e aplicação de membranas à base de celulose bacteriana e do nanocompósito celulose bacteriana/policaprolactona na ceratoplastia lamelar experimental em coelhos
title_full_unstemmed Desenvolvimento, caracterização e aplicação de membranas à base de celulose bacteriana e do nanocompósito celulose bacteriana/policaprolactona na ceratoplastia lamelar experimental em coelhos
title_sort Desenvolvimento, caracterização e aplicação de membranas à base de celulose bacteriana e do nanocompósito celulose bacteriana/policaprolactona na ceratoplastia lamelar experimental em coelhos
author Queiroz, Paulo Victor da Silva
author_facet Queiroz, Paulo Victor da Silva
author_role author
dc.contributor.authorLattes.por.fl_str_mv http://lattes.cnpq.br/0612690489308163
dc.contributor.author.fl_str_mv Queiroz, Paulo Victor da Silva
dc.contributor.advisor-co1.fl_str_mv Pontes, Kelly Cristine de Sousa
dc.contributor.advisor-co1Lattes.fl_str_mv http://lattes.cnpq.br/2423435012302575
dc.contributor.advisor-co2.fl_str_mv Carlo, Ricardo Junqueira Del
dc.contributor.advisor-co2Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788143A9
dc.contributor.advisor1.fl_str_mv Borges, Andrea Pacheco Batista
dc.contributor.advisor1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4767373D8
dc.contributor.referee1.fl_str_mv Benjamin, Laércio dos Anjos
dc.contributor.referee1Lattes.fl_str_mv http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4797917E7
dc.contributor.referee2.fl_str_mv Barud, Hernane da Silva
dc.contributor.referee2Lattes.fl_str_mv http://lattes.cnpq.br/7020467292690112
contributor_str_mv Pontes, Kelly Cristine de Sousa
Carlo, Ricardo Junqueira Del
Borges, Andrea Pacheco Batista
Benjamin, Laércio dos Anjos
Barud, Hernane da Silva
dc.subject.por.fl_str_mv Úlcera de córnea
Biomaterial
Substituto tecidual
topic Úlcera de córnea
Biomaterial
Substituto tecidual
Corneal ulcer
Biomaterial
Tissue substitute
CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::CLINICA E CIRURGIA ANIMAL::CLINICA CIRURGICA ANIMAL
dc.subject.eng.fl_str_mv Corneal ulcer
Biomaterial
Tissue substitute
dc.subject.cnpq.fl_str_mv CNPQ::CIENCIAS AGRARIAS::MEDICINA VETERINARIA::CLINICA E CIRURGIA ANIMAL::CLINICA CIRURGICA ANIMAL
description Corneal conditions are among the main causes of blindness in both humans and animals. Many surgical techniques are described to recover the cornea s physical structure and refractive state; among them is the use of different types of biomaterials. Due to the cornea s transparency, viscoelasticity and lack of vessels, biomaterials to be applied on them must undergo a careful selection. Bacterial cellulose (BC) and polycaprolactone (PCL) are two well known biomaterials, studied extensively in separate. The purpose of the present study was to produce and characterize membranes made of BC and PCL to be tested as corneal tissue substitutes in rabbit s cornea. Membranes of BC were produced through static culture of the bacteria Gluconacetobacter xylinus. Membranes were then washed, dried and swelled in acetone for 48 hours, followed by immersion in 25ml of 3% PCL solution for 72 hours. Next, membranes of the BC/PCL composite underwent several washes in acetone and were then dried to be characterized by scanning electron microscopy (SEM), X-ray diffraction, thermal and mechanical trials and luminous transmittance. In vivo study was conducted with the BC/PCL composite and with pure BC to evaluate their interactions with corneal tissue. At first, membranes were cut in 8mm diameter to be then applied in corneal tisse. Thirty six female rabbits were separated in three groups of 12 animals each. An ulcer of 5mm in diameter and 0.2mm in depth was created on the right cornea of all rabbits. From the base of the ulcer, the stroma was separated creating an interlamellar pocket, and each group of 12 animals received 8mm diameter BC/PCL composite membrane and the other group received BC membranes on the same size, both adjusted in the pocket. The last group did not receive any membrane, used as the control. Animals were followed for 45 days, where clinical evaluations for signs of pain and inflammation were conducted. Three animals of each group were euthanized on days 3, 7, 21 and 45 after surgery to collect samples for histology (HE and Picrosirius) and SEM. BC/PCL presented 84% of light transmission while pure BC presented 45%. Lower tension resistance and Young s module was observed on the composite compared to pure BC, while the composite presented greater elongation at rupture what demonstrates a greater viscoelasticity of this material. Eyes that received membranes presented moderate inflammation until 30 days after surgery, decreasing progressively until 45 days. Histology revealed the presence of lymphohistiocytic inflammatory infiltrate at 21 and 45 days in implanted groups, at a moderate intensity, as well as giant cells, suggesting chronic inflammatory and foreign body reactions. Fibroplasia was observed around the membranes, a positive sign of biocompatibility. Epithelization was complete only in control group, what shows that membranes surfaces did not allow epithelium growth and maintenance. Picrosirius staining showed a greater proportion of type III collagen at the end of 45 days in both implanted groups, suggesting that the repair process around the membranes was incomplete. SEM showed extracellular matrix and cells adhered to implant surfaces, suggesting integration at 45 days. Both membranes made of BC/PCL composite and BC alone stimulated chronic inflammatory and foreign body reactions and did not allow complete epithelization on their surfaces. Based on these conclusions, we believe that implants made of BC/PCL and BC using the here presented process are not indicated as a substitute for corneal tissue in rabbits.
publishDate 2012
dc.date.issued.fl_str_mv 2012-09-04
dc.date.available.fl_str_mv 2013-07-04
2015-03-26T13:47:10Z
dc.date.accessioned.fl_str_mv 2015-03-26T13:47:10Z
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dc.identifier.citation.fl_str_mv QUEIROZ, Paulo Victor da Silva. Development, characterization and application of membranes made of bacterial cellulose and of the nanocomposite bacterial cellulose/polycaprolactone on experimental lamellar keratoplasty in rabbits. 2012. 83 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.
dc.identifier.uri.fl_str_mv http://locus.ufv.br/handle/123456789/5122
identifier_str_mv QUEIROZ, Paulo Victor da Silva. Development, characterization and application of membranes made of bacterial cellulose and of the nanocomposite bacterial cellulose/polycaprolactone on experimental lamellar keratoplasty in rabbits. 2012. 83 f. Dissertação (Mestrado em Biotecnologia, diagnóstico e controle de doenças; Epidemiologia e controle de qualidade de prod. de) - Universidade Federal de Viçosa, Viçosa, 2012.
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