Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa
| Ano de defesa: | 2020 |
|---|---|
| Autor(a) principal: | |
| Orientador(a): | |
| Banca de defesa: | |
| Tipo de documento: | Dissertação |
| Tipo de acesso: | Acesso aberto |
| Idioma: | por |
| Instituição de defesa: |
Não Informado pela instituição
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| Programa de Pós-Graduação: |
Não Informado pela instituição
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| Departamento: |
Não Informado pela instituição
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| País: |
Não Informado pela instituição
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| Palavras-chave em Português: | |
| Link de acesso: | http://www.repositorio.ufc.br/handle/riufc/49896 |
Resumo: | Scaffolds are biomaterials composed of decellularized extracellular matrix (MEC), widely used in developed countries. During the production of scaffolds, MEC is isolated from its native cells and genetic material to produce a biomaterial with structural, biochemical and biomechanical characteristics of the original tissue. Collagen is a major component of scaffolds. In this context, Nile Tilapia skin appears as a new biomaterial useful for bioengineering, due to its mechanical properties and similarity in histological composition. The objective of this work was to develop a decellularized extracellular matrix from the tilapia skin as a new biomaterial for application in regenerative medicine. In this work, different physical, chemical and enzymatic methods were studied for the decellularization of tilapia skin, in order to obtain a scaffold with the best possible quality. Twelve (12) decellularization protocols were developed and tested, using ionic and non-ionic surfactants, such as sodium Dodecyl sulfate, sodium deoxycholate, Triton X-100, Tween 20, CHAPS and Nonidet P-40, with the purpose of eliminate cells (evaluated by DAPI staining), maintain the structure (evaluated by HE) and do not cause cytotoxicity. The best protocols tried were Tween 20 for 24h, CHAPS 8 mM for 4h and Triton X-100 at 1% for 6h, with decellularization percentages of 100%, 96.35% ± 3.65% and 97.83% ± 2.17% and were approved in the indirect cytotoxicity test for having cell viability of 86.71% ± 2.24%, 108.3% ± 6.49% and 95.63% ± 3.62%; respectively. The ideal irradiation dose for radio-sterilization was determined using the scaffolds produced by the CHAPS 8 mM protocol and testing with doses of 15 25, 30 and 50 kGy. With these results we conclude that the scaffolds produced from Tween 20 for 24h, CHAPS 8 mM for 4h and Triton X-100 at 1% for 6h have potential for application as biomedical materials, as they are the best in decellularization, maintaining the structure of fibers and not causing cytotoxicity. Additionally, the 25 kGy dose was considered the best for irradiation. |
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Mendoza Hernández, Eleicy NathalyPaier, Carlos Roberto KosckyMoraes Filho, Manoel Odorico de2020-02-06T18:09:35Z2020-02-06T18:09:35Z2020-01-15MENDOZA HERNÁNDEZ, Eleicy Nathaly. Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa. 2020. 86 f. Dissertação (Mestrado em Farmacologia) – Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2020. Disponível em: http://www.repositorio.ufc.br/handle/riufc/ 49896. Acesso em: 06 fev. 2020.http://www.repositorio.ufc.br/handle/riufc/49896Scaffolds are biomaterials composed of decellularized extracellular matrix (MEC), widely used in developed countries. During the production of scaffolds, MEC is isolated from its native cells and genetic material to produce a biomaterial with structural, biochemical and biomechanical characteristics of the original tissue. Collagen is a major component of scaffolds. In this context, Nile Tilapia skin appears as a new biomaterial useful for bioengineering, due to its mechanical properties and similarity in histological composition. The objective of this work was to develop a decellularized extracellular matrix from the tilapia skin as a new biomaterial for application in regenerative medicine. In this work, different physical, chemical and enzymatic methods were studied for the decellularization of tilapia skin, in order to obtain a scaffold with the best possible quality. Twelve (12) decellularization protocols were developed and tested, using ionic and non-ionic surfactants, such as sodium Dodecyl sulfate, sodium deoxycholate, Triton X-100, Tween 20, CHAPS and Nonidet P-40, with the purpose of eliminate cells (evaluated by DAPI staining), maintain the structure (evaluated by HE) and do not cause cytotoxicity. The best protocols tried were Tween 20 for 24h, CHAPS 8 mM for 4h and Triton X-100 at 1% for 6h, with decellularization percentages of 100%, 96.35% ± 3.65% and 97.83% ± 2.17% and were approved in the indirect cytotoxicity test for having cell viability of 86.71% ± 2.24%, 108.3% ± 6.49% and 95.63% ± 3.62%; respectively. The ideal irradiation dose for radio-sterilization was determined using the scaffolds produced by the CHAPS 8 mM protocol and testing with doses of 15 25, 30 and 50 kGy. With these results we conclude that the scaffolds produced from Tween 20 for 24h, CHAPS 8 mM for 4h and Triton X-100 at 1% for 6h have potential for application as biomedical materials, as they are the best in decellularization, maintaining the structure of fibers and not causing cytotoxicity. Additionally, the 25 kGy dose was considered the best for irradiation.Os scaffolds são biomateriais compostos por matriz extracelular (MEC) descelularizada, amplamente utilizados em países desenvolvidos. Na produção dos scaffolds, a MEC é isolada de suas células nativas e de material genético para produzir um biomaterial com características estruturais, bioquímicas e biomecânicas semelhantes às do tecido original. O colágeno é um dos principais componentes dos scaffolds. Nesse contexto, a pele da Tilápia do Nilo surge como um novo biomaterial útil à bioengenharia, devido às suas propriedades mecânicas satisfatórias e similaridade à pele humana quanto à composição e arranjo das fibras colagênicas. O objetivo deste trabalho é desenvolver uma matriz extracelular descelularizada a partir da pele da tilápia como novo biomaterial para aplicação em medicina regenerativa. Foram estudados diferentes métodos físicos, químicos e enzimáticos para a descelularização da pele de tilápia, com a finalidade de se obter um scaffold com a melhor qualidade possível. Foram desenvolvidos e experimentados doze (12) protocolos de descelularização, que usaram surfactantes iônicos e não iônicos, como o Dodecil sulfato de sódio, Deoxicolato de sódio, Triton X-100, Tween 20, CHAPS (3-[(3-Colamidopropil)dimetilamônio]-1-propanosulfonato) e Nonidet P-40, com a finalidade de eliminar remanescentes celulares (avaliados pela coloração de DAPI), manter a estrutura morfológica (avaliada pela técnica de hematoxilina-eosina ou HE) e não causar citotoxicidade. Os melhores protocolos experimentados foram o Tween 20 por 24h, CHAPS 8,0 mM por 4h e o Triton X-100 a 1% por 6h, com porcentagens de descelularização de 100%, 96,35% ± 3,65% e 97,83% ± 2,17%, respectivamente. Esses métodos também foram aprovados no teste de citotoxicidade indireta por apresentarem viabilidade celular de 86,71% ± 2,24%, 108,3% ± 6,49% e 95,63% ± 3,62%; respectivamente. A dose ideal de irradiação para radioesterilização foi determinada usando os scaffolds produzidos pelo protocolo CHAPS 8,0 mM e experimentando doses de 15 kGy, 25 kGy, 30 kGy e 50 kGy. Com estes resultados concluímos que os scaffolds produzidos a partir de Tween 20 por 24h, CHAPS 8 mM por 4h e Triton X-100 a 1% por 6h têm potencial de aplicação como materiais biomédicos, por serem os melhores na descelularização, mantendo a estrutura das fibras e não causando citotoxicidade. Por sua vez, a irradiação com raios gama na dose de 25 kGy foi considerada como a melhor para a esterilização de scaffolds produzidos pelo método do CHAPS.Engenharia TecidualTilápiaMateriais BiocompatíveisDesenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativaDevelopment of a decellularized extracellular matriz from tilápia skin (scaffold) as new biomaterial for application in regenerative medicineinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessLICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/49896/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52ORIGINAL2020_dis_enmhernández.pdf2020_dis_enmhernández.pdfapplication/pdf147855http://repositorio.ufc.br/bitstream/riufc/49896/3/2020_dis_enmhern%c3%a1ndez.pdfe923a0eb5f9ca50f8866205d466f5664MD53riufc/498962024-03-14 15:05:07.543oai:repositorio.ufc.br:riufc/49896Tk9URTogUExBQ0UgWU9VUiBPV04gTElDRU5TRSBIRVJFClRoaXMgc2FtcGxlIGxpY2Vuc2UgaXMgcHJvdmlkZWQgZm9yIGluZm9ybWF0aW9uYWwgcHVycG9zZXMgb25seS4KCk5PTi1FWENMVVNJVkUgRElTVFJJQlVUSU9OIExJQ0VOU0UKCkJ5IHNpZ25pbmcgYW5kIHN1Ym1pdHRpbmcgdGhpcyBsaWNlbnNlLCB5b3UgKHRoZSBhdXRob3Iocykgb3IgY29weXJpZ2h0Cm93bmVyKSBncmFudHMgdG8gRFNwYWNlIFVuaXZlcnNpdHkgKERTVSkgdGhlIG5vbi1leGNsdXNpdmUgcmlnaHQgdG8gcmVwcm9kdWNlLAp0cmFuc2xhdGUgKGFzIGRlZmluZWQgYmVsb3cpLCBhbmQvb3IgZGlzdHJpYnV0ZSB5b3VyIHN1Ym1pc3Npb24gKGluY2x1ZGluZwp0aGUgYWJzdHJhY3QpIHdvcmxkd2lkZSBpbiBwcmludCBhbmQgZWxlY3Ryb25pYyBmb3JtYXQgYW5kIGluIGFueSBtZWRpdW0sCmluY2x1ZGluZyBidXQgbm90IGxpbWl0ZWQgdG8gYXVkaW8gb3IgdmlkZW8uCgpZb3UgYWdyZWUgdGhhdCBEU1UgbWF5LCB3aXRob3V0IGNoYW5naW5nIHRoZSBjb250ZW50LCB0cmFuc2xhdGUgdGhlCnN1Ym1pc3Npb24gdG8gYW55IG1lZGl1bSBvciBmb3JtYXQgZm9yIHRoZSBwdXJwb3NlIG9mIHByZXNlcnZhdGlvbi4KCllvdSBhbHNvIGFncmVlIHRoYXQgRFNVIG1heSBrZWVwIG1vcmUgdGhhbiBvbmUgY29weSBvZiB0aGlzIHN1Ym1pc3Npb24gZm9yCnB1cnBvc2VzIG9mIHNlY3VyaXR5LCBiYWNrLXVwIGFuZCBwcmVzZXJ2YXRpb24uCgpZb3UgcmVwcmVzZW50IHRoYXQgdGhlIHN1Ym1pc3Npb24gaXMgeW91ciBvcmlnaW5hbCB3b3JrLCBhbmQgdGhhdCB5b3UgaGF2ZQp0aGUgcmlnaHQgdG8gZ3JhbnQgdGhlIHJpZ2h0cyBjb250YWluZWQgaW4gdGhpcyBsaWNlbnNlLiBZb3UgYWxzbyByZXByZXNlbnQKdGhhdCB5b3VyIHN1Ym1pc3Npb24gZG9lcyBub3QsIHRvIHRoZSBiZXN0IG9mIHlvdXIga25vd2xlZGdlLCBpbmZyaW5nZSB1cG9uCmFueW9uZSdzIGNvcHlyaWdodC4KCklmIHRoZSBzdWJtaXNzaW9uIGNvbnRhaW5zIG1hdGVyaWFsIGZvciB3aGljaCB5b3UgZG8gbm90IGhvbGQgY29weXJpZ2h0LAp5b3UgcmVwcmVzZW50IHRoYXQgeW91IGhhdmUgb2J0YWluZWQgdGhlIHVucmVzdHJpY3RlZCBwZXJtaXNzaW9uIG9mIHRoZQpjb3B5cmlnaHQgb3duZXIgdG8gZ3JhbnQgRFNVIHRoZSByaWdodHMgcmVxdWlyZWQgYnkgdGhpcyBsaWNlbnNlLCBhbmQgdGhhdApzdWNoIHRoaXJkLXBhcnR5IG93bmVkIG1hdGVyaWFsIGlzIGNsZWFybHkgaWRlbnRpZmllZCBhbmQgYWNrbm93bGVkZ2VkCndpdGhpbiB0aGUgdGV4dCBvciBjb250ZW50IG9mIHRoZSBzdWJtaXNzaW9uLgoKSUYgVEhFIFNVQk1JU1NJT04gSVMgQkFTRUQgVVBPTiBXT1JLIFRIQVQgSEFTIEJFRU4gU1BPTlNPUkVEIE9SIFNVUFBPUlRFRApCWSBBTiBBR0VOQ1kgT1IgT1JHQU5JWkFUSU9OIE9USEVSIFRIQU4gRFNVLCBZT1UgUkVQUkVTRU5UIFRIQVQgWU9VIEhBVkUKRlVMRklMTEVEIEFOWSBSSUdIVCBPRiBSRVZJRVcgT1IgT1RIRVIgT0JMSUdBVElPTlMgUkVRVUlSRUQgQlkgU1VDSApDT05UUkFDVCBPUiBBR1JFRU1FTlQuCgpEU1Ugd2lsbCBjbGVhcmx5IGlkZW50aWZ5IHlvdXIgbmFtZShzKSBhcyB0aGUgYXV0aG9yKHMpIG9yIG93bmVyKHMpIG9mIHRoZQpzdWJtaXNzaW9uLCBhbmQgd2lsbCBub3QgbWFrZSBhbnkgYWx0ZXJhdGlvbiwgb3RoZXIgdGhhbiBhcyBhbGxvd2VkIGJ5IHRoaXMKbGljZW5zZSwgdG8geW91ciBzdWJtaXNzaW9uLgo=Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-03-14T18:05:07Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false |
| dc.title.pt_BR.fl_str_mv |
Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa |
| dc.title.en.pt_BR.fl_str_mv |
Development of a decellularized extracellular matriz from tilápia skin (scaffold) as new biomaterial for application in regenerative medicine |
| title |
Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa |
| spellingShingle |
Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa Mendoza Hernández, Eleicy Nathaly Engenharia Tecidual Tilápia Materiais Biocompatíveis |
| title_short |
Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa |
| title_full |
Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa |
| title_fullStr |
Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa |
| title_full_unstemmed |
Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa |
| title_sort |
Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa |
| author |
Mendoza Hernández, Eleicy Nathaly |
| author_facet |
Mendoza Hernández, Eleicy Nathaly |
| author_role |
author |
| dc.contributor.co-advisor.none.fl_str_mv |
Paier, Carlos Roberto Koscky |
| dc.contributor.author.fl_str_mv |
Mendoza Hernández, Eleicy Nathaly |
| dc.contributor.advisor1.fl_str_mv |
Moraes Filho, Manoel Odorico de |
| contributor_str_mv |
Moraes Filho, Manoel Odorico de |
| dc.subject.por.fl_str_mv |
Engenharia Tecidual Tilápia Materiais Biocompatíveis |
| topic |
Engenharia Tecidual Tilápia Materiais Biocompatíveis |
| description |
Scaffolds are biomaterials composed of decellularized extracellular matrix (MEC), widely used in developed countries. During the production of scaffolds, MEC is isolated from its native cells and genetic material to produce a biomaterial with structural, biochemical and biomechanical characteristics of the original tissue. Collagen is a major component of scaffolds. In this context, Nile Tilapia skin appears as a new biomaterial useful for bioengineering, due to its mechanical properties and similarity in histological composition. The objective of this work was to develop a decellularized extracellular matrix from the tilapia skin as a new biomaterial for application in regenerative medicine. In this work, different physical, chemical and enzymatic methods were studied for the decellularization of tilapia skin, in order to obtain a scaffold with the best possible quality. Twelve (12) decellularization protocols were developed and tested, using ionic and non-ionic surfactants, such as sodium Dodecyl sulfate, sodium deoxycholate, Triton X-100, Tween 20, CHAPS and Nonidet P-40, with the purpose of eliminate cells (evaluated by DAPI staining), maintain the structure (evaluated by HE) and do not cause cytotoxicity. The best protocols tried were Tween 20 for 24h, CHAPS 8 mM for 4h and Triton X-100 at 1% for 6h, with decellularization percentages of 100%, 96.35% ± 3.65% and 97.83% ± 2.17% and were approved in the indirect cytotoxicity test for having cell viability of 86.71% ± 2.24%, 108.3% ± 6.49% and 95.63% ± 3.62%; respectively. The ideal irradiation dose for radio-sterilization was determined using the scaffolds produced by the CHAPS 8 mM protocol and testing with doses of 15 25, 30 and 50 kGy. With these results we conclude that the scaffolds produced from Tween 20 for 24h, CHAPS 8 mM for 4h and Triton X-100 at 1% for 6h have potential for application as biomedical materials, as they are the best in decellularization, maintaining the structure of fibers and not causing cytotoxicity. Additionally, the 25 kGy dose was considered the best for irradiation. |
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2020 |
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2020-02-06T18:09:35Z |
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2020-02-06T18:09:35Z |
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2020-01-15 |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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MENDOZA HERNÁNDEZ, Eleicy Nathaly. Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa. 2020. 86 f. Dissertação (Mestrado em Farmacologia) – Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2020. Disponível em: http://www.repositorio.ufc.br/handle/riufc/ 49896. Acesso em: 06 fev. 2020. |
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http://www.repositorio.ufc.br/handle/riufc/49896 |
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MENDOZA HERNÁNDEZ, Eleicy Nathaly. Desenvolvimento de matriz extracelular descelularizada (Scaffold) de pele de tilápia como novo biomaterial para aplicação em medicina regenerativa. 2020. 86 f. Dissertação (Mestrado em Farmacologia) – Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2020. Disponível em: http://www.repositorio.ufc.br/handle/riufc/ 49896. Acesso em: 06 fev. 2020. |
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