Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo
| Ano de defesa: | 2025 |
|---|---|
| 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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| Área do conhecimento CNPq: | |
| Link de acesso: | http://repositorio.ufc.br/handle/riufc/81532 |
Resumo: | Frutapin is a lectin with affinity for glucose/mannose found in the seeds of Artocarpus altilis (breadfruit) and shares structural similarity with artocarpin, which is known for its immunostimulatory and wound-healing properties. Given its low natural extraction yield, the expression of its recombinant form in Escherichia coli was standardized, thereby increasing its production and enabling studies on its pharmacological potential. This study aimed to investigate the recombinant form of frutapin (rFTP) for its wound-healing potential in vitro and in vivo by evaluating the viability of fibroblasts (L929), keratinocytes (HaCaT), and macrophages (RAW 264.7) using the resazurin assay, as well as nitric oxide production (Griess assay) and cytokines TNF-α, IL-1β, IL-6, and IL-10 (ELISA) in RAW 264.7 cell supernatants. Subsequently, rFTP (0.1%, 0.5%, and 1%) was incorporated into xyloglucan-based biomembranes and tested in excisional wounds in BALB/c mice (n = 6 animals/group), compared with untreated controls (sham) and biomembrane without the active compound. The animals were euthanized on days 3, 7, and 14 post-surgery. rFTP (0.15–1 mg/mL) increased the proliferation of L929 and HaCaT cells and modulated the inflammatory response in RAW 264.7 macrophages compared to the CTRL group, significantly enhancing proliferation and IL-10 production while reducing the levels of pro-inflammatory cytokines TNF-α, IL-6, and IL-1β, both in the presence and absence of LPS. The xyloglucan biomembrane containing 0.1% rFTP accelerated wound contraction and reduced inflammatory cell infiltration on days 3 and 7. Moreover, epithelial thickness was significantly higher in the rFTP-treated groups (0.1%) compared to the sham and control groups, highlighting rFTP’s ability to stimulate tissue regeneration and extracellular matrix remodeling. Levels of IL-1β, TNF-α, and IL-6 were reduced, while IL-10 was considerably increased, suggesting that rFTP modulates inflammation. These results indicate that rFTP is a promising candidate for wound management by promoting inflammatory control and regenerative stimulation. Thus, this work encourages further studies to elucidate the specific molecular mechanisms of rFTP and to assess its efficacy and safety in clinical trials. |
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Rabelo, Liviane Maria AlvesSousa, Felipe Domingos deAlencar, Nylane Maria Nunes de2025-07-11T11:47:26Z2025-07-11T11:47:26Z2025RABELO, Liviane Maria Alves. Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo, 2025. Dissertação (Mestrado em Farmacologia) – Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2025. Disponível em: http://repositorio.ufc.br/handle/riufc/81532. Acesso em: 11 jul. 2025.http://repositorio.ufc.br/handle/riufc/81532Frutapin is a lectin with affinity for glucose/mannose found in the seeds of Artocarpus altilis (breadfruit) and shares structural similarity with artocarpin, which is known for its immunostimulatory and wound-healing properties. Given its low natural extraction yield, the expression of its recombinant form in Escherichia coli was standardized, thereby increasing its production and enabling studies on its pharmacological potential. This study aimed to investigate the recombinant form of frutapin (rFTP) for its wound-healing potential in vitro and in vivo by evaluating the viability of fibroblasts (L929), keratinocytes (HaCaT), and macrophages (RAW 264.7) using the resazurin assay, as well as nitric oxide production (Griess assay) and cytokines TNF-α, IL-1β, IL-6, and IL-10 (ELISA) in RAW 264.7 cell supernatants. Subsequently, rFTP (0.1%, 0.5%, and 1%) was incorporated into xyloglucan-based biomembranes and tested in excisional wounds in BALB/c mice (n = 6 animals/group), compared with untreated controls (sham) and biomembrane without the active compound. The animals were euthanized on days 3, 7, and 14 post-surgery. rFTP (0.15–1 mg/mL) increased the proliferation of L929 and HaCaT cells and modulated the inflammatory response in RAW 264.7 macrophages compared to the CTRL group, significantly enhancing proliferation and IL-10 production while reducing the levels of pro-inflammatory cytokines TNF-α, IL-6, and IL-1β, both in the presence and absence of LPS. The xyloglucan biomembrane containing 0.1% rFTP accelerated wound contraction and reduced inflammatory cell infiltration on days 3 and 7. Moreover, epithelial thickness was significantly higher in the rFTP-treated groups (0.1%) compared to the sham and control groups, highlighting rFTP’s ability to stimulate tissue regeneration and extracellular matrix remodeling. Levels of IL-1β, TNF-α, and IL-6 were reduced, while IL-10 was considerably increased, suggesting that rFTP modulates inflammation. These results indicate that rFTP is a promising candidate for wound management by promoting inflammatory control and regenerative stimulation. Thus, this work encourages further studies to elucidate the specific molecular mechanisms of rFTP and to assess its efficacy and safety in clinical trials.A Frutapina é uma lectina com afinidade por glicose/manose presente em sementes de Artocapus altilis (fruta-pão) e apresenta semelhança estrutural com Artocarpina, conhecida por suas propriedades imunoestimulantes e cicatrizantes. Considerando seu baixo rendimento de extração natural, a expressão da sua forma recombinante em Escherichia coli foi padronizada, e, assim, aumentou sua produção, viabilizando estudos sobre seu potencial farmacológico. Este trabalho teve como objetivo a investigar a forma recombinante da frutapina (rFTP) quanto ao seu potencial cicatrizante in vitro e in vivo através da avaliação da viabilidade de Fibroblastos (L929), Queratinócitos (HaCaT) e Macrófagos (RAW 264.7) (resazurina), bem como a produção de Óxido Nítrico (Griess) e Citocinas TNF-a, IL-1b, IL-6 e IL-10 (Elisa) no sobrenatante da RAW 264.7. Em seguida, a rFTP (0,1%, 0,5% e 1%) foi incorporada em biomembranas de Xiloglucana e testada em feridas excisionais de camundongos BALB/c (n=6animais/grupo), comparando-se com os controles sem tratamento (Sham) e biomembrana, sem o ativo. Os animais foram eutanasiados após 3, 7 e 14 dias após a cirurgia. A rFTP (0,15 – 1mg/mL) aumentou a proliferação da L929, HaCaT, bem como modulou a resposta inflamatória da RAW 264.7, em comparação ao grupo CTRL, aumentando significativamente a proliferação e a produção da citocina anti-inflamatória IL-10 e reduzindo os níveis das citocinas pró-inflamatórias TNF-α e IL-6 e IL1β com e sem LPS. A biomembrana de xiloglucana contendo rFTP na concentração de 0,1% acelerou a contração das lesões e diminuiu o infiltrado inflamatório no 3º e 7º dia. Além disso, a espessura epitelial foi significativamente maior nos grupos tratados com rFTP (0,1%) em comparação aos grupos sham e controle, evidenciando a capacidade da rFTP de estimular a regeneração tecidual e estruturação da matriz extracelular. Os níveis de IL-1β, TNF-α e IL-6 foram reduzidos e houve um aumento considerável de IL-10, sugerindo que a rFTP modula inflamação. Esses resultados indicam que a rFTP é uma candidata promissora para o manejo de feridas através do controle inflamatório e estimulação regenerativa. Dessa forma, este trabalho impulsiona estudos adicionais para elucidar os mecanismos moleculares específicos da rFTP e avaliar sua eficácia e segurança em ensaios clínicos.Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivoEffect of the recombinant lectin r-frutapin from Artocarpus altilis on the wound healing process: in vitro and in vivo studiesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisLectinas de PlantasProteínas RecombinantesReepitelizaçãoPlant LectinsRecombinant ProteinsRe-EpithelializationCNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIAinfo:eu-repo/semantics/openAccessporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFChttps://orcid.org/0000-0001-5779-839Xhttp://lattes.cnpq.br/5481922226880838http://lattes.cnpq.br/9219662256316695http://lattes.cnpq.br/9542077634881460ORIGINAL2025_dis_lmarabelo.pdf2025_dis_lmarabelo.pdfapplication/pdf3456903http://repositorio.ufc.br/bitstream/riufc/81532/1/2025_dis_lmarabelo.pdf8ff4ef22e184b79b5d0e9d5f0da176b9MD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/81532/3/license.txt8a4605be74aa9ea9d79846c1fba20a33MD53riufc/815322025-07-11 08:48:31.022oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2025-07-11T11:48:31Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false |
| dc.title.pt_BR.fl_str_mv |
Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo |
| dc.title.en.pt_BR.fl_str_mv |
Effect of the recombinant lectin r-frutapin from Artocarpus altilis on the wound healing process: in vitro and in vivo studies |
| title |
Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo |
| spellingShingle |
Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo Rabelo, Liviane Maria Alves CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA Lectinas de Plantas Proteínas Recombinantes Reepitelização Plant Lectins Recombinant Proteins Re-Epithelialization |
| title_short |
Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo |
| title_full |
Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo |
| title_fullStr |
Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo |
| title_full_unstemmed |
Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo |
| title_sort |
Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo |
| author |
Rabelo, Liviane Maria Alves |
| author_facet |
Rabelo, Liviane Maria Alves |
| author_role |
author |
| dc.contributor.co-advisor.none.fl_str_mv |
Sousa, Felipe Domingos de |
| dc.contributor.author.fl_str_mv |
Rabelo, Liviane Maria Alves |
| dc.contributor.advisor1.fl_str_mv |
Alencar, Nylane Maria Nunes de |
| contributor_str_mv |
Alencar, Nylane Maria Nunes de |
| dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA |
| topic |
CNPQ::CIENCIAS BIOLOGICAS::FARMACOLOGIA Lectinas de Plantas Proteínas Recombinantes Reepitelização Plant Lectins Recombinant Proteins Re-Epithelialization |
| dc.subject.ptbr.pt_BR.fl_str_mv |
Lectinas de Plantas Proteínas Recombinantes Reepitelização |
| dc.subject.en.pt_BR.fl_str_mv |
Plant Lectins Recombinant Proteins Re-Epithelialization |
| description |
Frutapin is a lectin with affinity for glucose/mannose found in the seeds of Artocarpus altilis (breadfruit) and shares structural similarity with artocarpin, which is known for its immunostimulatory and wound-healing properties. Given its low natural extraction yield, the expression of its recombinant form in Escherichia coli was standardized, thereby increasing its production and enabling studies on its pharmacological potential. This study aimed to investigate the recombinant form of frutapin (rFTP) for its wound-healing potential in vitro and in vivo by evaluating the viability of fibroblasts (L929), keratinocytes (HaCaT), and macrophages (RAW 264.7) using the resazurin assay, as well as nitric oxide production (Griess assay) and cytokines TNF-α, IL-1β, IL-6, and IL-10 (ELISA) in RAW 264.7 cell supernatants. Subsequently, rFTP (0.1%, 0.5%, and 1%) was incorporated into xyloglucan-based biomembranes and tested in excisional wounds in BALB/c mice (n = 6 animals/group), compared with untreated controls (sham) and biomembrane without the active compound. The animals were euthanized on days 3, 7, and 14 post-surgery. rFTP (0.15–1 mg/mL) increased the proliferation of L929 and HaCaT cells and modulated the inflammatory response in RAW 264.7 macrophages compared to the CTRL group, significantly enhancing proliferation and IL-10 production while reducing the levels of pro-inflammatory cytokines TNF-α, IL-6, and IL-1β, both in the presence and absence of LPS. The xyloglucan biomembrane containing 0.1% rFTP accelerated wound contraction and reduced inflammatory cell infiltration on days 3 and 7. Moreover, epithelial thickness was significantly higher in the rFTP-treated groups (0.1%) compared to the sham and control groups, highlighting rFTP’s ability to stimulate tissue regeneration and extracellular matrix remodeling. Levels of IL-1β, TNF-α, and IL-6 were reduced, while IL-10 was considerably increased, suggesting that rFTP modulates inflammation. These results indicate that rFTP is a promising candidate for wound management by promoting inflammatory control and regenerative stimulation. Thus, this work encourages further studies to elucidate the specific molecular mechanisms of rFTP and to assess its efficacy and safety in clinical trials. |
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2025 |
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2025-07-11T11:47:26Z |
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2025-07-11T11:47:26Z |
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2025 |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/masterThesis |
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masterThesis |
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publishedVersion |
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RABELO, Liviane Maria Alves. Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo, 2025. Dissertação (Mestrado em Farmacologia) – Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2025. Disponível em: http://repositorio.ufc.br/handle/riufc/81532. Acesso em: 11 jul. 2025. |
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http://repositorio.ufc.br/handle/riufc/81532 |
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RABELO, Liviane Maria Alves. Efeito da lectina recombinante r-frutapina de Artocarpus altilis sobre o processo de cicatrização: estudos in vitro e in vivo, 2025. Dissertação (Mestrado em Farmacologia) – Faculdade de Medicina, Universidade Federal do Ceará, Fortaleza, 2025. Disponível em: http://repositorio.ufc.br/handle/riufc/81532. Acesso em: 11 jul. 2025. |
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