3D-MSCs A151 ODN-Loaded Exosomes Are Immunomodulatory And Reveal A Proteomic Cargo That Sustains Wound Resolution
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10451/52308 |
Resumo: | Introduction Non-healing wounds remain a major burden due to the lack of effective treatments. Mesenchymal stem cell-derived exosomes (MSC-Exo) have emerged as therapeutic options given their pro-regenerative and immunomodulatory features. Still, little is known on the exact mechanisms mediated by MSC-Exo. Importantly, modulation of their efficacy through 3D-physiologic cultures together with loading strategies continues underexplored. Objectives To uncover the MSC-Exo-mediated mechanism via proteomic analyses, and to use 3D-culture and loading technologies to expand MSC-Exo efficacy for cutaneous wound healing. Methods MSC-Exo were produced in either 3D or 2D cultures (Exo3D/Exo2D) and loaded with an exogenous immunosuppressive oligodeoxynucleotide (A151 ODN). Both, loaded and naïve exosomes were characterised regarding size, morphology and the presence of specific protein markers; while IPA analyses enabled to correlate their protein content with the effects observed in vitro and in vivo. The Exo3D/Exo2D regenerative potential was evaluated in vitro by assessing keratinocyte and fibroblast mitogenicity, motogenicity, and cytokine secretion as well as using an in vivo wound splinting model. Accordingly, the modulation of inflammatory and immune responses by A151-loaded Exo3D/Exo2D was also assessed. Results Exo3D stimulated mitogenically and motogenically keratinocytes and fibroblasts in vitro, with upregulation of IL-1α and VEGF-α or increased secretion of TGF-β, TNF-α and IL-10. In vivo, Exo3D reduced the granulation tissue area and promoted complete re-epithelization of the wound. These observations were sustained by the proteomic profiling of the Exo3D cargo that identified wound healing-related proteins, such as TGF-β, ITGA1-3/5, IL-6, CDC151, S100A10 and Wnt5α. Moreover, when loaded with A151 ODN, Exo3D differentially mediated wound healing-related trophic factors reducing the systemic levels of IL-6 and TNF-α at the late stage of wound healing in vivo. Conclusion Our results support the potential of A151-loaded Exo3D for the treatment of chronic wounds by promoting skin regeneration, while modulating the systemic levels of the pro-inflammatory cytokines |
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3D-MSCs A151 ODN-Loaded Exosomes Are Immunomodulatory And Reveal A Proteomic Cargo That Sustains Wound ResolutionExosomesImmunosuppressive oligodeoxynucleotide loadingImmunomodulationProteomics3D-cultured mesenchymal stem/stromal cellsWound healingIntroduction Non-healing wounds remain a major burden due to the lack of effective treatments. Mesenchymal stem cell-derived exosomes (MSC-Exo) have emerged as therapeutic options given their pro-regenerative and immunomodulatory features. Still, little is known on the exact mechanisms mediated by MSC-Exo. Importantly, modulation of their efficacy through 3D-physiologic cultures together with loading strategies continues underexplored. Objectives To uncover the MSC-Exo-mediated mechanism via proteomic analyses, and to use 3D-culture and loading technologies to expand MSC-Exo efficacy for cutaneous wound healing. Methods MSC-Exo were produced in either 3D or 2D cultures (Exo3D/Exo2D) and loaded with an exogenous immunosuppressive oligodeoxynucleotide (A151 ODN). Both, loaded and naïve exosomes were characterised regarding size, morphology and the presence of specific protein markers; while IPA analyses enabled to correlate their protein content with the effects observed in vitro and in vivo. The Exo3D/Exo2D regenerative potential was evaluated in vitro by assessing keratinocyte and fibroblast mitogenicity, motogenicity, and cytokine secretion as well as using an in vivo wound splinting model. Accordingly, the modulation of inflammatory and immune responses by A151-loaded Exo3D/Exo2D was also assessed. Results Exo3D stimulated mitogenically and motogenically keratinocytes and fibroblasts in vitro, with upregulation of IL-1α and VEGF-α or increased secretion of TGF-β, TNF-α and IL-10. In vivo, Exo3D reduced the granulation tissue area and promoted complete re-epithelization of the wound. These observations were sustained by the proteomic profiling of the Exo3D cargo that identified wound healing-related proteins, such as TGF-β, ITGA1-3/5, IL-6, CDC151, S100A10 and Wnt5α. Moreover, when loaded with A151 ODN, Exo3D differentially mediated wound healing-related trophic factors reducing the systemic levels of IL-6 and TNF-α at the late stage of wound healing in vivo. Conclusion Our results support the potential of A151-loaded Exo3D for the treatment of chronic wounds by promoting skin regeneration, while modulating the systemic levels of the pro-inflammatory cytokinesThis study was partially supported by FCT (PTDC/MED-TOX/29183/2017, PTDC/MED-QUI/31721/2017 and PTDC/SAU-SER/30197/2017), by strategic funding for iMed.ULisboa (UIDB/04138/2020 and UIDP/04138/2020), for UnIC (UID/00051/2020) and for iBiMED (UID/BIM/04501/2020), and by European Union, Quadro de Referência Estratégico Nacional (QREN), Fundo Europeu de Desenvolvimento Regional (FEDER) and Programa Operacional Fatores de Competitividade (COMPETE) (POCI-01-0145-FEDER-007628 and LISBOA-01-0145- FEDER-030197). VIB Proteomics Core and COST Actions (EU Framework Programme Horizon 2020) CA16113 and CA16119 are also acknowledged. The authors thank Dr. Tânia Carvalho for her valuable assistance on histological analysis. SPC also acknowledges EFIS-IL short-term fellowship.ElsevierRepositório da Universidade de LisboaP Camões, SérgioBulut, OzlemYazar, VolkanGaspar, Maria ManuelaSimões, SandraFerreira, RitaVitorino, RuiSantos, Jorge M.Gursel, IhsanMiranda, Joana P2022-04-12T14:01:55Z2022-012022-02-24T12:13:51Z2022-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10451/52308engCamões SP, Bulut O, Yazar V, Gaspar MM, Simões S, Ferreira R, et al. 3D-MSCs A151 ODN-loaded exosomes are immunomodulatory and reveal a proteomic cargo that sustains wound resolution. Journal of Advanced Research [Internet]. 2022;S2090123222000261. Disponível em: https://linkinghub.elsevier.com/retrieve/pii/S20901232220002612090-1232cv-prod-2694205https://doi.org/10.1016/j.jare.2022.01.013info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-11-08T16:56:13Zoai:repositorio.ul.pt:10451/52308Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T22:02:46.441897Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
3D-MSCs A151 ODN-Loaded Exosomes Are Immunomodulatory And Reveal A Proteomic Cargo That Sustains Wound Resolution |
title |
3D-MSCs A151 ODN-Loaded Exosomes Are Immunomodulatory And Reveal A Proteomic Cargo That Sustains Wound Resolution |
spellingShingle |
3D-MSCs A151 ODN-Loaded Exosomes Are Immunomodulatory And Reveal A Proteomic Cargo That Sustains Wound Resolution P Camões, Sérgio Exosomes Immunosuppressive oligodeoxynucleotide loading Immunomodulation Proteomics 3D-cultured mesenchymal stem/stromal cells Wound healing |
title_short |
3D-MSCs A151 ODN-Loaded Exosomes Are Immunomodulatory And Reveal A Proteomic Cargo That Sustains Wound Resolution |
title_full |
3D-MSCs A151 ODN-Loaded Exosomes Are Immunomodulatory And Reveal A Proteomic Cargo That Sustains Wound Resolution |
title_fullStr |
3D-MSCs A151 ODN-Loaded Exosomes Are Immunomodulatory And Reveal A Proteomic Cargo That Sustains Wound Resolution |
title_full_unstemmed |
3D-MSCs A151 ODN-Loaded Exosomes Are Immunomodulatory And Reveal A Proteomic Cargo That Sustains Wound Resolution |
title_sort |
3D-MSCs A151 ODN-Loaded Exosomes Are Immunomodulatory And Reveal A Proteomic Cargo That Sustains Wound Resolution |
author |
P Camões, Sérgio |
author_facet |
P Camões, Sérgio Bulut, Ozlem Yazar, Volkan Gaspar, Maria Manuela Simões, Sandra Ferreira, Rita Vitorino, Rui Santos, Jorge M. Gursel, Ihsan Miranda, Joana P |
author_role |
author |
author2 |
Bulut, Ozlem Yazar, Volkan Gaspar, Maria Manuela Simões, Sandra Ferreira, Rita Vitorino, Rui Santos, Jorge M. Gursel, Ihsan Miranda, Joana P |
author2_role |
author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Repositório da Universidade de Lisboa |
dc.contributor.author.fl_str_mv |
P Camões, Sérgio Bulut, Ozlem Yazar, Volkan Gaspar, Maria Manuela Simões, Sandra Ferreira, Rita Vitorino, Rui Santos, Jorge M. Gursel, Ihsan Miranda, Joana P |
dc.subject.por.fl_str_mv |
Exosomes Immunosuppressive oligodeoxynucleotide loading Immunomodulation Proteomics 3D-cultured mesenchymal stem/stromal cells Wound healing |
topic |
Exosomes Immunosuppressive oligodeoxynucleotide loading Immunomodulation Proteomics 3D-cultured mesenchymal stem/stromal cells Wound healing |
description |
Introduction Non-healing wounds remain a major burden due to the lack of effective treatments. Mesenchymal stem cell-derived exosomes (MSC-Exo) have emerged as therapeutic options given their pro-regenerative and immunomodulatory features. Still, little is known on the exact mechanisms mediated by MSC-Exo. Importantly, modulation of their efficacy through 3D-physiologic cultures together with loading strategies continues underexplored. Objectives To uncover the MSC-Exo-mediated mechanism via proteomic analyses, and to use 3D-culture and loading technologies to expand MSC-Exo efficacy for cutaneous wound healing. Methods MSC-Exo were produced in either 3D or 2D cultures (Exo3D/Exo2D) and loaded with an exogenous immunosuppressive oligodeoxynucleotide (A151 ODN). Both, loaded and naïve exosomes were characterised regarding size, morphology and the presence of specific protein markers; while IPA analyses enabled to correlate their protein content with the effects observed in vitro and in vivo. The Exo3D/Exo2D regenerative potential was evaluated in vitro by assessing keratinocyte and fibroblast mitogenicity, motogenicity, and cytokine secretion as well as using an in vivo wound splinting model. Accordingly, the modulation of inflammatory and immune responses by A151-loaded Exo3D/Exo2D was also assessed. Results Exo3D stimulated mitogenically and motogenically keratinocytes and fibroblasts in vitro, with upregulation of IL-1α and VEGF-α or increased secretion of TGF-β, TNF-α and IL-10. In vivo, Exo3D reduced the granulation tissue area and promoted complete re-epithelization of the wound. These observations were sustained by the proteomic profiling of the Exo3D cargo that identified wound healing-related proteins, such as TGF-β, ITGA1-3/5, IL-6, CDC151, S100A10 and Wnt5α. Moreover, when loaded with A151 ODN, Exo3D differentially mediated wound healing-related trophic factors reducing the systemic levels of IL-6 and TNF-α at the late stage of wound healing in vivo. Conclusion Our results support the potential of A151-loaded Exo3D for the treatment of chronic wounds by promoting skin regeneration, while modulating the systemic levels of the pro-inflammatory cytokines |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-04-12T14:01:55Z 2022-01 2022-02-24T12:13:51Z 2022-01-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://hdl.handle.net/10451/52308 |
url |
http://hdl.handle.net/10451/52308 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Camões SP, Bulut O, Yazar V, Gaspar MM, Simões S, Ferreira R, et al. 3D-MSCs A151 ODN-loaded exosomes are immunomodulatory and reveal a proteomic cargo that sustains wound resolution. Journal of Advanced Research [Internet]. 2022;S2090123222000261. Disponível em: https://linkinghub.elsevier.com/retrieve/pii/S2090123222000261 2090-1232 cv-prod-2694205 https://doi.org/10.1016/j.jare.2022.01.013 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
dc.source.none.fl_str_mv |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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