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Magnetic bioassembly platforms towards the generation of extracellular vesicles from human salivary gland functional organoids for epithelial repair

Salivary glands (SG) are exocrine organs with secretory units commonly injured by radiotherapy. Bio-engineered organoids and extracellular vesicles (EV) are currently under investigation as potential strategies for SG repair. Herein, three-dimensional (3D) cultures of SG functional organoids (SGo) a...

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Autores principales: Chansaenroj, Ajjima, Adine, Christabella, Charoenlappanit, Sawanya, Roytrakul, Sittiruk, Sariya, Ladawan, Osathanon, Thanaphum, Rungarunlert, Sasitorn, Urkasemsin, Ganokon, Chaisuparat, Risa, Yodmuang, Supansa, Souza, Glauco R., Ferreira, João N.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: KeAi Publishing 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8961305/
https://www.ncbi.nlm.nih.gov/pubmed/35387159
http://dx.doi.org/10.1016/j.bioactmat.2022.02.007
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author Chansaenroj, Ajjima
Adine, Christabella
Charoenlappanit, Sawanya
Roytrakul, Sittiruk
Sariya, Ladawan
Osathanon, Thanaphum
Rungarunlert, Sasitorn
Urkasemsin, Ganokon
Chaisuparat, Risa
Yodmuang, Supansa
Souza, Glauco R.
Ferreira, João N.
author_facet Chansaenroj, Ajjima
Adine, Christabella
Charoenlappanit, Sawanya
Roytrakul, Sittiruk
Sariya, Ladawan
Osathanon, Thanaphum
Rungarunlert, Sasitorn
Urkasemsin, Ganokon
Chaisuparat, Risa
Yodmuang, Supansa
Souza, Glauco R.
Ferreira, João N.
author_sort Chansaenroj, Ajjima
collection PubMed
description Salivary glands (SG) are exocrine organs with secretory units commonly injured by radiotherapy. Bio-engineered organoids and extracellular vesicles (EV) are currently under investigation as potential strategies for SG repair. Herein, three-dimensional (3D) cultures of SG functional organoids (SGo) and human dental pulp stem cells (hDPSC) were generated by magnetic 3D bioassembly (M3DB) platforms. Fibroblast growth factor 10 (FGF10) was used to enrich the SGo in secretory epithelial units. After 11 culture days via M3DB, SGo displayed SG-specific acinar epithelial units with functional properties upon neurostimulation. To consistently develop 3D hDPSC in vitro, 3 culture days were sufficient to maintain hDPSC undifferentiated genotype and phenotype for EV generation. EV isolation was performed via sequential centrifugation of the conditioned media of hDPSC and SGo cultures. EV were characterized by nanoparticle tracking analysis, electron microscopy and immunoblotting. EV were in the exosome range for hDPSC (diameter: 88.03 ± 15.60 nm) and for SGo (123.15 ± 63.06 nm). Upon ex vivo administration, exosomes derived from SGo significantly stimulated epithelial growth (up to 60%), mitosis, epithelial progenitors and neuronal growth in injured SG; however, such biological effects were less distinctive with the ones derived from hDPSC. Next, these exosome biological effects were investigated by proteomic arrays. Mass spectrometry profiling of SGo exosomes predicted that cellular growth, development and signaling was due to known and undocumented molecular targets downstream of FGF10. Semaphorins were identified as one of the novel targets requiring further investigations. Thus, M3DB platforms can generate exosomes with potential to ameliorate SG epithelial damage.
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spelling pubmed-89613052022-04-05 Magnetic bioassembly platforms towards the generation of extracellular vesicles from human salivary gland functional organoids for epithelial repair Chansaenroj, Ajjima Adine, Christabella Charoenlappanit, Sawanya Roytrakul, Sittiruk Sariya, Ladawan Osathanon, Thanaphum Rungarunlert, Sasitorn Urkasemsin, Ganokon Chaisuparat, Risa Yodmuang, Supansa Souza, Glauco R. Ferreira, João N. Bioact Mater Article Salivary glands (SG) are exocrine organs with secretory units commonly injured by radiotherapy. Bio-engineered organoids and extracellular vesicles (EV) are currently under investigation as potential strategies for SG repair. Herein, three-dimensional (3D) cultures of SG functional organoids (SGo) and human dental pulp stem cells (hDPSC) were generated by magnetic 3D bioassembly (M3DB) platforms. Fibroblast growth factor 10 (FGF10) was used to enrich the SGo in secretory epithelial units. After 11 culture days via M3DB, SGo displayed SG-specific acinar epithelial units with functional properties upon neurostimulation. To consistently develop 3D hDPSC in vitro, 3 culture days were sufficient to maintain hDPSC undifferentiated genotype and phenotype for EV generation. EV isolation was performed via sequential centrifugation of the conditioned media of hDPSC and SGo cultures. EV were characterized by nanoparticle tracking analysis, electron microscopy and immunoblotting. EV were in the exosome range for hDPSC (diameter: 88.03 ± 15.60 nm) and for SGo (123.15 ± 63.06 nm). Upon ex vivo administration, exosomes derived from SGo significantly stimulated epithelial growth (up to 60%), mitosis, epithelial progenitors and neuronal growth in injured SG; however, such biological effects were less distinctive with the ones derived from hDPSC. Next, these exosome biological effects were investigated by proteomic arrays. Mass spectrometry profiling of SGo exosomes predicted that cellular growth, development and signaling was due to known and undocumented molecular targets downstream of FGF10. Semaphorins were identified as one of the novel targets requiring further investigations. Thus, M3DB platforms can generate exosomes with potential to ameliorate SG epithelial damage. KeAi Publishing 2022-02-16 /pmc/articles/PMC8961305/ /pubmed/35387159 http://dx.doi.org/10.1016/j.bioactmat.2022.02.007 Text en © 2022 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Chansaenroj, Ajjima
Adine, Christabella
Charoenlappanit, Sawanya
Roytrakul, Sittiruk
Sariya, Ladawan
Osathanon, Thanaphum
Rungarunlert, Sasitorn
Urkasemsin, Ganokon
Chaisuparat, Risa
Yodmuang, Supansa
Souza, Glauco R.
Ferreira, João N.
Magnetic bioassembly platforms towards the generation of extracellular vesicles from human salivary gland functional organoids for epithelial repair
title Magnetic bioassembly platforms towards the generation of extracellular vesicles from human salivary gland functional organoids for epithelial repair
title_full Magnetic bioassembly platforms towards the generation of extracellular vesicles from human salivary gland functional organoids for epithelial repair
title_fullStr Magnetic bioassembly platforms towards the generation of extracellular vesicles from human salivary gland functional organoids for epithelial repair
title_full_unstemmed Magnetic bioassembly platforms towards the generation of extracellular vesicles from human salivary gland functional organoids for epithelial repair
title_short Magnetic bioassembly platforms towards the generation of extracellular vesicles from human salivary gland functional organoids for epithelial repair
title_sort magnetic bioassembly platforms towards the generation of extracellular vesicles from human salivary gland functional organoids for epithelial repair
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8961305/
https://www.ncbi.nlm.nih.gov/pubmed/35387159
http://dx.doi.org/10.1016/j.bioactmat.2022.02.007
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