Cargando…

Synergistic effect of chitosan-alginate composite hydrogel enriched with ascorbic acid and alpha-tocopherol under hypoxic conditions on the behavior of mesenchymal stem cells for wound healing

BACKGROUND: In regenerative medicine, especially skin tissue engineering, the focus is on enhancing the quality of wound healing. Also, several constructs with different regeneration potentials have been used for skin tissue engineering. In this study, the regenerative properties of chitosan-alginat...

Descripción completa

Detalles Bibliográficos
Autores principales: Ghahremani-nasab, Maryam, Akbari‑Gharalari, Naeimeh, Rahmani Del Bakhshayesh, Azizeh, Ghotaslou, Armita, Ebrahimi-kalan, Abbas, Mahdipour, Mahdi, Mehdipour, Ahmad
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10642036/
https://www.ncbi.nlm.nih.gov/pubmed/37953287
http://dx.doi.org/10.1186/s13287-023-03567-2
_version_ 1785146879527354368
author Ghahremani-nasab, Maryam
Akbari‑Gharalari, Naeimeh
Rahmani Del Bakhshayesh, Azizeh
Ghotaslou, Armita
Ebrahimi-kalan, Abbas
Mahdipour, Mahdi
Mehdipour, Ahmad
author_facet Ghahremani-nasab, Maryam
Akbari‑Gharalari, Naeimeh
Rahmani Del Bakhshayesh, Azizeh
Ghotaslou, Armita
Ebrahimi-kalan, Abbas
Mahdipour, Mahdi
Mehdipour, Ahmad
author_sort Ghahremani-nasab, Maryam
collection PubMed
description BACKGROUND: In regenerative medicine, especially skin tissue engineering, the focus is on enhancing the quality of wound healing. Also, several constructs with different regeneration potentials have been used for skin tissue engineering. In this study, the regenerative properties of chitosan-alginate composite hydrogels in skin wound healing under normoxic and hypoxic conditions were investigated in vitro. METHODS: The ionic gelation method was used to prepare chitosan/alginate (CA) hydrogel containing CA microparticles and bioactive agents [ascorbic acid (AA) and α-tocopherol (TP)]. After preparing composite hydrogels loaded with AA and TP, the physicochemical properties such as porosity, pore size, swelling, weight loss, wettability, drug release, and functional groups were analyzed. Also, the hemo-biocompatibility of composite hydrogels was evaluated by a hemolysis test. Then, the rat bone marrow mesenchymal stem cells (rMSCs) were seeded onto the hydrogels after characterization by flow cytometry. The survival rate was analyzed using MTT assay test. The hydrogels were also investigated by DAPI and H&E staining to monitor cell proliferation and viability. To induce hypoxia, the cells were exposed to CoCl(2). To evaluate the regenerative potential of rMSCs cultured on CA/AA/TP hydrogels under hypoxic conditions, the expression of the main genes involved in the healing of skin wounds, including HIF-1α, VEGF-A, and TGF-β1, was investigated by real-time PCR. RESULTS: The results demonstrated that the prepared composite hydrogels were highly porous, with interconnected pores that ranged in sizes from 20 to 188 μm. The evaluation of weight loss showed that the prepared hydrogels have the ability to biodegrade according to the goals of wound healing. The reduction percentage of CA/AA/TP mass in 21 days was reported as 21.09 ± 0.52%. Also, based on wettability and hemolysis tests of the CA/AA/TP, hydrophilicity (θ = 55.6° and 53.7°) and hemocompatibility with a hemolysis ratio of 1.36 ± 0.19 were evident for them. Besides, MTT assay, DAPI, and H&E staining also showed that the prepared hydrogels provide a suitable substrate for cell growth and proliferation. Finally, based on real-time PCR, increased expression levels of VEGF and TGF-β1 were observed in rMSCs in hypoxic conditions cultured on the prepared hydrogels. CONCLUSIONS: In conclusion, this study provides evidence that 3D CA/AA/TP composite hydrogels seeded by rMSCs in hypoxic conditions have great potential to improve wound healing. GRAPHICAL ABSTRACT: [Image: see text]
format Online
Article
Text
id pubmed-10642036
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-106420362023-11-14 Synergistic effect of chitosan-alginate composite hydrogel enriched with ascorbic acid and alpha-tocopherol under hypoxic conditions on the behavior of mesenchymal stem cells for wound healing Ghahremani-nasab, Maryam Akbari‑Gharalari, Naeimeh Rahmani Del Bakhshayesh, Azizeh Ghotaslou, Armita Ebrahimi-kalan, Abbas Mahdipour, Mahdi Mehdipour, Ahmad Stem Cell Res Ther Research BACKGROUND: In regenerative medicine, especially skin tissue engineering, the focus is on enhancing the quality of wound healing. Also, several constructs with different regeneration potentials have been used for skin tissue engineering. In this study, the regenerative properties of chitosan-alginate composite hydrogels in skin wound healing under normoxic and hypoxic conditions were investigated in vitro. METHODS: The ionic gelation method was used to prepare chitosan/alginate (CA) hydrogel containing CA microparticles and bioactive agents [ascorbic acid (AA) and α-tocopherol (TP)]. After preparing composite hydrogels loaded with AA and TP, the physicochemical properties such as porosity, pore size, swelling, weight loss, wettability, drug release, and functional groups were analyzed. Also, the hemo-biocompatibility of composite hydrogels was evaluated by a hemolysis test. Then, the rat bone marrow mesenchymal stem cells (rMSCs) were seeded onto the hydrogels after characterization by flow cytometry. The survival rate was analyzed using MTT assay test. The hydrogels were also investigated by DAPI and H&E staining to monitor cell proliferation and viability. To induce hypoxia, the cells were exposed to CoCl(2). To evaluate the regenerative potential of rMSCs cultured on CA/AA/TP hydrogels under hypoxic conditions, the expression of the main genes involved in the healing of skin wounds, including HIF-1α, VEGF-A, and TGF-β1, was investigated by real-time PCR. RESULTS: The results demonstrated that the prepared composite hydrogels were highly porous, with interconnected pores that ranged in sizes from 20 to 188 μm. The evaluation of weight loss showed that the prepared hydrogels have the ability to biodegrade according to the goals of wound healing. The reduction percentage of CA/AA/TP mass in 21 days was reported as 21.09 ± 0.52%. Also, based on wettability and hemolysis tests of the CA/AA/TP, hydrophilicity (θ = 55.6° and 53.7°) and hemocompatibility with a hemolysis ratio of 1.36 ± 0.19 were evident for them. Besides, MTT assay, DAPI, and H&E staining also showed that the prepared hydrogels provide a suitable substrate for cell growth and proliferation. Finally, based on real-time PCR, increased expression levels of VEGF and TGF-β1 were observed in rMSCs in hypoxic conditions cultured on the prepared hydrogels. CONCLUSIONS: In conclusion, this study provides evidence that 3D CA/AA/TP composite hydrogels seeded by rMSCs in hypoxic conditions have great potential to improve wound healing. GRAPHICAL ABSTRACT: [Image: see text] BioMed Central 2023-11-13 /pmc/articles/PMC10642036/ /pubmed/37953287 http://dx.doi.org/10.1186/s13287-023-03567-2 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Ghahremani-nasab, Maryam
Akbari‑Gharalari, Naeimeh
Rahmani Del Bakhshayesh, Azizeh
Ghotaslou, Armita
Ebrahimi-kalan, Abbas
Mahdipour, Mahdi
Mehdipour, Ahmad
Synergistic effect of chitosan-alginate composite hydrogel enriched with ascorbic acid and alpha-tocopherol under hypoxic conditions on the behavior of mesenchymal stem cells for wound healing
title Synergistic effect of chitosan-alginate composite hydrogel enriched with ascorbic acid and alpha-tocopherol under hypoxic conditions on the behavior of mesenchymal stem cells for wound healing
title_full Synergistic effect of chitosan-alginate composite hydrogel enriched with ascorbic acid and alpha-tocopherol under hypoxic conditions on the behavior of mesenchymal stem cells for wound healing
title_fullStr Synergistic effect of chitosan-alginate composite hydrogel enriched with ascorbic acid and alpha-tocopherol under hypoxic conditions on the behavior of mesenchymal stem cells for wound healing
title_full_unstemmed Synergistic effect of chitosan-alginate composite hydrogel enriched with ascorbic acid and alpha-tocopherol under hypoxic conditions on the behavior of mesenchymal stem cells for wound healing
title_short Synergistic effect of chitosan-alginate composite hydrogel enriched with ascorbic acid and alpha-tocopherol under hypoxic conditions on the behavior of mesenchymal stem cells for wound healing
title_sort synergistic effect of chitosan-alginate composite hydrogel enriched with ascorbic acid and alpha-tocopherol under hypoxic conditions on the behavior of mesenchymal stem cells for wound healing
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10642036/
https://www.ncbi.nlm.nih.gov/pubmed/37953287
http://dx.doi.org/10.1186/s13287-023-03567-2
work_keys_str_mv AT ghahremaninasabmaryam synergisticeffectofchitosanalginatecompositehydrogelenrichedwithascorbicacidandalphatocopherolunderhypoxicconditionsonthebehaviorofmesenchymalstemcellsforwoundhealing
AT akbarigharalarinaeimeh synergisticeffectofchitosanalginatecompositehydrogelenrichedwithascorbicacidandalphatocopherolunderhypoxicconditionsonthebehaviorofmesenchymalstemcellsforwoundhealing
AT rahmanidelbakhshayeshazizeh synergisticeffectofchitosanalginatecompositehydrogelenrichedwithascorbicacidandalphatocopherolunderhypoxicconditionsonthebehaviorofmesenchymalstemcellsforwoundhealing
AT ghotaslouarmita synergisticeffectofchitosanalginatecompositehydrogelenrichedwithascorbicacidandalphatocopherolunderhypoxicconditionsonthebehaviorofmesenchymalstemcellsforwoundhealing
AT ebrahimikalanabbas synergisticeffectofchitosanalginatecompositehydrogelenrichedwithascorbicacidandalphatocopherolunderhypoxicconditionsonthebehaviorofmesenchymalstemcellsforwoundhealing
AT mahdipourmahdi synergisticeffectofchitosanalginatecompositehydrogelenrichedwithascorbicacidandalphatocopherolunderhypoxicconditionsonthebehaviorofmesenchymalstemcellsforwoundhealing
AT mehdipourahmad synergisticeffectofchitosanalginatecompositehydrogelenrichedwithascorbicacidandalphatocopherolunderhypoxicconditionsonthebehaviorofmesenchymalstemcellsforwoundhealing