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Skin wound healing improvement in diabetic mice through FTIR microspectroscopy after implanting pluripotent stem cells

Diabetes is a chronic degenerative disease that carries multiple complications. One of the most important complications is the diabetic cutaneous complications, such as skin lesions, ulcerations, and diabetic foot, which are present in 30%–70% of the patients. Currently, the treatments for wound hea...

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Autores principales: Vazquez-Zapien, Gustavo J., Martinez-Cuazitl, Adriana, Granados-Jimenez, Alejandra, Sanchez-Brito, Miguel, Guerrero-Ruiz, Melissa, Camacho-Ibarra, Alejandro, Miranda-Ruiz, Misael A., Dox-Aguillón, Ian S., Ramirez-Torres, Jesus A., Mata-Miranda, Monica M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AIP Publishing LLC 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9908300/
https://www.ncbi.nlm.nih.gov/pubmed/36779176
http://dx.doi.org/10.1063/5.0130383
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author Vazquez-Zapien, Gustavo J.
Martinez-Cuazitl, Adriana
Granados-Jimenez, Alejandra
Sanchez-Brito, Miguel
Guerrero-Ruiz, Melissa
Camacho-Ibarra, Alejandro
Miranda-Ruiz, Misael A.
Dox-Aguillón, Ian S.
Ramirez-Torres, Jesus A.
Mata-Miranda, Monica M.
author_facet Vazquez-Zapien, Gustavo J.
Martinez-Cuazitl, Adriana
Granados-Jimenez, Alejandra
Sanchez-Brito, Miguel
Guerrero-Ruiz, Melissa
Camacho-Ibarra, Alejandro
Miranda-Ruiz, Misael A.
Dox-Aguillón, Ian S.
Ramirez-Torres, Jesus A.
Mata-Miranda, Monica M.
author_sort Vazquez-Zapien, Gustavo J.
collection PubMed
description Diabetes is a chronic degenerative disease that carries multiple complications. One of the most important complications is the diabetic cutaneous complications, such as skin lesions, ulcerations, and diabetic foot, which are present in 30%–70% of the patients. Currently, the treatments for wound healing include growth factors and cytokines, skin substitutes, hyperbaric oxygen therapy, and skin grafts. However, these treatments are ineffective due to the complex mechanisms involved in developing unhealed wounds. Considering the aforementioned complications, regenerative medicine has focused on this pathology using stem cells to improve these complications. However, it is essential to mention that there is a poor biomolecular understanding of diabetic skin and the effects of treating it with stem cells. For this reason, herein, we investigated the employment of pluripotent stem cells (PSC) in the wound healing process by carrying out morphometric, histological, and Fourier-transform infrared microspectroscopy (FTIRM) analysis. The morphometric analysis was done through a photographic follow-up, measuring the lesion areas. For the histological analysis, hematoxylin & eosin and picrosirius red stains were used to examine the thickness of the epidermis and the cellularity index in the dermis as well as the content and arrangement of collagen type I and III fibers. Finally, for the FTIRM analysis, skin cryosections were obtained and analyzed by employing a Cassegrain objective of 16× of an FTIR microscope coupled to an FTIR spectrometer. For this purpose, 20 mice were divided into two groups according to the treatment they received: the Isotonic Salt Solution (ISS) group and the PSCs group (n = 10). Both groups were induced to diabetes, and six days after diabetes induction, an excisional lesion was made in the dorsal area. Furthermore, using microscopy and FTIRM analysis, the skin healing process on days 7 and 15 post-skin lesion excision was examined. The results showed that the wound healing process over time, considering the lesion size, was similar in both groups; however, the PSCs group evidenced hair follicles in the wound. Moreover, the histological analysis evidenced that the PSCs group exhibited granulation tissue, new vessels, and better polarity of the keratinocytes. In addition, the amount of collagen increased with a good deposition and orientation, highlighting that type III collagen fibers were more abundant in the PSCs. Finally, the FTIR analysis evidenced that the PSCs group exhibited a faster wound healing process. In conclusion, the wounds treated with PSCs showed a more rapid wound healing process, less inflammatory cellular infiltration, and more ordered structures than the ISS group.
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spelling pubmed-99083002023-02-09 Skin wound healing improvement in diabetic mice through FTIR microspectroscopy after implanting pluripotent stem cells Vazquez-Zapien, Gustavo J. Martinez-Cuazitl, Adriana Granados-Jimenez, Alejandra Sanchez-Brito, Miguel Guerrero-Ruiz, Melissa Camacho-Ibarra, Alejandro Miranda-Ruiz, Misael A. Dox-Aguillón, Ian S. Ramirez-Torres, Jesus A. Mata-Miranda, Monica M. APL Bioeng Articles Diabetes is a chronic degenerative disease that carries multiple complications. One of the most important complications is the diabetic cutaneous complications, such as skin lesions, ulcerations, and diabetic foot, which are present in 30%–70% of the patients. Currently, the treatments for wound healing include growth factors and cytokines, skin substitutes, hyperbaric oxygen therapy, and skin grafts. However, these treatments are ineffective due to the complex mechanisms involved in developing unhealed wounds. Considering the aforementioned complications, regenerative medicine has focused on this pathology using stem cells to improve these complications. However, it is essential to mention that there is a poor biomolecular understanding of diabetic skin and the effects of treating it with stem cells. For this reason, herein, we investigated the employment of pluripotent stem cells (PSC) in the wound healing process by carrying out morphometric, histological, and Fourier-transform infrared microspectroscopy (FTIRM) analysis. The morphometric analysis was done through a photographic follow-up, measuring the lesion areas. For the histological analysis, hematoxylin & eosin and picrosirius red stains were used to examine the thickness of the epidermis and the cellularity index in the dermis as well as the content and arrangement of collagen type I and III fibers. Finally, for the FTIRM analysis, skin cryosections were obtained and analyzed by employing a Cassegrain objective of 16× of an FTIR microscope coupled to an FTIR spectrometer. For this purpose, 20 mice were divided into two groups according to the treatment they received: the Isotonic Salt Solution (ISS) group and the PSCs group (n = 10). Both groups were induced to diabetes, and six days after diabetes induction, an excisional lesion was made in the dorsal area. Furthermore, using microscopy and FTIRM analysis, the skin healing process on days 7 and 15 post-skin lesion excision was examined. The results showed that the wound healing process over time, considering the lesion size, was similar in both groups; however, the PSCs group evidenced hair follicles in the wound. Moreover, the histological analysis evidenced that the PSCs group exhibited granulation tissue, new vessels, and better polarity of the keratinocytes. In addition, the amount of collagen increased with a good deposition and orientation, highlighting that type III collagen fibers were more abundant in the PSCs. Finally, the FTIR analysis evidenced that the PSCs group exhibited a faster wound healing process. In conclusion, the wounds treated with PSCs showed a more rapid wound healing process, less inflammatory cellular infiltration, and more ordered structures than the ISS group. AIP Publishing LLC 2023-02-07 /pmc/articles/PMC9908300/ /pubmed/36779176 http://dx.doi.org/10.1063/5.0130383 Text en © 2023 Author(s). https://creativecommons.org/licenses/by/4.0/All article content, except where otherwise noted, is licensed under a Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Articles
Vazquez-Zapien, Gustavo J.
Martinez-Cuazitl, Adriana
Granados-Jimenez, Alejandra
Sanchez-Brito, Miguel
Guerrero-Ruiz, Melissa
Camacho-Ibarra, Alejandro
Miranda-Ruiz, Misael A.
Dox-Aguillón, Ian S.
Ramirez-Torres, Jesus A.
Mata-Miranda, Monica M.
Skin wound healing improvement in diabetic mice through FTIR microspectroscopy after implanting pluripotent stem cells
title Skin wound healing improvement in diabetic mice through FTIR microspectroscopy after implanting pluripotent stem cells
title_full Skin wound healing improvement in diabetic mice through FTIR microspectroscopy after implanting pluripotent stem cells
title_fullStr Skin wound healing improvement in diabetic mice through FTIR microspectroscopy after implanting pluripotent stem cells
title_full_unstemmed Skin wound healing improvement in diabetic mice through FTIR microspectroscopy after implanting pluripotent stem cells
title_short Skin wound healing improvement in diabetic mice through FTIR microspectroscopy after implanting pluripotent stem cells
title_sort skin wound healing improvement in diabetic mice through ftir microspectroscopy after implanting pluripotent stem cells
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9908300/
https://www.ncbi.nlm.nih.gov/pubmed/36779176
http://dx.doi.org/10.1063/5.0130383
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