Cargando…

A Combination of Physical and Chemical Treatments Is More Effective in The Preparation of Acellular Uterine Scaffolds

OBJECTIVE: Decellularized uterine scaffold, as a new achievement in tissue engineering, enables recellularization and regeneration of uterine tissues and supports pregnancy in a fashion comparable to the intact uterus. The acellular methods are methods preferred in many respects due to their similar...

Descripción completa

Detalles Bibliográficos
Autores principales: Masoomikarimi, Masoomeh, Salehi, Majid, Noorbakhsh, Farshid, Rajaei, Samira
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Royan Institute 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9868431/
https://www.ncbi.nlm.nih.gov/pubmed/36680481
http://dx.doi.org/10.22074/cellj.2022.8396
_version_ 1784876535858069504
author Masoomikarimi, Masoomeh
Salehi, Majid
Noorbakhsh, Farshid
Rajaei, Samira
author_facet Masoomikarimi, Masoomeh
Salehi, Majid
Noorbakhsh, Farshid
Rajaei, Samira
author_sort Masoomikarimi, Masoomeh
collection PubMed
description OBJECTIVE: Decellularized uterine scaffold, as a new achievement in tissue engineering, enables recellularization and regeneration of uterine tissues and supports pregnancy in a fashion comparable to the intact uterus. The acellular methods are methods preferred in many respects due to their similarity to normal tissue, so it is necessary to try to introduce an acellularization protocol with minimum disadvantages and maximum advantages. Therefore, this study aimed to compare different protocols to achieve the optimal uterus decellularization method for future in vitro and in vivo bioengineering experiments. MATERIALS AND METHODS: In this experimental study, rat uteri were decellularized by four different protocols (P) using sodium dodecyl sulfate (SDS), with different doses and time incubations (P1 and P2), SDS/Triton-X100 sequentially (P3), and a combination of physical (freeze/thaw) and chemical reagents (SDS/Triton X-100). The scaffolds were examined by histopathological staining, DNA quantification, MTT assay, blood compatibility assay, FESEM, and mechanical studies. RESULTS: Histology assessment showed that only in P4, cell residues were completely removed. Masson’s trichrome staining demonstrated that in P3, collagen fibers were decreased; however, no damage was observed in the collagen bundles using other protocols. In indirect MTT assays, cell viabilities achieved by all used protocols were significantly higher than the native samples. The percentage of red blood cell (RBC) hemolysis in the presence of prepared scaffolds from all 4 protocols was less than 2%. The mechanical properties of none of the obtained scaffolds were significantly different from the native sample except for P3. CONCLUSION: Uteri decellularized with a combination of physical and chemical treatments (P4) was the most favorable treatment in our study with the complete removal of cell residue, preservation of the three-dimensional structure, complete removal of detergents, and preservation of the mechanical property of the scaffolds.
format Online
Article
Text
id pubmed-9868431
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Royan Institute
record_format MEDLINE/PubMed
spelling pubmed-98684312023-01-23 A Combination of Physical and Chemical Treatments Is More Effective in The Preparation of Acellular Uterine Scaffolds Masoomikarimi, Masoomeh Salehi, Majid Noorbakhsh, Farshid Rajaei, Samira Cell J Original Article OBJECTIVE: Decellularized uterine scaffold, as a new achievement in tissue engineering, enables recellularization and regeneration of uterine tissues and supports pregnancy in a fashion comparable to the intact uterus. The acellular methods are methods preferred in many respects due to their similarity to normal tissue, so it is necessary to try to introduce an acellularization protocol with minimum disadvantages and maximum advantages. Therefore, this study aimed to compare different protocols to achieve the optimal uterus decellularization method for future in vitro and in vivo bioengineering experiments. MATERIALS AND METHODS: In this experimental study, rat uteri were decellularized by four different protocols (P) using sodium dodecyl sulfate (SDS), with different doses and time incubations (P1 and P2), SDS/Triton-X100 sequentially (P3), and a combination of physical (freeze/thaw) and chemical reagents (SDS/Triton X-100). The scaffolds were examined by histopathological staining, DNA quantification, MTT assay, blood compatibility assay, FESEM, and mechanical studies. RESULTS: Histology assessment showed that only in P4, cell residues were completely removed. Masson’s trichrome staining demonstrated that in P3, collagen fibers were decreased; however, no damage was observed in the collagen bundles using other protocols. In indirect MTT assays, cell viabilities achieved by all used protocols were significantly higher than the native samples. The percentage of red blood cell (RBC) hemolysis in the presence of prepared scaffolds from all 4 protocols was less than 2%. The mechanical properties of none of the obtained scaffolds were significantly different from the native sample except for P3. CONCLUSION: Uteri decellularized with a combination of physical and chemical treatments (P4) was the most favorable treatment in our study with the complete removal of cell residue, preservation of the three-dimensional structure, complete removal of detergents, and preservation of the mechanical property of the scaffolds. Royan Institute 2023-01 2022-12-28 /pmc/articles/PMC9868431/ /pubmed/36680481 http://dx.doi.org/10.22074/cellj.2022.8396 Text en Any use, distribution, reproduction or abstract of this publication in any medium, with the exception of commercial purposes, is permitted provided the original work is properly cited. https://creativecommons.org/licenses/by-nc/3.0/This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial 3.0 (CC BY-NC 3.0) License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Original Article
Masoomikarimi, Masoomeh
Salehi, Majid
Noorbakhsh, Farshid
Rajaei, Samira
A Combination of Physical and Chemical Treatments Is More Effective in The Preparation of Acellular Uterine Scaffolds
title A Combination of Physical and Chemical Treatments Is More Effective in The Preparation of Acellular Uterine Scaffolds
title_full A Combination of Physical and Chemical Treatments Is More Effective in The Preparation of Acellular Uterine Scaffolds
title_fullStr A Combination of Physical and Chemical Treatments Is More Effective in The Preparation of Acellular Uterine Scaffolds
title_full_unstemmed A Combination of Physical and Chemical Treatments Is More Effective in The Preparation of Acellular Uterine Scaffolds
title_short A Combination of Physical and Chemical Treatments Is More Effective in The Preparation of Acellular Uterine Scaffolds
title_sort combination of physical and chemical treatments is more effective in the preparation of acellular uterine scaffolds
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9868431/
https://www.ncbi.nlm.nih.gov/pubmed/36680481
http://dx.doi.org/10.22074/cellj.2022.8396
work_keys_str_mv AT masoomikarimimasoomeh acombinationofphysicalandchemicaltreatmentsismoreeffectiveinthepreparationofacellularuterinescaffolds
AT salehimajid acombinationofphysicalandchemicaltreatmentsismoreeffectiveinthepreparationofacellularuterinescaffolds
AT noorbakhshfarshid acombinationofphysicalandchemicaltreatmentsismoreeffectiveinthepreparationofacellularuterinescaffolds
AT rajaeisamira acombinationofphysicalandchemicaltreatmentsismoreeffectiveinthepreparationofacellularuterinescaffolds
AT masoomikarimimasoomeh combinationofphysicalandchemicaltreatmentsismoreeffectiveinthepreparationofacellularuterinescaffolds
AT salehimajid combinationofphysicalandchemicaltreatmentsismoreeffectiveinthepreparationofacellularuterinescaffolds
AT noorbakhshfarshid combinationofphysicalandchemicaltreatmentsismoreeffectiveinthepreparationofacellularuterinescaffolds
AT rajaeisamira combinationofphysicalandchemicaltreatmentsismoreeffectiveinthepreparationofacellularuterinescaffolds