Biocompatible and functional properties of a microdispersed tissue-specific 3D matrix from decellularized porcine cartilage

In contrast to decellularization of soft tissues for use as tissue-specific matrices in the creation of tissue-engineered constructs, decellularization of cartilage tissue requires several processing techniques, which can negatively affect the biocompatibility and functional properties of the native...

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Main Authors: E. A. Nemets, A. E. Lazhko, A. M. Grigoriev, Yu. B. Basok, A. D. Kirillova, V. I. Sevastianov
Format: Article
Language:Russian
Published: Federal Research Center of Transplantology and Artificial Organs named after V.I.Shumakov 2022-12-01
Series:Вестник трансплантологии и искусственных органов
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Online Access:https://journal.transpl.ru/vtio/article/view/1553
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author E. A. Nemets
A. E. Lazhko
A. M. Grigoriev
Yu. B. Basok
A. D. Kirillova
V. I. Sevastianov
author_facet E. A. Nemets
A. E. Lazhko
A. M. Grigoriev
Yu. B. Basok
A. D. Kirillova
V. I. Sevastianov
author_sort E. A. Nemets
collection DOAJ
description In contrast to decellularization of soft tissues for use as tissue-specific matrices in the creation of tissue-engineered constructs, decellularization of cartilage tissue requires several processing techniques, which can negatively affect the biocompatibility and functional properties of the native extracellular matrix (ECM).Objective: to study the biocompatible and functional properties of microdispersed tissue-specific 3D matrix from a porcine cartilage that is decellularized by sequential use of chemical, physical and enzymatic techniques.Materials and methods. For decellularization, microdispersed cartilage particles (MCPs), obtained by cryomilling, were incubated in detergent solutions (sodium dodecyl sulfate and Triton X-100), then treated with supercritical carbon dioxide (scCO2) with 10% ethanol and DNase I. The Ames test (Salmonella typhimurium reverse mutation assay) was used to determine the genotoxicity of decellularized microdispersed cartilage particles (dMCPs). Local and general toxic effects, as well as resorption of dMCPs were studied in vivo on sexually mature outbred rats. Decellularized MCP specimens (10 mg) were implanted into the thigh muscle tissue. Viability of human adipose-derived mesenchymal stem/stromal cells (hAdMSCs), when cultured on dMCPs, was analyzed by in vivo microscopy, stained with fluorescent Calcein AM dye. Cell metabolic activity was assessed using PrestoBlue™ Cell Viability Reagent.Results. It has been proven that porcine dMCPs implanted in rat muscle after treatment with scCO2 do not exhibit local and general toxic effects, and do not show genotoxicity and negative effects on the reproductive system of animals. After 6 months of in vivo experiment, most (87%) of the implanted decellularized cartilage was resorbed. It was shown that the resulting matrices are able to support adhesion and proliferation of hAdMSCs. Conclusion. Porcine dMCP specimens are suitable for biocompatible medical products in terms of local and general toxic effects, genotoxicity and reproductive toxicity, and can be used as a matrix for creating cell- and tissue-engineered cartilage constructs.
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institution Kabale University
issn 1995-1191
language Russian
publishDate 2022-12-01
publisher Federal Research Center of Transplantology and Artificial Organs named after V.I.Shumakov
record_format Article
series Вестник трансплантологии и искусственных органов
spelling doaj-art-b2c83101bbd1467fa7985dff80bb072f2025-08-20T03:44:23ZrusFederal Research Center of Transplantology and Artificial Organs named after V.I.ShumakovВестник трансплантологии и искусственных органов1995-11912022-12-01244738410.15825/1995-1191-2022-4-73-841130Biocompatible and functional properties of a microdispersed tissue-specific 3D matrix from decellularized porcine cartilageE. A. Nemets0A. E. Lazhko1A. M. Grigoriev2Yu. B. Basok3A. D. Kirillova4V. I. Sevastianov5Shumakov National Medical Research Center of Transplantology and Artificial OrgansMoscow State UniversityShumakov National Medical Research Center of Transplantology and Artificial OrgansShumakov National Medical Research Center of Transplantology and Artificial OrgansShumakov National Medical Research Center of Transplantology and Artificial OrgansShumakov National Medical Research Center of Transplantology and Artificial Organs; Institute of Biomedical Research and TechnologyIn contrast to decellularization of soft tissues for use as tissue-specific matrices in the creation of tissue-engineered constructs, decellularization of cartilage tissue requires several processing techniques, which can negatively affect the biocompatibility and functional properties of the native extracellular matrix (ECM).Objective: to study the biocompatible and functional properties of microdispersed tissue-specific 3D matrix from a porcine cartilage that is decellularized by sequential use of chemical, physical and enzymatic techniques.Materials and methods. For decellularization, microdispersed cartilage particles (MCPs), obtained by cryomilling, were incubated in detergent solutions (sodium dodecyl sulfate and Triton X-100), then treated with supercritical carbon dioxide (scCO2) with 10% ethanol and DNase I. The Ames test (Salmonella typhimurium reverse mutation assay) was used to determine the genotoxicity of decellularized microdispersed cartilage particles (dMCPs). Local and general toxic effects, as well as resorption of dMCPs were studied in vivo on sexually mature outbred rats. Decellularized MCP specimens (10 mg) were implanted into the thigh muscle tissue. Viability of human adipose-derived mesenchymal stem/stromal cells (hAdMSCs), when cultured on dMCPs, was analyzed by in vivo microscopy, stained with fluorescent Calcein AM dye. Cell metabolic activity was assessed using PrestoBlue™ Cell Viability Reagent.Results. It has been proven that porcine dMCPs implanted in rat muscle after treatment with scCO2 do not exhibit local and general toxic effects, and do not show genotoxicity and negative effects on the reproductive system of animals. After 6 months of in vivo experiment, most (87%) of the implanted decellularized cartilage was resorbed. It was shown that the resulting matrices are able to support adhesion and proliferation of hAdMSCs. Conclusion. Porcine dMCP specimens are suitable for biocompatible medical products in terms of local and general toxic effects, genotoxicity and reproductive toxicity, and can be used as a matrix for creating cell- and tissue-engineered cartilage constructs.https://journal.transpl.ru/vtio/article/view/1553articular cartilagedecellularization3d matrixresorptionbiocompatibilityadipose-derived mscsadhesionproliferationtissue-engineered construct
spellingShingle E. A. Nemets
A. E. Lazhko
A. M. Grigoriev
Yu. B. Basok
A. D. Kirillova
V. I. Sevastianov
Biocompatible and functional properties of a microdispersed tissue-specific 3D matrix from decellularized porcine cartilage
Вестник трансплантологии и искусственных органов
articular cartilage
decellularization
3d matrix
resorption
biocompatibility
adipose-derived mscs
adhesion
proliferation
tissue-engineered construct
title Biocompatible and functional properties of a microdispersed tissue-specific 3D matrix from decellularized porcine cartilage
title_full Biocompatible and functional properties of a microdispersed tissue-specific 3D matrix from decellularized porcine cartilage
title_fullStr Biocompatible and functional properties of a microdispersed tissue-specific 3D matrix from decellularized porcine cartilage
title_full_unstemmed Biocompatible and functional properties of a microdispersed tissue-specific 3D matrix from decellularized porcine cartilage
title_short Biocompatible and functional properties of a microdispersed tissue-specific 3D matrix from decellularized porcine cartilage
title_sort biocompatible and functional properties of a microdispersed tissue specific 3d matrix from decellularized porcine cartilage
topic articular cartilage
decellularization
3d matrix
resorption
biocompatibility
adipose-derived mscs
adhesion
proliferation
tissue-engineered construct
url https://journal.transpl.ru/vtio/article/view/1553
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AT amgrigoriev biocompatibleandfunctionalpropertiesofamicrodispersedtissuespecific3dmatrixfromdecellularizedporcinecartilage
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