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Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes

BACKGROUND: Articular cartilage (AC) injuries and malformations are commonly noticed because of trauma or age-related degeneration. Many methods have been adopted for replacing or repairing the damaged tissue. Currently available AC repair methods, in several cases, fail to yield good-quality long-l...

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Autores principales: Nanduri, Vibudha, Tattikota, Surendra Mohan, T., Avinash Raj, Sriramagiri, Vijaya Rama Rao, Kantipudi, Suma, Pande, Gopal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2014
Materias:
45
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4555603/
https://www.ncbi.nlm.nih.gov/pubmed/26535340
http://dx.doi.org/10.1177/2325967114539122
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author Nanduri, Vibudha
Tattikota, Surendra Mohan
T., Avinash Raj
Sriramagiri, Vijaya Rama Rao
Kantipudi, Suma
Pande, Gopal
author_facet Nanduri, Vibudha
Tattikota, Surendra Mohan
T., Avinash Raj
Sriramagiri, Vijaya Rama Rao
Kantipudi, Suma
Pande, Gopal
author_sort Nanduri, Vibudha
collection PubMed
description BACKGROUND: Articular cartilage (AC) injuries and malformations are commonly noticed because of trauma or age-related degeneration. Many methods have been adopted for replacing or repairing the damaged tissue. Currently available AC repair methods, in several cases, fail to yield good-quality long-lasting results, perhaps because the reconstructed tissue lacks the cellular and matrix properties seen in hyaline cartilage (HC). PURPOSE: To reconstruct HC tissue from 2-dimensional (2D) and 3-dimensional (3D) cultures of AC-derived human chondrocytes that would specifically exhibit the cellular and biochemical properties of the deep layer of HC. STUDY DESIGN: Descriptive laboratory study. METHODS: Two-dimensional cultures of human AC–derived chondrocytes were established in classical medium (CM) and newly defined medium (NDM) and maintained for a period of 6 weeks. These cells were suspended in 2 mm–thick collagen I gels, placed in 24-well culture inserts, and further cultured up to 30 days. Properties of chondrocytes, grown in 2D cultures and the reconstructed 3D cartilage tissue, were studied by optical and scanning electron microscopic techniques, immunohistochemistry, and cartilage-specific gene expression profiling by reverse transcription polymerase chain reaction and were compared with those of the deep layer of native human AC. RESULTS: Two-dimensional chondrocyte cultures grown in NDM, in comparison with those grown in CM, showed more chondrocyte-specific gene activity and matrix properties. The NDM-grown chondrocytes in 3D cultures also showed better reproduction of deep layer properties of HC, as confirmed by microscopic and gene expression analysis. The method used in this study can yield cartilage tissue up to approximately 1.6 cm in diameter and 2 mm in thickness that satisfies the very low cell density and matrix composition properties present in the deep layer of normal HC. CONCLUSION: This study presents a novel and reproducible method for long-term culture of AC-derived chondrocytes and reconstruction of cartilage tissue with properties similar to the deep layer of HC in vitro. CLINICAL RELEVANCE: The HC tissue obtained by the method described can be used to develop an implantable product for the replacement of damaged or malformed AC, especially in younger patients where the lesions are caused by trauma or mechanical stress.
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spelling pubmed-45556032015-11-03 Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes Nanduri, Vibudha Tattikota, Surendra Mohan T., Avinash Raj Sriramagiri, Vijaya Rama Rao Kantipudi, Suma Pande, Gopal Orthop J Sports Med 45 BACKGROUND: Articular cartilage (AC) injuries and malformations are commonly noticed because of trauma or age-related degeneration. Many methods have been adopted for replacing or repairing the damaged tissue. Currently available AC repair methods, in several cases, fail to yield good-quality long-lasting results, perhaps because the reconstructed tissue lacks the cellular and matrix properties seen in hyaline cartilage (HC). PURPOSE: To reconstruct HC tissue from 2-dimensional (2D) and 3-dimensional (3D) cultures of AC-derived human chondrocytes that would specifically exhibit the cellular and biochemical properties of the deep layer of HC. STUDY DESIGN: Descriptive laboratory study. METHODS: Two-dimensional cultures of human AC–derived chondrocytes were established in classical medium (CM) and newly defined medium (NDM) and maintained for a period of 6 weeks. These cells were suspended in 2 mm–thick collagen I gels, placed in 24-well culture inserts, and further cultured up to 30 days. Properties of chondrocytes, grown in 2D cultures and the reconstructed 3D cartilage tissue, were studied by optical and scanning electron microscopic techniques, immunohistochemistry, and cartilage-specific gene expression profiling by reverse transcription polymerase chain reaction and were compared with those of the deep layer of native human AC. RESULTS: Two-dimensional chondrocyte cultures grown in NDM, in comparison with those grown in CM, showed more chondrocyte-specific gene activity and matrix properties. The NDM-grown chondrocytes in 3D cultures also showed better reproduction of deep layer properties of HC, as confirmed by microscopic and gene expression analysis. The method used in this study can yield cartilage tissue up to approximately 1.6 cm in diameter and 2 mm in thickness that satisfies the very low cell density and matrix composition properties present in the deep layer of normal HC. CONCLUSION: This study presents a novel and reproducible method for long-term culture of AC-derived chondrocytes and reconstruction of cartilage tissue with properties similar to the deep layer of HC in vitro. CLINICAL RELEVANCE: The HC tissue obtained by the method described can be used to develop an implantable product for the replacement of damaged or malformed AC, especially in younger patients where the lesions are caused by trauma or mechanical stress. SAGE Publications 2014-06-27 /pmc/articles/PMC4555603/ /pubmed/26535340 http://dx.doi.org/10.1177/2325967114539122 Text en © The Author(s) 2014 http://creativecommons.org/licenses/by-nc-nd/3.0/ This article is distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License (http://www.creativecommons.org/licenses/by-nc-nd/3.0/) which permits non-commercial use, reproduction and distribution of the work as published without adaptation or alteration, without further permission provided the original work is attributed as specified on the SAGE and Open Access page (http://www.uk.sagepub.com/aboutus/openaccess.htm).
spellingShingle 45
Nanduri, Vibudha
Tattikota, Surendra Mohan
T., Avinash Raj
Sriramagiri, Vijaya Rama Rao
Kantipudi, Suma
Pande, Gopal
Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes
title Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes
title_full Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes
title_fullStr Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes
title_full_unstemmed Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes
title_short Reconstruction of Hyaline Cartilage Deep Layer Properties in 3-Dimensional Cultures of Human Articular Chondrocytes
title_sort reconstruction of hyaline cartilage deep layer properties in 3-dimensional cultures of human articular chondrocytes
topic 45
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4555603/
https://www.ncbi.nlm.nih.gov/pubmed/26535340
http://dx.doi.org/10.1177/2325967114539122
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