Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes
Background & objectives: Seeding density is one of the major parameters affecting the quality of tissue-engineered cartilage. The objective of this study was to evaluate different seeding densities of osteoarthritis chondrocytes (OACs) to obtain the highest quality cartilage. Methods: The OACs w...
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Wolters Kluwer Medknow Publications
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doaj-beb4ac21003d4e128d20ee44203aa3312020-11-25T02:30:40ZengWolters Kluwer Medknow PublicationsIndian Journal of Medical Research0971-59162019-01-01149564164910.4103/ijmr.IJMR_45_17Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytesB S ShamsulShiplu Roy ChowdhuryM Y HamdanB. H. I. RuszymahBackground & objectives: Seeding density is one of the major parameters affecting the quality of tissue-engineered cartilage. The objective of this study was to evaluate different seeding densities of osteoarthritis chondrocytes (OACs) to obtain the highest quality cartilage. Methods: The OACs were expanded from passage 0 (P0) to P3, and cells in each passage were analyzed for gross morphology, growth rate, RNA expression and immunochemistry (IHC). The harvested OACs were assigned into two groups: low (1×10[7] cells/ml) and high (3×10[7] cells/ml) cell density. Three-dimensional (3D) constructs for each group were created using polymerised fibrin and cultured for 7, 14 and 21 days in vitro using chondrocyte growth medium. OAC constructs were analyzed with gross assessments and microscopic evaluation using standard histology, IHC and immunofluorescence staining, in addition to gene expression and biochemical analyses to evaluate tissue development. Results: Constructs with a high seeding density of 3×10[7] cells/ml were associated with better quality cartilage-like tissue than those seeded with 1×10[7] cells/ml based on overall tissue formation, cell association and extracellular matrix distribution. The chondrogenic properties of the constructs were further confirmed by the expression of genes encoding aggrecan core protein and collagen type II. Interpretation & conclusions: Our results confirmed that cell density was a significant factor affecting cell behaviour and aggregate production, and this was important for establishing good quality cartilage.http://www.ijmr.org.in/article.asp?issn=0971-5916;year=2019;volume=149;issue=5;spage=641;epage=649;aulast=ShamsulCartilage - chondrocytes - collagen-fibrin - osteoarthritis - seeding density |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
B S Shamsul Shiplu Roy Chowdhury M Y Hamdan B. H. I. Ruszymah |
spellingShingle |
B S Shamsul Shiplu Roy Chowdhury M Y Hamdan B. H. I. Ruszymah Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes Indian Journal of Medical Research Cartilage - chondrocytes - collagen-fibrin - osteoarthritis - seeding density |
author_facet |
B S Shamsul Shiplu Roy Chowdhury M Y Hamdan B. H. I. Ruszymah |
author_sort |
B S Shamsul |
title |
Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes |
title_short |
Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes |
title_full |
Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes |
title_fullStr |
Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes |
title_full_unstemmed |
Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes |
title_sort |
effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes |
publisher |
Wolters Kluwer Medknow Publications |
series |
Indian Journal of Medical Research |
issn |
0971-5916 |
publishDate |
2019-01-01 |
description |
Background & objectives: Seeding density is one of the major parameters affecting the quality of tissue-engineered cartilage. The objective of this study was to evaluate different seeding densities of osteoarthritis chondrocytes (OACs) to obtain the highest quality cartilage.
Methods: The OACs were expanded from passage 0 (P0) to P3, and cells in each passage were analyzed for gross morphology, growth rate, RNA expression and immunochemistry (IHC). The harvested OACs were assigned into two groups: low (1×10[7] cells/ml) and high (3×10[7] cells/ml) cell density. Three-dimensional (3D) constructs for each group were created using polymerised fibrin and cultured for 7, 14 and 21 days in vitro using chondrocyte growth medium. OAC constructs were analyzed with gross assessments and microscopic evaluation using standard histology, IHC and immunofluorescence staining, in addition to gene expression and biochemical analyses to evaluate tissue development.
Results: Constructs with a high seeding density of 3×10[7] cells/ml were associated with better quality cartilage-like tissue than those seeded with 1×10[7] cells/ml based on overall tissue formation, cell association and extracellular matrix distribution. The chondrogenic properties of the constructs were further confirmed by the expression of genes encoding aggrecan core protein and collagen type II.
Interpretation & conclusions: Our results confirmed that cell density was a significant factor affecting cell behaviour and aggregate production, and this was important for establishing good quality cartilage. |
topic |
Cartilage - chondrocytes - collagen-fibrin - osteoarthritis - seeding density |
url |
http://www.ijmr.org.in/article.asp?issn=0971-5916;year=2019;volume=149;issue=5;spage=641;epage=649;aulast=Shamsul |
work_keys_str_mv |
AT bsshamsul effectofcelldensityonformationofthreedimensionalcartilaginousconstructsusingfibrinhumanosteoarthriticchondrocytes AT shipluroychowdhury effectofcelldensityonformationofthreedimensionalcartilaginousconstructsusingfibrinhumanosteoarthriticchondrocytes AT myhamdan effectofcelldensityonformationofthreedimensionalcartilaginousconstructsusingfibrinhumanosteoarthriticchondrocytes AT bhiruszymah effectofcelldensityonformationofthreedimensionalcartilaginousconstructsusingfibrinhumanosteoarthriticchondrocytes |
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