Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes

B. Shamsul, Shiplu Chowdhury, M. Y. Hamdan, Ruszymah Idrus

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish
Pages (from-to)641-649
Number of pages9
JournalIndian Journal of Medical Research
Volume149
Issue number5
DOIs
Publication statusPublished - 1 May 2019

Fingerprint

Cartilage
Chondrocytes
Fibrin
Cell Count
Tissue
Osteoarthritis
Aggrecans
Histology
Gene encoding
Collagen Type II
Gene expression
Immunochemistry
Gene Expression
RNA
Growth
Fluorescent Antibody Technique
Extracellular Matrix
Proteins
Staining and Labeling

Keywords

  • Cartilage
  • chondrocytes
  • collagen-fibrin
  • osteoarthritis
  • seeding density

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes. / Shamsul, B.; Chowdhury, Shiplu; Hamdan, M. Y.; Idrus, Ruszymah.

In: Indian Journal of Medical Research, Vol. 149, No. 5, 01.05.2019, p. 641-649.

Research output: Contribution to journalArticle

@article{d32d068ad6e84ed19ed2c154519f59ac,
title = "Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes",
abstract = "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.",
keywords = "Cartilage, chondrocytes, collagen-fibrin, osteoarthritis, seeding density",
author = "B. Shamsul and Shiplu Chowdhury and Hamdan, {M. Y.} and Ruszymah Idrus",
year = "2019",
month = "5",
day = "1",
doi = "10.4103/ijmr.IJMR_45_17",
language = "English",
volume = "149",
pages = "641--649",
journal = "Indian Journal of Medical Research",
issn = "0971-5916",
publisher = "Indian Council of Medical Research",
number = "5",

}

TY - JOUR

T1 - Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes

AU - Shamsul, B.

AU - Chowdhury, Shiplu

AU - Hamdan, M. Y.

AU - Idrus, Ruszymah

PY - 2019/5/1

Y1 - 2019/5/1

N2 - 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.

AB - 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.

KW - Cartilage

KW - chondrocytes

KW - collagen-fibrin

KW - osteoarthritis

KW - seeding density

UR - http://www.scopus.com/inward/record.url?scp=85070961481&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85070961481&partnerID=8YFLogxK

U2 - 10.4103/ijmr.IJMR_45_17

DO - 10.4103/ijmr.IJMR_45_17

M3 - Article

C2 - 31417032

AN - SCOPUS:85070961481

VL - 149

SP - 641

EP - 649

JO - Indian Journal of Medical Research

JF - Indian Journal of Medical Research

SN - 0971-5916

IS - 5

ER -