Effect of cell density on formation of three-dimensional cartilaginous constructs using fibrin & human osteoarthritic chondrocytes
| Autor: | Shiplu Roy Chowdhury, M Y Hamdan, B.S. Shamsul, Bt Hj Idrus Ruszymah |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Cartilage Articular 030106 microbiology Cell Cell Culture Techniques chondrocytes lcsh:Medicine Cell Count General Biochemistry Genetics and Molecular Biology Fibrin Chondrocyte Cartilage - chondrocytes - collagen-fibrin - osteoarthritis - seeding density Extracellular matrix 03 medical and health sciences 0302 clinical medicine Osteogenesis medicine Humans 030212 general & internal medicine Aggrecan seeding density Cell Proliferation biology Chemistry Cartilage lcsh:R Histology General Medicine Chondrogenesis Molecular biology osteoarthritis medicine.anatomical_structure Gene Expression Regulation biology.protein RNA Original Article collagen-fibrin |
| Zdroj: | Indian Journal of Medical Research, Vol 149, Iss 5, Pp 641-649 (2019) The Indian Journal of Medical Research |
| ISSN: | 0971-5916 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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