Objective: The aim of this study was to evaluate the growth characteristics of freshly isolated porcine chondrocytes in high-density pellet cultures and to preliminary investigate their use in an interactive in vitro model with synovial fibroblast cell lines to study rheumatoid arthritis (RA).
Design: 1.8x10(6) chondrocytes/cm2 were seeded in 48-multiwell plates. Thickness, cell number and cell distribution in pellet cross sections were documented over a 22-day-long period. Alcian blue staining, type I and type II collagen staining, real-time reverse transcriptase polymerase polymerase chain reaction (RT-PCR) and high performance liquid chromatography (HPLC) were used to characterize cartilage extracellular matrix (ECM) formation, and cell proliferation was demonstrated by Ki67 staining. Furthermore, 2-week-old chondrocyte pellets were co-cultured for additional 2 weeks with two human synovial fibroblast cell lines derived from a normal donor (non-invasive cell line) and a RA patient (invasive-aggressive (IA) cell line), respectively.
Results: Chondrocyte pellets from 11 individual preparations showed a significant increase in pellet thickness from 44+/-19 microm (day 3) to 282+/-19 microm (day 22). Calculation of chondrocyte distribution, cell number and pellet thickness indicated that pellet growth was due to ECM formation and not cell proliferation. This was also confirmed by low numbers of Ki67 positive chondrocytes and absence of cell clusters. HPLC, messenger RNA-analysis, histochemistry and antibody staining verified the expression of ECM components such as type II collagen, whereas type I collagen expression was very low. In contrast to the non-aggressive synovial fibroblast cell line, the IA synovial fibroblast cell line clearly showed cartilage invasion.
Conclusion: Pellet formation of freshly isolated chondrocytes followed a reproducible developmental kinetics and showed typical immature hyaline cartilage properties. Such uniform cartilage pellets are very useful as a substrate for interactive cell culture models that simulate diseases like RA.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.joca.2004.01.009 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
LncRNA-MEG3/miR-93-5p/SMAD7 axis mediates proliferative and inflammatory phenotypes of fibroblast-like synoviocytes in rheumatoid arthritis.
Int J Biol Macromol
January 2025
Department of Pharmacy, The First Affiliated Hospital of Anhui Medical University, Hefei 230022, China; The Grade 3 Pharmaceutical Chemistry Laboratory of State Administration of Traditional Chinese Medicine, Hefei 230022, China. Electronic address:
Synovial hyperplasia, inflammation and immune cell infiltration are the central pathological basis of rheumatoid arthritis (RA). Nonetheless, the cellular, molecular and immunological mechanisms of RA remain poorly understood. An integrated analysis of single-cell RNA (scRNA) and bulk RNA sequencing datasets aimed to unravel the cellular landscape, differentiation trajectory, transcriptome signature, and immunoinfiltration feature of RA synovium.
View Article and Find Full Text PDF[Xinfeng Capsule alleviates RA-FLS-induced angiogenesis in HUVEC cells by inhibiting the lncRNA HOTAIR/PI3K/AKT pathway].
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi
December 2024
Department of Rheumatology, First Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230031, China.
Objective To investigate the effect of serum containing Xinfeng capsule (XFC) on the angiogenesis of human umbilical vein endothelial cells (HUVEC) induced by rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS) and its mechanism of action. Methods An in vitro co-culture model of RA-FLS and HUVEC was established. Serum containing XFC was prepared by oral gavage of SD rats.
View Article and Find Full Text PDFReceptor interacting serine/threonine kinase 2 promotes rheumatoid arthritis progression and partially regulates nuclear factor kappa B pathway.
Cytojournal
November 2024
Department of Hand and Foot Surgery, Qilu Hospital, Cheeloo College of Medicine, Shandong University, Qingdao, China.
Objective: Rheumatoid arthritis (RA) is a disabling systemic autoimmune disease worldwide; however, its molecular pathway remains largely unknown. Thus, this study aimed to explore the effects of receptor-interacting serine/threonine kinase 2 (RIPK2) on RA progression and its underlying mechanism.
Material And Methods: RIPK2 expression was analyzed using real-time quantitative polymerase chain reaction, immunohistochemical staining, and Western blot (WB) analysis in RA synovial tissues or cells.
The role and mechanism of triptolide, a potential new DMARD, in the treatment of rheumatoid arthritis.
Ageing Res Rev
December 2024
The First Clinical Medical College, Nanjing University of Chinese Medicine, No. 138 Xianlin Avenue, Nanjing 210023, China. Electronic address:
Triptolide (TP) is the primary pharmacological component of Tripterygium Glycosides (TG), which has anti-inflammatory, antiproliferative, and immunosuppressive properties, among other pharmacological actions, and has excellent potential for developing into a new DMARD. We have reviewed the effects and mechanisms of TP on immunosuppression, inhibiting synovial proliferation, and preventing articular bone destruction in the treatment of rheumatoid arthritis (RA), which is a common disease in the elderly in this paper. We have found that TP has regulatory effects on multiple vital cells in the above-mentioned pathological process of RA, such as monocytes/macrophages, dendritic cells, T cells, fibroblast-like synoviocytes, and osteoclasts.
View Article and Find Full Text PDFSingle-cell profiling uncovers synovial fibroblast subpopulations associated with chondrocyte injury in osteoarthritis.
Front Endocrinol (Lausanne)
December 2024
Spinal Surgery, Zhejiang Chinese Medical University Affiliated Wenzhou Hospital of Integrated Traditional Chinese and Western Medicine, Wenzhou, Zhejiang, China.
Article Synopsis
- This study focuses on understanding how synovial cells and chondrocytes interact in osteoarthritis (OA), aiming to clarify their roles in OA development.
- Using single-cell sequencing, researchers analyzed the characteristics of synovial fibroblasts and their interactions with chondrocytes, revealing significant correlations between these cell types in both damaged and healthy cartilage.
- The findings indicated that some genes in synovial fibroblasts promote chondrocyte health, while others contribute to chondrocyte degradation and inflammation, highlighting a complex balance that affects OA progression.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!