Morphological changes of bovine nasal chondrocytes induced by interleukin-1α (original) (raw)
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Iranian biomedical journal, 2013
Previous studies have shown that some cytokines have protective effects on cartilage in joint diseases. In the current study, effects of IL-4 against morphological changes and tissue degradation induced by IL-1α on bovine nasal cartilage (BNC) explants were investigated. Fresh BNC samples were prepared from a slaughterhouse under sterile conditions. BNC explants culture was treated with both IL-lα (10 ng/ml) and IL-4 (50 ng/ml) at the same time for 28 days. The morphological characteristics of explants were assessed by using histology techniques and invert microscopy. Matrix metalloproteinase-1 (MMP-1) production was assessed within different days by using Western blotting. IL-lα induced prominent cartilage morphology degradation. The pro and active form of MMP-1 band substantially increased at day 21 of culture. In the presence of both IL-lα and IL-4, chondrocytes preserved their ordinary normal phenotype with intact extracellular matrix. In addition, a significant reduction in pro...
Iranian biomedical journal, 2011
Current treatments for joint diseases are moderately successful, but unfortunately are associated with significant side effects. This study was undertaken to investigate the combination effects of IL-4 and prednisolone on tissue characteristics and production of matrix metalloproteinase-1(MMP-1) in IL-lα-treated bovine nasal cartilage (BNC) explants. BNC explants were cultured in DMEM with IL-lα (10 ng/ml), IL-4 (50 ng/ml) and prednisolone (1 or 1,000 nM) at the same time for 28 days. At days 3, 7, 14, 21 and 28, the media were collected and replaced with fresh media, and the removed media were stored at -20°C. The alterations of tissue characteristics were assessed by using histology techniques. Western-blot method was used to determine the effects of IL-4 and prednisolone combination on MMP-1 production. The cell viability was evaluated by using lactate dehydrogenase assay test. In the presence of IL-lα alone, most chondrocytes were transformed into fibroblast-like morphology with...
Tissue Engineering Part A, 2012
Previous studies showed that human nasal chondrocytes (HNC) exhibit higher proliferation and chondrogenic capacity as compared to human articular chondrocytes (HAC). To consider HNC as a relevant alternative cell source for the repair of articular cartilage defects it is necessary to test how these cells react when exposed to environmental factors typical of an injured joint. We thus aimed this study at investigating the responses of HNC and HAC to exposure to interleukin (IL)-1b and low oxygen. For this purpose HAC and HNC harvested from the same donors (N = 5) were expanded in vitro and then cultured in pellets or collagen-based scaffolds at standard (19%) or low oxygen (5%) conditions. Resulting tissues were analyzed after a short (3 days) exposure to IL-1b, mimicking the initially inflammatory implantation site, or following a recovery time (1 or 2 weeks for pellets and scaffolds, respectively). After IL-1b treatment, constructs generated by both HAC and HNC displayed a transient loss of GAG (up to 21.8% and 36.8%, respectively) and, consistently, an increased production of metalloproteases (MMP)-1 and -13. Collagen type II and the cryptic fragment of aggrecan (DIPEN), both evaluated immunohistochemically, displayed a trend consistent with GAG and MMPs production. HNC-based constructs exhibited a more efficient recovery upon IL-1b withdrawal, resulting in a higher accumulation of GAG (up to 2.6-fold) compared to the corresponding HAC-based tissues. On the other hand, HAC displayed a positive response to low oxygen culture, while HNC were only slightly affected by oxygen percentage. Collectively, under the conditions tested mimicking the postsurgery articular environment, HNC retained a tissueforming capacity, similar or even better than HAC. These results represent a step forward in validating HNC as a cell source for cartilage tissue engineering strategies.
Russian Journal of Transplantology and Artificial Organs, 2019
Revitalization of decellularized or devitalized matrix scaffolds in tracheal tissue engineering typically involves seeding the autologous recipient cells or allogeneic cells under long-term cultivation. Objective: to study the capability of human nasal chondrocytes for colonization of devitalized scaffolds based on native human tracheal cartilage, with proinflammatory stimulation (cytokine) by adding Interleukin-1-beta (IL-1β) to the culture medium. Materials and methods. Scaffolds for tracheal tissue engineering were obtained from native human tracheal cartilage through devitalization and laser etching. The scaffold was revitalized by seeding the human nasal chondrocytes. Histological examination was performed after staining with hematoxylin and safranin-O, with further microscopy using a Nikon Eclipse L200 light microscope. X-ray microtomography was performed on a Phoenix nanotom m apparatus. Electron microscopy was performed on a Nova NanoSEM 230 setup. Results. There was statistically significant increase in the intensity of colonization (p = 0.0008) with nasal chondrocytes and stimulation of their migration activity (p < 0.0001) in the presence of IL-1β compared with the control groups. Conclusion. Addition of proinflammatory cytokine IL-1β (1 μg/ml) to the culture medium enhances volumetric seeding of devitalized cartilage scaffold with human nasal chondrocytes, allowing to create highly revitalized materials for tracheal tissue engineering.
Journal of Clinical Investigation, 1988
In inflammatory diseases such as rheumatoid arthritis, functions of chondrocytes including synthesis of matrix proteins and proteinases are altered through interactions with cells of the infiltrating pannus. One of the major secreted products of mononuclear inflammatory cells is IL-1. In this study we found that recombinant human IL-1f suppressed synthesis of cartilage-specific type II collagen by cultured human costal chondrocytes associated with decreased steady state levels ofal(II) and al(IX) procollagen mRNAs. In contrast, IL-1 increased synthesis of types I and III collagens and levels of al(I), a2(I), and al(III) procollagen mRNAs, as we described previously using human articular chondrocytes and synovial fibroblasts. This stimulatory effect of IL-1 was observed only when IL-1stimulated PGE2 synthesis was blocked by the cyclooxygenase inhibitor indomethacin. The suppression of type II collagen mRNA levels by IL-1 alone was not due to IL-i-stimulated PGE2, since addition of indomethacin did not reverse, but actually potentiated, this inhibition. Continuous exposure of freshly isolated chondrocytes from day 2 of culture to approximately half-maximal concentrations of IL-1 (2.5 pM) completely suppressed levels of type II collagen mRNA and increased levels of types I and III collagen mRNAs, thereby reversing the ratio of al(II)/al(I) procollagen mRNAs from > 6.0 to < 1.0 by day 7. IL-i, therefore, can modify, at a pretranslational level, the relative amounts of the different types of collagen synthesized in cartilage and thereby could be responsible for the inappropriate repair of cartilage matrix in inflammatory conditions.
Optimization method for isolation and culture of chondrocytes in human nasal cartilage tissue
Advances in cartilage tissue engineering were demonstrated cell therapy capabilities in various damages. Due to the increasing use of cell therapy, cells from different sources require to be associated with the desired tissue. Chondrocytes is one of the most widely used cell based repair strategies for cartilage. These cells are components of cartilage tissue and optimization of their isolation protocol with high efficiency and safety properties seems valuable issues. In this study, human nasal biopsy transferred to the laboratory, and was divided into small pieces after being washed two times. Then the pieces were digested with collagenase and incubated for 4 h and then centrifuged and the precipitate obtained by centrifugation was cultured in cell culture medium. In this study, chondrocyte cells obtained with normal morphology and the growth rate after one week in culture was 90%. This study aimed to optimize the human nasal chondrocyte isolation protocol and the results of this study showed that this method could be suitable for isolation of the nasal chondrocytes in therapeutic purposes.
Connective Tissue Research, 2011
The effects of high mobility group box protein (HMGB)-1, interleukin (IL)-1β, and IL-6 on equine articular chondrocytes were investigated, with emphasis on detecting differences between anatomical sites exposed to different loading in vivo, using three-dimensional (3D) cell cultures established with chondrocytes from dorsal radial facet (DRF, highly loaded) and palmar condyle (PC, less loaded) of the third carpal bone (C3). Expression of important genes involved in cartilage metabolism, presence of glycosaminoglycans and cartilage oligomeric matrix protein (COMP) in pellets, and concentrations of matrix metalloproteinase (MMP)-13 and aggrecan epitope CS 846 were evaluated. Compared to controls, IL-1β treatment increased gene expression of versican, matrix-degrading enzymes, and tissue inhibitor of metalloproteinase (TIMP)-1, and decreased aggrecan and collagen type I and type II expression. In addition, IL-1βtreated pellets showed decreased safranin O staining and increased COMP immunostaining and MMP-13 concentrations in culture supernatants. Effects of IL-6 and HMGB-1 on gene expression were variable, although upregulation of Sry-related high-mobility group box 9 (Sox9) was often present and statistically increased in HMGB-1-treated pellets. Response to cytokines rarely differed between DRF and PC pellets. Thus, site-associated cartilage deterioration in equine carpal osteoarthritis (OA) is not explained by topographically different responses to inflammatory mediators. Differences in gene expressions of structural matrix proteins in untreated DRF and PC pellets were noted in the youngest horses, which may indicate differences in the chondrocytes potential to produce matrix in vivo. Overall, a strong catabolic response was induced by IL-1β, whereas slight anabolic effects were induced by IL-6 and HMGB-1.
Rheumatology International, 1995
Inflammatory mediators such as the cytokines interleukin-1 (IL-1) or (TNFα), and prostaglandins [predominantly prostaglandin E2 (PGE2)] are generally considered to be involved in the breakdown of cartilage matrix in chondrodestructive diseases, especially rheumatoid arthritis and osteoarthritis. Their mode of action is not yet completely understood. Blastemal cells or differentiated chondroblasts/chondrocytes of limb buds from mouse embryos (day 12) in organoid cultures provide an efficient system to investigate the mechanism of action of these substances. Using recombinant human IL-1β, TNFα and PGE2 alone or together (in pairs) in this culture system, we found that none of these substances alone could affect chondrogenesis. TNFα, however, when combined with IL-1β, proved to be the more potent cytokine causing a transformation of embryonal chondrogenic cells into fibroblast-like cells and thus inhibiting the expression of the cartilage cell phenotype. This might be due to inhibition of both the morphgenetic and cytodifferentiation phases of chondrogenesis. The well-known synergistic interaction between both cytokines seems to be phase limited and may not occur in the postchondrogenesis phase. In addition, our results showed that TNFα alone or combined with PGE2 caused a marked breakdown of the cartilage matrix. These in vitro findings might be useful to elucidate the complexity of interactions between different cytokines and PGE2 involved in cartilage destruction processes in vivo.