scholarly journals In Vitro Effects of Low Doses of β-Caryophyllene, Ascorbic Acid and d-Glucosamine on Human Chondrocyte Viability and Inflammation

2021 ◽  
Vol 14 (3) ◽  
pp. 286
Author(s):  
Elena Mattiuzzo ◽  
Alessia Faggian ◽  
Rina Venerando ◽  
Andrea Benetti ◽  
Elisa Belluzzi ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Collagen Type ◽  
Protective Action ◽  
Human Chondrocytes ◽  
Low Doses ◽  
Type Ii ◽  
Collagen Type Ii ◽  
Human Chondrocyte ◽  
Anti Inflammatory

β-caryophyllene (BCP), a plant-derived sesquiterpene, has been reported to have anti-inflammatory and antioxidant effects. The purpose of this study is to evaluate the effects of BCP in combination with ascorbic acid (AA) and d-glucosamine (GlcN) against macrophage-mediated inflammation on in vitro primary human chondrocytes. Changes in cell viability, intracellular ROS generation, gene expression of pro-inflammatory mediators, metalloproteinases (MMPs), collagen type II and aggrecan were analyzed in primary human chondrocytes exposed to the conditioned medium (CM) of activated U937 monocytes and subsequently treated with BCP alone or in combination with AA and GlcN. The CM-induced chondrocyte cytotoxicity was reduced by the presence of low doses of BCP alone or in combination with AA and GlcN. The exposure of cells to CM significantly increased IL-1β, NF-κB1 and MMP-13 expression, but when BCP was added to the inflamed cells, alone or in combination with AA and GlcN, gene transcription for all these molecules was restored to near baseline values. Moreover, chondrocytes increased the expression of collagen type II and aggrecan when stimulated with AA and GlcN alone or in combination with BCP. This study showed the synergistic anti-inflammatory and antioxidative effects of BCP, AA and GlcN at low doses on human chondrocyte cultures treated with the CM of activated U937 cells. Moreover, the combination of the three molecules was able to promote the expression of collagen type II and aggrecan. All together, these data could suggest that BCP, AA and GlcN exert a chondro-protective action.

Phenotypic maintenance of articular chondrocytes in vitro requires BMP activity

2007 ◽  
Vol 20 (03) ◽  
pp. 185-191 ◽  
Author(s):  
A. O. Oshin ◽  
E. Caporali ◽  
C. R. Byron ◽  
A. A. Stewart ◽  
M. C. Stewart
Keyword(s):  
Articular Cartilage ◽  
Collagen Type ◽  
Articular Chondrocytes ◽  
Soluble Form ◽  
Mrna Levels ◽  
Type Ii ◽  
Collagen Type Ii ◽  
Endogenous Expression ◽  
Chondrocytic Phenotype

SummaryArticular chondrocytes are phenotypically unique cells that are responsible for the maintenance of articular cartilage. The articular chondrocytic phenotype is influenced by a range of soluble factors. In particular, members of the bone morphogenetic protein (BMP) family support the articular chondrocytic phenotype and stimulate synthesis of cartilaginous matrix. This study was carried out to determine the importance of BMPs in supporting the differentiated phenotype of articular chondrocytes in vitro. Exogenous BMP-2 supported expression of collagen type II and aggrecan in monolayer chondrocyte cultures, slowing the dedifferentiation process that occurs under these conditions. In contrast, BMP-2 had little effect on expression of these genes in three-dimensional aggregate cultures. Endogenous BMP-2 expression was lost in monolayer cultures, coincident with the down-regulation of collagen type II and aggrecan mRNAs, whereas BMP-2 mRNA levels were stable in aggregate cultures. Antagonism of endogenous BMP activity in aggregate cultures by Noggin or a soluble form of the BMP receptor resulted in reduced expression of collagen type II and aggrecan mRNAs, reduced collagen type II protein and sulfated glycosaminoglycan (GAG) deposition into the aggregate matrices and reduced secretion of GAGs into the culture media. These results indicate that endogenous BMPs are required for maintenance of the differentiated articular chondrocytic phenotype in vitro. These findings are of importance to cell-based strategies designed to repair articular cartilage. Articular chondrocytes require conditions that will support endogenous expression of BMPs to maintain the specialized phenotype of these cells.

scholarly journals Expression of the Heparan Sulfate Proteoglycan, Perlecan, during Mouse Embryogenesis and Perlecan Chondrogenic Activity In Vitro

1999 ◽  
Vol 145 (5) ◽  
pp. 1103-1115 ◽  
Author(s):  
M.M. French ◽  
S.E. Smith ◽  
K. Akanbi ◽  
T. Sanford ◽  
J. Hecht ◽  
...  
Keyword(s):  
Heparan Sulfate ◽  
Collagen Type ◽  
Heparan Sulfate Proteoglycan ◽  
Surrounding Tissue ◽  
Collagen Type Iv ◽  
Type Ii ◽  
Sulfate Proteoglycan ◽  
Collagen Type Ii ◽  
Murine Embryonic Fibroblast

Expression of the basement membrane heparan sulfate proteoglycan (HSPG), perlecan (Pln), mRNA, and protein has been examined during murine development. Both Pln mRNA and protein are highly expressed in cartilaginous regions of developing mouse embryos, but not in areas of membranous bone formation. Initially detected at low levels in precartilaginous areas of d 12.5 embryos, Pln protein accumulates in these regions through d 15.5 at which time high levels are detected in the cartilage primordia. Laminin and collagen type IV, other basal lamina proteins commonly found colocalized with Pln, are absent from the cartilage primordia. Accumulation of Pln mRNA, detected by in situ hybridization, was increased in d 14.5 embryos. Cartilage primordia expression decreased to levels similar to that of the surrounding tissue at d 15.5. Pln accumulation in developing cartilage is preceded by that of collagen type II. To gain insight into Pln function in chondrogenesis, an assay was developed to assess the potential inductive activity of Pln using multipotential 10T1/2 murine embryonic fibroblast cells. Culture on Pln, but not on a variety of other matrices, stimulated extensive formation of dense nodules reminiscent of embryonic cartilaginous condensations. These nodules stained intensely with Alcian blue and collagen type II antibodies. mRNA encoding chondrocyte markers including collagen type II, aggrecan, and Pln was elevated in 10T1/2 cells cultured on Pln. Human chondrocytes that otherwise rapidly dedifferentiate during in vitro culture also formed nodules and expressed high levels of chondrocytic marker proteins when cultured on Pln. Collectively, these studies demonstrate that Pln is not only a marker of chondrogenesis, but also strongly potentiates chondrogenic differentiation in vitro.

scholarly journals Increased TGF-β and BMP Levels and Improved Chondrocyte-Specific Marker Expression In Vitro under Cartilage-Specific Physiological Osmolarity

2019 ◽  
Vol 20 (4) ◽  
pp. 795 ◽  
Author(s):  
Ufuk Tan Timur ◽  
Marjolein Caron ◽  
Guus van den Akker ◽  
Anna van der Windt ◽  
Jenny Visser ◽  
...  
Keyword(s):  
Gene Expression ◽  
Mrna Expression ◽  
Protein Secretion ◽  
Collagen Type ◽  
Articular Chondrocytes ◽  
Human Articular Chondrocytes ◽  
Type Ii ◽  
Collagen Type Ii ◽  
Reporter Assay

During standard expansion culture (i.e., plasma osmolarity, 280 mOsm) human articular chondrocytes dedifferentiate, making them inappropriate for autologous chondrocyte implantation to treat cartilage defects. Increasing the osmolarity of culture media to physiological osmolarity levels of cartilage (i.e., 380 mOsm), increases collagen type II (COL2A1) expression of human articular chondrocytes in vitro, but the underlying molecular mechanism is not fully understood. We hypothesized that TGF-β superfamily signaling may drive expression of COL2A1 under physiological osmolarity culture conditions. Human articular chondrocytes were cultured in cytokine-free medium of 280 or 380 mOsm with or without siRNA mediated TGF-β2 knockdown (RNAi). Expression of TGF-β isoforms, and collagen type II was evaluated by RT-qPCR and immunoblotting. TGF-β2 protein secretion was evaluated using ELISA and TGF-β bioactivity was determined using an established reporter assay. Involvement of BMP signaling was investigated by culturing human articular chondrocytes in the presence or absence of BMP inhibitor dorsomorphin and BMP bioactivity was determined using an established reporter assay. Physiological cartilage osmolarity (i.e., physosmolarity) most prominently increased TGF-β2 mRNA expression and protein secretion as well as TGF-β bioactivity. Upon TGF-β2 isoform-specific knockdown, gene expression of chondrocyte marker COL2A1 was induced. TGF-β2 RNAi under physosmolarity enhanced TGF-β bioactivity. BMP bioactivity increased upon physosmotic treatment, but was not related to TGF-β2 RNAi. In contrast, dorsomorphin inhibited COL2A1 mRNA expression in human articular chondrocytes independent of the osmotic condition. Our data suggest a role for TGF-β superfamily member signaling in physosmolarity-induced mRNA expression of collagen type II. As physosmotic conditions favor the expression of COL2A1 independent of our manipulations, contribution of other metabolic, post-transcriptional or epigenetic factors cannot be excluded in the underlying complex and interdependent regulation of marker gene expression. Dissecting these molecular mechanisms holds potential to further improve future cell-based chondral repair strategies.

scholarly journals Δ8(14)-Ergostenol Glycoside Derivatives Inhibit the Expression of Inflammatory Mediators and Matrix Metalloproteinase

Molecules ◽  
2021 ◽  
Vol 26 (15) ◽  
pp. 4547
Author(s):  
Hyejin Moon ◽  
Myoungsil Ko ◽  
Yujin Park ◽  
Jeonguk Kim ◽  
Dowon Yoon ◽  
...  
Keyword(s):  
Side Effects ◽  
Matrix Metalloproteinase ◽  
Inflammatory Mediators ◽  
Inflammatory Disease ◽  
Collagen Type ◽  
Chronic Inflammatory Disease ◽  
Human Chondrocytes ◽  
Type Ii ◽  
Collagen Type Ii ◽  
Tnf Α

Arthritis is a chronic inflammatory disease accompanied by pathological reactions such as swelling, redness, fever, and pain in various joint areas. The drugs currently available to treat arthritis are associated with diverse side-effects. Therefore, there is a need for safer and more effective treatments to alleviate the inflammation of arthritis with fewer side-effects. In this study, a new sterol, Δ8(14)-ergostenol, was discovered, and its glycosides were synthesized and found to be more efficient in terms of synthesis or anti-inflammatory activity than either spinasterol or 5,6-dihydroergosterol is. Among these synthetic glycosides, galactosyl ergostenol inhibited the expression of inflammatory mediators in TNF-α-stimulated FLS and TNF-α-induced MMPs and collagen type II A1 degradation in human chondrocytes. These results suggest the new galactosyl ergostenol as a treatment candidate for arthritis.

Hypoxia promotes nucleus pulposus phenotype in 3D scaffolds in vitro and in vivo

2014 ◽  
Vol 21 (2) ◽  
pp. 303-309 ◽  
Author(s):  
Ganjun Feng ◽  
Li Li ◽  
Ying Hong ◽  
Hao Liu ◽  
Yueming Song ◽  
...  
Keyword(s):  
Nucleus Pulposus ◽  
Collagen Type ◽  
Type Ii ◽  
Collagen Type Ii ◽  
Low Oxygen ◽  
Nucleus Pulposus Cells ◽  
Experimental Conditions ◽  
3D Scaffolds

Object The role of oxygen in disc metabolism remains a matter of debate. Whether the effect of hypoxic priming on the nucleus pulposus phenotype can be maintained in vivo is not clear. The goal of the present study was to test the hypothesis that priming in a low oxygen tension in vitro could promote a nucleus pulposus phenotype in vivo. Methods Bovine nucleus pulposus cells were seeded in 3D scaffolds and subjected to varying oxygen tensions (2% and 20%) for 3 weeks. The constructs were then implanted subcutaneously for 8 weeks. Changes in the extracellular matrix were evaluated using quantitative real-time reverse transcriptase polymerase chain reaction, glycosaminoglycan (GAG) assay, DNA assay, collagen quantification, and histological and immunohistological analyses. Results Hypoxia resulted in greater production of sulfated glycosaminoglycan and higher levels of gene expression for collagen Type II, aggrecan, and SOX-9. Furthermore, after hypoxic priming, the subcutaneously implanted constructs maintained the nucleus pulposus phenotype, which was indicated by a significantly higher amount of glycosaminoglycan and collagen Type II. Conclusions Hypoxia enhanced the nucleus pulposus phenotype under experimental conditions both in vitro and in vivo. When used in combination with appropriate scaffold material, nucleus pulposus cells could be regenerated for tissue-engineering applications.

Effect of Cell Sheet Manipulation Techniques on the Expression of Collagen Type II and Stress Fiber Formation in Human Chondrocyte Sheets

2018 ◽  
Vol 24 (5-6) ◽  
pp. 469-478 ◽  
Author(s):  
Sopita Wongin ◽  
Saranatra Waikakul ◽  
Pojchong Chotiyarnwong ◽  
Wanwipa Siriwatwechakul ◽  
Kwanchanok Viravaidya-Pasuwat
Keyword(s):  
Collagen Type ◽  
Cell Sheet ◽  
Stress Fiber ◽  
Fiber Formation ◽  
Type Ii ◽  
Collagen Type Ii ◽  
Human Chondrocyte ◽  
Stress Fiber Formation

scholarly journals Effect of Decellularized Cartilage Bovine Scaffold and Hypoxic Condition on Stem Cell Differentiation to Chondrocyte: An In Vitro Study

2018 ◽  
Vol 35 ◽  
pp. 67-76
Author(s):  
Ferdiansyah Mahyudin ◽  
Dwikora Novembri Utomo ◽  
Tri Wahyu Martanto ◽  
Aries Rakhmat Hidayat ◽  
Linta Meyla Putri
Keyword(s):  
Tissue Engineering ◽  
Stem Cell ◽  
Cell Differentiation ◽  
Growth Factor ◽  
Collagen Type ◽  
Hypoxic Condition ◽  
Stem Cell Differentiation ◽  
Type Ii ◽  
Collagen Type Ii

Autologous Chondrocyte Implantation (ACI) has been established for years to treat cartilage defect. Application of tissue engineering has advantages over ACI as tissue engineering requires simpler procedures without leaving morbidity at the donor site. Decellularized bovine cartilage scaffold and hypoxic stem cell differentiation were used in this in vitro experimental study. Comparative test was done between three study groups using bone marrow mesenchymal stem cells treated in three different conditions: growth factor-rich chondrogenic medium, scaffold without growth factor, and combination of both. Each group was given two oxygen tension conditions of normoxia and hypoxic within phase of stem cell differentiation. Immunohistochemical examinations on SOX9, RUNX2, and collagen type II were done for evaluation. After 5-week treatment, the result showed that the highest expression SOX9 and collagen type II were found within the group that used the combination of both scaffold and chondrogenic medium in hypoxic condition. Collagen type II expression in scaffold without additional growth factor showed no statistically significant difference compared with the combination group in hypoxic condition. Cartilage tissue engineering has proven its effectiveness for cartilage regeneration. Decellularized biomaterial scaffold limited the use of growth factor resulting in better cost and resource efficiency.

Sign in / Sign up

Export Citation Format

Share Document