scholarly journals Osthole Inhibits Proliferation and Induces Catabolism in Rat Chondrocytes and Cartilage Tissue

2015 ◽  
Vol 36 (6) ◽  
pp. 2480-2493 ◽  
Author(s):  
Guoqing Du ◽  
Yi Song ◽  
Lei Wei ◽  
Linghui Li ◽  
Xuezong Wang ◽  
...  
Keyword(s):  
Type Ii Collagen ◽  
Cartilage Explants ◽  
Cartilage Tissue ◽  
Blue Staining ◽  
Marker Genes ◽  
Dependent Manner ◽  
Type Ii ◽  
Expression Levels ◽  
Safranine O ◽  
And Cartilage

Background/Aims: Cartilage destruction is thought to be the major mediator of osteoarthritis. Recent studies suggest that inhibition of subchrondral bone loss by anti-osteoporosis (OP) drug can protect cartilige erosion. Osthole, as a promising agent for treating osteoporosis, may show potential in treating osteoarthritis. The purpose of this study was to investigate whether Osthole affects the proliferation and catabolism of rat chondrocytes, and the degeneration of cartilage explants. Methods: Rat chondrocytes were treated with Osthole (0 μM, 6.25 μM, 12.5 μM, and 25 μM) with or without IL1-β (10ng/ml) for 24 hours. The expression levels of type II collagen and MMP13 were detected by western Blot. Marker genes for chondrocytes (A-can and Sox9), matrix metalloproteinases (MMPs), aggrecanases (ADAMTS5) and genes implicated in extracellular matrix catabolism were evaluated by qPCR. Cell proliferation was assessed by measuring proliferating cell nuclear antigen (PCNA) expression and fluorescence activated cell sorter. Wnt7b/β-catenin signaling was also investigated. Cartilage explants from two-week old SD rats were cultured with IL-1β, Osthole and Osthole plus IL-1β for four days and glycosaminoglycan (GAG) synthesis was assessed with toluidine blue staining and Safranine O/Fast Green FCF staining, collagen type II expression was detected by immunofuorescence. Results: Osthole reduced expression of chondrocyte markers and increased expression of MMP13, ADAMTS5 and MMP9 in a dose-dependent manner. Catabolic gene expression levels were further improved by Osthole plus IL-1β. Osthole inhibited chondrocyte proliferation. GAG synthesis and type II collagen were decreased in both the IL-1β groups and the Osthole groups, and significantly reduced by Osthole plus IL-1β. Conclusions: Our data suggested that Osthole increases the catabolism of rat chondrocytes and cartilage explants, this effect might be mediated through inhibiting Wnt7b/β-catenin pathway.

Detection of MMP-13 Activity on Intentionally Strain-Released Type-II Collagen Network in Bovine Articular Cartilage

2011 ◽  
Author(s):  
Nazli Caner ◽  
Jeffrey W. Ruberti
Keyword(s):  
Articular Cartilage ◽  
Degradation Kinetics ◽  
Type Ii Collagen ◽  
Healthy Tissue ◽  
Cartilage Explants ◽  
Cartilage Tissue ◽  
Mechanical Stimuli ◽  
Type Ii ◽  
Collagen Network ◽  
Mechanical Tension

Articular cartilage is a specialized avascular connective tissue found at the contact regions of diarthrodial joints. Cartilage has few cells (< 5% of the volume), though these cells can maintain the balance of turnover in healthy tissue, when the tissue is damaged, they are not able to repair the defects [1–3]. Extra cellular matrix (ECM) in cartilage comprises water, collagen (principally type II), proteoglycans and noncollagenous proteins. The type II collagen network, which is the dominant structural protein in cartilage ECM, constrains the expansion of the resident PGs and is generally held in mechanical tension. In osteoarthritis (OA), the balance of cartilage tissue production/degradation is thought to be affected by abnormal mechanical stimuli leading to net matrix resorption through production of excess degradative enzymes (e.g. matrix metalloproteinases (MMP) and aggrecanases) [4–8]. In OA tissue the amount of MMP-13 is thought to be increased relative to healthy tissue. OA typically occurs in older adults where, as cartilage ages, there is a marked decrease in the fixed charge density (FCD), the hydration and, consequently, mechanical tension on the collagen type II network [9–11]. We have hypothesized that loss of tension on the collagen network accelerates degradation by MMP. Detection of the effect of MMP on loaded, native cartilage could lead to insight about cartilage degradation kinetics in OA. However, it is quite difficult to controllably deliver MMP to cartilage, to activate the MMP during detensioning of the collagen network and to detect the effect on the cartilage mechanics (because cost limits the amount of MMP used). We have developed a transpirational enzyme loading method which is capable of precisely dosing bovine cartilage explants with a small, known quantity of MMP-13. Following enzyme insertion, we are able to detect the activity of the MMP on osmotically compressed cartilage (i.e. cartilage with a detensioned collagen network) via a simple hydration measurement.

scholarly journals The Extract of Fructus Psoraleae Promotes Viability and Cartilaginous Formation of Rat Chondrocytes In Vitro

2016 ◽  
Vol 2016 ◽  
pp. 1-8
Author(s):  
Xin Pan ◽  
Kang Xu ◽  
Xuefeng Qiu ◽  
Wen Zhao ◽  
Dong Wang ◽  
...  
Keyword(s):  
Petroleum Ether Extract ◽  
Cell Aggregation ◽  
Type Ii Collagen ◽  
Cartilage Degeneration ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Type Ii ◽  
Sprague Dawley ◽  
Expression Levels

This study aimed to investigate the extract components of FP on rat chondrocyte function and cartilaginous formation in vitro. Petroleum ether extract (P-e) of FP extract components was selected to treat Sprague-Dawley rat chondrocytes. Cell viability was tested with different concentrations (0.1, 1, 10, and 100 μg/mL) of P-e treatment. Concentrations of 0.1 and 1 μg/mL P-e conditioned culture mediums were used for treating chondrocytes in experiments. Cell proliferation was measured via DNA incorporation assay. Type II collagen, aggrecan, and Sox-9 genes expression levels were measured with RT-PCR. Additionally, cartilaginous formation was analyzed with type II collagen immunofluorescence, H&E, and alcian blue staining. Concentrations of 0.1 and 1 μg/mL P-e showed low cytotoxicity and demonstrated stimulatory effects on chondrocyte proliferation in early stages. Following 6 days of P-e culture, aggrecan and Sox-9 gene expression levels of the 1 μg/mL P-e group were upregulated by 1.82- (p<0.05) and 2.06-fold (p<0.05), respectively, versus controls. Moreover, 1 μg/mL P-e significantly stimulated cell aggregation and type II collagen deposits after 1 week of treatment. Noteworthy, tight cartilaginous structures formed in the 10-day 1 μg/mL P-e conditioned culture. These findings suggest that P-e has the potential to treat cartilage degeneration induced by chondrocyte failure.

Effects of Artesunate on the Expressions of Insulin-Like Growth Factor-1, Osteopontin and C-Telopeptides of Type II Collagen in a Rat Model of Osteoarthritis

Pharmacology ◽  
2017 ◽  
Vol 101 (1-2) ◽  
pp. 1-8 ◽  
Author(s):  
Zhe Bai ◽  
Xiao-Hui Guo ◽  
Chi Tang ◽  
Si-Tong Yue ◽  
Long Shi ◽  
...  
Keyword(s):  
Growth Factor ◽  
Type Ii Collagen ◽  
Cartilage Tissue ◽  
Enzyme Linked Immunosorbent Assay ◽  
Insulin Like Growth Factor ◽  
Type Ii ◽  
Pathological Changes ◽  
Model Group ◽  
Mankin Score ◽  
And Cartilage

Objective: The study aims to explore the effects of artesunate on insulin-like growth factor-1 (IGF-1), Osteopontin (OPN), and C-telopeptides of type II collagen (CTX-II) in serum, synovial fluid (SF), and cartilage tissues of rats with osteoarthritis (OA). Methods: OA models were established. Normal model, artesunate, and Viatril-S groups (20 rats respectively) were set. Enzyme-linked immunosorbent assay, IHC staining, and quantitative real-time polymerase chain reaction were conducted to calculate IGF-1, OPN, and CTX-II levels in serum, SF, and cartilage tissues of rats. The pathological changes in cartilage tissues were evaluated with Mankin score and Hematoxylin-Eosin staining. Results: Compared with the normal group, the model group showed increased IGF-1 level; decreased OPN, CTX-II levels in the serum and SF; and contrary results were seen in the cartilage tissues. A gradual ascending IGF-1 level and descending OPN and CTX-II levels existed in the serum and SF in the artesunate and Viatril-S groups after 2 weeks. The model group showed the most obvious pathological changes and highest Mankin score compared with the other groups. Higher IGF-1 level and lower OPN, CTX-II levels were exhibited in the cartilage tissue in the artesunate and Viatril-S groups but not in the model group. Conclusion: Artesunate and Viatril-S inhibit OA development by elevating IGF-1 level and reducing OPN and CTX-II levels.

The Effect of Crosslinking Agents on the Properties of Type II Collagen Biomaterials

2021 ◽  
Vol 57 (4) ◽  
pp. 166-180
Author(s):  
Maria-Minodora Marin ◽  
Madalina Georgiana Albu Kaya ◽  
George Mihail Vlasceanu ◽  
Jana Ghitman ◽  
Ionut Cristian Radu ◽  
...  
Keyword(s):  
Research Work ◽  
Cartilage Tissue Engineering ◽  
Chemical Properties ◽  
Cartilage Regeneration ◽  
Type Ii Collagen ◽  
Biomechanical Properties ◽  
Cartilage Tissue ◽  
Cross Linking ◽  
Type Ii ◽  
Crosslinking Agents

Type II collagen has been perceived as the indispensable element and plays a crucial role in cartilage tissue engineering. Thus, materials based on type II collagen have drawn farther attention in both academic and research for developing new systems for the cartilage regeneration. The disadvantage of using type II collagen as a biomaterial for tissue repairing is its reduced biomechanical properties. This can be solved by physical, enzymatic or chemical cross-linking processes, which provide biomaterials with the required mechanical properties for medical applications. To enhance type II collagen properties, crosslinked collagen scaffolds with different cross-linking agents were prepared by freeze-drying technique. The present research work studied the synthesis of type II collagen biomaterials with and without crosslinking agents. Scaffolds morphology was observed by MicroCT, showing in all cases an appropriate microstructure for biological applications, and the mechanical studies were performed using compressive tests. DSC showed an increase in denaturation temperature with an increase in cross-linking agent concentration. FTIR suggested that the secondary structure of collagen is not affected after the cross-linking; supplementary, to confirm the characteristic triple-helix conformation of collagen, the CD investigation was performed. The results showed that the physical-chemical properties of type II collagen were improved by cross-linking treatments.

scholarly journals A Novel High Sensitivity Type II Collagen Blood-Based Biomarker, PRO-C2, for Assessment of Cartilage Formation

2018 ◽  
Vol 19 (11) ◽  
pp. 3485 ◽  
Author(s):  
Yunyun Luo ◽  
Yi He ◽  
Ditte Reker ◽  
Natasja Gudmann ◽  
Kim Henriksen ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Growth Factor ◽  
Knee Osteoarthritis ◽  
High Sensitivity ◽  
Type Ii Collagen ◽  
Cartilage Explants ◽  
Type Ii ◽  
Serum Samples ◽  
Human Cartilage ◽  
Cartilage Formation

N-terminal propeptide of type II collagen (PIINP) is a biomarker reflecting cartilage formation. PIINP exists in two main splice variants termed as type IIA and type IIB collagen NH2-propeptide (PIIANP, PIIBNP). PIIANP has been widely recognized as a cartilage formation biomarker. However, the utility of PIIBNP as a marker in preclinical and clinical settings has not been fully investigated yet. In this study, we aimed to characterize an antibody targeting human PIIBNP and to develop an immunoassay assessing type II collagen synthesis in human blood samples. A high sensitivity electrochemiluminescence immunoassay, hsPRO-C2, was developed using a well-characterized antibody against human PIIBNP. Human cartilage explants from replaced osteoarthritis knees were cultured for ten weeks in the presence of growth factors, insulin-like growth factor 1 (IGF-1) or recombinant human fibroblast growth factor 18 (rhFGF-18). The culture medium was changed every seven days, and levels of PIIBNP, PIIANP, and matrix metalloproteinase 9-mediated degradation of type II collagen (C2M) were analyzed herein. Serum samples from a cross-sectional knee osteoarthritis cohort, as well as pediatric and rheumatoid arthritis samples, were assayed for PIIBNP and PIIANP. Western blot showed that the antibody recognized PIIBNP either as a free fragment or attached to the main molecule. Immunohistochemistry demonstrated that PIIBNP was predominately located in the extracellular matrix of the superficial and deep zones and chondrocytes in both normal and osteoarthritic articular cartilage. In addition, the hsPRO-C2 immunoassay exhibits acceptable technical performances. In the human cartilage explants model, levels of PIIBNP, but not PIIANP and C2M, were increased (2 to 7-fold) time-dependently in response to IGF-1. Moreover, there was no significant correlation between PIIBNP and PIIANP levels when measured in knee osteoarthritis, rheumatoid arthritis, and pediatric serum samples. Serum PIIBNP was significantly higher in controls (KL0/1) compared to OA groups (KL2/3/4, p = 0.012). The hsPRO-C2 assay shows completely different biological and clinical patterns than PIIANP ELISA, suggesting that it may be a promising biomarker of cartilage formation.

scholarly journals Paradoxical effects of cyclosporin A on collagen arthritis in rats.

1983 ◽  
Vol 158 (6) ◽  
pp. 2007-2015 ◽  
Author(s):  
N Kaibara ◽  
T Hotokebuchi ◽  
K Takagishi ◽  
I Katsuki
Keyword(s):  
T Cells ◽  
Cyclosporin A ◽  
Type Ii Collagen ◽  
Delayed Type Hypersensitivity ◽  
Induction Phase ◽  
Dependent Manner ◽  
Type Ii ◽  
Skin Reactions ◽  
Collagen Arthritis ◽  
Paradoxical Effects

The effect of the immunosuppressive agent cyclosporin A (CS-A) on collagen arthritis in Sprague-Dawley rats is investigated. A 14-d course of CS-A treatment at doses of 15 mg/kg per day or more, begun on the same day as type II collagen immunization, suppressed the development of arthritis as well as humoral and delayed-type hypersensitivity (DTH) skin test responses to type II collagen, possibly by interfering with helper T cells. Additional studies demonstrated that CS-A treatment only during the induction phase of immunity proved to be successful. When CS-A treatment was started only during the immediately preclinical phase of arthritis or after the disease onset, a significant enhancement of the disease was obtained in a dose-dependent manner. This enhancement was accompanied by an augmentation of DTH skin reactions, while antibody responses were either suppressed or unaffected. These results appear to be attributable at least in part to a suppressive effect of CS-A on a population of suppressor T cells, thus resulting in a T cell-mediated helper effect. It is therefore reasonable to assume that the paradoxical effects of CS-A on collagen arthritis in rats might be caused by an altering of the sensitive balance of the two regulatory subpopulations of T cells. It is also possible that cell-mediated immune responses may play an important role in influencing the course of the disease.

scholarly journals The discrimination of type I and type II collagen and the label-free imaging of engineered cartilage tissue

Biomaterials ◽  
2010 ◽  
Vol 31 (36) ◽  
pp. 9415-9421 ◽  
Author(s):  
Ping-Jung Su ◽  
Wei-Liang Chen ◽  
Tsung-Hsien Li ◽  
Chen-Kuan Chou ◽  
Te-Hsuen Chen ◽  
...  
Keyword(s):  
Type Ii Collagen ◽  
Cartilage Tissue ◽  
Type I ◽  
Type Ii ◽  
Label Free ◽  
Engineered Cartilage

scholarly journals Anisotropic Shape-Memory Alginate Scaffolds Functionalized with Either Type I or Type II Collagen for Cartilage Tissue Engineering

2017 ◽  
Vol 23 (1-2) ◽  
pp. 55-68 ◽  
Author(s):  
Henrique V. Almeida ◽  
Binulal N. Sathy ◽  
Ivan Dudurych ◽  
Conor T. Buckley ◽  
Fergal J. O'Brien ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Shape Memory ◽  
Cartilage Tissue Engineering ◽  
Type Ii Collagen ◽  
Cartilage Tissue ◽  
Type I ◽  
Type Ii

scholarly journals Cartilage Protective and Chondrogenic Capacity of WIN-34B, a New Herbal Agent, in the Collagenase-Induced Osteoarthritis Rabbit Model and in Progenitor Cells from Subchondral Bone

2013 ◽  
Vol 2013 ◽  
pp. 1-12 ◽  
Author(s):  
Jeong-Eun Huh ◽  
Yeon-Cheol Park ◽  
Byung-Kwan Seo ◽  
Jae-Dong Lee ◽  
Yong-Hyeon Baek ◽  
...  
Keyword(s):  
Progenitor Cells ◽  
Cartilage Repair ◽  
Subchondral Bone ◽  
Chondrogenic Differentiation ◽  
Rabbit Model ◽  
Type Ii Collagen ◽  
Blue Staining ◽  
Vehicle Group ◽  
Potential Candidate ◽  
Type Ii

We sought to determine the cartilage repair capacity of WIN-34B in the collagenase-induced osteoarthritis rabbit model and in progenitor cells from subchondral bone. The cartilage protective effect of WIN-34B was measured by clinical and histological scores, cartilage area, and proteoglycan and collagen contents in the collagenase-induced osteoarthritis rabbit model. The efficacy of chondrogenic differentiation of WIN-34B was assessed by expression of CD105, CD73, type II collagen, and aggrecanin vivoand was analyzed by the surface markers of progenitor cells, the mRNA levels of chondrogenic marker genes, and the level of proteoglycan, GAG, and type II collagenin vitro. Oral administration of WIN-34B significantly increased cartilage area, and this was associated with the recovery of proteoglycan and collagen content. Moreover, WIN-34B at 200 mg/kg significantly increased the expression of CD105, CD73, type II collagen, and aggrecan compared to the vehicle group. WIN-34B markedly enhanced the chondrogenic differentiation of CD105 and type II collagen in the progenitor cells from subchondral bone. Also, we confirmed that treatment with WIN-34B strongly increased the number of SH-2(CD105) cells and expression type II collagen in subchondral progenitor cells. Moreover, WIN-34B significantly increased proteoglycan, as measured by alcian blue staining; the mRNA level of type IIα1 collagen, cartilage link protein, and aggrecan; and the inhibition of cartilage matrix molecules, such as GAG and type II collagen, in IL-1β-treated progenitor cells. These findings suggest that WIN-34B could be a potential candidate for effective anti-osteoarthritic therapy with cartilage repair as well as cartilage protection via enhancement of chondrogenic differentiation in the collagenase-induced osteoarthritis rabbit model and progenitor cells from subchondral bone.

Human Cartilage Tissue Engineering with Pluronic and Cultured Chondrocyte Sheet

2007 ◽  
Vol 342-343 ◽  
pp. 89-92 ◽  
Author(s):  
Jae Ho Jeong ◽  
Y.M. Moon ◽  
S.O. Kim ◽  
S.S. Yun ◽  
Hong In Shin
Keyword(s):  
Tissue Engineering ◽  
Cell Sheet ◽  
Cartilage Tissue Engineering ◽  
Type Ii Collagen ◽  
New Method ◽  
Cartilage Tissue ◽  
Type Ii ◽  
Human Cartilage ◽  
Covered Group ◽  
Chondrocyte Sheet

Despite many outstanding research works on cartilage tissue engineering, actual clinical application is not quite successful because of the absorption and progressive distortion of tissue engineered cartilage. We have developed a new method of cartilage tissue engineering comprising chondrocyte mixed Pluronic F-127 and cultured chondrocyte cell sheet which entirely cover the cell-Pluronic complex. We believe the addition of cultured chondrocyte cell sheet enhances the efficacy of chondrogenesis in vivo. Human ear cartilage piece was enzymatically dissociated and chondrocyte suspension was acquired. Chondrocytes were cultured and expanded as the routine manner. Cultured chondrocytes were plated in high-density monolayer and cultured with Chondrogenic media in 5% CO2 incubator. After 3 weeks of culture, chondrocyte cell sheet was formed and complete single sheet of chondrocyte could be harvested by gentle manipulation of culture plate with a cell scraper. Chondrocyte-Pluronic complex was established by mixing 1x 106 cells with 0.5 of Pluronic F- 127. Chondrocyte-Pluronic complex was completely covered with a sheet of cultured chondrocyte. The completed tissue engineered constructs were implanted into the subcutaneous tissue pocket of nude mice on the back. Tissue engineered constructs without cultured cell sheet were used as control. Samples were harvested at 8 weeks postoperatively and they were subjected to histological analysis and assayed for glycosaminoglycan (GAG), and type II collagen. Grossly, the size of cartilage specimen of cultured chondrocyte cell sheet covered group was larger than that of the control. On histologic examination, the specimen of cultured chondrocyte cell sheet covered group showed lacunae-containing cells embedded in a basophilic matrix. The chondrocyte cell sheet covered group specimen resembled mature or immature cartilage. The result of measurement of GAG and type II collagen of cartilage specimen of cultured chondrocyte sheet covered group was higher than that of the control. In conclusion, the new method of cartilage tissue engineering using chondrocyte cell sheet seems to be an effective method providing higher cartilage tissue gain and reliable success rate for cartilage tissue engineering.

Sign in / Sign up

Export Citation Format

Share Document