scholarly journals Mummy Prevents IL-1β-Induced Inflammatory Responses and Cartilage Matrix Degradation via Inhibition of NF-қB Subunits Gene Expression in Pellet Culture System

2018 ◽  
Vol 8 (2) ◽  
pp. 283-289 ◽  
Author(s):  
Fereshteh Morrovati ◽  
Nahid Karimian Fathi ◽  
Jafar Soleimani Rad ◽  
Azadeh Montaseri
Keyword(s):  
Gene Expression ◽  
Culture System ◽  
Inflammatory Responses ◽  
Cartilage Matrix ◽  
Matrix Degradation ◽  
Pellet Culture ◽  
And Cartilage

scholarly journals Sanmiao formula inhibits chondrocyte apoptosis and cartilage matrix degradation in a rat model of osteoarthritis

2014 ◽  
Vol 8 (4) ◽  
pp. 1065-1074 ◽  
Author(s):  
YING XU ◽  
GUO-JING DAI ◽  
QIAN LIU ◽  
ZHEN-LI LIU ◽  
ZHI-QIAN SONG ◽  
...  
Keyword(s):  
Rat Model ◽  
Cartilage Matrix ◽  
Chondrocyte Apoptosis ◽  
Matrix Degradation ◽  
And Cartilage

scholarly journals Acupuncture Delays Cartilage Degeneration through Upregulating SIRT1 Expression in Rats with Osteoarthritis

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Hui Liu ◽  
Tingting Zhang ◽  
Min Liu ◽  
Chunhong Wang ◽  
Jinfeng Yan
Keyword(s):  
Cartilage Degeneration ◽  
Cartilage Matrix ◽  
Enzyme Linked Immunosorbent Assay ◽  
Matrix Degradation ◽  
Matrix Synthesis ◽  
Sirt1 Expression ◽  
Tnf Α ◽  
Related Proteins ◽  
And Cartilage

Silent mating type information regulation 2 homolog 1 (SIRT1) has been reported to inhibit osteoarthritic gene expression in chondrocytes. Here, efforts in this study were made to unveil the specific role of SIRT1 in the therapy of acupuncture on cartilage degeneration in osteoarthritis (OA). Specifically, OA was established by the anterior cruciate ligament transection method in the right knee joint of rats, subsequent to which acupuncture was performed on two acupoints. Injection with shSIRT1 sequence–inserted lentiviruses was conducted to investigate the role of SIRT1 in acupuncture-mediated OA. Morphological changes and cell apoptosis in rat OA cartilages were examined by safranin-O staining and terminal deoxynucleotidyl transferase-mediated nick-end labeling (TUNEL) assay, respectively. The serum levels of tumor necrosis factor (TNF)-α and interleukin (IL)-2 in OA rats were assessed by enzyme-linked immunosorbent assay (ELISA). The expressions of SIRT1, cartilage matrix degradation-related proteins (matrix metalloproteinase (MMP)-9 and ADAMTS5), NF-κB signaling-related markers (p-p65/p65 and p-IκBα/IκBα), and cartilage matrix synthesis-related proteins (collagen II and aggrecan) in the OA cartilage were analyzed by western blot. As a result, acupuncture counteracted OA-associated upregulation of TNF-α, IL-2, cartilage matrix degradation-related proteins, and NF-κB signaling-related markers, morphological damage, apoptosis, SIRT1 downregulation, and loss of cartilage matrix synthesis-related proteins in rat articular cartilages. SIRT1 silencing reversed acupuncture-induced counteractive effects on the aforementioned OA-associated phenomena (except apoptosis, the experiment regarding which under SIRT1 silencing was not performed). Collectively, acupuncture inhibited chondrocyte apoptosis, inflammation, NF-κB signaling activation, and cartilage matrix degradation by upregulating SIRT1 expression to delay OA-associated cartilage degeneration.

329 CHONDROGENIC POTENTIAL OF PORCINE ADIPOSE-DERIVED STEM CELLS, CHONDROCYTES, PERIOSTEAL CELLS, AND FIBROBLASTS IN A PELLET CULTURE SYSTEM

2015 ◽  
Vol 27 (1) ◽  
pp. 253 ◽  
Author(s):  
K. K. Herzog ◽  
D. J. Milner ◽  
S. J. Johnson ◽  
M. B. Wheeler
Keyword(s):  
Alcian Blue ◽  
Culture System ◽  
Negative Control ◽  
Cartilage Matrix ◽  
Pellet Culture ◽  
Chondrogenic Potential ◽  
Periosteal Cell ◽  
Periosteal Cells ◽  
Collagen Ii ◽  
Safranin O

Regenerative medicine has long sought to develop therapies for articular cartilage repair and for enhancing endochondral ossification to address complications of long bone healing. The objective of this study was to determine the chondrogenic potential of porcine primary cell cultures for possible utility in orthopedic tissue engineering applications. Adipose-derived mesenchymal stem cell (ASC), chondrocyte (positive control), periosteal cell, and fibroblast (negative control) primary cell cultures from 8- to 12-month-old Yorkshire pigs were plated at 5000 to 10 000 cells cm–2 in 75-cm2 cell culture flasks using high-glucose Dulbecco's Modified Eagle's Medium (DMEM) supplemented with NaHCO3, 10% fetal bovine serum (FBS), and antimicrobials (penicillin-streptomycin, gentamicin sulfate, and amphotericin B), then incubated at 37°C, 5% CO2, and 18% O2. Cells were trypsinized at ~80% confluency and transferred into 15-mL conical tubes at 500 000 cells tube–1. Suspensions were washed twice by centrifugation in DPBS then condensed via a final centrifugation in 1.0 mL of negative control media (DMEM + 10% FBS) or chondrogenic base media consisting of high-glucose DMEM with 40 µg mL–1 of proline, 50 µM ascorbic acid-2-phosphate, 100 nM dexamethasone, antimicrobials, and 1× insulin-transferrin-selenium solution added. Chondrogenic additives tested were 2% FBS, Kartogenin (200 nM, 400 nM, or 4 µM), 10 ng mL–1 of BMP-4, or a combination of 10 ng mL–1 of BMP-6 + 10 ng mL–1 of TGFβ-3. Pellets were incubated with media changed every 2 to 3 days for a period of 2 to 4 weeks, fixed with 4% paraformaldehyde in DPBS, and then frozen at –80°C in Neg 50 Frozen Section Medium. Eight-micrometer sections were cut using a cryostat onto charged slides. Histochemical staining was performed using hematoxylin and eosin (H&E) for cell morphology and Safranin O and Alcian Blue for cartilage matrix markers. Immunofluorescent staining was done to detect collagen II and aggrecan with the nuclear marker lamin-C used to assess cell viability. Chondrocyte pellets grown in chondrogenic media, regardless of additive, exhibited cartilage matrix formation with H&E and stained strongly positive with Safranin O, Alcian Blue, and collagen II. The ASC pellets grown in chondrogenic media showed mixed cell morphology and areas of early cartilage matrix formation with H&E and stained faintly positive with Safranin O, Alcian Blue, and collagen II at 3 weeks in culture. Periosteal cell pellets had similar morphology in all conditions and did not stain positive for cartilage matrix markers. Fibroblast pellets did not survive any condition for processing. In conclusion, porcine chondrocytes and ASC were able to form cartilage matrix in a pellet culture system with chondrogenic media regardless of additive, while porcine fibroblasts and periosteal cells showed limited to no chondrogenic potential in the conditions tested.

miR-211 Reduces the Degradation of Articular Cartilage Matrix by Targeting Matrix Metalloproteninase 9

2019 ◽  
Vol 9 (9) ◽  
pp. 1292-1297
Author(s):  
Yumei Tian ◽  
Jian Wang
Keyword(s):  
Cartilage Damage ◽  
Cartilage Matrix ◽  
Cartilage Tissue ◽  
Control Group ◽  
Matrix Degradation ◽  
Mankin Score ◽  
Joint Cavity ◽  
Mmp9 Expression ◽  
Col2a1 Expression ◽  
And Cartilage

Matrix metalloproteninase 9 (MMP9) promotes osteoarthritis (OA) cartilage matrix degradation. Abnormal miR-211 expression is associated with OA. Bioinformatics analysis showed a targeted relationship between miR-211 and MMP9 mRNA 3′-UTR. Our study intends to evaluate miR-211’s role in the destruction of chondrocyte matrix in OA model rats. The OA model rats were divided into OA group, OA group+agomir-NC group, OA group+agomir-211 group, followed by analysis of the arthritis Mankin score, Hyp and IL-1β content by ELISA, MMP-9 and COL2A1 expression by western blot and miR-211 level by qRT-PCR. The cartilage tissues were divided into control group, IL-1β+ agomir-NC group, IL-1β+ agomir-211 group, and the expression of MMP-9 and COL2A1 in chondrocytes were detected. Compared with Sham group, IL-1β and Hyp levels in the joint cavity fluid of the OA group were significantly increased and miR-211 expression was significantly decreased with increased MMP9 expression and decreased COL2A1 expression. Injection of agomir-211 into the joint cavity of OA rats significantly reduced MMP9 expression in cartilage tissue, decreased Hyp content in the joint cavity fluid, increased COL2A1 expression, and decreased Mankin score and cartilage damage. IL-1β treatment significantly up-regulated MMP9 expression and decreased COL2A1 expression; miR-211 overexpression attenuated IL-1β-induced cartilage matrix degradation and cartilage destruction. miR-211 plays a role in regulating MMP9 expression and promoting OA. miR-211 overexpression inhibits MMP9 expression, reduces the degradation of cartilage matrix and increases collagen synthesis, thus ameliorating OA development.

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