scholarly journals OPN Deficiency Increases the Severity of Osteoarthritis Associated with Aberrant Chondrocyte Senescence and Apoptosis and Upregulates the Expression of Osteoarthritis-Associated Genes

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Jian Tian ◽  
Chao Cheng ◽  
Shi-Da Kuang ◽  
Chao Su ◽  
Xin Zhao ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Synovial Fluid ◽  
Joint Damage ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Knee Oa ◽  
Normal Chondrocytes ◽  
Apoptosis Assay ◽  
Oa Chondrocytes

Objectives. A recent work has reported that the elevated osteopontin (OPN) levels in the articular cartilage and synovial fluid are correlated with the progressive osteoarthritis (OA) joint damage, and OPN has a protective effect against OA by suppressing the expressions of OA-associated genes. The present study examined whether the OPN deficiency was susceptible to OA through the regulation of chondrocyte senescence and apoptosis and the expressions of OA-associated genes. Methods. The mRNA levels of COL2A1 and OPN were compared between human OA chondrocytes and normal chondrocytes. The effects of OPN siRNA on the SA-β-Gal expressions and the percentage of apoptotic chondrocytes were examined by using SA-β-Gal staining and apoptosis assay, and the effects on the expressions of COL2A1 and OA-associated genes (COL10A1, IL-1β, TNF-ɑ, MMP-13, and ADAMTS5) were examined by western blot analysis and quantitative real-time RT-PCR. Furthermore, an in vivo OA model was established to examine the effects of OPN siRNA on the senescence and apoptosis of OA chondrocytes and the expressions of OA-associated genes. Results. The mRNA levels of COL2A1 and OPN were decreased in knee OA chondrocytes in comparison with those in normal chondrocytes. The OPN deficiency enhanced the senescence and apoptosis of OA chondrocytes and increased the expressions of COL10A1, IL-1β, TNF-ɑ, MMP-13, and ADAMTS5 but decreased the expression of COL2A1. Meanwhile, OPN deficiency could result in severe, accelerated OA in vivo, which was also associated with enhanced senescence and apoptosis of chondrocytes and elevated expressions of OA-associated genes. Conclusions. The findings of this study suggest that the OPN deficiency can result in accelerated OA, which is associated with enhanced senescence and apoptosis of OA chondrocytes and the upregulated expressions of OA-associated genes.

scholarly journals Quantitative assessment of CYP11B1 and CYP11B2 expression in aldosterone-producing adenomas

2002 ◽  
pp. 795-802 ◽  
Author(s):  
F Fallo ◽  
V Pezzi ◽  
L Barzon ◽  
P Mulatero ◽  
F Veglio ◽  
...  
Keyword(s):  
Real Time ◽  
Secretory Activity ◽  
Zona Glomerulosa ◽  
Zona Fasciculata ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Pathophysiological Role ◽  
Aldosterone Production

BACKGROUND: The presence and pathophysiological role of CYP11B1 (11beta-hydroxylase) gene in the zona glomerulosa of human adrenal cortex is still controversial. METHODS: In order to specifically quantify CYP11B1, CYP11B2 (aldosterone synthase) and CYP17(17alpha-hydroxylase) mRNA levels, we developed a real-time RT-PCR assay and examined the expression in a series of adrenal tIssues, including six normal adrenals from patients adrenalectomized for renal cancer and twelve aldosterone-producing adenomas (APA) from patients with primary aldosteronism. RESULTS: CYP11B1 mRNA levels were clearly detected in normal adrenals, which comprised both zona glomerulosa and fasciculata/reticularis cells, but were also measured at a lower range (P<0.05) in APA. The levels of CYP11B2 mRNA were lower (P<0.005) in normal adrenals than in APA. CYP17 mRNAlevels were similar in normal adrenals and in APA. In patients with APA, CYP11B2 and CYP11B1 mRNA levels were not correlated either with basal aldosterone or with the change from basal aldosterone in response to posture or to dexamethasone. No correlation between CYP11B1 mRNA or CYP11B2 mRNA and the percentage of zona fasciculata-like cells was observed in APA. CONCLUSIONS: Real-time RT-PCR can be reliably used to quantify CYP11B1 and CYP11B2 mRNA levels in adrenal tIssues. Expression of CYP11B1 in hyperfunctioning zona glomerulosa suggests an additional formation of corticosterone via 11beta-hydroxylase, providing further substrate for aldosterone biosynthesis. CYP11B1 and CYP11B2 mRNA levels in APA are not related to the in vivo secretory activity of glomerulosa cells, where post-transcriptional factors might ultimately regulate aldosterone production.

scholarly journals Xenogenic Implantation of Equine Synovial Fluid-Derived Mesenchymal Stem Cells Leads to Articular Cartilage Regeneration

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Mohammed Zayed ◽  
Steven Newby ◽  
Nabil Misk ◽  
Robert Donnell ◽  
Madhu Dhar
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Articular Cartilage ◽  
Synovial Fluid ◽  
Cartilage Repair ◽  
Cartilage Regeneration ◽  
Large Animal

Horses are widely used as large animal preclinical models for cartilage repair studies, and hence, there is an interest in using equine synovial fluid-derived mesenchymal stem cells (SFMSCs) in research and clinical applications. Since, we have previously reported that similar to bone marrow-derived MSCs (BMMSCs), SFMSCs may also exhibit donor-to-donor variations in their stem cell properties; the current study was carried out as a proof-of-concept study, to compare the in vivo potential of equine BMMSCs and SFMSCs in articular cartilage repair. MSCs from these two sources were isolated from the same equine donor. In vitro analyses confirmed a significant increase in COMP expression in SFMSCs at day 14. The cells were then encapsulated in neutral agarose scaffold constructs and were implanted into two mm diameter full-thickness articular cartilage defect in trochlear grooves of the rat femur. MSCs were fluorescently labeled, and one week after treatment, the knee joints were evaluated for the presence of MSCs to the injured site and at 12 weeks were evaluated macroscopically, histologically, and then by immunofluorescence for healing of the defect. The macroscopic and histological evaluations showed better healing of the articular cartilage in the MSCs’ treated knee than in the control. Interestingly, SFMSC-treated knees showed a significantly higher Col II expression, suggesting the presence of hyaline cartilage in the healed defect. Data suggests that equine SFMSCs may be a viable option for treating osteochondral defects; however, their stem cell properties require prior testing before application.

Feasibility and Efficacy of Mobilization of BCR/ABL- PBSC in CML Patients Receiving Imatinib.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2858-2858
Author(s):  
Ravi Bhatia ◽  
Helen Xu ◽  
David Snyder ◽  
Tinisha McDonald ◽  
Marilyn Slovak ◽  
...  
Keyword(s):  
Stem Cells ◽  
Median Time ◽  
Autologous Transplantation ◽  
Disease Stage ◽  
Imatinib Treatment ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Target Number ◽  
Cd34 Cells

Abstract Although imatinib is highly effective in inducing remissions in CML patients, the long-term durability of response is not clear. Here we report updated results of a clinical trial investigating the feasibility and efficacy of collection and storage of PBSC from patients in complete cytogenetic remission (CCR) on imatinib, for use in autologous transplantation in the event of subsequent relapse. PBSC were collected from 36 patients [31 CP, 5 AP (at start of imatinib); median age 45 years (range 22–70); 21 males, 15 females; median time from diagnosis 25 months (mos) (6–90); median duration of imatinib treatment 13 mos (6–41); median time from CCR 7 mos (1–26)]. Patients were administered G-CSF (10μg/kg/day), and PBSC collection initiated on day +5 with a targeted minimum of 2x106 CD34+ cells/kg. Imatinib was continued during G-CSF administration and PBSC collection. The G-CSF dose was escalated in case of poor collection. The median number of CD34+ cells (106/kg) collected was 2.56 (0.31–6.19) with a median of 3 phereses (1–13). Five patients failed to collect the target number of CD34+ cells, achieving a median of 0.87x106 CD34+ cells after a median 5 collections. Seven patients required &gt;6 collections to reach the target cell dose. There was no significant relationship between rapid (≤3) or slow (&gt;3) collection, or failure to collect, and clinical characteristics such as age, sex, disease stage, prior interferon, time from diagnosis, time on imatinib, and duration of CCR. PBSC collections were evaluated for BCR/ABL contamination by cytogenetics and PCR (Q-PCR and nested RT-PCR). Ph+ cells were detected on cytogenetic examination in 1 or more collections from 5 patients and Ph- abnormal clones were detected in 4 patients. Patients with Ph+ PBSC were slower collectors than those with Ph- PBSC (median 8 vs. 3 collections, p=0.04). BCR/ABL mRNA was detected by PCR in 1 or more collection from 30 of 32 patients evaluated. Two patients, both with BCR/ABL mRNA detected in pre-mobilzation marrow, collected with a single PCR negative collection. Three patients had collections with low levels of BCR/ABL mRNA detected only with more sensitive nested RT-PCR. Of 134 separate collections analyzed, BCR/ABL mRNA was detected by Q-PCR in 113 (84%), by nested RT-PCR in 11 (8%), while 10 (8%) were PCR negative. Rapid collectors had significantly lower BCR/ABL mRNA levels in their collections compared to pre-mobilization marrow (p&lt;0.05), whereas slow collectors did not show significant change in BCR/ABL levels. CD34+ cells isolated from PBSC showed significantly increased BCR/ABL mRNA levels compared with total nucleated cells (BCR/ABL:B2M ratio of 0.0006±0.0002 for NC vs. 0.03±0.01 for CD34+ cells, p=0.002). PBSC were injected into NOD/SCID mice to evaluate for presence of BCR/ABL+ progenitors capable of in vivo engraftment. Human cell engraftment was confirmed by flow cytometry in 3 of 4 patients, and BCR/ABL mRNA was detected in engrafted cells by Q-PCR. Our results indicate that cytogenetically negative PBSC collections can be obtained from CML patients receiving imatinib, but that mobilization is relatively poor. Rapid collectors have reduced BCR/ABL+ cell contamination in PBSC collections, and PCR negative collections are possible. However, the majority of PBSC products show evidence of persistent malignant stem cells. Additional strategies to enhance reliability and rapidity of collection and further deplete BCR/ABL+ stem cells in the PBSC product need to be explored.

Endothelin-1 concentrations are correlated with the severity of knee osteoarthritis

2016 ◽  
Vol 64 (4) ◽  
pp. 872-874 ◽  
Author(s):  
Zhe Zhao ◽  
Enqi Li ◽  
Qing Cao ◽  
Jie Sun ◽  
Baotong Ma
Keyword(s):  
Articular Cartilage ◽  
Synovial Fluid ◽  
Knee Osteoarthritis ◽  
Elevated Serum ◽  
Cartilage Destruction ◽  
Case Group ◽  
Endothelin 1 ◽  
Knee Oa ◽  
Potent Vasoconstrictor

Endothelin-1, a potent vasoconstrictor regulator, contributes to articular cartilage destruction. Therefore, we aim to assess the correlation of endothelin-1 concentrations with the development and severity of knee osteoarthritis (OA). This study included a population of 209 patients with knee OA. Kellgren-Lawrence (KL) grading was utilized to score the severity of OA. The case group had higher serum endothelin-1 concentrations than controls. Patients with knee OA with a relatively higher grade showed significantly elevated serum and synovial fluid (SF) endothelin-1 concentrations compared with those with lower KL grades. A significant correlation was found between serum and SF endothelin-1 concentrations and KL grades. Serum and SF endothelin-1 concentrations are correlated with the development and progression of knee OA.

MUSCLE-INDUCED PATELLOFEMORAL JOINT LOADING RAPIDLY AFFECTS CARTILAGE mRNA LEVELS IN A SITE SPECIFIC MANNER

2004 ◽  
Vol 08 (01) ◽  
pp. 1-12 ◽  
Author(s):  
Andrea L. Clark ◽  
Linda Mills ◽  
David A Hart ◽  
Walter Herzog
Keyword(s):  
Articular Cartilage ◽  
Mechanical Loading ◽  
Patellofemoral Joint ◽  
Mrna Level ◽  
Recovery Period ◽  
Cartilage Explants ◽  
Mrna Levels ◽  
Biological Responses

Mechanical loading of articular cartilage affects the synthesis and degradation of matrix macromolecules. Much of the work in this area has involved mechanical loading of articular cartilage explants or cells in vitro and assessing biological responses at the mRNA and protein levels. In this study, we developed a new experimental technique to load an intact patellofemoral joint in vivo using muscle stimulation. The articular cartilages were cyclically loaded for one hour in a repeatable and measurable manner. Cartilage was harvested from central and peripheral regions of the femoral groove and patella, either immediately after loading or after a three hour recovery period. Total RNA was isolated from the articular cartilage and biological responses were assessed on the mRNA level using the reverse transcriptase-polymerase chain reaction. Articular cartilage from intact patellofemoral joints demonstrated heterogeneity at the mRNA level for six of the genes assessed independent of the loading protocol. Cyclical loading of cartilage in its native environment led to alterations in mRNA levels for a subset of molecules when assessed immediately after the loading period. However, the increases in TIMP-1 and decreases in bFGF mRNA levels were transient; being present immediately after load application but not after a three hour recovery period.

scholarly journals Expression of glucose-dependent insulinotropic polypeptide in the zebrafish

2009 ◽  
Vol 297 (6) ◽  
pp. R1803-R1812 ◽  
Author(s):  
Michelle C. Musson ◽  
Lisa I. Jepeal ◽  
Patrick D. Mabray ◽  
Irina V. Zhdanova ◽  
Wellington V. Cardoso ◽  
...  
Keyword(s):  
Early Stage ◽  
Model Organism ◽  
Receptor Activation ◽  
Evolutionary Development ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Glucagon Receptor ◽  
Nutrient Deposition ◽  
Zebrafish Larvae

In mammals, glucose-dependent insulinotropic polypeptide (GIP) is synthesized predominately in the small intestine and functions in conjunction with insulin to promote nutrient deposition. However, little is known regarding GIP expression and function in early vertebrates like the zebrafish, a model organism representing an early stage in the evolutionary development of the compound vertebrate pancreas. Analysis of GIP and insulin ( insa) expression in zebrafish larvae by RT-PCR demonstrated that although insa was detected as early as 24 h postfertilization (hpf), GIP expression was not demonstrated until 72 hpf, shortly after the completion of endocrine pancreatic development but prior to the commencement of independent feeding. Furthermore, whole mount in situ hybridization of zebrafish larvae showed expression of GIP and insa in the same tissues, and in adult zebrafish, RT-PCR and immunohistochemistry demonstrated GIP expression in both the intestine and the pancreas. Receptor activation studies showed that zebrafish GIP was capable of activating the rat GIP receptor. Although previous studies have identified four receptors with glucagon receptor-like sequences in the zebrafish, one of which possesses the capacity to bind GIP, a functional analysis of these receptors has not been performed. This study demonstrates interactions between the latter receptor and zebrafish GIP, identifying it as a potential in vivo target for the ligand. Finally, food deprivation studies in larvae demonstrated an increase in GIP and proglucagon II mRNA levels in response to fasting. In conclusion, the results of these studies suggest that although the zebrafish appears to be a model of an early stage of evolutionary development of GIP expression, the peptide may not possess incretin properties in this species.

Wear of Articular Cartilage: The Effect of Crystals

1993 ◽  
Vol 207 (1) ◽  
pp. 41-58 ◽  
Author(s):  
A Hayes ◽  
B Harris ◽  
P A Dieppe ◽  
S E Clift
Keyword(s):  
Articular Cartilage ◽  
Synovial Fluid ◽  
Crystal Size ◽  
Articular Surface ◽  
Normal Cartilage ◽  
Pin On Disc ◽  
Size And Morphology ◽  
Arthritic Joints

An investigation of the effect of crystals in a lubricant on the wear of articular cartilage in vitro was carried out in order to examine the hypothesis that crystals present in synovial fluid could cause abrasive damage of the articular surface. Plugs of cartilage were worn against a stainless steel counterface in a pin-on-disc wear rig. The concentration of cartilage debris present in the lubricant was assessed by measuring the bound sulphate originating from the glycosaminoglycans by ion chromatography. Results indicated that the presence of crystals in the lubricant significantly increased the concentration of wear debris and that the crystal size and morphology influenced the type of damage sustained by the cartilage. Other experimental evidence suggested that cartilage scratched in vivo was no more susceptible to further in vitro damage in this experimental model than normal cartilage. These results implied that crystals present in the synovial fluid of arthritic joints have the potential to cause excessive wear of the articular surface, but that if such crystals are removed the scratched cartilage may not be susceptible to any further damage by abrasive wear.

RT-PCR Analysis of MMP-9 Expression in Human Articular Cartilage Chondrocytes and Synovial Fluid Cells

1996 ◽  
Vol 71 (4) ◽  
pp. 208-213 ◽  
Author(s):  
K. Tsuchiya ◽  
W. J. Matoney ◽  
T. Vu ◽  
A. R. Hoffman ◽  
D. J. Schurman ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Synovial Fluid ◽  
Pcr Analysis ◽  
Human Articular Cartilage ◽  
Rt Pcr ◽  
Synovial Fluid Cells

The Response of Articular Cartilage to the In Vivo Replacement of Synovial Fluid with Saline

1983 ◽  
Vol &NA; (174) ◽  
pp. 285???292
Author(s):  
ROBERT G. JOHNSON ◽  
MORLEY A. HERBERT ◽  
STEWART WRIGHT ◽  
CHRIS OFFIERSKI ◽  
JIM KELLAM ◽  
...  
Keyword(s):  
Articular Cartilage ◽  
Synovial Fluid
Sign in / Sign up

Export Citation Format

Share Document