Percentage Of Apoptotic Chondrocytes Research Articles

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

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.

Procaine and saline have similar effects on articular cartilage and synovium in rat knee

BackgroundIntra-articular local anaesthetics are widely used for providing postoperative analgesia and decreasing the need for opioids. Procaine has proven positive effects in carpal tunnel syndrome and chondromalacia patella. However, the effect of procaine on articular cartilage has not yet been studied. The aim of this study was to evaluate the effects of intra-articular procaine injection on the articular cartilage and the synovium.MethodsTwenty adult Sprague-Dawley rats were enrolled in the study. After providing anaesthesia and aseptic conditions, 0.25 ml of 10% procaine was injected to the right knee joint, and 0.25 ml of normal saline (as control group) was injected to the left knee joint. Knee joint samples were obtained from four rats in each group after appropriate euthanasia on days 1, 2, 7, 14 and 21. The histological sections of the articular and periarticular regions and the synovium were evaluated by two histologists, and inflammatory changes were graded according to a five-point scale in a blinded manner. The apoptosis of chondrocytes was determined by the caspase-3 indirect immunoperoxidase method.ResultsThere were no significant differences in inflammation between procaine and saline groups at any of the time intervals. Slight inflammatory infiltration due to injection was seen in both groups on the 1st day. Haemorrhage was observed in both groups at days 1 and 2, and the difference between groups was not found to be significant. No significant difference was detected in the percentage of apoptotic chondrocytes between groups at any of the time intervals.ConclusionsInjection of procaine seems safe to use intra-articularly based on this in vivo study on rat knee cartilage. However, further studies investigating both the analgesic and histopathological effects of procaine on damaged articular cartilage and synovium models are needed.

Effect of sodium levothyroxine on histomorphometry, histopathology, histochemistry, and immunohistochemistry of articular cartilage in female mice

Sodium levothyroxine is one of the common medicines used for treatment of hypothyroidism. The aim of the present study was to assess the effect of sodium levothyroxine on the middle part of tibia plateau in knee joint and related hormones in individuals affected by hypothyroidism. Fifty female mice were randomized into five groups. The first group was considered as control; the second and third groups were received different doses of levothyroxine; the forth group was undergone hypothyroidism; and the fifth group, the animals with hypothyroidism, was received levothyroxine orally. After 8 weeks, serum samples were taken to determine TSH, T4, FT4, T3, FT3, T3RU, calcium, and phosphorus. The cartilage of tibia plateau was evaluated using hematoxylin–eosin (histomorphometry of tibia plateau), toluidine blue (proteoglycans of matrix), TUNEL (chondrocytes apoptosis), and immunohistochemistry (rate of Collagen-II). The results showed that the number of chondrocytes, cartilage thickness, proteoglycans, and Collagen-II and the rate of T3 in the third group were significantly decreased; whereas, the percentage of apoptotic chondrocytes in the third group was significantly increased compared to those of other experimental groups (P < 0.05). FT3, FT4, TSH, calcium, and phosphorus values in most groups were also significantly different (P < 0.05). Regarding widely consumption of levothyroxine in hypothyroidism and its effects on knee joint cartilage, levothyroxine at high doses should be administered with care.

Resveratrol inhibits apoptosis by increase in the proportion of chondrocytes in the S phase of cell cycle in articular cartilage of ACLT plus Mmx rats.

The current study was aimed to investigate the effect of resveratrol on apoptosis inhibition in chondrocytes in ACLT plus Mmx rat model. TUNEL assay revealed a markedly higher level of apoptotic chondrocytes in the cartilage of untreated ACLT plus Mmx rats. The percentage of apoptotic chondrocytes was found to be 29.5 and 40.75%, respectively at 21 and 45 days. The percentage of apoptotic chondrocytes at 21 and 45 days in resveratrol (5 mg/kg) treated ACLT plus Mmx rats was found to be 13% and 2%, respectively. Real-time PCR analysis revealed that treatment of the ACLT plus Mmx rats with resveratrol for 45 days caused a significant increase in the expression of miR-18a compared to that in untreated rats. Flow cytometry and BrdUrd incorporation assay revealed that the proportion of chondrocytes in the S phase was increased to 51.4% in resveratrol treatment group compared to 25.3% in the untreated ACLT plus Mmx rats. Western blot analysis showed that treatment of the ACLT plus Mmx rats with resveratrol decreased the expression of ATM protein kinase and GFP protein without any effect on the expression of GFP-ϒ-tubulin in chondrocytes. In addition, resveratrol treatment also led to reduction in the activity of luciferase in the chondrocytes of ACLT plus Mmx rats. Resveratrol treatment of the ACLT plus Mmx rats decreases the expression level of ATM protein and checkpoint kinase 2 (CHK2) phosphorylation in chondrocytes. H2AX and 53BP1 phosphorylation was decreased in ACLT plus Mmx rats on treatment with resveratrol for 45 days. Immunofluorescence results revealed a markedly lower level of H2AX and 53BP1 nuclear foci in the chondrocytes of ACLT plus Mmx rats treated with resveratrol. Thus resveratrol treatment of the ACLT plus Mmx rats inhibits chondrocyte apoptosis and increases proportion of cells in the S phase of cell cycle which may be through the increase in expression of miR18a. A significant relation appears between resveratrol and miR-18a expression in the chondrocytes.

Nitric oxide-associated chondrocyte apoptosis in trauma patients after high-energy lower extremity intra-articular fractures.

BackgroundThe primary goal of this study was to identify nitric oxide (NO)-induced apoptosis in traumatized chondrocytes in intra-articular lower extremity fractures and the secondary goal was to identify the timeline of NO-induced apoptosis after injury.Materials and methodsThis is a prospective collection of samples of human cartilage harvested at the time of surgery to measure apoptotic cell death and the presence of NO by immunohistochemistry. Three patients met the criteria for control subjects and eight patients sustained high-energy intra-articular fractures and were included in the study. Subjects who sustained intra-articular acetabular, tibial, calcaneal and talus fracture had articular cartilage harvested at the time of surgical intervention. All 8 patients underwent open reduction and internal fixation of the displaced intra-articular fractures. The main outcome measures were rate of apoptosis, degree of NO-induced apoptosis in chondrocytes, and the timeline of NO-induced apoptosis after high-energy trauma.ResultsThe percentage of apoptotic chondrocytes was higher in impacted samples than in normal cartilage (56 vs 4 %), confirming the presence of apoptosis after intra-articular fracture. The percentage of cells with NO was greater in apoptotic cells than in normal cells (59 vs 20 %), implicating NO-induction of apoptosis. The correlation between chondrocyte apoptosis and increasing time from injury was found to be −0.615, indicating a decreasing rate of apoptosis post injury.ConclusionsThe data showed the involvement of NO-induced apoptosis of chondrocytes after high-energy trauma, which decreased with time from injury.

Experimental Osteoarthritis in Rats Is Attenuated by ABC294640, a Selective Inhibitor of Sphingosine Kinase-2

Background/Aims: Osteoarthritis (OA) is a progressive degenerative disease characterized by cartilage degradation and chondrocyte apoptosis, which may involve aberrant sphingolipid metabolism. ABC294640 is a compound that selectively inhibits sphingosine kinase-2, a key enzyme in the sphingolipid pathway. Our goal was to assess the pharmacological effects of ABC294640 in the monosodium iodoacetate (MIA) model of OA. Methods: MIA (3 mg) was injected into the right knee joint to induce osteoarthritis in rats. Subsequently, the rats were treated with vehicle, ABC294640 or tramadol over a 28-day period. To assess pain, incapacitance readings were obtained weekly. MIA-injected knee joints were evaluated for histological damage, cartilage degradation and chondrocyte apoptosis (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling histochemistry). Results: The percent weight bearing in vehicle/MIA rats significantly (p < 0.01) decreased from 48.8 ±0.8 (day 0) to 41.9 ±2.9 (day 28). In contrast, these values in ABC294640-treated rats were virtually the same on days 0 and 28. Knee joint histology scores were less severe in ABC294640-treated rats. Cartilage proteoglycan staining was more prominent in ABC294640/MIA animals than in vehicle/MIA rats. The percentage of apoptotic chondrocytes was decreased from 39.5% (vehicle treatment) to 25.8% (ABC294640 treatment). Conclusion: ABC294640 attenuated the knee joint histological damage and pain associated with MIA-induced OA in rats.

Suppressive Effect of Hyaluronan on Chondrocyte Apoptosis in Experimentally Induced Acute Ostaoarthritis in Dogs

Dogs receiving anterior cruciate ligament transection (ACLT) were treated with either intravenous (IV) or intraarticular (IA) administration of hyaluronan (HA), and differences in appearance of chondrocyte apoptosis of the stifle joint were investigated. Chondrocyte apoptosis was detected using flow cytometry as well as by staining with TdT-mediated dUTP nick end labeling (TUNEL). The percentage of apoptotic chondrocytes in dogs with ACLT was significantly higher than that in intact (non-ACLT) dogs. Dogs treated with IA or IV injection of HA after ACLT had fewer apoptotic chondrocytes than non-treated dogs after ACLT. It was suggested that ACLT-induced apoptosis of chondrocytes was suppressed by HA administration of either IA or IV.