Evaluation of Selected Parameters of Lipid Peroxidation and Paraoxonase Activity in Blood of Patients with Joint Osteoarthritis.

BACKGROUND: Oxidative stress constitutes an important risk factor for tissue/cell damage and is involved in the occurrence and progression of Osteoarthritis (OA). OBJECTIVE: The aim of this study was to evaluate the concentration of Malondialdehyde (MDA) in Synovial Fluid (SF) and blood as well as the activity of serum and SF paraoxonase (PON1) of patients with OA. Our study aimed to assess local and systemic responses to oxidative stress. METHODS: The study included 43 subjects with OA qualified to total joint replacement. The control group consisted of 58 subjects with no musculoskeletal disorders. The malondialdehyde level were measured in venous blood plasma, erythrocytes (MDAe) and Synovial Fluid (SF) while PON1 activities were estimated in serum and SF. RESULTS: No differences in MDA concentrations in erythrocytes and blood plasma between healthy subjects and OA patients were observed. The significant higher level of MDA in SF compared with blood serum was seen in both genders (in woman about 46.2%, in men - 75.0%) and in both assessed joints (in hip about 53.8%, in knee - 115.9%). Total protein concentration and PON1 activity in serum were significantly higher in OA patients (about 14.6% and 87.9% respectively). The systemic response to osteoarthritis (serum PON1 activity) was higher about 33.6% in men than women and not dependent on affected joint. CONCLUSION: The changes observed in osteoarthritic SF show that the process of antioxidant enzymatic defence is intensified locally, not systemically. The lipid hydroperoxides elimination may lead to restore joints homeostasis.

Lysosomal enzymes in serum and synovial fluid in patients with osteoarthritis.

OBJECTIVE: To assess the activity of arylsulfatase (AS), acid phosphatase (ACP), cathepsin D (CAT-D) and alpha-1 antitrypsin (AAT) in blood serum and synovial fluid (SF) of patients with hip or knee osteoarthritis (OA). METHODS: The study included 43 subjects with OA (35 hip OA, 8 knee OA), submitted total joint replacement. The control group consisted of 58 subjects with no past history of musculoskeletal disorders. RESULTS: The OA blood serum samples showed a significantly higher level of lysosomal enzymes activity than in the control group (AS by 17.8%, AAT by 42.4%); only the CAT-D activity decreased by 50%). AS, ACP and CAT-D activities were about two-fold higher in SF when compared with blood of OA patients. The differences between the genders were visible in the SF: Total protein concentration, activity of ACP (both higher in OA men) and activity of CAT-D (higher in OA women). Between the involved hip and knee, there were no significant differences in all estimated parameters in the blood serum of the OA group. In regard to the SF, only ACP activity was significantly higher in patients with a hip involved. CONCLUSIONS: The osteoarthritic SF enzymatic profile differs from that in normal joints. The OA in joints is not reflected in the systemic response. Our preliminary results suggest further studies on role of lysosomal enzymes (ACP and AS) as biomarkers for the detection of osteoarthritis.

The activity of cathepsin D and alpha-1 antitrypsin in hip and knee osteoarthritis.

The progress of cartilage decay during joint degeneration is not well monitored with biochemical methods. The role of cathepsin D (CAT-D) in articular cartilage deterioration remains unclear. The aim of this study is to assess the activity of CAT-D and alpha-1 antitrypsin (AAT) in blood in patients with hip or knee osteoarthritis. The activity of CAT-D and AAT in blood serum of 40 women and 21 men with hip or knee osteoarthritis was determined before total joint replacement, on the tenth day after surgery, and once in 54 healthy patients. The preoperative activity of CAT-D in patients with osteoarthritis was lower by 53.6% (11.00 ± 4.54 10(-2) nM released tyrosine/mg protein/min, P < 0.001) and after surgery by 55.0% (10.67 ± 4.64 10(-2) nM released tyrosine/mg protein/min, P < 0.001) when compared to its activity in healthy patients. There was no significant statistical difference between CAT-D activity before the surgery and its activity on the tenth day after it in the analyzed group (P< 0.496). Simultaneously, the preoperative activity of AAT in the OA (osteoarthritis) patients was by 25.5% (0.93 ± 0.32 mg inhibited trypsin/ml blood serum, P < 0.001) and postoperative was by 44.9% higher (1.26 ± 0.36 mg inhibited trypsin/ml blood serum, P < 0.001) than in healthy patients. The low CAT-D activity in osteoarthritis of big joints is associated with a decrease of cartilage cells during the degenerative process. The higher activity of acute phase protein AAT in OA patients' blood serum confirms the inflammatory component in the osteoarthritis process.

The concentration of thiobarbituric acid reactive substances (TBARS) and paraoxonase activity in blood of patients with osteoarthrosis after endoprosthesis implantation.

BACKGROUND: The aim of this study was to evaluate the concentration of malondialdehyde (MDA) in erythrocytes and in blood plasma and the activity of blood paraoxonase (PON1) of patients with osteoarthrosis (OA) submitted to endoprosthesis implantation for evaluating oxidative stress. MATERIAL/METHODS: Study was conducted on 55 patients with OA and on 54 total movement-efficient volunteers. The material for the study was venous blood plasma, serum and erythrocytes. RESULTS: Increased concentration of MDAe before surgery was observed in the group of men and in patients with a degenerative process affecting hip joints. After an implantation of endoprosthesis, MDAe decreased to the level observed in the control groups. In the study group MDA concentration in plasma was slightly lower before surgery, and after an operation it reached the value of the parameter of the reference groups. Regardless of sex or age, paraoxonase activity was almost twice as high in almost all subgroups as in the reference group. A positive correlation between PON 1 activity and MDAe concentration was demonstrated both before and after surgery in the group of men. CONCLUSIONS: The increase of PON1 activity in patients' serum in relation to the control groups indicates a probable pathogenic role of the increased formation of reactive oxygen species in the course of OA and may suggest acute inflammation of the synovial joint. The high level of PON 1 activity after endoprosthesis implantation indicates that surgical treatment may additionally stimulate ROS generation. MDAe concentration indicate more intensive process of lipid peroxidation in the elderly.

Oxidative equilibrium in the prophylaxis of degenerative joint changes: an analysis of pre- and postoperative activity of antioxidant enzymes in patients with hip and knee osteoarthritis.

BACKGROUND: Osteoarthritis is often accompanied by disturbance of oxidative equilibrium. The aim of the study was to analyze antioxidant defense system function in patients with hip and knee osteoarthritis by assessing superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) activity in erythrocytes. MATERIAL/METHODS: The study included 26 women and 18 men who underwent hip or knee arthroplasty. The pre- and postoperative activities of the antioxidant enzymes were evaluated. SOD activity was determined using the Misra/Fridovich method, CAT by the Beer/Seizer method, and the activity of GPx by the Paglia/Valentine method. The results were compared with those of a healthy control group. RESULTS: SOD activity of the patients was insignificantly lower than in the control group. However, after arthroplasty it increased in subgroups: in males by 13% (p<0.04), in females by 7% (p<0.05), in the group <69 years of age by ca. 14%) (p<0.01), and in the group with idiopathic OA by 12% (p<0.005). The activities of CAT and GPx in all the patient groups were significantly higher than in the controls. It increased even more on the 10th day after arthroplasty with the exception of the patients with rheumatoid osteoarthritis, in whom a decrease by 65.5% (p<0.008) were found, and in the patients > or =69 years old by 9.3% (NS). CONCLUSIONS: Knowledge of antioxidant enzyme activity might offer new targets for future therapeutic methods for the treatment of degenerative joint disease.

B16 and cloudman S91 mouse melanoma cells susceptibility to apoptosis after dacarbazine treatment.

Considering the necessity of an individual choice of cytostatic drugs for patients with cancer disease and tumor cells' resistance to these compounds, their ability to induction of apoptosis should be investigated. The aim of this study was to determine the influence of dacarbazine (DTIC) on morphology and kinetics of proliferation of B16 and Cloudman S91 cells. It is important to determine the kind of death induced by the DTIC and the effect of a specific concentration. The evaluation of apoptosis and necrosis in these two mouse melanoma cell lines in vitro was performed. Induction of apoptosis was estimated in annexin V binding assay by flow cytometry. DNA content and cell cycle phases were determined by propidium iodide staining. DTIC induced morphological changes typical for apoptosis and necrosis in both cell lines. DTIC caused cell cycle arrest in S and G2/M phase of both cell lines which showed hypertetraploidy. The highest induction of apoptosis was observed in DTIC concentration of 200 microg/mL for B16 cells (11%) and 100 microg/mL for apoptosis Cloudman S91 cells (22.2%). Higher doses of DTIC caused intensification of necrotic process. The B16 melanoma cells are more sensitive to DTIC than the Cloudman S91 cells, however more intensive apoptotic process was detected in Cloudman S91 cells already at lower concentration of DTIC.