Tomasz Mazurkiewicz

Prevalence of genetic risk factors related with thrombophilia and hypofibrinolysis in patients with osteonecrosis of the femoral head in Poland

BackgroundThe etiology of osteonecrosis of femoral head (ONFH) has not been fully elucidated. Increased intravascular coagulation and/or hypofibrinolysis have been proposed as pathogenic mechanisms. Previous reports demonstrated significant association between incidence of ONFH and polymorphisms of genes related with thrombophilia especially in Caucasian subjects. The aim of our study was to evaluate the relationship between genetic mutations leading to coagulation disorders and ONFH in Polish patients.MethodsWe have investigated the frequencies of four markers among 68 unrelated individuals with clinically and radiographically documented ONFH and among 100 healthy unrelated blood donors in Eastern part of Poland. The three genes were involved in thrombophilia: factor V Leiden (G1691A), prothrombin (G20210A), Methylenetetrahydrofolate Reductase (MTHFR C677T) and one in hypofibrinolysis: Tissue Plasminogen Activator (PLAT TPA25 I/D). The samples were genotyped with polymerase chain reaction followed by restriction enzyme analysis for the restriction fragment length polymorphisms. The allele and genotype frequencies were analyzed in the relation to ONFH etiology (idiopathic and secondary), gender, age (patients younger or older than 50 years) and the number of affected joints (unilateral or bilateral ONFH).ResultsNo significant difference in allele frequencies between patients and control groups were observed in genes involved in thrombophilia. We have found a statistically significant increased frequency of D allele of PLAT TPA 25 I/D polymorphism between the entire group of patients with ONFH and controls (p=0,026, OR=1,54, CI 0,99-2,4). D allele frequency was also significantly increased in patients with primary ONFH (p=0,009, OR=1,81 CI 1,1-3,01), in males (p= 0,013; OR 1,74; 95% CIs 1,08-2,78), patients older than 50 years (p= 0,018, OR= 2,04; 95% CIs 1,09-3,82) and in cases with bilateral ONFH (p= 0,01; OR= 1,92; 95% CIs 1,13-3,27) (Table 9). The differences in DD homozygous genotype frequency were statistically significant for patients with idiopathic ONFH compared with control group (p=0,023, OR=2,75, CI 0,99-7,9) and in cases of bilateral ONFH (p=0,034; OR 3,12; 95% CIs 1,06-9,18) (Table 10). The frequencies of ID heterozygous genotype were statistically significantly higher in entire group of patients with ONFH (p=0,004 OR 2,71; 95% CIs 1,32-5,57), idiopathic ONFH (p= 0,01; OR 2,91; 95% CIs 1,24-6,87), males (p=0,0007; OR 3,75; 95% CIs 1,67-8,42), patients older than 50 years (p=0,001; OR 6,89; 95% CIs 1,87-25,84) and in cases with bilateral ONFH (p=0,009; OR 3,19; 95% CIs 1,26-8,03).ConclusionThe results suggest that inherited hypofibrinolysis is a risk factor of idiopathic ONFH in Polish population.

Determination of pamidronate in bisphosphonate-enriched bone cement by ion-pair hplc and capillary electrophoresis

Abstract The presence of pamidronate during local use of bisphosphonates (BP)-enriched bone cement was determined. The question was whether pamidronate implanted into the bone cement is eluted. The study was performed on 10 probes of BP-enriched bone cement located in 0.9% NaCl. The probes were incubated for 3 and 6 weeks. Ion-pair HPLC was used for the detection of pamidronate. Then, capillary electrophoresis was applied for quantitative analysis of pamidronate in the 3rd and 6th week after incubation. The presence of pamidronate, eluted from BP-enriched bone cement into 0.9% NaCl solution 3 and 6 weeks after incubation, was demonstrated. These results may explain the changes in the level of cytokine RANKL and bone turnover marker osteoprotegrin in rats’ serum treated with BP-enriched bone cement 3 and 6 weeks after surgery. The possibility of effective local use of BP-enriched bone cement in veterinary medicine was underlined. The results, and the former conducted research, point out that the clinical applications of BP-enriched bone cement in vivo may have some validity in the future.

The presence of pamidronate in bone cement affects serum biochemical markers in the rat

Abstract The main aim of the study was to assess whether the presence of biphosphate pamidronate (PA) in the cement implanted into the tibial bones had any effect on the chosen biochemical markers in rat’s serum characterising homeostasis. Forty adult male Wistar rats were divided into two control groups and two experimental groups. Tibial bone of rats in the experimental groups was implanted with PA-enriched cement, whereas the bone in control-group’s rats was implanted with cement without PA. Serum activities of alanine aminotransferase (ALT), aspartate aminotransferase (AST), and creatine kinase (CK) were determined three and six weeks after the surgery. Statistically significant differences in the activities of AST and CK of the rats after implantation with non-enriched cement when compared to rats given PA-enriched cement implantation, were found. Six weeks after treatment, AST levels decreased significantly in rats with PA-enriched cement, whereas rats in the control group (implanted with non-enriched cement) demonstrated a significant increase in AST activity in comparison to the same values determined after three weeks and values of PA-enriched cement rats determined after six weeks. The activities of CK were higher in rats with PA-enriched implants than in the control group three weeks after surgery, but six weeks after the treatment, rats implanted with enriched cement reached lower values than animals implanted with non-enriched cement. The use of PA in the cement had also some positive effect on the homeostasis of the rats after the surgery and a positive influence on the post operative muscle regeneration process.