Adam Kamiński

The association of IL-1β, IL-2, and IL-6 gene polymorphisms with bone mineral density and osteoporosis in postmenopausal women

OBJECTIVE Osteoporosis is a common disorder with a strong genetic component. The genetics of osteoporosis impacts on the prediction, diagnosis, prognosis, and treatment of the disease. STUDY DESIGN The aim of the present study was to examine associations between cytokine gene polymorphisms (IL-1beta, IL-2, IL-6) and bone mineral density (BMD) values in postmenopausal women. The study included 226 postmenopausal women with a diagnosed BMD T-score lower than -2.5 SD (mean: -3.02+/-.053) and 224 postmenopausal women with a BMD T-score greater than -2.5 SD (mean: -1.33+/-0.51). RESULTS Among the women with T-scores below -2.5 SD, the BMD values were significantly lower in the carriers of the IL-6 GG genotype compared with those with the CC and GC genotypes (0.70+/-0.38 vs. 0.73+/-0.25 and 0.74+/-0.23 for the lumbar spine, 0.54+/-0.18 vs. 0.56+/-0.15 and 0.58+/-0.22 for the femoral neck). There were no statistically significant associations between the IL-1beta and IL-2 genotypes and BMD values in the group of women with T-scores below -2.5 SD. CONCLUSION The results of the present study suggest an association of the IL-6 -174 G/C polymorphism with osteoporosis in postmenopausal women.

The RANKL/RANK/OPG signal trail: significance of genetic polymorphisms in the etiology of postmenopausal osteoporosis

OBJECTIVES Recent studies have demonstrated that disorders of bone metabolism, which is regulated by RANK/RANKL/OPG signaling pathway, are the cause of osteoporosis. The aim of the study was to investigate the distribution of genotypes of the RANK 575C>T and RANKL -643C>T polymorphisms and to analyze their relationship with bone parameters in postmenopausal women. MATERIAL AND METHODS A total of 310 postmenopausal Caucasian women (139 with osteoporosis, 107 with osteopenia, and 64 healthy postmenopausal controls) were included. Bone mineral density (BMD) at the lumbar region of the spine (L2-L4) was measured by dual energy X-ray absorptiometry (DXA). Genetic analysis was performed using the PCR-RFLP method. RESULTS Analysis of the frequency of genotypes and alleles of the RANK 575C>T and RANKL -643C>T polymorphisms did not show any statistically significant differences between the study groups (osteoporosis and osteopenia) and postmenopausal women with normal t-score value (ns). Notably, a significant association between the RANKL -643C>T polymorphism and body mass, such as BMI values in osteoporotic women (p<0.05), was observed. CONCLUSIONS Our results suggest lack of association between the 575C>T RANK polymorphism and the development of osteoporosis. The -643C>T RANKL polymorphism, through its significant influence on body weight and BMI value, may contribute to the development of osteoporosis in postmenopausal women.

TNF-α concentrations in pre-operative synovial fluid for predicting early post-operative function and pain after fast-track total knee arthroplasty

BACKGROUND Tumor necrosis factor-alpha (TNF-α) helps regulate neuroinflammation and anxiety and could conceivable predict early post-operative pain and function after fast-track total knee arthroplasty (TKA). METHODS In patients with severe osteoarthritic knees undergoing TKA, we assessed: the correlations between pre-operative concentrations of TNF-α in synovial fluid; pre- and six-week post-operative knee function and pain; pre- and post-operative anxiety; pre- and post-operative synovial fluid concentrations of cartilage oligomeric matrix protein (COMP); age and body mass index (BMI). RESULTS Of 100 enrolled patients, 78 had evaluable TNF-α data, and 58 had evaluable COMP data. Pre-operative TNF-α concentrations were inversely correlated with post-operative pain scores during walking (rS=-0.26, P=0.03) and with change of pain at rest during six weeks after TKA (rs=-0.28, P=0.03) and were directly correlated with a higher post-operative Knee Society score (KSS) (rS=0.43, P<0.001) and with greater increases in this score during six weeks after TKA (rS=0.33, P=0.001). Mean TNF-α concentrations were higher in the 39 patients reporting any pre-operative pain at rest than in 36 patients reporting no pre-operative pain (P=0.015) and were the only independent predictor of pre-operative pain at rest (OR=13, P=0.02). Independent predictors of better post-operative knee function were higher log-transformed TNF-α concentrations (β=0.38, P=0.002) and male sex (β=0.28, P=0.02). CONCLUSIONS High levels of pre-operative TNF-α concentrations could be used as an independent predictor of better knee function at six weeks of follow-up. In patients with lower pre-operative TNF-α concentrations, post-operative pain management may improve the early outcome of the operated joint.

Polymorphisms of OPG and their relation to the mineral density of bones in pre- and postmenopausal women.

OBJECTIVES The aim of the study was to evaluate the frequency of gene polymorphisms OPG -163A/G, -950T/C and 1181G/C, assessing their relations with the clinical parameters of osseous turnover and the degree of postmenopausal osteoporosis. STUDY DESIGN The study included 800 women of postmenopausal (505) and reproductive (295) age from Poland. The postmenopausal group included women with osteoporosis and osteopenia, as well as healthy individuals. All the women of reproductive age were healthy. The frequency of the tested gene polymorphisms was evaluated within the group where BMD (bone mineral density) was marked and also in the control group. MAIN OUTCOME MEASURES The frequencies of the polymorphisms of OPG genotypes in the women were characteristic of the population. RESULTS OPG -950T/C polymorphism has been associated with body weight and birth weight. OPG 1181G/C and OPG -163A/G polymorphisms have been associated not only with body weight and birth weight, but also with reduced bone density and an increased risk of postmenopausal osteoporosis. CONCLUSIONS Evaluation of the polymorphism -950T/C of the OPG gene showed that the CC genotype may appear as an increased risk factor for the faster loss of bone mass and the onset of osteoporosis in Polish postmenopausal women. This polymorphism may be a genetic marker that is responsible for the development of osteoporosis. The homozygous genotypes of polymorphisms 1181G/C and -163A/G of the OPG gene may play a role in increased risks of osteoporosis and may be linked to the birth weights of women.

The effect of smoking on posttraumatic pseudoarthrosis healing after internal stabilization, treated with platelet rich plasma (PRP)

Abstract Disturbed or delayed healing remains one of the most serious fracture-related complications, despite bone capacity for internal regeneration and reabsorption. Considerable progress in the understanding and treatment of fractures has been noted. The aim of our study was to evaluate treatment outcome in patients (smokers and non-smokers) with post-traumatic pseudoarthrosis. Hypothesis Determinate when administration of growth factors is most beneficial, and whether it accelerates bone union. Material and methods The study included patients treated for post-traumatic pseudoarthrosis resulting from multiple bone fractures. The study group and controls were further subdivided into: non-smokers, non-smokers >2 years after quitting, and smokers. Independent tests were performed for men and women. The study group, apart from other methods of treatment, received concentrated PRP (platelet-rich plasma) to aid the process of bone healing, or in cases of delayed healing confirmed by radiological assessment on follow-up visits. Results Mean time of fracture healing was 8 weeks for non-smokers and non-smokers >2 years after quitting, whereas in smokers the healing process was significantly prolonged (18 weeks in both, men and women). Conclusions The risk for infection is smaller in non-smokers as compared to smokers, with the latter being at an elevated risk for bone inflammation and delayed union.

Expression of genes modulated by epigallocatechin-3-gallate in breast cancer cells

Summary Introduction: Breast cancer is the most common malignant cancer among women. Both drug resistance and metastasis are major problems in the treatment of breast cancer. Therefore, adjuvant therapy may improve patients’ survival and affect their quality of life. It is suggested that epigallocatechin gallate (EGCG) which is well known for its chemopreventive activity and acts on numerous molecular targets may inhibit the growth and metastasis of some cancers. Hence, discovering the metastatic molecular mechanisms for breast cancer may be useful for therapy. Objective: The aim of the study was to determine the effect of EGGC on the mRNA expression level of genes such as ZEB1, ABCB1, MDM2, TWIST1 and PTEN in MCF-7 breast cancer cells. Methods: MCF7/DOX were cultured in the presence of 0.2 μM DOX and EGCG (20-50 μM). The mRNA expression level was determined by real-time quantitative PCR using RealTime ready Custom Panel 96 kit. Results: Our results showed an important increase (about 2-fold for 20 μM EGCG + 0.2 μM DOX and 2.5-fold for 50 μM EGCG + 0.2 μM DOX, p<0.05) in ZEB1 expression levels. In case of ABCB1 gene lack of influence on the mRNA level was observed (p>0.05). We also observed significant decrease of ZEB1 expression in MCF7 cells with 20 μM and 50 μM EGCG (p<0.05). In addition, EGCG (20 μM) caused an increase of MDM2 and PTEN mRNA levels in almost 100% (p<0.05) and 40% (p>0.05), respectively. Lack of the influence of EGCG was noted for the TWIST1 gene expression. In case of MCF7/DOX we showed an increase of mRNA level of PTEN gene about 50% (p<0.05). Conclusions: These results suggest that EGCG may be potentially used in adjuvant therapy in the breast cancer treatment.

The rs1256044 polymorphism in the ESR2 gene and the risk for osteoporosis in Polish postmenopausal women

Abstract Population association studies have demonstrated a strong association between ESR2 SNPs and BMD, indicating that ESR2 may influence attainment of bone mass. The aim of the study was to investigate the ESR2 gene, located on chromosome 14q linked with BMD, which demonstrates a correlation with changes in bone mass in healthy Caucasian women. The study included 675 unrelated Polish postmenopausal women, including 109 with osteopenia, 333 with osteoporosis and 233 healthy women. The women were classified into the following groups: osteopenia, osteoporosis and normal T-score. Analysis of genotype frequency for the ESR2 rs1256044 polymorphism revealed no statistically significant differences. No statistically significant differences were noted for the allele frequency. However, it is noticeable that the CT genotype occurred more often in women with osteopenia (50.4%, OR = 1.14) and osteoporosis (54.7%, OR = 1.33) than controls (47.7%). There were statistically significant differences between the clinical parameters and distribution of genotypes in patients with osteopenia but not osteoporosis. ESR2 polymorphisms demonstrate minimal influence on BMD changes in women. Identification of various genes with little impact on BMD, such as ESR2, might help design a screening panel for osteoporosis risk assessment in healthy subjects.

Association betweenGREM2 gene polymorphism with osteoporosis and osteopenia in postmenopausal women

OBJECTIVE Osteoporosis is a civilization disease, in which the dominant symptoms are the loss of bone mass and disturbances in bone structure. Gremlin-2 is one of the BMP (bone morphogenetic proteins) antagonists and participate in osteogenesis and osteoblast differentiation. The aim of the study was to analyze whether the GREM2 gene polymorphism is significantly more common in postmenopausal women than in healthy women and whether it is a predisposing factor for the osteoporosis development. STUDY DESIGN The study consisted of 675 unrelated Polish postmenopausal women, including 109 with osteopenia, 333 with osteoporosis and 233 healthy women. The effect of the GREM2 polymorphism on T-score, Z-score, L2L4AM, L2L4YA, L2L4BMD, body mass, BMI, birth weight was statistically evaluated. RESULTS Statistical significance was observed between the TT and TC genotypes and also between TT and CC genotypes in the case of birth weight in the control group and the group of women with osteoporosis. Analysis of body mass in women with osteoporosis showed the statistical significance between genotypes TT and CC, TT and TC. Analysis of the frequencies of TT, TC and CC genotypes of the rs4454537 polymorphism of the GREM2 gene showed no statistical significance between studied groups. CONCLUSION Our study found that the most frequent genotype in the group of women with osteopenia and osteoporosis was TC while in the group of healthy women the protective TT genotype was dominant. Hence, it can be postulated that the TT genotype is a protective factor against the development of osteoporosis.

The importance of the Wnt/β-catenin pathwayand LRP5 protein in bone metabolism of postmenopausal women

BACKGROUND Postmenopausal osteoporosis is the most common metabolic bone disease among women. The Wnt signaling pathway has been known to be the critical regulator of osteoblastogenesis. Alterations in this mechanism may have consequences for bone remodeling in humans. OBJECTIVES The aim of the study was to evaluate the frequency of genotypes and alleles of single nucleotide polymorphism (SNP) rs4988321 and rs312009 of LRP5 in Polish postmenopausal women with osteopenia (n = 109) and osteoporosis (n = 333). Potential correlations between genetic polymorphisms, bone mineral density (BMD), risk for bone fractures, and other clinical parameters were analyzed. MATERIAL AND METHODS Genomic DNA was extracted from the blood samples and the sequence polymorphisms of LRP5 gene were detected using real-time polymerase chain reaction (RT-PCR) methods with melting curve analysis. We also calculated the odds ratio (OR) for the LRP5 genotypes and the alleles. Then, we evaluated the effect of the LRP5 polymorphism on T-score, Z-score, L2L4AM, L2L4YA, L2L4BMD, body mass index (BMI), and other clinical parameters. RESULTS No statistically significant differences in the distribution of LRP5 rs312009 genotypes between the groups were observed. Furthermore, our findings indicate that there is no correlation between LRP5 genotypes and the clinical characteristics of women with osteopenia/osteoporosis. In contrast, there was an increased value of OR in heterozygotes for rs4988321, both in patients with osteopenia (OR = 1.47) and in those with osteoporosis (OR = 1.33). In our study, we were not able to calculate the OR parameter for the AA genotype due to its low prevalence in the population. CONCLUSIONS Our results suggest that the Val667Met LRP5 (rs312009) polymorphism may contribute to an elevated risk for fractures in postmenopausal Polish women.

The diagnostic potential of glutathione S-transferase (GST)polymorphisms in patients with colorectal cancer

BACKGROUND Colorectal cancer (CRC) is one of the most common cancers in the world. Despite improvements in screening for early diagnosis, CRC is one of the leading causes of cancer deaths. OBJECTIVES The aim of the study was to determine a potential association between the frequency of GSTM1 and GSTT1 null genotypes and the risk of CRC in the Polish population. Moreover, we analyzed the clinical parameters with the glutathione S-transferase (GST) gene polymorphisms in patients with CRC. MATERIAL AND METHODS The study was conducted on 512 Caucasians, including 279 patients (105 women and 174 men) with CRC. DNA from peripheral blood was extracted and the multiplex polymerase chain reaction (PCR) technique was used for glutathione S-transferase theta (GSTT1) and mu (GSTM1) gene deletion genotyping. RESULTS We found no statistically significant differences in the frequency of the GST gene polymorphisms in patients with CRC and controls. The prevalence of the GSTM1*0 variant in the test subjects was higher than in controls (45.9% vs 42.9%; p > 0.05). The frequency of the GSTT1*0 variant was also higher in patients with CRC compared to the control population (21.1% vs 18.9%; p > 0.05). In addition, the effect of the GSTM1 and GSTT1 polymorphisms on the incidence of CRC was also analyzed. There was a slight, but not statistically significant, increase of the risk of colon cancer for the GSTM1*0 and GSTT1*0 variants. Moreover, we examined the GST genotype due to the cancer TNM classification and the location of the primary tumor. Statistically significant differences in the distribution of the GSTT1*0 and GSTT1*1 genotypes in both the stage and the location of the primary tumor were observed. CONCLUSIONS It is suggested that the GSTT1 polymorphism may have an impact on the severity of the tumor and its location.