Conference Papers

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This collection contains abstracts of conference papers, presented at local and international conferences by the staff of the Faculty of Medicine

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2011 - 20122

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Author: search.filters.author.Silva, N.Has files: No

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    Establishment of human endometriai stromal cell cultures as in-vitro models to test the effects of metalloestrogens in women with or without endometriosis
    (Sri Lanka Medical Association, 2012) Silva, N.; Tennekoon, K.; Senanayake, H.; Samarakoon, S.; Fernando, S.; Wickremasinghe, A.R.; John, R.P.
    INTRODUCTION: Effects of metalloestrogens like cadmium (Cd), lead (PbJ and nickel (Ni) are difficult to demonstrate in-vivo due to toxicity and therefore requires suitable in-vitro models. AIMS: To establish endometriai stromal cell (ESC) cultures as in-vitro models to demonstrate the effects of metalloestrogens in women with and without endometriosis. METHODS: Stromal cells were isolated from eutopic endometriai samples from five women with endometriosis (patients) and five women without endometriosis (controlsj. ESC cultures were established and maintained in RPMI medium. Cultures were treated with Cd, Pb and Ni at concentrations of 10-6M, 10-9M and 10-9M respectively. At 24 h and 48 h, cell number was counted using the Neubauer haemocytometer. Sulphorhodamine(SRB) cytotoxicity assay was used to test the effect of different concentrations of metals on ESC cultures. After 24 h of treatment, caspase levels in ESC cultures were evaluated with a commercially available ELISA kit Relative cell proliferation, SRB assay results and caspase levels were analyzed with ANOVA. RESULTS: ESC cultures were maintained up to 30 days. In both patients and controls Cd, but not Pb or Ni increased the relative proliferation in ESC cultures (p<0.05). At 48 h, Cd induced ESC proliferation was higher in patients than in controls (p=0.02). SRB assay results and caspase levels were similar in the two groups. CONCLUSIONS: Established ESC cultures served as stable in-vitro models. Cd induced the proliferation of ESC from women with endometriosis which appears to be independent of reduced apoptosis. Similar effects of Pb and Ni could not be demonstrated in this in vitro model.
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    Accumulation of toxic metals in ectopic endometrial tissue
    (Sri Lanka Medical Association, 2011) Silva, N.; Senanayaka, H.; Peiris-John, R.; Wickremasinghe, R.; Waduge, V.
    INTRODUCTION AND OBJECTIVES: Aim of this study was to determine the association between heavy metals and endometriosis. METHODS: A case-control study was conducted in a tertiary care hospital comparing 50 cases of endometriosis with 50 age matched controls confirmed by laparoscopy or laparotomy. Blood samples and ectopic endometrial tissue samples were obtained and digested with supra pure 65% HNO3. Samples were analysed for cadmium by graphite furnace atomic absorption spectroscopy (GFASS). In a subset of cases (n=30) and controls (n=40) samples were analyzed for nickel, lead and zinc by Total Reflection X-ray Fluorescence (TXRF) technique. T-tests and Spearman's correlation coefficient were used for analysis. RESULTS: Cases had significantly higher (p=0.011) mean (+SD) blood nickel levels (1.69+1.18 ug/1) as compared to controls (0.96+0.94 ug/1). Blood levels of cadmium (0.75±0.46 ug/I vs 0.77 ±0.37 ug/1) lead (5.8S±1.25 ug/1 vs 3.45±1.25 ug/1) and zinc (123.02±2.49 ug/1 vs151.35 ±2.46 ug/1) were similar in the two groups (p=0.289, p=0.123s and p=0.339, respectively). Tissue samples had significantly higher (p=0.001, p<0.001, p=0.049 respectively) mean (+SD) levels of cadmium (2.86±0.61 ug/1 vs 0.8±0.48 ug/1) nickel (8.64±0.4.12 ug/Lvsl.58±1.21ug/l) and lead (11.22±4.67 ug/1 vs 5.86±3.38 ug/1) compared to blood levels. Mean (+SD) blood levels of zinc (117.48 ±2.64 ug/1) was higher (p<0.001) than that in tissues (27.54±6.65 ug/1). There was a positive correlation between blood zinc levels and tissue zinc levels (r=0.417;p= 0.034) but there was no correlation between blood and tissue levels of cadmium, nickel and lead (r=0.237,p=0.085; r=0.372,^=0.113; r=0.70,/?=0.067, respectively). CONCLUSIONS: Cadmium, nickel and lead but not zinc accumulate in ectopic endometrial tissue.