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Magnesium Research 2013; 26 (3): 120-4 ORIGINAL ARTICLE Salivary mineral composition in patients with oral cancer Copyright © 2017 John Libbey Eurotext. Téléchargé par un robot venant de 88.99.165.207 le 12/08/2017. Anna Dziewulska1 , Joanna Janiszewska-Olszowska1 , Teresa Bachanek2 , Katarzyna Grocholewicz1 1 Chair and Department of General Dentistry Pomeranian Medical University of Szczecin, Al. Powstancow Wlkp. 72, 70-111 Szczecin; 2 Department of Conservative Dentistry with Endodontics Medical University in Lublin, 7 Karmelicka Street, 20-081 Lublin Correspondence: Joanna Janiszewska-Olszowska. Chair and Department of General Dentistry Pomeranian Medical University of Szczecin. Al. Powstancow Wlkp. 72, 70-111 Szczecin <[email protected]> Abstract. Aim. To analyse the mineral content of saliva in patients with oral cancer in order to identify possible markers that might aid the diagnosis of oral cancer. Subjects and methods.The study group consisted of 34 patients, aged 35-72 years with a diagnosis of oral cancer, including seven women and 27 men, before the start of treatment. Samples of unstimulated saliva were collected in plastic containers. The concentrations of sodium and potassium were assessed using ion selective electrodes, and the concentrations of calcium, magnesium, iron and phosphorus were assessed using colorimetric methods. Results. Statistically significant differences between the study and control groups were found only for the concentration of sodium – higher concentrations were found in the study group. When comparing different cancer localisations, the highest levels of salivary sodium were found in cases of cancer of the floor of the oral cavity, and the lowest levels in tongue or parotid gland cancer. The highest calcium levels were found in cancer of the floor of the oral cavity, and the lowest levels in tongue cancer. The highest levels of magnesium were found in cancer of the floor of the oral cavity, and the lowest in tongue cancer. As regards the different histological types, higher sodium and calcium levels were found in squamous cell carcinomas than in other types. Conclusion. Salivary mineral content in patients with oral squamous cell carcinoma is indicative of oral dehydration; however, we found no evidence of any salivary mineral markers that would be useful for the diagnosis of oral cancer. Key words: oral cancer, squamous cell carcinoma, salivary minerals, sodium, calcium, magnesium as well as local factors (periodontal and dental status) [2, 3]. Sodium, potassium and calcium are the major cations in unstimulated, whole saliva [4]. The calcium phosphate found dissolved in saliva, influences the process of demineralisation and remineralisation of the enamel [4], as well as the formation of dental calculus [5, 6]. Other inorganic components of dental calculus include magnesium and sodium [6]. There is no evidence that the level of magnesium in saliva reflects the general status, neither is it increased after supplementation [8]; however, 120 To cite this article: Dziewulska A, Janiszewska-Olszowska J, Bachanek T, Grocholewicz K. Salivary mineral composition in patients with oral cancer. Magnes Res 2013; 26(3): 120-4 doi:10.1684/mrh.2013.0346 doi:10.1684/mrh.2013.0346 Saliva is part of the natural environment of the oral cavity. It has a fundamental role in the integrity of mineralised dental tissues, as well as of the soft tissues of the oral mucosa. The inorganic components of dental calculus are predominantly calcium and phosphorus [1]. Apart from calcium, the role of salivary minerals has not been described in detail.– In fact, little evidence exists concerning the association between salivary electrolytes and oral health. The levels of salivary minerals are a consequence of the general health status (including age, smoking and alcohol use), Salivary mineral composition in oral cancer Table 1. Distribution of the study group according to cancer localisation and smoking status in the study and control groups. Copyright © 2017 John Libbey Eurotext. Téléchargé par un robot venant de 88.99.165.207 le 12/08/2017. Localisation Tongue Oral cavity floor Tonsil Parotid gland Other localisation* Total Smoking status Present smokers Past smokers Non-smokers Total Females n % 1 14.29 1 14.29 2 28.57 1 14.29 2 28.57 7 100 Study group n % 13 38.24 13 38.24 8 23.52 34 100 Males n % 10 37.04 7 25.93 4 14.81 3 11.11 3 11.11 27 100 Control group n % 5 16.67 12 40 13 43.33 30 100 * Mandibular alveolus, maxillary alveolus, velum and uvula, cheek, throat lower salivary magnesium concentrations have been demonstrated in migraine patients as compared to young, healthy individuals [9]. The iron content is a consequence of desquamation of the epithelial cells as well as possible gingival bleeding [10, 11]. The pathological cell proliferation seen in oral cancer is associated with tissue destruction, and loss of cell electrolytes and macromolecules. Inflammation and bleeding may also be present. The aim of the study was to analyse the mineral content of saliva in patients with oral cancer, in order to search for markers that might help in the diagnosis of oral cancer. Subjects and methods The study group consisted of 34 patients with oral cancer, aged 35-72 years, including seven women and 27 men, before treatment initiation. All patients were examined shortly after the diagnosis of cancer, and had no detectable metastases. The control group consisted of generally healthy dental patients, aged 37-70 years, including five women and 25 men. The distribution of the study group according to cancer localisation is presented in table 1. The most frequently diagnosed oral cancer in women was tonsil cancer, whereas in men, tongue cancer was the most common. The smoking status in the study and control groups is presented in table 1. After ethical approval (Ref No. KE0254/121/2006), informed consent was obtained from every participant. A detailed questionnaire concerning oral hygiene, habits, history and family interview was completed in addition to the examination. Unstimulated saliva was collected in plastic containers in a separate room, in a comfortable sitting position. The concentrations of sodium and potassium were assessed using ion selective electrodes by Konelab (Thermo Electron Corporation, Vantaa, Finland). The concentrations of calcium, magnesium, iron and phosphorus were assessed using colorimetric methods in a Konelab analyser (Thermo Electron Corporation). The concentration of calcium was assessed using the Calcium test by Konelab, with Arsenazo III (SigmaAldrich, St.Louis, USA) at a wavelength of 600 nm. Magnesium content was analysed using an Mg-Kit (bioMerieux, Craponne, France), with Camalgite (Sigma-Aldrich) at 520 nm, iron, using the Ferentest with Feren S (bioMerieux) at 593 nm, and phosphorus with phosphore UV with ammonium molybdate at 340 nm. Data normality was assessed using the ShapiroWilk test. Variance homogeneity was assessed using the Fischer F-test. Comparisons were made between the study and control groups, as well as within the study group between subgroups of different cancer localisation or histological type. The Mann-Whitney test was used for comparisons between the groups and subgroups. The Spearman rank coefficient was used to assess correlations between parametric variables. For the latter, cross-tabulation and 2 homogeneity or the independence test with Yates’s correction were used for subgroups of less than five subjects. The level of significance p<0.05 was used as a reference. All the tests were performed using statistical package Statistica 8.0 (StatSoft, Poland). Results No statistically significant differences concerning age and sex, smoking status or alcohol use were found between the study and control groups. The general diseases diagnosed were hypertension (14 patients in the study group and 15 in the control group), and hyperthyroidism (two patients in the study and two in the control groups). Apart from 121 A. DZIEWULSKA, ET AL. Copyright © 2017 John Libbey Eurotext. Téléchargé par un robot venant de 88.99.165.207 le 12/08/2017. mild gingival inflammation in both the study and control groups, no other concomitant pathology that might have had a known or probable effect on salivary mineral composition was seen in any of the patients examined. The mineral composition of saliva in the study and control groups is presented in table 2. Statistically significant differences between the study and control groups were found only regarding the concentration of sodium – higher concentrations were found in the study group (p = 0.01). There were no statistically significant differences in salivary mineral composition as a function of age, although the level of phosphorus was higher in men than in women (p = 0.03). The mineral salivary composition was not significantly different in tobacco smokers or alcohol abusers. When comparing the different localisations, statistically significant differences were found involving the concentrations of sodium, calcium and magnesium. The highest levels of salivary sodium were found in patients with cancer of the floor of the oral cavity, and the lowest levels in tongue and parotid gland cancer (p = 0.001). The highest calcium levels were found in those with cancer of the floor of the oral cavity, and the lowest levels in patients with tongue cancer (p = 0.05). The highest levels of magnesium were found in patients with cancer of the floor of the oral cavity, and the lowest levels in tongue cancer patients (p = 0.04). As regards the histological types of oral cancer, statistically significant differences were found in sodium and calcium levels. Higher sodium (p = 0.01) and calcium (p = 0.01) levels were found in squamous cell carcinomas than in other histological types. Discussion Salivary mineral composition in smokers and non-smokers has seldom been studied and conflicting results have been obtained. Kolte et al. [7], have concluded that smoking influences the mineral content of saliva, with higher levels of salivary magnesium, but lower levels of calcium and phosphorus having been found in healthy smokers than in non-smokers. Different results were reported by Grocholewicz [12] as well as by Sewon et al. [13], who analysed salivary calcium levels in 603 female patients aged 50-62 years, 122 including 90 smokers, and found that smokers had lower bone density and higher salivary calcium than non-smokers. Contrary to these findings, in the present study, no significant differences were found between smokers and non-smokers. This fact may be due to the high frequency of smoking in present study. Similar results have been obtained by Erdemir et al. [1] in a study of 12 smokers and 12 non-smokers with chronic periodontitis. Higher concentration of sodium in the study group supports an earlier study by Fuchs et al. [14]. It might be supposed that an elevated sodium level could reflect dehydration due to the oral carcinoma itself, as well as smoking and alcohol consumption. However, the salivary rate was not analysed in the present study. Salivary mineral composition in patients with oral cancer has rarely been studied. The results from the few studies found are summarised in table 2. The concentration of potassium in the present investigation is similar to that reported by Shpitzer et al. [15] as well as by Błoniarz et al. [16]. The lack of statistically significant differences between patients with oral cancer and the control group also supports earlier findings. The results involving magnesium concentration in saliva contrast with those found in the study by Grandinaru et al. [17] who found higher salivary magnesium levels in patients with parotid gland tumors. No statistically significant difference in calcium levels were found which contrasts with the study by Grandinaru et al.[17] who found significantly lower salivary calcium concentrations in patients with malignant parotid tumours. This might have been due to tumour localisation where there was higher variability than in the present study. Shpitzer et al. [15] found a lower mean salivary concentration of potassium and higher concentrations of sodium, calcium, phosphorus and magnesium in patients with oral squamous cell cancer of the lateral aspect of the mobile tongue, compared to a control group of healthy individuals, which is generally in agreement with the present study. The presence of iron ions in saliva results from epithelial desquamation and gingival bleeding. The fact that no differences were seen between the study and control groups may reflect a similar prevalence of gingivitis and associated gingival bleeding in both groups. Extremely high iron concentrations have been reported by Błoniarz Salivary mineral composition in oral cancer Table 2. Mineral composition of saliva in the study and control groups in the present study and in the literature. Mineral Sodium [mmol/L] Potassium [mmol/L] Calcium [mmol/L] Copyright © 2017 John Libbey Eurotext. Téléchargé par un robot venant de 88.99.165.207 le 12/08/2017. Magnesium [mmol/L] Iron [mmol/L] Phosphorus [mmol/L] Sodium [mmol/L] Potassium [mmol/L] Calcium [mmol/L] Magnesium [mmol/L] Iron [mmol/L] Phosphorus [mmol/L] Group Study Control Study Control Study Control Study Control Study Control Study Control Study Control Study Control Study Control Study Control Study Control Study Control Present study SD Median 13.61 20.5 8.09 18 8.14 25 9.27 25.63 0.66 0.77 0.55 0.75 0.26 0.38 0.39 0.36 1.26 0.36 0.92 0.36 11.2 4.97 9.31 6.61 Data from literature Błoniarz et al., 2003 47.55 8.35 23.42 18.51 2.96 1.03 0.79 0.21 22.9 4.47 no data no data Mean 27.29 20 26.96 25.93 0.99 0.82 0.44 0.42 0.74 0.57 10.3 9.7 Q3 17 15 22.7 21.56 0.46 0.35 0.22 0.2 0.18 0.18 3.46 4.77 Q1 35 21 34.04 31.13 1.55 1.04 0.66 0.5 0.72 0.36 8.53 8.13 Range (min.-max.) 13.00-61.00 12.00-49.00 6.23-44.52 5.37-44.95 0.02-2.72 0.18-2.43 0.08-0.96 0.08-2.20 0,00-7.2 0.00-4.14 1.90-35.22 0.77-34.93 Sphitzer et al., 2007 22.5 19.8 20.7 24.3 1.47 0.92 0.37 0.29 no data no data 7.52 5.43 et al. [16], both in the study and control groups, which is difficult to explain. It can be concluded that the comparison of salivary mineral content in patients with oral cancer and a control group provided no evidence of any putative salivary mineral markers that could constitute a diagnostic tool. 2. Bales CW, Freeland-Graves JH, Askey S, Behmardi F, Pobocik RS, Fickel JJ, Greenlee P. Zinc, magnesium, copper and protein concentrations in human saliva: age- and sex-related differences. Am J Clin Nutr 1990; 51: 462-9. Disclosure 4. Rehak NN, Cecco SA, Csako G. Biochemical Composition and Electrolyte Balance of “Unstimulated whole Human Saliva. Clin Chem Lab Med 2000; 38: 335-43. Financial support: none. Conflict of interest: none. References 1. Erdemir EO, Erdemir A. The detection of salivary minerals in smokers and non-smokers with chronic periodontitis by the inductively coupled plasmaatomic emission spectrophotometry technique. J Periodontol 2006; 77: 990-5. 3. Skomro P, Opalko K, Bohdziewicz O, Nocen I, Janiszewska-Olszowska J. Limited effect of low frequency magnetic Fields on the concentrations of calcium, magnesium and fluoride in saliva. Magnes Res 2009; 22: 89-92. 5. Bachanek T., Mielnik-Błaszczak M. Ślina a stan zdrowia jamy ustnej – wybrane aspekty. Prz Stomatol Wieku Rozw 1995; 3-4 (11-12): 41-43. 6. Moss S. Rola śliny w utrzymaniu zdrowia jamy ustnej. Stomatol Współcz 1994; 2: 154-8. 7. Kolte AP, Kolte RA, Laddha RK. Effect of smoking on salivary composition and periodontal status. J Indian Soc Periodontol 2012; 16: 350-3. 123 A. DZIEWULSKA, ET AL. 8. Witkowski M, Hubert J, Mazur A. Methods of assessment of magnesium status in humans: a systematic review. Magnes Res 2011; 24: 163-80. 9. Sarchielli P, Coata G, Firenze C, Morucci P, Abbritti G, Gallai V. Serum and salivary magnesium levels in migraine and tension-type headache. Results in a group of adult patients. Cephalalgia 1992; 12: 21-7. 10. Cieślak M, J˛edrzejewska T, Zgirski A. Determinations of magnesium, iron and copper in the saliva of healthy subjects. Czas Stomatol 1990; 43: 202-6. Copyright © 2017 John Libbey Eurotext. Téléchargé par un robot venant de 88.99.165.207 le 12/08/2017. 11. Zarzecka J. Zawartość wybranych pierwiastków w ślinie mieszanej, niestymulowanej u wytapiaczy surówki Huty im. Tadeusza Sendzimira. Czas Stomatol 2002; LV; 12: 789-798. 12. Grocholewicz K. The evaluation of selected health parameters of the stomatognathic system in postmenopausal women and their analysis with reference to the skeletal status determined by the ultrasound method. Habilitation Thesis, Pomeranian Medical University, 2008. 124 13. Sewón L, Laine M, Karjalainen S, Doroguinskaia A, Lehtonen-Veromaa M. Salivary calcium reflects skeletal bone density of heavy smokers. Arch Oral Biol 2004; 49: 355-8. 14. Fuchs PN, Rogić D, Vidović-Juras D, Susić M, Milenović A, Brailo V, Boras VV. Salivary analytes in patients with oral squamous cell carcinoma. Coll Antropol 2011; 35: 359-62. 15. Shpitzer T, Bahar G, Feinmesser R, Nagler RM. A comprehensive salivary analysis for oral cancer diagnosis. J Cancer Res Clin Oncol 2007; 133: 613-7. 16. Błoniarz J., Rahnama M., Zar˛eba S. The influence of carcinogenesis in the oral cavity on the level of some bioelements in the saliva. Rocz Państ Zakł Hig 2003; 54;3: 295-300. 17. Grădinaru I, Ghiciuc CM, Popescu E, Nechifor C, Mândreci I, Nechifor M. Blood plasma and saliva levels of magnesium and other bivalent cations in patients with parotid gland tumors. Magnes Res 2007; 20: 254-8.