1919 Research Report Dental Journal (Majalah Kedokteran Gigi) 2017 March; 50(1): 19–22 The correlation between the use of personal protective equipment and level wild-type p53 of dental technicians in Surabaya Puspa Dila Rohmaniar,1 Titiek Berniyanti,2 and Retno Pudji Rahayu3 1Department of Dental Public Health, Faculty of Dentistry, Insitut Ilmu Kesehatan Bhakti Wiyata, Kediri - Indonesia 2Department of Dental Public Health, Faculty of Dental Medicine, Universitas Airlangga, Surabaya – Indonesia 3Department of Oral Pathology and Maxillofacial, Faculty of Dental Medicine, Universitas Airlangga, Surabaya - Indonesia abstract Background: Exposure of metals among dental technicians that come from the working environment can lead to the formation reactive oxygen species (ROS). ROS can cause mutations in the p53 gene (p53). The mutation is transversion mutation Guanine- Thymine. p53 mutations can lead to low expression of the wild-type p53 protein (p53). Wild-type p53 involved in many biological processes such as regulation of genes involved in cell cycle, cell growth after DNA damage, and apoptosis. However, exposure to metals among dental technicians can be prevented through the use of personal protective equipment (PPE) during work. Purpose: The purpose of this study was to analyze the correlation between the use of personal protective equipment to wild-type p53 protein levels among dental technicians in Surabaya. Method: This study was observational analytic with cross sectional approach. 40 samples were taken by random sampling. Data were retrieved through interviews and observations. Wild-type p53 was analyzed from saliva with indirect ELISA method. Analysis of data used Kolmogorov Smirnov normality test and a Pearson correlation test. Value significance was p<0.05 (95% confidence level). Result: There was a significant association between the use of personal protective equipment with wild-type p53 levels with p=0.002 Conclusion: The use PPE properly is positively correlated with the wild-type p53 protein levels of dental technicians in Surabaya. Keywords: personal protective equipment; dental technician; P53 wild Correspondence: Titiek Berniyanti, Department of Dental Public Health, Faculty of Dental Medicine, Universitas Airlangga. Jl. Mayjend. Prof. Dr. Moestopo no. 47 Surabaya 60132, Indonesia. E-mail: berniyanti@gmail.com introduction Dental technician should be exposed to various physical agents, chemical, and biological derived from the work environment. That exposure by inhalation, ingestion or skin contact.1 Exposure of dust or smoke to the dental technician are coming from grinding of dental restoration materials during processing.2 There is a study that reported high concentrations of cobalt metal, nickel, chromium in blood of dental technicians in Surabaya, such as: levels of cobalt: 27 g/ L, nickel 37 g/ L, and chromium 117 mcg/ L.3 Other research in northern Jordan also reported high levels of cobalt and chromium in blood of dental technicians. 2 Exposure may result in potential lung diseases such as bronchial asthma, cancer, mesothelioma and pneumoconiosis depends on the duration of exposure.4 The prevalences of contact dermatitis among dental technicians are 22% in Australia and 43% in Denmark.5 The prevalence of pnumoconis among dental technicians in Ankara is 10.1% and the prevalence of dermatitis kontakta of the dental technicians in Germany is 16%.4 It is important for dental technicians to obey the standards and safety procedures. Dental technicians must wear personal protective equipment (PPE) including work wear, protective mask, protective gloves and goggles, and ventilate the workplace. If ventilation, exhauster, adequate and adequately filter will reduce the level of chromium, cobalt, and nickel in the air.1 Genotoxic metal exposure may increase the number of reactive oxygen species (ROS). The metal ions of chromium, cobalt, and nickel can produce hydroxyl radicals (OH) through the Fenton and Haber-Weis reaction. The Dental Journal (Majalah Kedokteran Gigi) p-ISSN: 1978-3728; e-ISSN: 2442-9740. Accredited No. 56/DIKTI/Kep./2012. Open access under CC-BY-SA license. Available at http://e-journal.unair.ac.id/index.php/MKG DOI: 10.20473/j.djmkg.v50.i1.p19-22 http://dx.doi.org/10.20473/j.djmkg.v50.i1.p19-22 20 Rohmaniar, et al./Dent. J. (Majalah Kedokteran Gigi) 2017 March; 50(1): 19–22 hydroxyl radicals can cause DNA damage.6 Typical damage caused by DNA oxidation is a transversion mutations in guanine.7 p53 protein is involved in various biological processes such as regulation of genes involved in cell cycle, cell growth after DNA damage, and apoptosis.8 p53 is considered as the gene most commonly mutated in human malignant tumors. p53 tumor suppressor gene is mutated in 50% of human tumors in various organs. With the development of molecular biology techniques can explain that one of the causes of the malignant process is a failure or inactivation of tumor suppressor genes p53.7,9 Cells that have mutations or loss of wild-type p53 gene, the p53 protein expression wild does not occur or occurs wild-type p53 protein expression but can not function as activating gene transcription in some target.10 p53 protein expressed by tumor suppressor p53 gene can be detected through saliva. Saliva is a diagnostic medium for the detection of various molecules contained in the blood; saliva can provide similar information about the human status as obtained from a blood test without invasive procedures.11 The purpose of this study was to analyze the correlation between the use of personal protective equipment and wild-type p53 protein levels among the dental technicians in Surabaya. material and methods The Ethics Committee of the Faculty of Dental Medicine, Universitas Airlangga has approved the implementation of this study, and all respondents in this study had signed a written consent. This study was conducted among 40 dental technicians in Surabaya. Sampling was done by random sampling with sample criteria namely; working on dentures with metal-containing material mixed Ni, Co and, Cr for more than 3 years.3 All the participants were accepted all procedures such as saliva examination, answering questions and using PPE through a questionnaire. The criteria gender are not differentiated because gender differences have no significant effects on the levels of p53.12,13 The use of PPE during working hours by participants is via observations and questions using questionnaires and then do the scoring. Examinations of PPE include the frequency and manner of use of masks, gloves, goggles, laboratories work clothes, and shoes by a dental technician. Before taking saliva samples, subjects were instructed to not eat, smoke, antiseptic gargle one hour earlier.14 Each saliva sample was taken between at 10.00 a.m -13.00 p.m.12 The participants were asked to first collect their saliva and accumulated it in their mouth then instructed to spit 3cc into a tube. Each sample of saliva collected was centrifuged 2000 rpm for 10 minutes in order to obtain supernatan.12 The level of p53 wild was analysed at the Institute of Tropical Disease, Universitas Airlangga. p53 saliva levels were checked by indirect ELISA method using Human TP53 (tumor protein p53) ELISA kit (Elabscience Biotechnology Co., Wuhan, Hubei, China ). Rating score of PPE based on exposure categories main lines of metal that enters the body. Masks and gloves have the highest percentage due to exposure to the metal enters the body through three main channels; respiratory, oral, and skin.15 How to score PPE is based on research conducted by Risdayanti.16 Categories score of PPE usage frequenty score questionnaires are divided into: always, rarely, and never with respective scores of 100, 50, and 0. Categories score of PPE usage procedures are divided into: true, is not always true, and not true with respective scores of 100, 50, and 0. Summing scores PPE usage frequency and usage procedures PPE score. The score was multiplied by the percentage weighting of PPE. The percentages of weighting were 30% for mask , 25% gloves, 20% for goggles, 15% for laboratories work clothes and 10% for shoes. results Based on observations and questionnaires the use of PPE and p53 levels among dental technician in Surabaya. In Table 1 shows the average score PPE dental technicians in Surabaya is 86.19 ± 27.41. The average P53 wild score 10 Table 1. Mean and standard deviation score the use of PPE and levels of P53 wild Score PPE P53 N 40 40 Mean ± SD 86.19 ± 27.41 0.27 ± 0.28 Figure 1. Relationship P53wild levels with a score of use PPE. Table 2. Relationship scores the use of PPE and levels of wild P53 protein among dental technicians in Surabaya Independent variable Dependent variable P R Score PPE Level P53 0,002 0,466 * P<0.05 = significant correlation Figure 1. Relationship p53wild levels with a score of use PPE. To ta l s co re A PD Dental Journal (Majalah Kedokteran Gigi) p-ISSN: 1978-3728; e-ISSN: 2442-9740. Accredited No. 56/DIKTI/Kep./2012. Open access under CC-BY-SA license. Available at http://e-journal.unair.ac.id/index.php/MKG DOI: 10.20473/j.djmkg.v50.i1.p19-22 http://dx.doi.org/10.20473/j.djmkg.v50.i1.p19-22 2121Rohmaniar, et al./Dent. J. (Majalah Kedokteran Gigi) 2017 March; 50(1): 19–22 dental technicians is 0.27 ± 0.28. Figure 1 shows that levels p53 increase as increasing score of PPE. Table 2 shows the significance correlation test Pearson between score of PPE and level wild-type p53 saliva among dental technicians is p= 0,002 with significance value of p<0.05. the correlation Pearson value is R = 0.446 discussion Based on the results of this study showed there were positive correlation between the levels of wild-type p53 with a score of use of personal protective equipment at the dental technicians in Surabaya. If dental technician used PPE appropriately during work, so the levels of wild-type p53 was higher too. Because protective clothing, protective mask, protective gloves and goggles. can prevent metal exposure during work.1 This metal exposure can increase the number ROS endogenous.17 Increasing ROS can oxidize DNA and produce 8-hidroksi-deoksiguanosin (8-OH-dG). This products induce transversions mutation guanine into thymin,18 the same typical mutations occurs in p53 gene. 7 Cells that have mutations or loss of the p53 gene, the expression of p53 (p53 protein) might or might not occur p53 protein expression but if occurred it could not be functioning as an activating gene transcription in some target.10 The dental technicians in Surabaya, on average, did not use PPE properly during the work, so the wild-type p53 levels in saliva became lower than normal.12,19 This occurs because of exposure to the metal derived from a dental technician working environment can not be prevented through the use of personal protective equipments which include the use of masks, gloves, goggles, clothing and lab work, special shoes. When the use of personal protective equipment is adequate then it will reduce the level of chromium, cobalt, and nickel in blood.1 Masks can minimize the chemical exposure entering the respiratory tract. The results showed that the efficiency of face masks appropriate to reduce exposure to chemicals are inhaled to reach 70% -95%.20 Disposible latex gloves can inhibit the penetration of chemicals through the skin.21 .Goggles, protective clothing and shoes can reduce chemical exposure. 22,23,24 Nickel, chromium, and cobalt can enter to the body through of inhalation, oral and skin contact. Inhalation exposure is exposure to the metal through the respiratory tract. Inhalation exposure become the main route of exposure entry of metal into the human body.15,25 Digestion and inhalation exposure can accumulate in the oral cavity. The trachea and bronchi are covered by the ciliated epithelium and coated by a thin layer of mucus secreted from goblet cells. Cilia and mucus in the bronchi and trachea epithelial layer can be pushing up the particles that accumulated toward to the mouth surface. Particles that containing mucus then discharged from the respiratory tract with spit or swallow. Some particles fagosited by macrophages, but some are absorbed through the epithelial tissue and then diffuses and circulating into the blood vessel.26,27 Molecular components in the blood vessels can entry into saliva through transcellular pathway (passive diffusion and active transport) and paracellular pathways (ultrafiltration). The composition of saliva and then is secreted into transport molecules from blood to saliva.11 The other hand, nickel may activate hypoxia-signaling pathways by mediating transcription factor hypoxia induced factor-1 (HIF-1). Nickel plays a role in inactivating the enzyme prolyl hydroxylase, resulting in hipoksia.28 HIF-1 is downregulated by tumor suppressor protein p53 homeodomain-interacting protein kinase-2 (HIPK2).29 Genotoxic metal exposure can increase the number ROS endogenous. The metal ions of chromium, cobalt, and nickel can produce hydroxyl radicals (OH) through the Fenton and Haber- Weis reaction. Fenton reaction is the reaction of the transition metal ion with H2O2 to generate OH radicals and metal ions are oxidized. Chromium, cobalt and nickel are type reagent Fenton. Haber- Weiss reaction is a reaction consisting of oxidation of the metal ions are reduced by O2 and then react with H2O2 to produce OH- radicals. 17 The hydroxyl radical Fenton reaction and results from Haber-weiss can cause DNA damage.6 Typical damage caused by oxidation of DNA are guanine transversion into thymine in p53 (p53 gene).7 Cells that have mutations or loss of the p53 gene, the expression of p53 (p53 protein) does not occur or occurs p53 protein expression but can not function as activating gene transcription in some target.10 It can be concluded that the use PPE properly was positively correlated with the wild-type p53 protein levels in dental technicians, the use of PPE appropriately during work, prevented metal exposure, and decreased levels of p53. The contribution of PPE on the level of p53 wild on dental technicians was really significant. Table 1. Mean and standard deviation score the use of PPE and levels of wild-type p53 Score PPE p53 n 40 40 Mean ± SD 86.19 ± 27.41 0.27 ± 0.28 Table 2. Relationship scores the use of PPE and levels of wild-type p53 protein among dental technicians in Surabaya Independent variable Dependent variable P R Score PPE Level p53 0,002 0,466 * P<0.05 = significant correlation Dental Journal (Majalah Kedokteran Gigi) p-ISSN: 1978-3728; e-ISSN: 2442-9740. Accredited No. 56/DIKTI/Kep./2012. Open access under CC-BY-SA license. Available at http://e-journal.unair.ac.id/index.php/MKG DOI: 10.20473/j.djmkg.v50.i1.p19-22 http://dx.doi.org/10.20473/j.djmkg.v50.i1.p19-22 22 Rohmaniar, et al./Dent. J. 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Cross-talk between HIF and p53 as mediators of molecular responses to physiological and genotoxic stresses. Mol Cancer 2013; 12(93): p 2-7 Dental Journal (Majalah Kedokteran Gigi) p-ISSN: 1978-3728; e-ISSN: 2442-9740. Accredited No. 56/DIKTI/Kep./2012. Open access under CC-BY-SA license. Available at http://e-journal.unair.ac.id/index.php/MKG DOI: 10.20473/j.djmkg.v50.i1.p19-22 http://dx.doi.org/10.20473/j.djmkg.v50.i1.p19-22