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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Quantitative analysis of leaching of different metals in human saliva from dental casting alloys: An <italic>in vivo</italic>
study</title>
<author><name sortKey="Siddharth, Ramashanker" sort="Siddharth, Ramashanker" uniqKey="Siddharth R" first="Ramashanker" last="Siddharth">Ramashanker Siddharth</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Gautam, Roopali" sort="Gautam, Roopali" uniqKey="Gautam R" first="Roopali" last="Gautam">Roopali Gautam</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Chand, Pooran" sort="Chand, Pooran" uniqKey="Chand P" first="Pooran" last="Chand">Pooran Chand</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Agrawal, Kaushal Kishor" sort="Agrawal, Kaushal Kishor" uniqKey="Agrawal K" first="Kaushal Kishor" last="Agrawal">Kaushal Kishor Agrawal</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Singh, Raghuwar Dayal" sort="Singh, Raghuwar Dayal" uniqKey="Singh R" first="Raghuwar Dayal" last="Singh">Raghuwar Dayal Singh</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Singh, Balendra Pratap" sort="Singh, Balendra Pratap" uniqKey="Singh B" first="Balendra Pratap" last="Singh">Balendra Pratap Singh</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">26929514</idno>
<idno type="pmc">4762334</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4762334</idno>
<idno type="RBID">PMC:4762334</idno>
<idno type="doi">10.4103/0972-4052.164906</idno>
<date when="2015">2015</date>
<idno type="wicri:Area/Pmc/Corpus">000695</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000695</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Quantitative analysis of leaching of different metals in human saliva from dental casting alloys: An <italic>in vivo</italic>
study</title>
<author><name sortKey="Siddharth, Ramashanker" sort="Siddharth, Ramashanker" uniqKey="Siddharth R" first="Ramashanker" last="Siddharth">Ramashanker Siddharth</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Gautam, Roopali" sort="Gautam, Roopali" uniqKey="Gautam R" first="Roopali" last="Gautam">Roopali Gautam</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Chand, Pooran" sort="Chand, Pooran" uniqKey="Chand P" first="Pooran" last="Chand">Pooran Chand</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Agrawal, Kaushal Kishor" sort="Agrawal, Kaushal Kishor" uniqKey="Agrawal K" first="Kaushal Kishor" last="Agrawal">Kaushal Kishor Agrawal</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Singh, Raghuwar Dayal" sort="Singh, Raghuwar Dayal" uniqKey="Singh R" first="Raghuwar Dayal" last="Singh">Raghuwar Dayal Singh</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Singh, Balendra Pratap" sort="Singh, Balendra Pratap" uniqKey="Singh B" first="Balendra Pratap" last="Singh">Balendra Pratap Singh</name>
<affiliation><nlm:aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</nlm:aff>
</affiliation>
</author>
</analytic>
<series><title level="j">The Journal of the Indian Prosthodontic Society</title>
<idno type="ISSN">0972-4052</idno>
<idno type="eISSN">1998-4057</idno>
<imprint><date when="2015">2015</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><sec id="st1"><title>Background:</title>
<p>The issue of biomaterial-derived ionic release in various sites of the human body has attracted the interest of many investigators because of the possibility that debris or degradation products elicit a foreign body reaction or have a role in the induction of pathological processes.</p>
</sec>
<sec id="st2"><title>Purpose:</title>
<p>The purpose was to evaluate the saliva of denture wearers after insertion of the prosthesis for leaching of metals from metallic denture.</p>
</sec>
<sec id="st3"><title>Materials and Methods:</title>
<p>Total 20 subjects of age group of 40–60 years including both males (10) and females (10) were selected for the study. Total subjects were divided into 2 groups each containing 10 subjects, Group I (control group): Subjects having dentition intact up to second molar and free of any dental restoration; Group II (study group): Partially edentulous subjects rehabilitated with cast-metal removable partial denture. Saliva samples were taken at three stages that is, 1 h, 24 h and 72 h after the denture insertion from subjects of study group as well as from the control group. Atomic absorption spectroscopy (AAS) was used to estimate the concentration of elemental ions. Obtained data's were analyzed using SPSS (Statistical Package for Social Sciences) version 15.0 statistical analysis software. The values were represented in a number (%) and mean ± standard deviation.</p>
</sec>
<sec id="st4"><title>Results:</title>
<p>At 1 h, 24 h and 72 h after the denture insertion in study group, chromium (Cr) had statistically significant higher mean concentration as compared to manganese (Mn) (<italic>P</italic>
< 0.001). Cr had maximum concentration (0.1479 + 0.0052) immediately after denture insertion while maximum concentration of Mn (0.1479 + 0.0052) was found 24 h after denture insertion.</p>
</sec>
<sec id="st5"><title>Conclusion:</title>
<p>Metal-based dentures show maximum leaching immediately after wearing of the prosthesis which decreased significantly over the period of 3 days. Cr and Mn were the metal ions mainly found in saliva of cast partial denture wearer. No concentration of cobalt, molybdenum (Mo) and iron (Fe) was found in saliva of metal base denture wearer. There was a significant change in concentration of elutes in saliva in first 72 h/3 days making time an effective variable was observed.</p>
</sec>
</div>
</front>
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</author>
</analytic>
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<biblStruct><analytic><author><name sortKey="Anusance, Kj" uniqKey="Anusance K">KJ Anusance</name>
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</back>
</TEI>
<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Indian Prosthodont Soc</journal-id>
<journal-id journal-id-type="iso-abbrev">J Indian Prosthodont Soc</journal-id>
<journal-id journal-id-type="publisher-id">JIPS</journal-id>
<journal-title-group><journal-title>The Journal of the Indian Prosthodontic Society</journal-title>
</journal-title-group>
<issn pub-type="ppub">0972-4052</issn>
<issn pub-type="epub">1998-4057</issn>
<publisher><publisher-name>Medknow Publications & Media Pvt Ltd</publisher-name>
<publisher-loc>India</publisher-loc>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">26929514</article-id>
<article-id pub-id-type="pmc">4762334</article-id>
<article-id pub-id-type="publisher-id">JIPS-15-206</article-id>
<article-id pub-id-type="doi">10.4103/0972-4052.164906</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject>
</subj-group>
</article-categories>
<title-group><article-title>Quantitative analysis of leaching of different metals in human saliva from dental casting alloys: An <italic>in vivo</italic>
study</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Siddharth</surname>
<given-names>Ramashanker</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
<xref ref-type="corresp" rid="cor1"></xref>
</contrib>
<contrib contrib-type="author"><name><surname>Gautam</surname>
<given-names>Roopali</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
</contrib>
<contrib contrib-type="author"><name><surname>Chand</surname>
<given-names>Pooran</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
</contrib>
<contrib contrib-type="author"><name><surname>Agrawal</surname>
<given-names>Kaushal Kishor</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
</contrib>
<contrib contrib-type="author"><name><surname>Singh</surname>
<given-names>Raghuwar Dayal</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
</contrib>
<contrib contrib-type="author"><name><surname>Singh</surname>
<given-names>Balendra Pratap</given-names>
</name>
<xref ref-type="aff" rid="aff1"></xref>
</contrib>
</contrib-group>
<aff id="aff1">Department of Prosthodontics, King George Medical University, Lucknow, Uttar Pradesh, India</aff>
<author-notes><corresp id="cor1"><bold>Address for correspondence:</bold>
Dr. Ramashanker Siddharth, King George Medical University, Lucknow, Uttar Pradesh - 226 003, India. E-mail: <email xlink:href="ramashanker09@rediffmail.com">ramashanker09@rediffmail.com</email>
</corresp>
</author-notes>
<pub-date pub-type="ppub"><season>Jul-Sep</season>
<year>2015</year>
</pub-date>
<volume>15</volume>
<issue>3</issue>
<fpage>206</fpage>
<lpage>210</lpage>
<history><date date-type="received"><day>18</day>
<month>7</month>
<year>2014</year>
</date>
<date date-type="accepted"><day>21</day>
<month>12</month>
<year>2014</year>
</date>
</history>
<permissions><copyright-statement>Copyright: © 2015 The Journal of Indian Prosthodontic Society</copyright-statement>
<copyright-year>2015</copyright-year>
<license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc-sa/3.0"><license-p>This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.</license-p>
</license>
</permissions>
<abstract><sec id="st1"><title>Background:</title>
<p>The issue of biomaterial-derived ionic release in various sites of the human body has attracted the interest of many investigators because of the possibility that debris or degradation products elicit a foreign body reaction or have a role in the induction of pathological processes.</p>
</sec>
<sec id="st2"><title>Purpose:</title>
<p>The purpose was to evaluate the saliva of denture wearers after insertion of the prosthesis for leaching of metals from metallic denture.</p>
</sec>
<sec id="st3"><title>Materials and Methods:</title>
<p>Total 20 subjects of age group of 40–60 years including both males (10) and females (10) were selected for the study. Total subjects were divided into 2 groups each containing 10 subjects, Group I (control group): Subjects having dentition intact up to second molar and free of any dental restoration; Group II (study group): Partially edentulous subjects rehabilitated with cast-metal removable partial denture. Saliva samples were taken at three stages that is, 1 h, 24 h and 72 h after the denture insertion from subjects of study group as well as from the control group. Atomic absorption spectroscopy (AAS) was used to estimate the concentration of elemental ions. Obtained data's were analyzed using SPSS (Statistical Package for Social Sciences) version 15.0 statistical analysis software. The values were represented in a number (%) and mean ± standard deviation.</p>
</sec>
<sec id="st4"><title>Results:</title>
<p>At 1 h, 24 h and 72 h after the denture insertion in study group, chromium (Cr) had statistically significant higher mean concentration as compared to manganese (Mn) (<italic>P</italic>
< 0.001). Cr had maximum concentration (0.1479 + 0.0052) immediately after denture insertion while maximum concentration of Mn (0.1479 + 0.0052) was found 24 h after denture insertion.</p>
</sec>
<sec id="st5"><title>Conclusion:</title>
<p>Metal-based dentures show maximum leaching immediately after wearing of the prosthesis which decreased significantly over the period of 3 days. Cr and Mn were the metal ions mainly found in saliva of cast partial denture wearer. No concentration of cobalt, molybdenum (Mo) and iron (Fe) was found in saliva of metal base denture wearer. There was a significant change in concentration of elutes in saliva in first 72 h/3 days making time an effective variable was observed.</p>
</sec>
</abstract>
<kwd-group><title>Key Words</title>
<kwd>Dental casting alloys</kwd>
<kwd>leaching</kwd>
<kwd>saliva</kwd>
</kwd-group>
</article-meta>
</front>
<body><sec sec-type="intro" id="sec1-1"><title>INTRODUCTION</title>
<p>The issue of biomaterial-derived ionic release in various sites of the human body has attracted the interest of many investigators because of the possibility that debris or degradation products elicit a foreign body reaction or have a role in the induction of pathological processes. Thus, research in the broader biomedical literature has focused on the release of substances from various devices including orthopedic and dental materials.[<xref rid="ref1" ref-type="bibr">1</xref>
] Faber (1957) suggested that a cast metal base is suitable particularly for lower denture-based.[<xref rid="ref2" ref-type="bibr">2</xref>
] Taylor (1962) claimed that metal denture bases accurately reproduce the details that have been recorded in the impression without distortion or warpage.[<xref rid="ref3" ref-type="bibr">3</xref>
]</p>
<p>Traditionally gold alloys have been most popular for the fabrication of metal denture bases. They are well tolerated by the patient and do not corrode in the oral environment. However, search for the economic alternatives lead to the evolution of base metal alloys. The use of cobalt-chromium (Co-Cr) alloys is increasing rapidly especially in the field of partial dentures. The composition of the various alloys varied considerably, but the basic pattern is cobalt (Co) 63–68% and chromium (Cr) 25–30% with addition of minor elements such as molybedinium (Mo), manganese (Mn), carbon (C), iron (Fe), silicon (Si), etc.[<xref rid="ref4" ref-type="bibr">4</xref>
]</p>
<p>The complexity and diversity of these alloys make understanding their biocompatibility difficult because any element in an alloy may be released and may influence the body. Release of elements from the alloy is nearly always necessary for adverse biologic effects such as toxicity, allergy, or mutagenicity. The biologic response to released elements depends on which element is released, the quantity released, and the duration of exposure to tissues.</p>
<p>Literature regarding the biocompatibility of denture-based materials mainly consists of <italic>in vitro</italic>
studies. Although, intra-orally different factors such as saliva characteristics, chewing or thermal and chemical dietary changes may influence the biological behavior of denture base materials. Therefore, this study was done to evaluate the saliva of cast partial denture wearers after insertion of the prosthesis for leaching of metals from Co-Cr base metal alloys. It was hypothesized that there should not be leaching of metallic ions from cast partial denture base in human saliva environment.</p>
</sec>
<sec sec-type="materials|methods" id="sec1-2"><title>MATERIALS AND METHODS</title>
<p>This <italic>in vivo</italic>
study was conducted in Department of Prosthodontics, faculty of dental sciences, King George Medical University Lucknow in collaboration with Indian Institute of Toxicology Research (IITR), Lucknow. Total 20 subjects of age group of 40–60 years including both males (10) and females (10) were selected for the study. Subjects had good oral hygiene (S-OHI < 1.3–3) and were free of any dental restoration. Subjects are having history of using any dental prosthesis in past or presently, working in industries related to any metal work, and/or taking any kind of medications affecting salivary flow rate were excluded from the study. Total subjects were divided into two groups each containing 10 subjects:</p>
<p><list list-type="bullet"><list-item><p>Group I (control group): Dentulous subjects without any dental restoration</p>
</list-item>
<list-item><p>Group II (study group): Partially edentulous subjects with Kennedy's class II in the mandibular arch and rehabilitated with cast-metal removable partial denture (RPD) [<xref ref-type="fig" rid="F1">Figure 1</xref>
]. Ethical clearance and informed consent were taken from each subject participating in the study.</p>
</list-item>
</list>
</p>
<fig id="F1" position="float"><label>Figure 1</label>
<caption><p>Cast Framework try-in done in patient's mouth</p>
</caption>
<graphic xlink:href="JIPS-15-206-g001"></graphic>
</fig>
<p>Cast partial RPD framework for each subject of Group II was fabricated using Co-Cr alloy (Wironit, BEGO Bremer Goldschlägerei Wilh. Herbst GmbH and Co., Germany) by the conventional technique.</p>
<p>After recovering the castings from the investment, they were blasted with 50 μ aluminum oxide particles at 80 psi and were ultrasonically cleaned. Polishing of cast partial RPD frameworks was first done electrolytically in an electrolytic polishing device. Oxides are removed evenly from all surfaces, but it did not result in high gloss. Finally, mechanical polishing was done using rubbers and brushes for polishing with paste to obtain a high gloss. For the purpose of standardization, it was confirmed that the weight of the cast partial RPD framework was kept same (20 g) for all the subjects. Cast Framework try-in was done in patient's mouth [<xref ref-type="fig" rid="F1">Figure 1</xref>
]. Jaw relations were recorded. Teeth setting and try-in were done. After flasking and curing with heat-cured resin, cast partial RPD was finished, polished and inserted in patient's mouth [<xref ref-type="fig" rid="F2">Figure 2</xref>
]. Postoperative instructions were given. All the subjects were instructed to wear the prosthesis 12 h/day.</p>
<p><inline-graphic xlink:href="JIPS-15-206-g002.jpg"></inline-graphic>
</p>
<fig id="F2" position="float"><label>Figure 2</label>
<caption><p>Cast partial RPD inserted in patient's mouth</p>
</caption>
<graphic xlink:href="JIPS-15-206-g003"></graphic>
</fig>
<p>Patients were asked to spit every 30 s into preweighed screw-capped containers for 5-min period.[<xref rid="ref5" ref-type="bibr">5</xref>
] The containers were re-weighed to estimate the salivary flow rate, assuming that 1 ml of saliva weighed 1 g. Samples were taken at three stages:</p>
<p><list list-type="bullet"><list-item><p>Stage I: 1 h after the insertion – sample x</p>
</list-item>
<list-item><p>Stage II: 1-day after the insertion – sample y</p>
</list-item>
<list-item><p>Stage III: 3 days after the insertion – sample z</p>
</list-item>
</list>
</p>
<p>All the saliva samples were collected at the same time every day. Three samples were taken for each patient. Samples were sent immediately after collection to IITR Lucknow in an insulated box containing ice cubes to maintain the temperature below 0°C and were stored at −20°C in a refrigerator before starting the analysis.</p>
<p>Saliva sample was transferred to a conical flask and digested on a hot plate with 1% of HNO<sub>3</sub>
. After dry-up, 1% HNO<sub>3</sub>
was added to make up 10 ml volume. After that, sample was transferred in a test tube to atomic absorption spectrophotometer (Awanta sigma; GBC, Australia), as shown in <xref ref-type="fig" rid="F3">Figure 3</xref>
, for estimation of metal ion concentration.[<xref rid="ref6" ref-type="bibr">6</xref>
] Atomic absorption spectroscopy was used to estimate the concentration of elemental ions that is, chromium (Cr), manganese (Mn), cobalt (Co), molybdenum (Mo) and iron (Fe) leaching from Co-Cr alloy in saliva of cast partial denture wearer. With AAS, it is possible to determine about 70 elements (mainly metals) at very low concentrations. The sample is atomized at a very high temperature (2500°C–3000°C), and the free atoms have line spectrum. It means that they can only absorb the energy of light at discrete energy levels according to the excitations of electrons. Only a light with a concrete wavelength belongs to each of these excitation energies and when this light is absorbed it is missing from the continuous spectra of the electromagnetic radiation that is, a black line appears in the absorption spectrum of the atom. Only the examined atoms can absorb it. As a result of absorption, the intensity of light decreases, this is proportional to the number of the examined atoms being present. That makes very sensitive quantitative measurements possible. Following the table shows the metal ions for which saliva sample was tested and their corresponding wavelength of detection:</p>
<fig id="F3" position="float"><label>Figure 3</label>
<caption><p>Atomic absorption spectrophotometer</p>
</caption>
<graphic xlink:href="JIPS-15-206-g004"></graphic>
</fig>
<p>The statistical analysis was done using SPSS (Statistical Package for Social Sciences) version 15.0 statistical analysis software. The values were represented in a number (%) and mean ± standard deviation.</p>
</sec>
<sec id="sec1-3"><title>OBSERVATION AND RESULT</title>
<p>Only Cr and Mn were isolated; none of the other trace elements could be isolated on any time interval in saliva of subjects rehabilitated with cast partial denture. Furthermore, none of these ions that is, Cr and Mn were detected in saliva of subjects acting as control. Before the assessment of data, the distribution was checked for normality using Kolmogorov–Smirnov test and was found to be asymmetric and not normal (<italic>P</italic>
> 0.05). Hence, a nonparametric analytical plan was used.</p>
<p><xref ref-type="table" rid="T1">Table 1</xref>
compares the pattern of change in Cr and Mn metal levels at different time intervals during the study. At 1 h, 24 h and 72 h after the denture insertion in study group, Chromium (Cr) had statistically significant higher mean concentration as compared to Manganese (Mn) (<italic>P</italic>
< 0.001). A gradual decrease in mean Cr levels was observed from 1 h after denture insertion to day 3 intervals with a maximum value at 1 h and minimum value at day 3 intervals. However, for Mn, the mean value was maximum at day 1 and minimum at day 3 intervals. <xref ref-type="table" rid="T2">Table 2</xref>
shows a comparison of change in mean levels of Cr and Mn with time using Kruskall Wallis test (nonparametric ANOVA). It indicates a significant difference in metal ion concentrations at different time intervals for both Cr (χ<sup>2</sup>
= 20.589; <italic>P</italic>
< 0.001) as well as Mn (χ<sup>2</sup>
= 23.396; <italic>P</italic>
< 0.001).</p>
<p><inline-graphic xlink:href="JIPS-15-206-g005.jpg"></inline-graphic>
</p>
<table-wrap id="T1" position="float"><label>Table 1</label>
<caption><p>Comparison of change in metal levels (microgram/milliliter) on different time intervals</p>
</caption>
<graphic xlink:href="JIPS-15-206-g006"></graphic>
</table-wrap>
<table-wrap id="T2" position="float"><label>Table 2</label>
<caption><p>Analysis of variance (Kruskall–Wallis test) for concentration of metal ions at different time intervals</p>
</caption>
<graphic xlink:href="JIPS-15-206-g007"></graphic>
</table-wrap>
<p>Between time-interval comparison using Mann–Whitney U-test [<xref ref-type="table" rid="T3">Table 3</xref>
] revealed that except for day 1 and day 3 comparisons, all the between time interval comparisons for Mn was found to be significant statistically (<italic>P</italic>
< 0.05). The order of concentration for two metals was as follows: Cr: 1 h > day 1 ≃ day 3 Mn: Day 1 > 1 h > day 3.</p>
<table-wrap id="T3" position="float"><label>Table 3</label>
<caption><p>Between time-interval comparison of metal concentrations (Mann–Whitney U-test)</p>
</caption>
<graphic xlink:href="JIPS-15-206-g008"></graphic>
</table-wrap>
</sec>
<sec sec-type="discussion" id="sec1-4"><title>DISCUSSION</title>
<p>There are sizeable literatures on <italic>in vitro</italic>
release studies of dental casting alloys, but experiments using human saliva are rare, and few <italic>in vivo</italic>
studies have been reported.[<xref rid="ref7" ref-type="bibr">7</xref>
] Obtained results from this <italic>in vivo</italic>
study rejected the null hypothesis and showed that there is leaching of metal ions from cast partial denture frameworks in human saliva. Cr and Mn were the metal ions mainly found in saliva of the denture wearer.</p>
<p>Biting stresses on dentures can be extremely high, temperatures may fluctuate between 25°C and 45°C and pH may change instantaneously from acidic to alkaline. The warm and moist oral environment, which is also enzyme and bacteria rich, is conducive to further decay.[<xref rid="ref8" ref-type="bibr">8</xref>
] Allergic and/or inflammatory reactions from dental casting alloys have been ascribed mainly due to leaching of metal ions from denture-based material; for sensitization to occur, these metal ions must presumably be released from the prosthesis into saliva, in which it is conveyed to the oral mucosa or, after being swallowed, to the gastro-intestinal tract.[<xref rid="ref9" ref-type="bibr">9</xref>
] Prosthesis was inserted on the same day of curing to minimize any changes (physical and chemical) occurring due to storage of prosthesis.[<xref rid="ref10" ref-type="bibr">10</xref>
] Saliva samples were collected 3 times that is, 1 h, 1 day and 3 days after insertion of the denture as these timings coincide with postinsertion recall visits of the patients.</p>
<p>For the detection of release of elemental ions, atomic absorption spectrophotometry was employed. Atomic absorption spectrophotometer (AAS)[<xref rid="ref7" ref-type="bibr">7</xref>
] was used as opposed to inductively coupled plasma mass spectroscopy because of simplicity, speed and economy of AAS technique. This technique has following advantage over other techniques: High sensitivity (10–10 g [flame], 10–14 g [nonflame]), good accuracy (relative error 0.1% ~ 0.5%), and high selectivity. A resonance line source is required for each element to be determined.[<xref rid="ref11" ref-type="bibr">11</xref>
]</p>
<p>After running the saliva sample in AAS, release of Cr, Mn was found primarily over the period of 3 days while Co and Mo despite being one of the main components were not found in the samples [<xref ref-type="table" rid="T1">Table 1</xref>
]. It has been seen that two factors affect the degree of release of one or more metal ions: Alloy type and saliva pH.[<xref rid="ref12" ref-type="bibr">12</xref>
] When exposed to the oral environment for prolonged periods of time dental alloys are susceptible, in varying degrees, to tarnish. This tendency to tarnish is usually attributed to chemical or electrochemical corrosion and is responsible for the release of elemental ions.[<xref rid="ref13" ref-type="bibr">13</xref>
] <italic>In vivo</italic>
, numerous biological factors, including organic acids and enzymes which are produced by oral micro-organisms or are contained in food, may contribute to alloy corrosion. It must be emphasized that the <italic>in vitro</italic>
data do not necessarily reflect the <italic>in vivo</italic>
clinical conditions.[<xref rid="ref14" ref-type="bibr">14</xref>
]</p>
<p>Release of Cr was maximum (0.1479 μg/ml) in sample taken 1 h after insertion and it decreased further over 1-day (0.0622 μg/ml) and was minimum (0.0586 pg/ml) on the 3<sup>rd</sup>
day of prosthesis wearing. The difference in the metal ion concentration of Cr was significant (<italic>P</italic>
< 0.001) over the period of 3 days [<xref ref-type="table" rid="T1">Table 1</xref>
]. As most of the Cr ions have leached immediately after wearing of the prosthesis, this may be because surface layer of these alloy mainly have oxides of Cr as protective layer causing passivation and as time of prosthesis wearing increases this layer decreases.[<xref rid="ref14" ref-type="bibr">14</xref>
] However, Cr is not mutagenic, it is carcinogenic.[<xref rid="ref9" ref-type="bibr">9</xref>
] However, even the maximum value of Cr concentration, as found in study, does not exceed daily dietary intake of Cr (Wataha, 2000).[<xref rid="ref15" ref-type="bibr">15</xref>
]</p>
<p>It was noted that the release of Mn was maximum (0.0065pg/ml) after 1-day as seen and it decreased significantly (<italic>P</italic>
< 0.001) after 3<sup>rd</sup>
day [<xref ref-type="table" rid="T1">Table 1</xref>
]. Despite being only 1% of total alloy composition, it was found in saliva. Although the literature shows that the relative amount of released metals does not reflect their relative weight or volume portion in the alloy.[<xref rid="ref15" ref-type="bibr">15</xref>
]</p>
<p>It was found in the study that the overall concentration of both the metals that is, Cr and Mn decreased significantly after 3 days of prosthesis wearing. However, the concentration of chromium was significantly higher than Mn at all the time intervals, making Cr the main leaching metal ion. This may be due to much higher content of the Cr in alloy than Mn and Cr being more reactive than Mn.[<xref rid="ref16" ref-type="bibr">16</xref>
]</p>
<p>Despite being main component, Co and Mo were not found in saliva over 3 days period. It has been shown that for Co-Cr- Mo dental alloys Cr forms the protective oxide layer that prevents the corrosion of alloy. This passivating effect is further enhanced by the Mo.[<xref rid="ref14" ref-type="bibr">14</xref>
] As seen in study concentration of Co and Mo is not found initially it may be found as the usage period of partial denture increases.[<xref rid="ref12" ref-type="bibr">12</xref>
<xref rid="ref17" ref-type="bibr">17</xref>
]</p>
<p>Maximum release of elemental ions from cast partial denture was seen 1 h after insertion [<xref ref-type="table" rid="T3">Table 3</xref>
]. Conditioning cast dental restorations before placing them intra-orally might reduce the release of elements inside the oral cavity.[<xref rid="ref14" ref-type="bibr">14</xref>
] <italic>In vitro</italic>
studies have shown some conditioning procedures to be useful because they remove the initial (labile) elements.[<xref rid="ref13" ref-type="bibr">13</xref>
] Previous <italic>in vitro</italic>
studies have used various conditioning times and types of conditioning media.[<xref rid="ref17" ref-type="bibr">17</xref>
] The release of elements into conditioning media varies among alloys and elements. Previous work showed that in alloys conditioned for longer periods, the degree of element release becomes lower over time.[<xref rid="ref14" ref-type="bibr">14</xref>
<xref rid="ref18" ref-type="bibr">18</xref>
]</p>
<p>It was found in the study that significant leaching was found in saliva from denture base materials, but as for every study, this study also has certain limitations. The time period over which leaching was studied that is, 72 h was less. As because of biodegradation of metal denture-based over the time, certain elutes which were not found in saliva initially may become detectable with further use of the prosthesis. A further cytotoxic evaluation of elutes leaching in the concentration found in the study is required to complete the study.</p>
</sec>
<sec sec-type="conclusion" id="sec1-5"><title>CONCLUSION</title>
<p>Metal-based dentures show maximum leaching immediately after wearing of the prosthesis which decreased significantly over the period of 3 days. Cr and Mn were the metal ions mainly found in saliva of cast partial denture wearer. No concentration of cobalt (Co), molybdenum (Mo) and iron (Fe) was found in saliva of metal base denture wearer. There was a significant change in concentration of elutes in saliva in first 72 h/3 days making time an effective variable was observed.</p>
</sec>
</body>
<back><fn-group><fn fn-type="supported-by"><p><bold>Source of Support:</bold>
Nil</p>
</fn>
<fn fn-type="conflict"><p><bold>Conflict of Interest:</bold>
None.</p>
</fn>
</fn-group>
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