Adherence of Candida to complete denture surfaces in vitro: A comparison of conventional and CAD/CAM complete dentures
Identifieur interne : 002F34 ( Pmc/Corpus ); précédent : 002F33; suivant : 002F35Adherence of Candida to complete denture surfaces in vitro: A comparison of conventional and CAD/CAM complete dentures
Auteurs : Afnan F. Al-Fouzan ; Lamya A. Al-Mejrad ; Ahmed M. AlbarragSource :
- The Journal of Advanced Prosthodontics [ 2005-7806 ] ; 2017.
Abstract
The goal of this study was to compare the adhesion of
Twenty discs of acrylic resin poly (methyl methacrylate) were fabricated with CAD/CAM and conventional procedures (heat-polymerized acrylic resin). The specimens were divided into two groups: 10 discs were fabricated using the CAD/CAM procedure (Wieland Digital Denture Ivoclar Vivadent), and 10 discs were fabricated using a conventional flasking and pressure-pack technique.
There was a significant difference in the adhesion of
The CAD/CAM procedure for fabricating complete dentures showed promising potential for reducing the adherence of
Url:
DOI: 10.4047/jap.2017.9.5.402
PubMed: 29142649
PubMed Central: 5673618
Links to Exploration step
PMC:5673618Le document en format XML
<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Adherence of <italic>Candida</italic> to complete denture surfaces <italic>in vitro</italic>: A comparison of conventional and CAD/CAM complete dentures</title><author><name sortKey="Al Fouzan, Afnan F" sort="Al Fouzan, Afnan F" uniqKey="Al Fouzan A" first="Afnan F." last="Al-Fouzan">Afnan F. Al-Fouzan</name><affiliation><nlm:aff id="A1">Prosthetic Dental Science, College of Dentistry, King Saud University, Riyadh, Saudi Arabia.</nlm:aff></affiliation></author><author><name sortKey="Al Mejrad, Lamya A" sort="Al Mejrad, Lamya A" uniqKey="Al Mejrad L" first="Lamya A." last="Al-Mejrad">Lamya A. Al-Mejrad</name><affiliation><nlm:aff id="A1">Prosthetic Dental Science, College of Dentistry, King Saud University, Riyadh, Saudi Arabia.</nlm:aff></affiliation></author><author><name sortKey="Albarrag, Ahmed M" sort="Albarrag, Ahmed M" uniqKey="Albarrag A" first="Ahmed M." last="Albarrag">Ahmed M. Albarrag</name><affiliation><nlm:aff id="A2">Department of Pathology, School of Medicine, King Saud University, Riyadh, Saudi Arabia.</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">29142649</idno><idno type="pmc">5673618</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5673618</idno><idno type="RBID">PMC:5673618</idno><idno type="doi">10.4047/jap.2017.9.5.402</idno><date when="2017">2017</date><idno type="wicri:Area/Pmc/Corpus">002F34</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">002F34</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Adherence of <italic>Candida</italic> to complete denture surfaces <italic>in vitro</italic>: A comparison of conventional and CAD/CAM complete dentures</title><author><name sortKey="Al Fouzan, Afnan F" sort="Al Fouzan, Afnan F" uniqKey="Al Fouzan A" first="Afnan F." last="Al-Fouzan">Afnan F. Al-Fouzan</name><affiliation><nlm:aff id="A1">Prosthetic Dental Science, College of Dentistry, King Saud University, Riyadh, Saudi Arabia.</nlm:aff></affiliation></author><author><name sortKey="Al Mejrad, Lamya A" sort="Al Mejrad, Lamya A" uniqKey="Al Mejrad L" first="Lamya A." last="Al-Mejrad">Lamya A. Al-Mejrad</name><affiliation><nlm:aff id="A1">Prosthetic Dental Science, College of Dentistry, King Saud University, Riyadh, Saudi Arabia.</nlm:aff></affiliation></author><author><name sortKey="Albarrag, Ahmed M" sort="Albarrag, Ahmed M" uniqKey="Albarrag A" first="Ahmed M." last="Albarrag">Ahmed M. Albarrag</name><affiliation><nlm:aff id="A2">Department of Pathology, School of Medicine, King Saud University, Riyadh, Saudi Arabia.</nlm:aff></affiliation></author></analytic><series><title level="j">The Journal of Advanced Prosthodontics</title><idno type="ISSN">2005-7806</idno><idno type="eISSN">2005-7814</idno><imprint><date when="2017">2017</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><sec><title>PURPOSE</title><p>The goal of this study was to compare the adhesion of <italic>Candida albicans</italic> to the surfaces of CAD/CAM and conventionally fabricated complete denture bases.</p></sec><sec><title>MATERIALS AND METHODS</title><p>Twenty discs of acrylic resin poly (methyl methacrylate) were fabricated with CAD/CAM and conventional procedures (heat-polymerized acrylic resin). The specimens were divided into two groups: 10 discs were fabricated using the CAD/CAM procedure (Wieland Digital Denture Ivoclar Vivadent), and 10 discs were fabricated using a conventional flasking and pressure-pack technique. <italic>Candida</italic> colonization was performed on all the specimens using four <italic>Candida albicans</italic> isolates. The difference in <italic>Candida albicans</italic> adhesion on the discs was evaluated. The number of adherent yeast cells was calculated by the colony-forming units (CFU) and by Fluorescence microscopy.</p></sec><sec><title>RESULTS</title><p>There was a significant difference in the adhesion of <italic>Candida albicans</italic> to the complete denture bases created with CAD/CAM and the adhesion to those created with the conventional procedure. The CAD/CAM denture bases exhibited less adhesion of <italic>Candida albicans</italic> than did the denture bases created with the conventional procedure (<italic>P</italic><.05).</p></sec><sec><title>CONCLUSION</title><p>The CAD/CAM procedure for fabricating complete dentures showed promising potential for reducing the adherence of <italic>Candida</italic> to the denture base surface. Clinical Implications. Complete dentures made with the CAD/CAM procedure might decrease the incidence of denture stomatitis compared with conventional dentures.</p></sec></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Cunha Cruz, J" uniqKey="Cunha Cruz J">J Cunha-Cruz</name></author><author><name sortKey="Hujoel, Pp" uniqKey="Hujoel P">PP Hujoel</name></author><author><name sortKey="Nadanovsky, P" uniqKey="Nadanovsky P">P Nadanovsky</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Emami, E" uniqKey="Emami E">E Emami</name></author><author><name sortKey="De Souza, Rf" uniqKey="De Souza R">RF de Souza</name></author><author><name sortKey="Kabawat, M" uniqKey="Kabawat M">M Kabawat</name></author><author><name sortKey="Feine, Js" uniqKey="Feine J">JS Feine</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Dye, Ba" uniqKey="Dye B">BA Dye</name></author><author><name sortKey="Li, X" uniqKey="Li 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<front><journal-meta><journal-id journal-id-type="nlm-ta">J Adv Prosthodont</journal-id><journal-id journal-id-type="iso-abbrev">J Adv Prosthodont</journal-id><journal-id journal-id-type="publisher-id">JAP</journal-id><journal-title-group><journal-title>The Journal of Advanced Prosthodontics</journal-title></journal-title-group><issn pub-type="ppub">2005-7806</issn><issn pub-type="epub">2005-7814</issn><publisher><publisher-name>The Korean Academy of Prosthodontics</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">29142649</article-id><article-id pub-id-type="pmc">5673618</article-id><article-id pub-id-type="doi">10.4047/jap.2017.9.5.402</article-id><article-categories><subj-group subj-group-type="heading"><subject>Original Article</subject></subj-group></article-categories><title-group><article-title>Adherence of <italic>Candida</italic> to complete denture surfaces <italic>in vitro</italic>: A comparison of conventional and CAD/CAM complete dentures</article-title></title-group><contrib-group><contrib contrib-type="author" corresp="yes"><contrib-id contrib-id-type="orcid" authenticated="true">https://orcid.org/0000-0003-2535-4641</contrib-id><name><surname>Al-Fouzan</surname><given-names>Afnan F.</given-names></name><xref ref-type="aff" rid="A1">1</xref></contrib><contrib contrib-type="author"><name><surname>Al-mejrad</surname><given-names>Lamya A.</given-names></name><xref ref-type="aff" rid="A1">1</xref></contrib><contrib contrib-type="author"><name><surname>Albarrag</surname><given-names>Ahmed M.</given-names></name><xref ref-type="aff" rid="A2">2</xref></contrib></contrib-group><aff id="A1"><label>1</label>Prosthetic Dental Science, College of Dentistry, King Saud University, Riyadh, Saudi Arabia.</aff><aff id="A2"><label>2</label>Department of Pathology, School of Medicine, King Saud University, Riyadh, Saudi Arabia.</aff><author-notes><corresp>Corresponding author: Afnan F. Al-Fouzan. Department of Prosthodontics, College of Dentistry, King Saud University P O Box 89300, Riyadh 11682, Saudi Arabia. Tel. +966118055645: <email>aalfouzan@ksu.edu.sa</email></corresp></author-notes><pub-date pub-type="ppub"><month>10</month><year>2017</year></pub-date><pub-date pub-type="epub"><day>16</day><month>10</month><year>2017</year></pub-date><volume>9</volume><issue>5</issue><fpage>402</fpage><lpage>408</lpage><history><date date-type="received"><day>07</day><month>2</month><year>2017</year></date><date date-type="rev-recd"><day>22</day><month>4</month><year>2017</year></date><date date-type="accepted"><day>04</day><month>7</month><year>2017</year></date></history><permissions><copyright-statement>© 2017 The Korean Academy of Prosthodontics</copyright-statement><copyright-year>2017</copyright-year><copyright-holder>The Korean Academy of Prosthodontics</copyright-holder><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by-nc/3.0/"><license-p>This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by-nc/3.0">http://creativecommons.org/licenses/by-nc/3.0</ext-link>) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p></license></permissions><abstract><sec><title>PURPOSE</title><p>The goal of this study was to compare the adhesion of <italic>Candida albicans</italic> to the surfaces of CAD/CAM and conventionally fabricated complete denture bases.</p></sec><sec><title>MATERIALS AND METHODS</title><p>Twenty discs of acrylic resin poly (methyl methacrylate) were fabricated with CAD/CAM and conventional procedures (heat-polymerized acrylic resin). The specimens were divided into two groups: 10 discs were fabricated using the CAD/CAM procedure (Wieland Digital Denture Ivoclar Vivadent), and 10 discs were fabricated using a conventional flasking and pressure-pack technique. <italic>Candida</italic> colonization was performed on all the specimens using four <italic>Candida albicans</italic> isolates. The difference in <italic>Candida albicans</italic> adhesion on the discs was evaluated. The number of adherent yeast cells was calculated by the colony-forming units (CFU) and by Fluorescence microscopy.</p></sec><sec><title>RESULTS</title><p>There was a significant difference in the adhesion of <italic>Candida albicans</italic> to the complete denture bases created with CAD/CAM and the adhesion to those created with the conventional procedure. The CAD/CAM denture bases exhibited less adhesion of <italic>Candida albicans</italic> than did the denture bases created with the conventional procedure (<italic>P</italic><.05).</p></sec><sec><title>CONCLUSION</title><p>The CAD/CAM procedure for fabricating complete dentures showed promising potential for reducing the adherence of <italic>Candida</italic> to the denture base surface. Clinical Implications. Complete dentures made with the CAD/CAM procedure might decrease the incidence of denture stomatitis compared with conventional dentures.</p></sec></abstract><kwd-group><kwd>Complete denture</kwd><kwd>CAD/CAM</kwd><kwd><italic>Candida</italic></kwd><kwd>Adhesion</kwd><kwd>Edentulous</kwd><kwd>Roughness</kwd></kwd-group><funding-group><award-group><funding-source><institution-wrap><institution>Female Center for Scientific and Medical Colleges, King Saud University</institution><institution-id institution-id-type="CrossRef">http://dx.doi.org/10.13039/501100005328</institution-id></institution-wrap></funding-source></award-group></funding-group></article-meta></front><body><sec sec-type="intro"><title>INTRODUCTION</title><p>Edentulism is a “debilitating and irreversible condition and is described as the final marker of disease burden for oral health.”<xref rid="B1" ref-type="bibr">1</xref><xref rid="B2" ref-type="bibr">2</xref> Among older adults, edentulism is considered one of the most prevalent condition worldwide<xref rid="B3" ref-type="bibr">3</xref> although the percentage of tooth loss has decreased in recent years.<xref rid="B4" ref-type="bibr">4</xref> In the U.S., the prevalence of edentulism is 15% in patients between the ages of 65 and 74 years old and 22% in patients older than 75 years according to the National Health and Nutrition Examination Survey.<xref rid="B3" ref-type="bibr">3</xref> Emami et al.<xref rid="B2" ref-type="bibr">2</xref> reported that the majority of edentulous patients wear one or two complete dentures. The use of removable prostheses has increased due to the increasing number of older patients, who are the primary wearers of dentures, in the general population.<xref rid="B4" ref-type="bibr">4</xref> Being edentulous influences oral health and general well-being.<xref rid="B2" ref-type="bibr">2</xref> The effects on oral health include impaired masticatory efficiency and denture-related oral lesions, such as angular cheilitis, traumatic ulcers and denture stomatitis (DS).<xref rid="B5" ref-type="bibr">5</xref><xref rid="B6" ref-type="bibr">6</xref> DS is an inflammation of the mucosa underlying a removable prosthesis.<xref rid="B7" ref-type="bibr">7</xref> Shulman et al.<xref rid="B8" ref-type="bibr">8</xref> conducted a study in the U.S. and found that the prevalence of DS among denture wearers was 28%.<xref rid="B8" ref-type="bibr">8</xref> Another epidemiological study reported that the prevalence of DS was highest among elderly and female denture wearers.<xref rid="B9" ref-type="bibr">9</xref> In addition, that study stated that the prevalence of DS ranged from 15% to over 70%.<xref rid="B9" ref-type="bibr">9</xref> Sadig<xref rid="B10" ref-type="bibr">10</xref> conducted a study in 2010 that reported the incidence of DS to be approximately 62%. Many factors might lead to the development of DS.<xref rid="B11" ref-type="bibr">11</xref> Some of these are related to systemic and immune diseases and impaired salivary flow, and others are related to the dentures themselves, such as poor denture hygiene, denture-induced trauma, roughness and the presence of pores in the acrylic surface.<xref rid="B11" ref-type="bibr">11</xref> Among these factors, <italic>Candida</italic> colonization is well established as a predisposing factor for the denture wearers experiencing development of denture stomatitis.<xref rid="B12" ref-type="bibr">12</xref><xref rid="B13" ref-type="bibr">13</xref> All the previously mentioned factors contribute to <italic>Candida</italic> colonization on denture and mucosal surfaces.<xref rid="B10" ref-type="bibr">10</xref><italic>Candida</italic> is not harmful by itself, and studies have demonstrated that it is part of the normal flora.<xref rid="B11" ref-type="bibr">11</xref> There are 20 species of <italic>Candida</italic> among the 300 to 400 species of microorganisms in the oral cavity.<xref rid="B14" ref-type="bibr">14</xref> The presence of <italic>Candida</italic> ranges from 20% to 50% in healthy dentate individuals.<xref rid="B15" ref-type="bibr">15</xref><xref rid="B16" ref-type="bibr">16</xref> Becoming edentulous and wearing dentures both cause changes in the oral microbial flora, which lead to an increase in <italic>Candida</italic> colonization as high as 60% to 100%.<xref rid="B11" ref-type="bibr">11</xref><xref rid="B17" ref-type="bibr">17</xref><italic>Candida</italic> has the ability to grow on targeted surfaces in several ways.<xref rid="B18" ref-type="bibr">18</xref> One of the ways is the formation of biofilms, which are significant causes of infection.<xref rid="B18" ref-type="bibr">18</xref> The adhesion of <italic>Candida</italic> to the oral mucosa is an important factor in the resistance to host clearance mechanisms in the oral cavity. Among other <italic>Candida</italic> species, <italic>Candida albicans</italic> is the most prevalent isolated species in DS, followed by <italic>Candida glabrata</italic>.<xref rid="B9" ref-type="bibr">9</xref><xref rid="B11" ref-type="bibr">11</xref><xref rid="B12" ref-type="bibr">12</xref><xref rid="B19" ref-type="bibr">19</xref> Other <italic>Candida</italic> species, such as <italic>Candida krusei</italic>, <italic>Candida Kefyr</italic>, <italic>Candida parapsilosis</italic>, and <italic>Candida tropicalis</italic>, occur at lower prevalences.<xref rid="B9" ref-type="bibr">9</xref><xref rid="B11" ref-type="bibr">11</xref><xref rid="B12" ref-type="bibr">12</xref> In addition to adhesion to the oral mucosa, the adhesion of <italic>Candida</italic> to acrylic is considered to be a critical factor in the development of DS.<xref rid="B20" ref-type="bibr">20</xref><xref rid="B21" ref-type="bibr">21</xref> This adhesion occurs through the formation of biofilms on the denture surface that act as protective reservoirs that prevent <italic>Candida</italic> from being washed away by saliva or dislodgment forces.<xref rid="B19" ref-type="bibr">19</xref> Many studies have proven that both <italic>Candida albicans</italic> and <italic>Candida glabrata</italic> form these biofilms on denture surfaces.<xref rid="B11" ref-type="bibr">11</xref><xref rid="B22" ref-type="bibr">22</xref><xref rid="B23" ref-type="bibr">23</xref><xref rid="B24" ref-type="bibr">24</xref> Several studies have linked the surface characteristics of denture acrylics to the amount of <italic>Candida</italic> biofilm adhesion.<xref rid="B25" ref-type="bibr">25</xref><xref rid="B26" ref-type="bibr">26</xref> Ramage et al.<xref rid="B21" ref-type="bibr">21</xref> reported that imperfections on the denture surface contribute to an increase in the adhesion of <italic>Candida</italic>, which becomes imbedded within these imperfections. Other studies have also confirmed that surface roughness and surface crevices facilitate <italic>Candida</italic> colonization on denture surfaces.<xref rid="B27" ref-type="bibr">27</xref> In a review, Bidra et al.<xref rid="B28" ref-type="bibr">28</xref> reported that decreasing the porosity decreases <italic>Candida</italic> adhesion. Conventional heat-cured poly(methyl methacrylate) (PMMA) is the most popular material for dentures.<xref rid="B29" ref-type="bibr">29</xref> It has been proven that this curing method increases the number of pores in the denture surface.<xref rid="B29" ref-type="bibr">29</xref></p><p>In 1994, the first report in English to discuss the use of computer-aided technology (CAD/CAM) in the fabrication of complete denture with rapid prototyping technology was published.<xref rid="B30" ref-type="bibr">30</xref> Numerous CAD/CAM denture systems appear in the market, the dentures are milled from pre-polymerized pucks of resin.<xref rid="B31" ref-type="bibr">31</xref><xref rid="B32" ref-type="bibr">32</xref> CAD/CAM-fabricated complete dentures have several advantages over conventionally fabricated complete dentures.<xref rid="B28" ref-type="bibr">28</xref> One of these is a decrease in porosity because with CAD/CAM, the denture base is formed from a prepolymerized block of acrylic resin.<xref rid="B28" ref-type="bibr">28</xref> This decrease in porosity might decrease the adhesion of <italic>Candida</italic> to the dentures.<xref rid="B28" ref-type="bibr">28</xref> Also CAD/CAM fabricated dentures release a small amount of monomer, which may affect microbial adhesion and trigger a mucosal allergy reaction, but this effect is not as statistically significantly as the conventional dentures.<xref rid="B32" ref-type="bibr">32</xref> Usually the internal surface of complete denture is not highly polished which may affect the roughness threshold of microbial adhesion.<xref rid="B33" ref-type="bibr">33</xref> Therefore, the aim of the present study was to compare the adhesion of <italic>Candida</italic> on the surfaces of CAD/CAM and conventionally fabricated complete denture bases. The null hypothesis was that there would be no difference in <italic>Candida</italic> adhesion between the CAD/CAM and conventionally fabricated complete denture bases.</p></sec><sec sec-type="materials|methods"><title>MATERIALS AND METHODS</title><p>Twenty discs of Pink acrylic resin denture bases were fabricated with CAD/CAM and conventional procedures. The specimens were divided into two groups: discs that were fabricated using the CAD/CAM procedure (Wieland Digital Denture Ivoclar Vivadent, Schaan, Liechtenstein) and discs that were fabricated using a conventional flasking and pressure-pack technique.</p><p>Ten discs were fabricated from PMMA acrylic resin denture base material (major.base20 heat-processed PMMA, Moncalieri (TO), Italy) using a conventional flasking and pressure-pack technique. A stainless steel disc-shaped mold (3 × 10 mm) was used to make the discs according to the manufacturer's instructions.</p><p>The discs were cleaned with a steam jet (Wasi-Steam Classic, Wassermann Dental-Maschinen, Hamburg, Germany) after the deflasking procedure. The excess flash was removed using carbide cutters (Black Hawk Cutter, Horeco, Hopf, Ringleb & Co. GmbH & CIE, Berlin, Germany). The surface was finished with waterproof paper. Next, polishing was performed with a polishing compact unit (Derotor, London, England) consisting of a polishing lathe, a 45-mm polishing brush, and a pleated buff nettle cloth (Renfert GmbH, Industrie-gebiet, Hilzingen, Germany) with pumice (Pumice CL 60, Coarse Grade, Whip Mix Corporation, Louisville, KY, USA). Finally, the discs were cleaned with water and soap using a regular toothbrush followed by a steam jet.</p><p>Ten discs of 10 × 3 mm were designed using Zenotec CAD software (Wieland Digital Denture Ivoclar Vivadent, Schaan, Liechtenstein). PMMA blocks were used (opera system, Principauté de Monaco, French). Milling was performed using Zenotec select ion (Wieland Digital Denture Ivoclar Vivadent, Schaan, Liechtenstein). These discs were finished and polished following the same procedures used for the discs made with the conventional method.</p><p>For both CAD/CAM and conventional denture materials, 30 readings were taken from randomly selected spots on the surface of each disc and subjected to surface roughness analysis. The measurements were made using a non-contact optical three-dimensional profilometer (Contour GT-I, Bruker) (<xref ref-type="fig" rid="F1">Fig. 1</xref>, <xref ref-type="fig" rid="F2">Fig. 2</xref>). Roughness is expressed as the surface area roughness (Sa, µm).</p><p><italic>Candida</italic> isolates were subcultured on Sabouraud dextrose agar (SDA) (Oxoid, Hampshire, UK) for 24 hours at 37℃. The isolates were identified to the species level using the germ tube test and API20C-AUX methods. For the preparation of the yeast suspensions, the cells were inoculated in Sabouraud dextrose broth (SDB) (Oxoid, UK) and incubated for 18 hours at 37℃ with shaking at 150 rpm. The yeast cells were harvested by centrifugation at 3000 g for 10 min and washed twice with phosphate-buffered saline (PBS; pH 7.0). The cell density was adjusted to 1 × 10<sup>7</sup> cells/mL in SDB.</p><p>Biofilm formation by the Candia albicans clinical isolates was investigated with the microtiter plate method, as described previously.<xref rid="B34" ref-type="bibr">34</xref> From the biofilm screening assays, four <italic>Candida albicans</italic> isolates (CA-1, CA-2, CA-3, and CA-4) were selected for the adherence assay based on their ability to form biofilms.</p><p>Biofilm-producing <italic>Candida albicans</italic> isolates were selected for the adherence assay. The discs were disinfected using 70% alcohol and washed with sterile distilled water before use in the adherence assay. For the adherence assay, yeast cells from a fresh culture were added to the SDB medium to a final concentration of 1 × 10<sup>7</sup> cells/mL. The discs of dental materials were placed in a 24-well tissue culture plate (Corning, St. Louis, MO, USA), and 1 mL of the yeast cell suspension was added. The plate was incubated for 90 min at 37℃ with shaking at 75 rpm. After incubation, the discs were transferred to new wells and washed three times with PBS to remove the non-adherent cells. The adherent <italic>Candida</italic> cells were dislodged from the surfaces of the dentures in 2 mL of PBS by scraping and vortexing for 5 minutes. The cell suspension was then gently sonicated for 3 minutes and plated on SDA. The colony-forming unit (CFU) counts were determined after 24 hours of incubation at 37℃. The determinations of the CFU counts were repeated on three different occasions separately for the four <italic>Candida albicans</italic> isolates with fresh cultures and adherence to the two different denture materials. The number of adherent cells is expressed as CFU/mL.</p><p><italic>Candida</italic> was applied to the two types of discs (20 discs) and allowed to adhere as described above. After washing, the discs were stained with acridine orange (AO) (Sigma Aldrich, St. Louis, MO, USA) for 5 minutes, and the excess stain was gently rinsed off with PBS. The discs were examined under a fluorescent microscope (SMZ25, Nikon, Tokyo, Japan). Images were captured and analyzed using NIS-Elements imaging and analysis software (Nikon, Tokyo, Japan) (<xref ref-type="fig" rid="F3">Fig. 3</xref>, <xref ref-type="fig" rid="F4">Fig. 4</xref>).</p><p>Descriptive statistics were used to describe the <italic>Candida albicans</italic> isolate adhesion values for the conventional and CAD/CAM complete dentures. Student's t test was used to compare the adhesion of the <italic>Candida albicans</italic> isolates and the surface roughnesses between the CAD/CAM and conventional complete denture bases. A <italic>P</italic> value of < 0.5 was considered statistically significant.</p></sec><sec sec-type="results"><title>RESULTS</title><p>This study assessed the adherence of <italic>Candida albicans</italic> to the surface of denture bases materials manufactured in two different ways. <xref ref-type="table" rid="T1">Table 1</xref> presents the mean <italic>Candida albicans</italic> isolate adhesion values and their standard deviations for the conventional and CAD/CAM complete dentures. The conventional complete denture surface exhibited a higher adherence of <italic>Candida albicans</italic> isolates than did the CAD/CAM surface. The percentage of reduction in the adhesion of <italic>Candida albicans</italic> (CA-1, CA-2 and CA-3) on the CAD/CAM complete dentures was significantly lower than on the conventional dentures (<italic>P</italic> < .05).</p><p><xref ref-type="table" rid="T2">Table 2</xref> presents the surface roughness of the complete denture surface. The conventional complete denture surface exhibited a surface roughness that was significantly greater than that of the CAD/CAM dentures.</p><p>The adherence ability was also assessed with acridin orange staining and observation under a fluorescent microscope. The number of yeast cells that adhered to the CAD/CAM discs was lower than the number that adhered to the conventional discs. This microscopic observation is in agreement with and confirms the CFU count. Four different <italic>Candida albicans</italic> isolates were used in this study to ensure that the outcome of the study was not influenced by biological differences in the <italic>Candida</italic> strains.</p></sec><sec sec-type="discussion"><title>DISCUSSION</title><p>The null hypothesis of this study was rejected because there was a difference in <italic>Candida</italic> adhesion between the CAD/CAM and the conventionally fabricated complete denture bases. In the present study, the adhesion of <italic>Candida albicans</italic> to the CAD/CAM and conventionally fabricated complete denture base surfaces was evaluated. The CFU count, which reflects cell adherence to the denture base, was used to assess the difference in adherence. Interestingly, <italic>Candida albicans</italic> exhibited differing adherence ability to the two different denture bases. <italic>Candida albicans</italic> adhered to the CAD/CAM base with a lower affinity than it did to the conventional denture base, which had greater surface roughness than CAD/CAM base.</p><p>The mechanism by which <italic>Candida</italic> species cause denture stomatitis can be summarized as follows. <italic>Candida</italic> can form biofilms on mucosal and denture surfaces that promote plaque deposition on the denture surface.<xref rid="B9" ref-type="bibr">9</xref> This deposition causes the mucosa to be continuously exposed to the microorganisms in the biofilm, which eventually causes DS.<xref rid="B9" ref-type="bibr">9</xref> These biofilms are important contributors to the development of DS.<xref rid="B35" ref-type="bibr">35</xref><italic>Candida albicans</italic> is the most frequently isolated species in such biofilms.<xref rid="B9" ref-type="bibr">9</xref><xref rid="B11" ref-type="bibr">11</xref><xref rid="B12" ref-type="bibr">12</xref><xref rid="B19" ref-type="bibr">19</xref> The capability of <italic>Candida albicans</italic> to adhere to and form biofilms on PMMA varies.<xref rid="B36" ref-type="bibr">36</xref> The presence of a denture in the oral cavity facilitates the adherence of <italic>Candida albicans</italic> and causes infection.<xref rid="B37" ref-type="bibr">37</xref></p><p>The reasons for the popularity of PMMA include its ease of handling, low cost, and esthetics.<xref rid="B38" ref-type="bibr">38</xref> However, it has many disadvantages, including its dimensional instability, residual monomer content, weak strength, water absorption, color instability, and porosity. Porosity is considered to be a shortcoming when it exceeds 11% because at this point, the mechanical properties and esthetics are compromised, and the material becomes a reservoir for microorganisms.<xref rid="B29" ref-type="bibr">29</xref> In 1968, the Academy of Denture Prosthetics stated that dentures should be free of pores to ensure adequate cleaning and resistance to stains and the adherence of microorganisms.<xref rid="B38" ref-type="bibr">38</xref> According to the American Dental Association's specifications for the porosity of denture base polymers, “there shall be no bubbles or voids when viewed without magnification”.<xref rid="B39" ref-type="bibr">39</xref> Conventional heat-cured PMMA is the most common curing technique.<xref rid="B29" ref-type="bibr">29</xref> Porosity in heat-cured PMMA denture bases is an unfavorable result. Porosity can be caused by : air trapped during mixing, monomer contraction during polymerization, monomer vaporization associated with the exothermic reaction and the presence of residual monomer, insufficient mixing of monomer and polymer, a processing temperature higher than 74℃, the way the mold is packed, and inadequate compression on the flask.<xref rid="B40" ref-type="bibr">40</xref><xref rid="B41" ref-type="bibr">41</xref><xref rid="B42" ref-type="bibr">42</xref><xref rid="B43" ref-type="bibr">43</xref> The surface characteristics of the denture might contribute to an increase in <italic>Candida</italic> colonization due to hydrophobicity and roughness.<xref rid="B25" ref-type="bibr">25</xref><xref rid="B26" ref-type="bibr">26</xref></p><p>Roughness is an important factor; the rougher the surface, the greater the <italic>Candida</italic> colonization will be.<xref rid="B25" ref-type="bibr">25</xref> There are several methods and devices available to measure the surface roughness, including visual assessments, scanning electron microscopy, profilometry, laser specular reflectance, and atomic force microscopy.<xref rid="B44" ref-type="bibr">44</xref> Profilometry is a common device for measuring surface roughness.<xref rid="B45" ref-type="bibr">45</xref> Profilometry can be performed with either contact or non-contact methods.<xref rid="B46" ref-type="bibr">46</xref> In this study, a non-contact optical three-dimensional profilometer was used. The non-contact method uses a laser or light beam to obtain a surface profile.<xref rid="B47" ref-type="bibr">47</xref> Kukiattrakoon et al.<xref rid="B47" ref-type="bibr">47</xref> claimed that one of the disadvantages of the contact method is that it can damage the surface by producing scratches. However, one study reported that not all irregularities in specimens can be penetrated by the sensor needle of a mechanical profilometer.<xref rid="B48" ref-type="bibr">48</xref> In the present study, an optical laser profilometer that provided a three-dimensional profile was used. According to Joniot et al.,<xref rid="B49" ref-type="bibr">49</xref> this method provides a non-contact, non-destructive, and rapid quantitative measurement of surface roughness.</p><p>As mentioned earlier, increased porosity has been proven to increase microorganism colonization.<xref rid="B48" ref-type="bibr">48</xref><xref rid="B50" ref-type="bibr">50</xref> Over the past several years, many attempts have been made to improve both the material properties and the curing and processing techniques.<xref rid="B29" ref-type="bibr">29</xref> The advantage of the CAD/CAM method over the conventional method is that a prepolymerized block of acrylic resin is used to mill the denture base.<xref rid="B28" ref-type="bibr">28</xref> The elimination of mixing used in the conventional procedures will decrease the porosity, which will ultimately decrease <italic>Candida</italic> adhesion. This study is considered one of the first studies that used microbiological essay comparing CAD/CAM and conventional procedure to fabricate dentures.</p></sec><sec sec-type="conclusions"><title>CONCLUSION</title><p>Alterations of denture surface characteristics, such as porosity and surface roughness, contribute to a decrease in <italic>Candida</italic> adhesion, which ultimately decreases the risk of denture stomatitis. The surface characteristics of complete dentures fabricated with the CAD/CAM procedure exhibited promising potential for reducing the adherence of <italic>Candida</italic> to the denture base surface. Moreover, the adhesion of <italic>Candida albicans</italic> to the surfaces is significantly affected by the interactions with other microorganisms in the oral cavity. Therefore, further research is needed to fully determine the difference in adhesion to the surface of these denture base materials in complex <italic>Candida</italic> and bacterial models beside <italic>in vivo</italic> studies. 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person-group-type="author"><name><surname>Kasina</surname><given-names>SP</given-names></name><name><surname>Ajaz</surname><given-names>T</given-names></name><name><surname>Attili</surname><given-names>S</given-names></name><name><surname>Surapaneni</surname><given-names>H</given-names></name><name><surname>Cherukuri</surname><given-names>M</given-names></name><name><surname>Srinath</surname><given-names>HP</given-names></name></person-group><article-title>To evaluate and compare the porosities in the acrylic mandibular denture bases processed by two different polymerization techniques, using two different brands of commercially available denture base resins - an in vitro study</article-title><source>J Int Oral Health</source><year>2014</year><volume>6</volume><fpage>72</fpage><lpage>77</lpage></element-citation></ref></ref-list></back><floats-group><fig id="F1" orientation="portrait" position="float"><label>Fig. 1</label><caption><title>Surface roughness of the CAD/CAM complete denture base.</title></caption><graphic xlink:href="jap-9-402-g001"></graphic></fig><fig id="F2" orientation="portrait" position="float"><label>Fig. 2</label><caption><title>Surface roughness of the conventional complete denture base.</title></caption><graphic xlink:href="jap-9-402-g002"></graphic></fig><fig id="F3" orientation="portrait" position="float"><label>Fig. 3</label><caption><title>Candida cells adhered to the conventional discs (stained with acridine orange).</title></caption><graphic xlink:href="jap-9-402-g003"></graphic></fig><fig id="F4" orientation="portrait" position="float"><label>Fig. 4</label><caption><title>Candida cells adhered to the CAD/CAM discs (stained with acridine orange).</title></caption><graphic xlink:href="jap-9-402-g004"></graphic></fig><table-wrap id="T1" orientation="portrait" position="float"><label>Table 1</label><caption><title>Adhesion of <italic>Candida albicans</italic> isolates (CA-1 to CA-4) to the conventional and CAD/CAM discs</title></caption><alternatives><graphic xlink:href="jap-9-402-i001"></graphic><table frame="hsides" rules="rows"><col width="14%" span="1"></col><col width="14%" span="1"></col><col width="14%" span="1"></col><col width="14%" span="1"></col><col width="14%" span="1"></col><col width="16%" span="1"></col><col width="14%" span="1"></col><thead><tr><th valign="middle" align="center" rowspan="2" colspan="1">Candida albicans isolates</th><th valign="middle" align="center" rowspan="1" colspan="2">Conventional CFU/mL</th><th valign="middle" align="center" rowspan="1" colspan="2">CAD/CAM CFU/mL</th><th valign="middle" align="center" rowspan="2" colspan="1">% Reduction in adherence (± SD)</th><th valign="middle" align="center" rowspan="2" colspan="1"><italic>P</italic> value</th></tr><tr><th valign="middle" align="center" rowspan="1" colspan="1">Mean</th><th valign="middle" align="center" rowspan="1" colspan="1">SD</th><th valign="middle" align="center" rowspan="1" colspan="1">Mean</th><th valign="middle" align="center" rowspan="1" colspan="1">SD</th></tr></thead><tbody><tr><td valign="top" align="center" rowspan="1" colspan="1">CA-1</td><td valign="top" align="center" rowspan="1" colspan="1">2.3 × 10<sup>3</sup></td><td valign="top" align="center" rowspan="1" colspan="1">8.4 × 10<sup>2</sup></td><td valign="top" align="center" rowspan="1" colspan="1">1.1 × 10<sup>3</sup></td><td valign="top" align="center" rowspan="1" colspan="1">6.0 × 10<sup>2</sup></td><td valign="top" align="center" rowspan="1" colspan="1">51.7 ± 17.1</td><td valign="top" align="center" rowspan="1" colspan="1">.0364</td></tr><tr><td valign="top" align="center" rowspan="1" colspan="1">CA-2</td><td valign="top" align="center" rowspan="1" colspan="1">5.4 × 10<sup>3</sup></td><td valign="top" align="center" rowspan="1" colspan="1">1.6 × 10<sup>2</sup></td><td valign="top" align="center" rowspan="1" colspan="1">2.1 × 10<sup>3</sup></td><td valign="top" align="center" rowspan="1" colspan="1">8.7 × 10<sup>2</sup></td><td valign="top" align="center" rowspan="1" colspan="1">60.2 ± 17.8</td><td valign="top" align="center" rowspan="1" colspan="1">.0041</td></tr><tr><td valign="top" align="center" rowspan="1" colspan="1">CA-3</td><td valign="top" align="center" rowspan="1" colspan="1">2.0 × 10<sup>3</sup></td><td valign="top" align="center" rowspan="1" colspan="1">9.7 × 10<sup>2</sup></td><td valign="top" align="center" rowspan="1" colspan="1">1.2 × 10<sup>3</sup></td><td valign="top" align="center" rowspan="1" colspan="1">8.8 × 10<sup>2</sup></td><td valign="top" align="center" rowspan="1" colspan="1">47.2 ± 24.8</td><td valign="top" align="center" rowspan="1" colspan="1">.0448</td></tr><tr><td valign="top" align="center" rowspan="1" colspan="1">CA-4</td><td valign="top" align="center" rowspan="1" colspan="1">2.4 × 10<sup>3</sup></td><td valign="top" align="center" rowspan="1" colspan="1">1.1 × 10<sup>3</sup></td><td valign="top" align="center" rowspan="1" colspan="1">1.5 × 10<sup>3</sup></td><td valign="top" align="center" rowspan="1" colspan="1">7.2 × 10<sup>2</sup></td><td valign="top" align="center" rowspan="1" colspan="1">35.7 ± 19.1</td><td valign="top" align="center" rowspan="1" colspan="1">.0648</td></tr></tbody></table></alternatives></table-wrap><table-wrap id="T2" orientation="portrait" position="float"><label>Table 2</label><caption><title>Comparison of the surface roughness of the complete denture surface (CAD/CAM vs. conventional)</title></caption><alternatives><graphic xlink:href="jap-9-402-i002"></graphic><table frame="hsides" rules="rows"><col width="25%" span="1"></col><col width="25%" span="1"></col><col width="25%" span="1"></col><col width="25%" span="1"></col><thead><tr><th valign="middle" align="center" rowspan="2" colspan="1">Complete denture surface</th><th valign="middle" align="center" rowspan="1" colspan="1">Mean (µm)</th><th valign="middle" align="center" rowspan="2" colspan="1">SD (µm)</th><th valign="middle" align="center" rowspan="2" colspan="1"><italic>P</italic> value</th></tr><tr><th valign="middle" align="center" rowspan="1" colspan="1">Sa</th></tr></thead><tbody><tr><td valign="top" align="center" rowspan="1" colspan="1">CAD/CAM</td><td valign="top" align="right" rowspan="1" colspan="1">0.037</td><td valign="top" align="right" rowspan="1" colspan="1">0.001</td><td valign="top" align="right" rowspan="1" colspan="1"></td></tr><tr><td valign="top" align="center" rowspan="1" colspan="1">Conventional</td><td valign="top" align="right" rowspan="1" colspan="1">0.073</td><td valign="top" align="right" rowspan="1" colspan="1">0.015</td><td valign="top" align="right" rowspan="1" colspan="1">.020</td></tr></tbody></table></alternatives></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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