<?xml version="1.0" encoding="UTF-8"?><!DOCTYPE article PUBLIC "-//NLM//DTD JATS (Z39.96) Journal Publishing DTD v1.2 20190208//EN" "http://jats.nlm.nih.gov/publishing/1.2/JATS-journalpublishing1.dtd"><article xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" article-type="research-article" dtd-version="1.2" xml:lang="en">
    <front>
        <journal-meta>
            <journal-id journal-id-type="pmc">F1000Research</journal-id>
            <journal-title-group>
                <journal-title>F1000Research</journal-title>
            </journal-title-group>
            <issn pub-type="epub">2046-1402</issn>
            <publisher>
                <publisher-name>F1000 Research Limited</publisher-name>
                <publisher-loc>London, UK</publisher-loc>
            </publisher>
        </journal-meta>
        <article-meta>
            <article-id pub-id-type="doi">10.12688/f1000research.171971.1</article-id>
            <article-categories>
                <subj-group subj-group-type="heading">
                    <subject>Research Article</subject>
                </subj-group>
                <subj-group>
                    <subject>Articles</subject>
                </subj-group>
            </article-categories>
            <title-group>
                <article-title>Comparison between the protective effect of applying sodium fluoride varnish alone and applying CO2 laser with it in permanent teeth. &#x00a0;In Vitro study</article-title>
                <fn-group content-type="pub-status">
                    <fn>
                        <p>[version 1; peer review: 1 approved, 1 not approved]</p>
                    </fn>
                </fn-group>
            </title-group>
            <contrib-group>
                <contrib contrib-type="author" corresp="no">
                    <name>
                        <surname>Alkhateeb</surname>
                        <given-names>Sham Mohamad Farouq</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Data Curation</role>
                    <role content-type="http://credit.niso.org/">Formal Analysis</role>
                    <role content-type="http://credit.niso.org/">Investigation</role>
                    <role content-type="http://credit.niso.org/">Methodology</role>
                    <role content-type="http://credit.niso.org/">Software</role>
                    <role content-type="http://credit.niso.org/">Validation</role>
                    <role content-type="http://credit.niso.org/">Visualization</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Original Draft Preparation</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Review &amp; Editing</role>
                    <xref ref-type="aff" rid="a1">1</xref>
                </contrib>
                <contrib contrib-type="author" corresp="no">
                    <name>
                        <surname>Kouchaji</surname>
                        <given-names>Chaza Nader</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Conceptualization</role>
                    <role content-type="http://credit.niso.org/">Formal Analysis</role>
                    <role content-type="http://credit.niso.org/">Investigation</role>
                    <role content-type="http://credit.niso.org/">Methodology</role>
                    <role content-type="http://credit.niso.org/">Software</role>
                    <role content-type="http://credit.niso.org/">Supervision</role>
                    <role content-type="http://credit.niso.org/">Validation</role>
                    <role content-type="http://credit.niso.org/">Visualization</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Original Draft Preparation</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Review &amp; Editing</role>
                    <xref ref-type="aff" rid="a2">2</xref>
                </contrib>
                <contrib contrib-type="author" corresp="yes">
                    <name>
                        <surname>Alkhaled</surname>
                        <given-names>Abdulrhman Ahmad</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Conceptualization</role>
                    <role content-type="http://credit.niso.org/">Data Curation</role>
                    <role content-type="http://credit.niso.org/">Formal Analysis</role>
                    <role content-type="http://credit.niso.org/">Investigation</role>
                    <role content-type="http://credit.niso.org/">Methodology</role>
                    <role content-type="http://credit.niso.org/">Resources</role>
                    <role content-type="http://credit.niso.org/">Software</role>
                    <role content-type="http://credit.niso.org/">Validation</role>
                    <role content-type="http://credit.niso.org/">Visualization</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Original Draft Preparation</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Review &amp; Editing</role>
                    <uri content-type="orcid">https://orcid.org/0000-0002-5510-4339</uri>
                    <xref ref-type="corresp" rid="c1">a</xref>
                    <xref ref-type="aff" rid="a2">2</xref>
                </contrib>
                <contrib contrib-type="author" corresp="no">
                    <name>
                        <surname>Hamadah</surname>
                        <given-names>Omar</given-names>
                    </name>
                    <role content-type="http://credit.niso.org/">Data Curation</role>
                    <role content-type="http://credit.niso.org/">Formal Analysis</role>
                    <role content-type="http://credit.niso.org/">Investigation</role>
                    <role content-type="http://credit.niso.org/">Methodology</role>
                    <role content-type="http://credit.niso.org/">Software</role>
                    <role content-type="http://credit.niso.org/">Validation</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Original Draft Preparation</role>
                    <role content-type="http://credit.niso.org/">Writing &#x2013; Review &amp; Editing</role>
                    <xref ref-type="aff" rid="a1">1</xref>
                    <xref ref-type="aff" rid="a3">3</xref>
                </contrib>
                <aff id="a1">
                    <label>1</label>Higher Institute of Laser Research and Applications, Damascus University, Damascus, Syria</aff>
                <aff id="a2">
                    <label>2</label>Department of Pediatric Dentistry, Damascus University Faculty of Dentistry, Damascus, Syria</aff>
                <aff id="a3">
                    <label>3</label>Oral Medicine Department, Damascus University Faculty of Dentistry, Damascus, Syria</aff>
            </contrib-group>
            <author-notes>
                <corresp id="c1">
                    <label>a</label>
                    <email xlink:href="mailto:abdulrhman.alkhaled@damascusuniversity.edu.sy">abdulrhman.alkhaled@damascusuniversity.edu.sy</email>
                </corresp>
                <fn fn-type="conflict">
                    <p>No competing interests were disclosed.</p>
                </fn>
            </author-notes>
            <pub-date pub-type="epub">
                <day>19</day>
                <month>4</month>
                <year>2026</year>
            </pub-date>
            <pub-date pub-type="collection">
                <year>2026</year>
            </pub-date>
            <volume>15</volume>
            <elocation-id>588</elocation-id>
            <history>
                <date date-type="accepted">
                    <day>1</day>
                    <month>4</month>
                    <year>2026</year>
                </date>
            </history>
            <permissions>
                <copyright-statement>Copyright: &#x00a9; 2026 Alkhateeb SMF et al.</copyright-statement>
                <copyright-year>2026</copyright-year>
                <license xlink:href="https://creativecommons.org/licenses/by/4.0/">
                    <license-p>This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
                </license>
            </permissions>
            <self-uri content-type="pdf" xlink:href="https://f1000research.com/articles/15-588/pdf"/>
            <abstract>
                <sec>
                    <title>Background</title>
                    <p>Fluoride administration plays a critical role in the management and prophylaxis of dental caries. Research indicates that laser irradiation can enhance the uptake of fluoride into dental enamel, an effect that is particularly pronounced when used as an adjunct to topical fluoride treatments. The present study aimed to evaluate the impact of CO&#x2082; laser irradiation on fluoride uptake in the enamel of permanent teeth.</p>
                </sec>
                <sec>
                    <title>Methods</title>
                    <p>Six human upper premolars, extracted for orthodontic reasons, were selected for this study. The roots were separated from the crowns, and each crown was then sectioned mesiodistally to create mesial and distal halves. These halves were randomly assigned to either an experimental or a control group (n&#x00a0;=&#x00a0;6 per group). All specimens received an application of 5% sodium fluoride (NaF) varnish. Subsequently, the samples in the experimental group were subjected to irradiation with a 1&#x00a0;W CO2 laser for a duration of 15&#x00a0;seconds. To assess the effects of the treatments, fluoride uptake was measured and the topographic characteristics of the enamel surface were examined using scanning electron microscopy (SEM) and energy-dispersive X-ray spectroscopy (EDX). Statistical analysis of the data was performed using SPSS software version 26 (IBM, USA).</p>
                </sec>
                <sec>
                    <title>Results</title>
                    <p>Energy-dispersive X-ray spectrometry analysis showed a significant increase in the percentage of fluoride with the application of the laser, The results of the mean ranks showed that the fluoride and CO&#x2082; laser group recorded significantly higher ranks in the fluoride element (8.75 vs. 4.25), reflecting a higher concentration and confirmed statistical significance. Statistically, in the Mann-Whitney U test, a significant statistical difference in fluoride ratios appeared between the two groups (P value&#x00a0;=&#x00a0;0.026) smaller than (0.05), which indicates the effectiveness of using CO&#x2082; lasers in enhancing fluoride absorption within the tooth structure.</p>
                </sec>
                <sec>
                    <title>Conclusion</title>
                    <p>The results showed CO&#x2082; laser irradiation of dental enamel resulted in increased fluoride uptake.</p>
                </sec>
            </abstract>
            <kwd-group kwd-group-type="author">
                <kwd>Laser; Gas; Dental Enamel; caries; Fluoride.</kwd>
            </kwd-group>
            <funding-group>
                <award-group id="fund-1">
                    <funding-source>None</funding-source>
                </award-group>
                <funding-statement>The author(s) declared that no grants were involved in supporting this work.</funding-statement>
            </funding-group>
        </article-meta>
    </front>
    <body>
        <sec id="sec5" sec-type="intro">
            <title>Introduction</title>
            <p>Dental caries deserves attention because it has become a chronic disease affecting children worldwide.
                <sup>
                    <xref ref-type="bibr" rid="ref1">1</xref>,
                    <xref ref-type="bibr" rid="ref2">2</xref>
                </sup>
            </p>
            <p>Dental caries is a multifactorial disease caused by the simultaneous interaction of various factors&#x2014; sugar and dental biofilms &#x2014;in the oral cavity.</p>
            <p>The introduction of fluoride into dental practice over seventy years ago is widely regarded as the principal reason for the global decline in dental caries. Despite this public health success, the sole confirmed adverse effect of fluoride use in dentistry is dental fluorosis, a condition resulting from excessive fluoride ingestion during tooth development. Recent epidemiological trends indicate a concurrent rise in the prevalence of dental fluorosis and a decline in caries rates.
                <sup>
                    <xref ref-type="bibr" rid="ref3">3</xref>
                </sup>
            </p>
            <p>The most common varnish used in dentistry to prevent tooth caries is sodium fluoride (NaF).
                <sup>
                    <xref ref-type="bibr" rid="ref4">4</xref>
                </sup>
            </p>
            <p>Topical application of NaF on teeth mitigates enamel demineralization while enhancing the remineralization process. The primary antimicrobial mechanism of fluoride is not direct bacterial destruction, but rather an indirect action through the inhibition of bacterial acid production, which results in a higher pH at the tooth surface.
                <sup>
                    <xref ref-type="bibr" rid="ref5">5</xref>,
                    <xref ref-type="bibr" rid="ref6">6</xref>
                </sup>
            </p>
            <p>The combined use of laser irradiation and topical fluoride represents a novel approach to caries prevention by increasing the enamel&#x2019;s affinity for and incorporation of fluoride. Zancope et al. were the first to report that CO2 lasers cause morphological changes on the surface of tooth enamel, such as fusion and dissolution.
                <sup>
                    <xref ref-type="bibr" rid="ref7">7</xref>
                </sup>
            </p>
            <p>On the other hand, studies were conducted to evaluate the effect of carbon dioxide laser on the structure of tooth enamel and dentin, and showed that its absorption by dental tissue was high.
                <sup>
                    <xref ref-type="bibr" rid="ref8">8</xref>
                </sup>
            </p>
            <p>The 10600&#x00a0;nm CO2 laser has an absorption coefficient in hydroxyapatite ten times lower than that of 9600&#x00a0;nm and this allows deeper penetration and higher flow by the 10600&#x00a0;nm CO2 laser to produce the desired thermal effects on caries inhibition compared to the CO2 laser at other wavelengths.
                <sup>
                    <xref ref-type="bibr" rid="ref9">9</xref>
                </sup>
            </p>
            <p>Calcium, phosphorus and fluoride ions play a crucial role in remineralizing tooth enamel.
                <sup>
                    <xref ref-type="bibr" rid="ref8">8</xref>
                </sup>
            </p>
            <p>A high level of calcium in the remineralization solution may enhance the rate of mineral deposition in the caries lesion and the calcium level greatly affects the defense mechanism of the hard tissues of the teeth and as the number of caries increases the calcium level decreases.
                <sup>
                    <xref ref-type="bibr" rid="ref10">10</xref>
                </sup>
            </p>
            <p>This study aimed to evaluate and compare the efficacy of CO
                <sub>2</sub> laser irradiation versus a CO
                <sub>2</sub> application in enhancing fluoride uptake by the surfaces of permanent teeth following treatment with NaF varnish.</p>
        </sec>
        <sec id="sec6">
            <title>Materials and methods</title>
            <sec id="sec7">
                <title>Source of teeth specimens</title>
                <p>The six human upper premolars were obtained from the Department of Oral and Maxillofacial Surgery in College of Dentistry, University of Damascus after routine extractions for orthodontic reasons and not specifically for this study. Teeth were stored in distilled water at 4&#x00a0;&#x00b0;C until use.</p>
                <p>Teeth were examined to ensure they were free of any caries, abrasions or fractures. They were divided into two sections, a mesial section and a distal section, so that each treatment was applied to a section of the tooth. The root was cut from the Cementoenamel junction and the crown were stored in distilled water to prevent dehydration (
                    <xref ref-type="fig" rid="f1">
Figure 1</xref>).</p>
                <fig fig-type="figure" id="f1" orientation="portrait" position="float">
                    <label>
Figure 1. </label>
                    <caption>
                        <title>A picture of the teeth used in the research before and after the section.</title>
                    </caption>
                    <graphic id="gr1" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/189644/2f02adc4-9fe4-4148-8777-a70b18df1eef_figure1.gif"/>
                </fig>
                <p>Mesial section was used as the experimental specimen and the distal as the control.</p>
                <p>All specimens were first dried and treated with 5% NaF varnish (VOCO&#x00ae;, Germany). A distinction was then made between the groups: the experimental specimens were laser irradiated, while the control specimens were not. Sodium fluoride varnish (5% NaF varnish (VOCO, Germany, Product Code: [190/10]) was applied to the mesial section of the tooth with a carbon dioxide laser with a wavelength of 10600&#x00a0;nm (power 1&#x00a0;W &#x2013; pulse duration 15&#x00a0;ms) while the control sample was in the distal section and sodium fluoride varnish alone was applied (
                    <xref ref-type="fig" rid="f2">
Figure 2</xref>).</p>
                <fig fig-type="figure" id="f2" orientation="portrait" position="float">
                    <label>
Figure 2. </label>
                    <caption>
                        <title>A picture of the sodium fluoride varnish used by VOCO and how it is applied.</title>
                    </caption>
                    <graphic id="gr2" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/189644/2f02adc4-9fe4-4148-8777-a70b18df1eef_figure2.gif"/>
                </fig>
                <p>
                    <xref ref-type="table" rid="T1">
Table 1</xref>: lists earlier research that used CO
                    <sub>2</sub> laser irradiation with comparable settings. so that the parameters used in this investigation may be compared to those documented in the literature. With many research concentrating on 2&#x00a0;W power output, it is clear that the power parameter (1&#x00a0;W) employed in this study fits within the range documented in earlier investigations (0.4&#x2013;10&#x00a0;W). The wavelength that is most frequently used in research assessing how CO2 lasers affect dental hard tissues is 10,600&#x00a0;nm.</p>
                <table-wrap id="T1" orientation="portrait" position="float">
                    <label>
Table 1. </label>
                    <caption>
                        <title>The earlier research that used CO
                            <sub>2</sub> laser irradiation with comparable settings.</title>
                    </caption>
                    <table content-type="article-table" frame="hsides">
                        <thead>
                            <tr>
                                <th align="left" colspan="1" rowspan="1" valign="top">Authors</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Year</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Journal</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Wavelength</th>
                                <th align="left" colspan="1" rowspan="1" valign="top">Power</th>
                            </tr>
                        </thead>
                        <tbody>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Chin-Ying SH, Xiaoli G, Jisheng P, Wefel JS.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2004</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">Journal of Dentistry</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">10600&#x00a0;nm (model LX-20, Luxar Corp., Bothell, WA, USA)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2&#x00a0;W - 4&#x00a0;W</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Tepper SA, Zehnder M, Pajarola GF, Schmidlin PR.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2004</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">Journal of Dentistry</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">10600&#x00a0;nm (Sharplan 15F, Lumenis Inc., Santa Clara, CA, USA).</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2&#x00a0;W</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Steiner-Oliveira C, Rodrigues LK, Soares LE, Martin AA, Zezell DM, Nobre-dos-Santos M.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2006</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">Dental Materials Journal</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">10600&#x00a0;nm (Model UM L30, Union Medical Engineering Co., Yangju-si, Gyeonggi-Do, Korea)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2&#x00a0;W, 4&#x00a0;W, 6&#x00a0;W, 8&#x00a0;W, and 10&#x00a0;W</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Souza-Gabriel AE, Colucci V, Turssi CP, Serra MC, Corona SA.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2010</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">Microscopy Research and Technique</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">10600&#x00a0;nm</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2&#x00a0;W</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Lepri TP, Colucci V, Turssi CP, Corona SA.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2013</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">
                                    <ext-link ext-link-type="uri" xlink:href="https://link.springer.com/journal/10103">Lasers in Medical Science</ext-link>
</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">10600&#x00a0;nm (Opus 20; Opus Dent, Tel Aviv, Israel)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2&#x00a0;W</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Bahrololoomi Z, Fotuhi Ardakani F, Sorouri M.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2015</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">Journal of Dentistry</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">10600&#x00a0;nm (Daeshin Enterprise Corp., Seoul, South Korea)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">1&#x00a0;W</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Soltanimehr E, Bahrampour E, Yousefvand Z.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2019</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">BMC Oral Health.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">10600&#x00a0;nm (Lasersat 15tm, Satelec, Merignac, France)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2&#x00a0;W</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Dehghan H, Mojarad F, Serajzadeh M, Fekrazad R.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2020</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">Oral Health &amp; Preventive Dentistry</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">10600&#x00a0;nm (Smart US 20D, Deka: Florence, Italy)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2&#x00a0;W</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Zhao IS, Xue VW, Yin IX, Niu JY, Lo ECM, Chu CH.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2021</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">ScienceDirect</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">9300&#x00a0;nm (Model DL-500; Access Laser Co, Everett, WA, USA)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.67&#x00a0;W</td>
                            </tr>
                            <tr>
                                <td align="left" colspan="1" rowspan="1" valign="top">Eissa NM, Elshourbagy EM, Gomaa NE.</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">2022</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">Heliyon Journal</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">10600&#x00a0;nm (DEKALaser Technologies, Florence, Italy)</td>
                                <td align="left" colspan="1" rowspan="1" valign="top">0.4&#x00a0;W</td>
                            </tr>
                        </tbody>
                    </table>
                </table-wrap>
                <p>The varnish was applied using a small brush according to the manufacturer&#x2019;s instructions by applying a thin layer over the entire occlusal surface of the tooth.</p>
                <p>The sample in the mesial section was exposed to CO2 laser radiation (10600&#x00a0;nm wavelength, 1&#x00a0;W power, pulsed mode, 15&#x00a0;ms pulse duration) with a wavelength of 10600&#x00a0;nm and a power of 1&#x00a0;W for 15&#x00a0;seconds with a pulsed mode and a pulse duration of 15&#x00a0;ms
                    <sup>
                        <xref ref-type="bibr" rid="ref11">11</xref>
                    </sup> at a distance of 1&#x00a0;cm. Irradiation was performed in a continuous scanning motion that allowed the entire surface to be irradiated and we re-preserved it with distilled water (
                    <xref ref-type="fig" rid="f3">
Figure 3</xref>).</p>
                <fig fig-type="figure" id="f3" orientation="portrait" position="float">
                    <label>
Figure 3. </label>
                    <caption>
                        <title>Image of the parameters used.</title>
                    </caption>
                    <graphic id="gr3" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/189644/2f02adc4-9fe4-4148-8777-a70b18df1eef_figure3.gif"/>
                </fig>
                <p>The test of the effect of the laser on the varnish was by means of a SEM (scanning electron microscope) (T-Scan company VEGA II XMU, Czech Republic). The amount of fluoride, calcium, phosphorus and carbon remaining on the two samples was tested using spectral analysis of EDX (Energy-dispersive X-ray spectroscopy, Oxford Instruments, UK) of the scanning electron microscope located in the Atomic Energy Commission (
                    <xref ref-type="fig" rid="f4">
Figure 4</xref>). The sample was analyzed by SEM to observe the topographic properties of the enamel surface after different treatments and SEM images of the central region of the samples were taken using different reagents with two different detectors SE (Secondary Electrons Detector) and BSE (Backscattered Electrons Detector). EDX analysis can provide information on the chemical composition/distribution of elements over a non-destructively large surface area.
                    <sup>
                        <xref ref-type="bibr" rid="ref12">12</xref>
                    </sup> The response variable was mainly a change in fluoride concentration and a change in the concentration of carbonate, calcium and phosphorus that make up hydroxyapatite, which was analyzed by EDX.</p>
                <fig fig-type="figure" id="f4" orientation="portrait" position="float">
                    <label>
Figure 4. </label>
                    <caption>
                        <title>Image of the sample after preparation for entry into the scanning electron microscope.</title>
                    </caption>
                    <graphic id="gr4" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/189644/2f02adc4-9fe4-4148-8777-a70b18df1eef_figure4.gif"/>
                </fig>
            </sec>
            <sec id="sec8">
                <title>Ethical approval and informed consent</title>
                <p>This 
                    <italic toggle="yes">in vitro</italic> study utilized extracted human teeth collected from patients who underwent orthodontic treatment. This study was from 24/8/2025 to 27/9/2025 The study protocol was reviewed and approved by the Institutional Review Board (IRB) of College of Dentistry, University of Damascus On August 21, 2025 under reference number (DN-210-125-3-9-1/21/8/2025) &#x201c;Not applicable as the study used discarded biological material&#x201d;]. All participants (or their guardians) provided written informed consent for the use of their extracted teeth for research purposes, in accordance with the principles of the Declaration of Helsinki. Patient identifiers were removed to ensure confidentiality.</p>
            </sec>
        </sec>
        <sec id="sec9" sec-type="results">
            <title>Results</title>
            <p>The results were evaluated according to the surface shape in electron microscope images and according to the concentration of fluoride, calcium, phosphorus and carbon.</p>
            <p>In this study, SEM images showed that CO2 laser irradiation fuses the surface, creating a smooth, recrystallized side. Fusion between enamel surface crystals was also observed after carbon dioxide laser treatment (
                <xref ref-type="fig" rid="f5">
Figure 5</xref>,
                <xref ref-type="fig" rid="f6">6</xref>).</p>
            <fig fig-type="figure" id="f5" orientation="portrait" position="float">
                <label>
Figure 5. </label>
                <caption>
                    <title>A: Image of the mesial section (fluoride + laser) with SE detector (Secondary Electrons Detector), x1000 measurement magnification and 20&#x00a0;kV acceleration voltage.</title>
                    <p>B: Image of the distal section (fluoride only) with SE detector, x1000 magnification and 20&#x00a0;kV acceleration voltage. C: Image of the mesial section (fluoride + laser) with SE detector, x500 magnification and 20&#x00a0;kV acceleration voltage. D: Image of the distal section (fluoride only) with SE detector, x500 magnification and 20&#x00a0;kV acceleration voltage.</p>
                </caption>
                <graphic id="gr5" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/189644/2f02adc4-9fe4-4148-8777-a70b18df1eef_figure5.gif"/>
            </fig>
            <fig fig-type="figure" id="f6" orientation="portrait" position="float">
                <label>
Figure 6. </label>
                <caption>
                    <title>A: Image of the mesial section (fluoride + laser) with a BSE detector (Backscattered Electrons Detector), x200 magnification, and 20&#x00a0;kV acceleration voltage.</title>
                    <p>B: Image of the lateral section (fluoride only) with a BSE detector, x200 magnification, and 20&#x00a0;kV acceleration voltage. C: Image of the medial section (fluoride + laser) with BSE detector, x500 magnification and 20&#x00a0;kV acceleration voltage. D: Image of the lateral section (fluoride only) with BSE detector, x500 magnification and 20&#x00a0;kV acceleration voltage.</p>
                </caption>
                <graphic id="gr6" orientation="portrait" position="float" xlink:href="https://f1000research-files.f1000.com/manuscripts/189644/2f02adc4-9fe4-4148-8777-a70b18df1eef_figure6.gif"/>
            </fig>
            <p>Comparing the apparent differences in the mean of the measured items between the two groups (
                <xref ref-type="table" rid="T2">
Table 2</xref>) and the descriptive statistics for fluoride, calcium, phosphorus, and carbon ratios in both study groups (
                <xref ref-type="table" rid="T3">
Table 3</xref>,
                <xref ref-type="table" rid="T4">4</xref>,
                <xref ref-type="table" rid="T5">5</xref>,
                <xref ref-type="table" rid="T6">6</xref>).</p>
            <table-wrap id="T2" orientation="portrait" position="float">
                <label>
Table 2. </label>
                <caption>
                    <title>The comparison between elements using the mean.</title>
                </caption>
                <table content-type="article-table" frame="hsides">
                    <thead>
                        <tr>
                            <th align="left" colspan="1" rowspan="1" valign="top">Element</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride group</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride + CO2 laser</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">The difference between averages</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Comments</th>
                        </tr>
                    </thead>
                    <tbody>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Fluoride</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.35</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">1.73</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">+1.38</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">Obvious apparent increase in fluoride with laser application</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Calcium</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">1.62</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">3.72</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">+2.10</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">Significant apparent increase in calcium</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Phosphorus</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.99</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.65</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">&#x2212;0.34</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">Slight decrease in phosphorus</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Carbon</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">72.89</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">67.30</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">&#x2212;5.59</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">Slight carbon reduction</td>
                        </tr>
                    </tbody>
                </table>
            </table-wrap>
            <table-wrap id="T3" orientation="portrait" position="float">
                <label>
Table 3. </label>
                <caption>
                    <title>The descriptive statistics of fluoride ratios.</title>
                </caption>
                <table content-type="article-table" frame="hsides">
                    <thead>
                        <tr>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride group</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride + CO2 laser group</th>
                        </tr>
                    </thead>
                    <tbody>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">
                                <bold>Mean</bold>
</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.4</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">1.7</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">
                                <bold>Std. Deviation</bold>
</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.3</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">1.4</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">
                                <bold>Median</bold>
</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.2</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">1.4</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">
                                <bold>Minimum</bold>
</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.1</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">0.2</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">
                                <bold>Maximum</bold>
</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">1.0</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">4.3</td>
                        </tr>
                    </tbody>
                </table>
            </table-wrap>
            <table-wrap id="T4" orientation="portrait" position="float">
                <label>
Table 4. </label>
                <caption>
                    <title>The descriptive statistics of calcium ratios.</title>
                </caption>
                <table content-type="article-table" frame="hsides">
                    <thead>
                        <tr>
                            <th align="left" colspan="1" rowspan="1" valign="top">Calcium</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride group</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride + CO2 laser group</th>
                        </tr>
                    </thead>
                    <tbody>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">Mean</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">1.62</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">3.72</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">Std. Deviation</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">1.56</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">6.28</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">Median</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">1.10</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">1.28</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">Minimum</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.50</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">0.60</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">Maximum</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">4.65</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">16.50</td>
                        </tr>
                    </tbody>
                </table>
            </table-wrap>
            <table-wrap id="T5" orientation="portrait" position="float">
                <label>
Table 5. </label>
                <caption>
                    <title>The descriptive statistics of phosphorus ratios.</title>
                </caption>
                <table content-type="article-table" frame="hsides">
                    <thead>
                        <tr>
                            <th align="left" colspan="1" rowspan="1" valign="top">Phosphorus</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride group</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride + CO2 laser group</th>
                        </tr>
                    </thead>
                    <tbody>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Mean</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.99</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.65</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Std. Deviation</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">1.13</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.92</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Median</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.39</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.35</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Minimum</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.11</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">0.00</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Maximum</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">2.44</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">2.50</td>
                        </tr>
                    </tbody>
                </table>
            </table-wrap>
            <table-wrap id="T6" orientation="portrait" position="float">
                <label>
Table 6. </label>
                <caption>
                    <title>The descriptive statistics of carbon ratios.</title>
                </caption>
                <table content-type="article-table" frame="hsides">
                    <thead>
                        <tr>
                            <th align="left" colspan="1" rowspan="1" valign="top">Carbon</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride group</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride + CO2 laser group</th>
                        </tr>
                    </thead>
                    <tbody>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">Mean</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">72.89</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">67.29</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">Std. Deviation</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">8.97</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">7.42</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">Median</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">74.24</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">68.75</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">Minimum</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">57.13</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">57.80</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="middle">Maximum</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">84.12</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">77.56</td>
                        </tr>
                    </tbody>
                </table>
            </table-wrap>
            <p>The results of the Mann-Whitney U test showed a significant statistical difference in fluoride ratios between the two groups (P value&#x00a0;=&#x00a0;0.026) smaller than 0.05, indicating the effectiveness of using CO
                <sub>2</sub> lasers in enhancing fluoride absorption within the tooth structure. In contrast, the remaining elements (calcium, phosphorus, carbon) did not show significant differences, although there were apparent changes in the arithmetic mean, in the rest of the descriptive statistics measures, and in the ranks of the averages (
                <xref ref-type="table" rid="T7">
Table 7</xref>).</p>
            <table-wrap id="T7" orientation="portrait" position="float">
                <label>
Table 7. </label>
                <caption>
                    <title>Mann-Whitney U test results.</title>
                </caption>
                <table content-type="article-table" frame="hsides">
                    <thead>
                        <tr>
                            <th align="left" colspan="1" rowspan="1" valign="top"/>
                            <th align="left" colspan="1" rowspan="1" valign="top">Fluoride</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Calcium</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">phosphorus</th>
                            <th align="left" colspan="1" rowspan="1" valign="top">Carbon</th>
                        </tr>
                    </thead>
                    <tbody>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Test value</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">4.5</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">15</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">17</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">11</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">P value</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">0.026</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">0.699</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">0.937</td>
                            <td align="left" colspan="1" rowspan="1" valign="middle">0.31</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Comparison with significance level</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">Less than 0.05</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">Greater than 0.05</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">Greater than 0.05</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">Greater than 0.05</td>
                        </tr>
                        <tr>
                            <td align="left" colspan="1" rowspan="1" valign="top">Result</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">There is a statistically significant difference between the two groups, indicating a clear effect of the laser in raising fluoride levels</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">no statistically significant difference</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">no statistically significant difference</td>
                            <td align="left" colspan="1" rowspan="1" valign="top">no statistically significant difference</td>
                        </tr>
                    </tbody>
                </table>
            </table-wrap>
        </sec>
        <sec id="sec10" sec-type="discussion">
            <title>Discussion</title>
            <p>This study evaluated the efficacy of a CO
                <sub>2</sub> laser for enhancing fluoride uptake in dental enamel. The combined application of a CO
                <sub>2</sub> laser with fluoride varnishes or gels, which can quadruple fluoride absorption, may significantly improve caries prevention. This synergistic effect could potentially reduce the required fluoride dosage and frequency of clinical applications. The mechanism is attributed to an increased formation of fluorapatite within the enamel, thereby elevating its resistance to acid demineralization.
                <sup>
                    <xref ref-type="bibr" rid="ref13">13</xref>
                </sup>
            </p>
            <p>In order to obtain a protective effect, a laser was used to raise the temperature of the tooth enamel to the melting point of its inorganic components or to the decomposition point of its organic components. On the other hand, non-thermal laser radiation was used after tooth fluoridation to achieve a slow fluoride release effect. This type of reaction is called laser-activated fluoride treatment.
                <sup>
                    <xref ref-type="bibr" rid="ref11">11</xref>
                </sup>
            </p>
            <p>Carbon dioxide laser irradiation of tooth enamel, within certain parameters, results in a specific temperature increase above the irreversible pulpitis threshold of 5.5&#x00a0;&#x00b0;C,
                <sup>
                    <xref ref-type="bibr" rid="ref14">14</xref>
                </sup> which changes the chemical composition of the tooth enamel surface and the morphological structure
                <sup>
                    <xref ref-type="bibr" rid="ref15">15</xref>
                </sup> and the increase may lead to permanent damage to the tooth pulp.</p>
            <p>Excessive temperature elevation during laser application poses a risk of injury to the dental pulp. A study addressing this risk compared the thermal effects of two laser systems: a CO
                <sub>2</sub> laser (at 1 and 2&#x00a0;W) and a diode laser (at 5 and 7&#x00a0;W). The results indicated that the mean temperature rise remained below the 5.5&#x00a0;&#x00b0;C critical threshold for pulp damage. Based on this evidence, the authors determined that both lasers could be used safely without compromising pulp health.
                <sup>
                    <xref ref-type="bibr" rid="ref16">16</xref>
                </sup>
            </p>
            <p>The absorption depth of port and ivory is 12&#x00a0;&#x03bc;m, compared to the absorption depth of water of 15&#x00a0;&#x03bc;m.
                <sup>
                    <xref ref-type="bibr" rid="ref17">17</xref>
                </sup>
            </p>
            <p>The absorption effects of tooth enamel and dentin at these depths depend on how the laser energy affects them and the laser energy depends on laser parameters such as power, mode of operation (continuous or pulsed wave mode), flow (energy density) and dose.</p>
            <p>The continuous wave mode resulted in carbonization, dissolution and cracking of the enamel and the development of the pulse mode allowed variations by frequency and pulse duration in milliseconds (ms) and microseconds (&#x03bc;s). This improved the thermal control of the laser in tissue. The dose depends on the radiation method: spot radiation or sweeping motion, the number of repeated radiations.
                <sup>
                    <xref ref-type="bibr" rid="ref9">9</xref>
                </sup>
            </p>
            <p>Several hypotheses have been advanced to explain the mechanisms by which combined laser and fluoride treatment enhances resistance to enamel demineralization. A principal theory posits that laser irradiation creates micro-spaces within the enamel structure, which subsequently serve to retain fluoride ions more effectively. These structural alterations are also believed to contribute directly to reducing enamel solubility. The observed morphological changes on the enamel surface following the sequential application of topical fluoride and CO
                <sub>2</sub> laser irradiation provide a rationale for the superior protective effect compared to fluoride treatment alone. This synergistic effect is further attributed to the laser-induced conversion of hydroxyapatite to the more acid-resistant fluorapatite, a process accompanied by the melting and recrystallization of hydroxyapatite crystals.
                <sup>
                    <xref ref-type="bibr" rid="ref18">18</xref>
                </sup>
            </p>
            <p>The results of previous studies showed that treating the surface of tooth enamel using a carbon dioxide laser reduces demineralization of tooth enamel compared to the control group. Studies have shown that wavelengths from 9 to 11 micrometers of CO2 lasers are effectively absorbed by dental hydroxyapatite, causing loss of carbonate mineral, which in turn reduces acid reactivity.
                <sup>
                    <xref ref-type="bibr" rid="ref19">19</xref>,
                    <xref ref-type="bibr" rid="ref20">20</xref>
                </sup>
            </p>
            <p>The integration of lasers in preventive dentistry reflects a modern paradigm shift within the field, moving away from a traditional focus on restorative treatment and toward a philosophy of disease prevention. This approach is consistent with the recommendations of major professional bodies and is part of a wider scientific consensus advocating for less invasive caries management strategies,
                <sup>
                    <xref ref-type="bibr" rid="ref21">21</xref>
                </sup> Which aims to preserve tissues to the maximum extent, and in this new approach, prevention has acquired an important role that it has not played before in the past.
                <sup>
                    <xref ref-type="bibr" rid="ref11">11</xref>
                </sup>
            </p>
            <p>All experiments combining CO2 and fluoride laser radiation have shown better results in preventing tooth caries when compared to a single treatment. Therefore, this &#x2018;combination therapy&#x2019; may be clinically effective, and at the same time, it may only involve moderate daily doses of fluoride and low levels of laser radiation energy, which is consistent with our current study.</p>
        </sec>
        <sec id="sec11" sec-type="conclusion">
            <title>Conclusion</title>
            <p>

                <list list-type="bullet">
                    <list-item>
                        <label>&#x2022;</label>
                        <p>Irradiation of tooth enamel at specific wavelengths and CO2 laser energy densities alters hydroxyapatite crystals, making the surface more resistant.</p>
                    </list-item>
                    <list-item>
                        <label>&#x2022;</label>
                        <p>Laser therapy can be an alternative or synergistic to topical fluoridation to prevent tooth enamel caries with a longer-lasting effect.</p>
                    </list-item>
                    <list-item>
                        <label>&#x2022;</label>
                        <p>When a carbon dioxide laser is combined with fluoride, it is possible to reduce the laser energy density and reduce the number of times fluoride is applied.</p>
                    </list-item>
                    <list-item>
                        <label>&#x2022;</label>
                        <p>If this CO2 laser technology becomes available at an affordable cost and the results can be applied in clinical practice, there will be a promising future for this laser in preventing tooth caries.</p>
                    </list-item>
                </list>
            </p>
            <sec id="sec12">
                <title>Recommendations</title>
                <p>Although these results are very promising, more studies should be developed in order to test these radiation conditions under different challenges, and simulate other clinical variables and therefore, future clinical studies are recommended to confirm these results.</p>
            </sec>
        </sec>
    </body>
    <back>
        <sec id="sec15" sec-type="data-availability">
            <title>Data availability</title>
            <p>The dataset includes raw EDX values for fluoride, calcium, phosphorus, and carbon, as well as the data used for statistical analyses and tables are It is available to the public at this link.</p>
            <p>Figshare: Comparison between the protective effect of applying sodium fluoride varnish alone and applying CO2 laser with it in permanent teeth. 
                <ext-link ext-link-type="uri" xlink:href="https://doi.org/10.6084/m9.figshare.31333387.v2">https://doi.org/10.6084/m9.figshare.31333387.v2</ext-link>
                <sup>
                    <xref ref-type="bibr" rid="ref22">22</xref>
                </sup>
            </p>
            <p>The dataset includes:
                <list list-type="bullet">
                    <list-item>
                        <label>&#x2022;</label>
                        <p>Research Data</p>
                    </list-item>
                    <list-item>
                        <label>&#x2022;</label>
                        <p>EDx images</p>
                    </list-item>
                </list>
            </p>
            <p>Data are available under the terms of the 
                <ext-link ext-link-type="uri" xlink:href="https://creativecommons.org/licenses/by/4.0/">CC BY 4.0</ext-link>
            </p>
            <sec id="sec16">
                <title>Extended data</title>
                <p>Extended data associated with this study, including EDX spectra, and detailed statistical output tables, are available in the same repository as the underlying data under a 
                    <ext-link ext-link-type="uri" xlink:href="https://creativecommons.org/licenses/by/4.0/">CC BY 4.0 license</ext-link>.</p>
            </sec>
        </sec>
        <ack>
            <title>Acknowledgments</title>
            <p>None.</p>
        </ack>
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                <aff id="r485241a1">
                    <label>1</label>Oral Pathology and MIicrobiology, Bharati Vidyapeeth (Deemed to be University) Dental College and Hospital Navi Mumbai, Navi Mumbai, Maharashtra, India</aff>
            </contrib-group>
            <author-notes>
                <fn fn-type="conflict">
                    <p>
                        <bold>Competing interests: </bold>No competing interests were disclosed.</p>
                </fn>
            </author-notes>
            <pub-date pub-type="epub">
                <day>4</day>
                <month>6</month>
                <year>2026</year>
            </pub-date>
            <permissions>
                <copyright-statement>Copyright: &#x00a9; 2026 Sachdev SS</copyright-statement>
                <copyright-year>2026</copyright-year>
                <license xlink:href="https://creativecommons.org/licenses/by/4.0/">
                    <license-p>This is an open access peer review report distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
                </license>
            </permissions>
            <related-article ext-link-type="doi" id="relatedArticleReport485241" related-article-type="peer-reviewed-article" xlink:href="10.12688/f1000research.171971.1"/>
            <custom-meta-group>
                <custom-meta>
                    <meta-name>recommendation</meta-name>
                    <meta-value>reject</meta-value>
                </custom-meta>
            </custom-meta-group>
        </front-stub>
        <body>
            <p>The introduction states the importance of fluoride, then abruptly moves to the harmful effects of fluoride, followed by the&#x00a0;enhancement using a laser. Nowhere is the need for the application of lasers specified.&#x00a0;Is the normal fluoride application insufficient such that additional laser application is required?</p>
            <p> The introduction is majorly unformatted and needs proper structuring in paragraphs. Needs better flow of the content.</p>
            <p> Table 1 not required. Non-contributory to manuscript. Authors can just cite the justification for their range. SEM and EDX full forms are repeated.&#x00a0;</p>
            <p> The AI generated placeholder [&#x201c;Not applicable as the study used discarded biological material&#x201d;] has got retained in the manuscript after the ethical approval number. The authors probably forgot to remove it. So currently there is a reference ethical approval number and 'not applicable' statement at the same time, which are contradicting each other.</p>
            <p> Indicate by arrows what is to be seen in the SEM images, and also mention that in the legends. Include SEM results in the abstract too.</p>
            <p> </p>
            <p> Discussion makes a strong claim 'The combined application of a CO
                <sub>2</sub>&#x00a0;laser with fluoride varnishes or gels, which can quadruple fluoride absorption, may significantly improve caries prevention. ' The authors need to specify that such a claim is based on their results or from existing literature. If existing literature, cite the references accordingly. But in this case, the present study is not required, if it is already established that lasers enhance the binding four-fold. If based on the present study's findings, it is too strong a claim based on an in vitro study on 12 samples.</p>
            <p> </p>
            <p> Justification for the sample size is required, otherwise the study is just a pilot study with a small sample.</p>
            <p> Citations for all the mechanisms stated for action of laser are required. Currently, only one reference is cited at the end of the paragraph discussing the mechanisms.</p>
            <p> </p>
            <p> The discussion flow is abrupt and inconsistent, requiring overall re-writing.</p>
            <p> </p>
            <p> Conclusion must have a modest tone, and organized as a paragraph rather than bulleted list.</p>
            <p>Is the work clearly and accurately presented and does it cite the current literature?</p>
            <p>No</p>
            <p>If applicable, is the statistical analysis and its interpretation appropriate?</p>
            <p>I cannot comment. A qualified statistician is required.</p>
            <p>Are all the source data underlying the results available to ensure full reproducibility?</p>
            <p>No source data required</p>
            <p>Is the study design appropriate and is the work technically sound?</p>
            <p>No</p>
            <p>Are the conclusions drawn adequately supported by the results?</p>
            <p>No</p>
            <p>Are sufficient details of methods and analysis provided to allow replication by others?</p>
            <p>No</p>
            <p>Reviewer Expertise:</p>
            <p>Oral Pathology, Dental Materials, Dental Innovations, Dental Anatomy, Dentistry</p>
            <p>I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above.</p>
        </body>
    </sub-article>
    <sub-article article-type="reviewer-report" id="report480471">
        <front-stub>
            <article-id pub-id-type="doi">10.5256/f1000research.189644.r480471</article-id>
            <title-group>
                <article-title>Reviewer response for version 1</article-title>
            </title-group>
            <contrib-group>
                <contrib contrib-type="author">
                    <name>
                        <surname>Qasim</surname>
                        <given-names>Alhan Ahmed</given-names>
                    </name>
                    <xref ref-type="aff" rid="r480471a1">1</xref>
                    <role>Referee</role>
                </contrib>
                <aff id="r480471a1">
                    <label>1</label>University of Baghdad, Baghdad, Baghdad Governorate, Iraq</aff>
            </contrib-group>
            <author-notes>
                <fn fn-type="conflict">
                    <p>
                        <bold>Competing interests: </bold>No competing interests were disclosed.</p>
                </fn>
            </author-notes>
            <pub-date pub-type="epub">
                <day>19</day>
                <month>5</month>
                <year>2026</year>
            </pub-date>
            <permissions>
                <copyright-statement>Copyright: &#x00a9; 2026 Qasim AA</copyright-statement>
                <copyright-year>2026</copyright-year>
                <license xlink:href="https://creativecommons.org/licenses/by/4.0/">
                    <license-p>This is an open access peer review report distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</license-p>
                </license>
            </permissions>
            <related-article ext-link-type="doi" id="relatedArticleReport480471" related-article-type="peer-reviewed-article" xlink:href="10.12688/f1000research.171971.1"/>
            <custom-meta-group>
                <custom-meta>
                    <meta-name>recommendation</meta-name>
                    <meta-value>approve</meta-value>
                </custom-meta>
            </custom-meta-group>
        </front-stub>
        <body>
            <p>Greetings :</p>
            <p> 1. Introduction: It needs to highlight the important of enamel protection role against acid dissolution.</p>
            <p> paragraph 3 "&#x00a0;Recent epidemiological trends indicate .............." the reference is old ; attention should be paid to this.</p>
            <p> 2. Discussion: the results of this study need further clarification and comparison&#x00a0; with other studies, and it is preferable recent.</p>
            <p> The authors did not mention the limitations of this study.</p>
            <p> 3.&#x00a0;Conclusion: second point"&#x00a0;&#x00a0;Laser therapy can be an alternative or synergistic to topical fluoridation to prevent tooth enamel caries with a longer-lasting effect" according to the result of this study laser is&#x00a0;synergistic to topical fluoride application to increase fluoride uptake by enamel surface.</p>
            <p>Is the work clearly and accurately presented and does it cite the current literature?</p>
            <p>Partly</p>
            <p>If applicable, is the statistical analysis and its interpretation appropriate?</p>
            <p>Yes</p>
            <p>Are all the source data underlying the results available to ensure full reproducibility?</p>
            <p>Yes</p>
            <p>Is the study design appropriate and is the work technically sound?</p>
            <p>Yes</p>
            <p>Are the conclusions drawn adequately supported by the results?</p>
            <p>Partly</p>
            <p>Are sufficient details of methods and analysis provided to allow replication by others?</p>
            <p>Yes</p>
            <p>Reviewer Expertise:</p>
            <p>Preventive Dentistry</p>
            <p>I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard.</p>
        </body>
        <back>
            <ref-list>
                <title>References</title>
                <ref id="rep-ref-480471-1">
                    <label>1</label>
                    <mixed-citation publication-type="journal">
                        <person-group person-group-type="author"/>:
                        <article-title>CO2 Laser and Topical Fluoride Therapy in the Control of Caries Lesions on Demineralized Primary Enamel</article-title>.
                        <source>
                            <italic>The Scientific World Journal</italic>
                        </source>.<year>2015</year>;<volume>2015</volume>(<issue>1</issue>) :
                        <elocation-id>10.1155/2015/547569</elocation-id>
                        <pub-id pub-id-type="doi">10.1155/2015/547569</pub-id>
                    </mixed-citation>
                </ref>
            </ref-list>
        </back>
    </sub-article>
</article>
