<clinical_study>
  <!-- This xml conforms to an XML Schema at:
    https://clinicaltrials.gov/ct2/html/images/info/public.xsd -->
  <required_header>
    <download_date>ClinicalTrials.gov processed this data on September 20, 2023</download_date>
    <link_text>Link to the current ClinicalTrials.gov record.</link_text>
    <url>https://clinicaltrials.gov/ct2/show/NCT01664871</url>
  </required_header>
  <id_info>
    <org_study_id>UC1110</org_study_id>
    <nct_id>NCT01664871</nct_id>
  </id_info>
  <brief_title>Pharmacokinetics of Diammine Silver Fluoride</brief_title>
  <official_title>Characterization of Fluoride and Silver Pharmacokinetics of Diammine Silver Fluoride Applied Topically to Teeth of Adults</official_title>
  <sponsors>
    <lead_sponsor>
      <agency>Universidad Peruana Cayetano Heredia</agency>
      <agency_class>Other</agency_class>
    </lead_sponsor>
  </sponsors>
  <source>Universidad Peruana Cayetano Heredia</source>
  <oversight_info>
    <has_dmc>No</has_dmc>
  </oversight_info>
  <brief_summary>
    <textblock>
      There is growing worldwide interest in the use of diammine silver fluoride (DSF) as a topical&#xD;
      agent to treat dentin hypersensitivity and dental caries. It has been available in various&#xD;
      formulations for many years, but its safety profile has never been fully characterized.&#xD;
&#xD;
      This preliminary study determined the applied doses (3 teeth treated), maximum serum&#xD;
      concentrations, and time to maximum serum concentration for fluoride and silver in 6 adults&#xD;
      over 4 h. Fluoride was determined using the indirect diffusion method with a fluoride&#xD;
      selective electrode, and silver was determined using inductively coupled plasma-mass&#xD;
      spectrometry. The mean amount of DSF solution applied to the 3 teeth was 7.57 mg (6.04 µL).&#xD;
&#xD;
      .&#xD;
    </textblock>
  </brief_summary>
  <detailed_description>
    <textblock>
      Participants Six healthy volunteers (4 females, mean age 36.2, range 23-52 yr) were recruited&#xD;
      among staff members of the Universidad Católica de Santa María, Arequipa, Peru. Participants&#xD;
      were taking no medications and had most permanent teeth present. Participants were asked to&#xD;
      avoid fish or tea for 12 h before the study and not use fluoridated toothpaste 4 h prior. The&#xD;
      Comité de Etica de Investigacion, Universidad Católica de Santa María approved the study and&#xD;
      informed consent was obtained.&#xD;
&#xD;
      Study Agent DSF [Ag(NH3)2F, CAS RN 33040-28-7, Saforide, Toyo Seiyaku Kasei Co. Ltd. Osaka,&#xD;
      Japan] was used. DSF is clear and colorless, with a weak odor of ammonia. The 38% solution&#xD;
      contains between 24.4-28.8% (w/v) silver (Ag), and 5.0-5.9% fluoride (F). Concentrations of&#xD;
      Ag and F in the DSF used for study were 24.9% and 5.5%, respectively, and the specific&#xD;
      gravity was 1.253 at 25°C (Certificate of Analysis Lot #155060). Diammine silver fluoride is&#xD;
      also referred to in the literature as silver diammine fluoride, silver diamine fluoride or&#xD;
      silver fluoride.&#xD;
&#xD;
      Procedures Three maxillary teeth (cuspid and premolars) in the same quadrant were treated.&#xD;
      Three teeth were treated because this was the average number of sensitive teeth reported in&#xD;
      an epidemiological study of adults [5]. The teeth were free of restorations or cavitation.&#xD;
      The facial surface was dried with cotton gauze and then DSF was applied to the cervical area&#xD;
      of each tooth. The teeth were isolated in order to avoid wetting the brush with saliva. Each&#xD;
      tooth was coated using a separate microbrush, which was dipped in the DSF and weighed before&#xD;
      and after the application (Model S2000, Kern &amp; Sohn GmbH, Balingen, Germany). The sum of the&#xD;
      differences in weight of material applied was calculated for each participant.&#xD;
&#xD;
      Blood Collection Immediately before application of DSF and then again at approximately 30&#xD;
      min, 1, 2,3 and 4 h after application, blood was obtained from a peripheral vein. The time&#xD;
      periods were picked based on a previous pharmacokinetics study of fluoride varnish [6]. The&#xD;
      blood was transferred to a fluoride-free plastic Vacutainer tube with clot activator (BD&#xD;
      Vacutainer Plus Plastic Serum Tube, BD Diagnostics, New Jersey). The tubes were spun at 1100&#xD;
      g for 10 min and serum transferred by plastic pipette to plastic cryogenic tubes. The samples&#xD;
      were frozen and transported to the University of Washington for analysis.&#xD;
&#xD;
      Analysis Fluoride. Samples were thawed and analyzed in duplicate or triplicate using the&#xD;
      diffusion and detection method with hexamethyldisiloxane (HMDS) and Orion F-sensitive&#xD;
      electrode [7-8]. To improve detection, the pre-diffusion step was omitted [9], and seals were&#xD;
      completed before mixing 1 mL serum with 1 mL HMDS, saturated 3 M H2SO4 [10]. The trapping&#xD;
      solution was 20 µmoles 1 M NaOH. It was dried down to eliminate variability of evaporation&#xD;
      and residual HMDS before adding 100 µL 3 M acetic acid to a final pH 3.4. Standards were 10&#xD;
      and 100 µM fluoride in the same final concentration of NaOH and acetic acid corresponding to&#xD;
      1 and 10 µM F in the samples. ORION F and sleeve type reference electrodes were coupled with&#xD;
      an Accumet Basic 0.1 mV meter (Thermo Fisher Scientific, Milwaukee WI).&#xD;
&#xD;
      Vacutainers were tested for fluoride by adding bovine plasma to 2 tubes and letting them sit&#xD;
      overnight before combining the 2 samples and comparing them to plasma directly placed in&#xD;
      diffusion dishes. Average concentrations (μM±SD) were 0.34±0.014 and 0.32±0.012,&#xD;
      respectively. Therefore, less than 2% F in the serum samples came from the Vacutainers and&#xD;
      clot enhancer. Recovery (mean±SD) of 1 nmole/ml F added to bovine serum was 101±4%. Fluoride&#xD;
      levels were averaged and are expressed as µM.&#xD;
&#xD;
      Silver. Samples were thawed and analyzed using Inductively Coupled Plasma-Mass Spectrometry&#xD;
      (EPA 6020a Rev.1 2007). Serum (0.5 mL) was brought to 2 mL with dilute acid (final&#xD;
      concentration: 2% HCl, 1% HNO3; trace metal grade, Fisher, Fairlawn, NJ). Samples were&#xD;
      centrifuged (1290 g) for 10 min and filtered (0.45 µm, PTFE; all polypropylene syringe) into&#xD;
      15 mL polypropylene centrifuge tubes. The samples were analyzed on an Agilent 7500CE (Santa&#xD;
      Clara, CA) inductively-coupled mass spectrometer (ICPMS) with He as the collision cell gas&#xD;
      and an ASX-510 autosampler (CETAC, Omaha, NB). A Micromist nebulizer (Glass Expansion,&#xD;
      Pocasset, MA) was used (1.15 L/min carrier gas; no makeup gas). Spray chamber temperature was&#xD;
      2°C. RF power was 1500 W. Ag was quantified using Y as internal standard. Calibrants were&#xD;
      diluted with 2% HCl and 1% HNO3 prepared from commercial ICPMS grade solutions (Ultra&#xD;
      Scientific, N. Kingstown, RI; BDH Aristar, VWR, Radnor, PA) and ranged from 0.5-50 ng/mL.&#xD;
      Calibration was by weighted (1/cps) linear regression. Data was corrected by procedure blank.&#xD;
      Values of silver less than 2 ng/mL were recoded to 2. Silver concentrations are expressed as&#xD;
      ng/mL.&#xD;
&#xD;
      Soft Tissue Assessment The gingiva adjacent to the application of DSF and the mucosa&#xD;
      generally were observed at baseline and 24 h after treatment. Erythema, bleeding, white&#xD;
      changes, ulceration and pigmentation were assessed using methods developed for an earlier&#xD;
      study [1].&#xD;
    </textblock>
  </detailed_description>
  <overall_status>Completed</overall_status>
  <start_date>July 2011</start_date>
  <completion_date type="Actual">September 2011</completion_date>
  <primary_completion_date type="Actual">August 2011</primary_completion_date>
  <phase>Phase 1</phase>
  <study_type>Interventional</study_type>
  <has_expanded_access>No</has_expanded_access>
  <study_design_info>
    <allocation>N/A</allocation>
    <intervention_model>Single Group Assignment</intervention_model>
    <primary_purpose>Treatment</primary_purpose>
    <masking>None (Open Label)</masking>
  </study_design_info>
  <primary_outcome>
    <measure>Level of Fluoride in Plasma</measure>
    <time_frame>baseline, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours</time_frame>
    <description>Samples were thawed and analyzed in duplicate or triplicate using the diffusion and detection method with hexamethyldisiloxane (HMDS) and Orion F-sensitive electrode</description>
  </primary_outcome>
  <primary_outcome>
    <measure>Level of Silver in Plasma</measure>
    <time_frame>baseline, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours</time_frame>
    <description>Samples were thawed and analyzed using Inductively Coupled Plasma-Mass Spectrometry</description>
  </primary_outcome>
  <secondary_outcome>
    <measure>Gingival alterations</measure>
    <time_frame>baseline, immediately after treatment, 24 hours after treatment</time_frame>
    <description>The gingiva adjacent to the application of DSF and the mucosa generally were observed at baseline and 24 h after treatment. Erythema, bleeding, white changes, ulceration and pigmentation were assessed using methods developed for an earlier study</description>
  </secondary_outcome>
  <number_of_arms>1</number_of_arms>
  <enrollment type="Actual">6</enrollment>
  <condition>Fluoride and Silver Pharmacokinetics of DSF Applied Topically to Teeth of Adults. Erythema, Bleeding, White Changes, Ulceration and Pigmentation Were Assessed</condition>
  <arm_group>
    <arm_group_label>Levels of SIlver and Fluoride in Plasma</arm_group_label>
    <arm_group_type>Experimental</arm_group_type>
  </arm_group>
  <intervention>
    <intervention_type>Device</intervention_type>
    <intervention_name>DIammine SIlver Fluoride</intervention_name>
    <description>Concentrations of Silver and Fluoride in the DSF used for study were 24.9% and 5.5%, respectively Average total weight (calculated volume) of Diammine Silver Fluoride applied was 7.57 mg (6.04 µL), corresponding to a mean application of 0.33 mg Fluoride based on the lot analysis. The mean total amount of Silver applied to the 3 teeth was 1.50 mg.</description>
    <arm_group_label>Levels of SIlver and Fluoride in Plasma</arm_group_label>
  </intervention>
  <eligibility>
    <criteria>
      <textblock>
        Inclusion Criteria:&#xD;
&#xD;
          -  Subjects with dental abfraction, erosion or root caries at least in one upper or lower&#xD;
             premolar or molar&#xD;
&#xD;
          -  Those teeth should have hypersensitivity with cold air&#xD;
&#xD;
        Exclusion Criteria:&#xD;
&#xD;
          -  Use of any type of hypersensitivity toothpaste&#xD;
&#xD;
          -  Use of pain medication&#xD;
&#xD;
          -  Pregnant women&#xD;
&#xD;
          -  Individuals with sensitivity to silver or other metal ions&#xD;
      </textblock>
    </criteria>
    <gender>All</gender>
    <minimum_age>10 Years</minimum_age>
    <maximum_age>60 Years</maximum_age>
    <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
  </eligibility>
  <overall_official>
    <last_name>Peter Milgrom, DDS</last_name>
    <role>Principal Investigator</role>
    <affiliation>University of Washington</affiliation>
  </overall_official>
  <overall_official>
    <last_name>Elsa Vasquez, DDS</last_name>
    <role>Study Director</role>
    <affiliation>Universidad Catolica Santa Maria Arequipa</affiliation>
  </overall_official>
  <overall_official>
    <last_name>Jorge Castillo, DDS MSD PhD</last_name>
    <role>Study Chair</role>
    <affiliation>Universidad Peruana Cayetano Heredia</affiliation>
  </overall_official>
  <location>
    <facility>
      <name>Universidad Catolica Santa Maria</name>
      <address>
        <city>Arequipa</city>
        <country>Peru</country>
      </address>
    </facility>
  </location>
  <location_countries>
    <country>Peru</country>
  </location_countries>
  <verification_date>August 2012</verification_date>
  <study_first_submitted>August 10, 2012</study_first_submitted>
  <study_first_submitted_qc>August 10, 2012</study_first_submitted_qc>
  <study_first_posted type="Estimate">August 14, 2012</study_first_posted>
  <last_update_submitted>August 10, 2012</last_update_submitted>
  <last_update_submitted_qc>August 10, 2012</last_update_submitted_qc>
  <last_update_posted type="Estimate">August 14, 2012</last_update_posted>
  <responsible_party>
    <responsible_party_type>Sponsor</responsible_party_type>
  </responsible_party>
  <condition_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Erythema</mesh_term>
  </condition_browse>
  <intervention_browse>
    <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
    <mesh_term>Fluorides</mesh_term>
  </intervention_browse>
  <pending_results>
    <submitted>May 20, 2013</submitted>
    <returned>August 9, 2013</returned>
    <submitted>December 10, 2015</submitted>
    <returned>January 14, 2016</returned>
    <submitted>December 3, 2021</submitted>
    <returned>February 17, 2022</returned>
  </pending_results>
  <!-- Results have not yet been posted for this study                                          -->
</clinical_study>


download date: ClinicalTrials.gov processed this data on September 20, 2023
link text: Link to the current ClinicalTrials.gov record.
url: https://clinicaltrials.gov/ct2/show/NCT01664871
org study id: UC1110
nct id: NCT01664871
lead sponsor: 
has dmc: No
There is growing worldwide interest in the use of diammine silver fluoride (DSF) as a topical
      agent to treat dentin hypersensitivity and dental caries. It has been available in various
      formulations for many years, but its safety profile has never been fully characterized.

      This preliminary study determined the applied doses (3 teeth treated), maximum serum
      concentrations, and time to maximum serum concentration for fluoride and silver in 6 adults
      over 4 h. Fluoride was determined using the indirect diffusion method with a fluoride
      selective electrode, and silver was determined using inductively coupled plasma-mass
      spectrometry. The mean amount of DSF solution applied to the 3 teeth was 7.57 mg (6.04 µL).

      .
Participants Six healthy volunteers (4 females, mean age 36.2, range 23-52 yr) were recruited
      among staff members of the Universidad Católica de Santa María, Arequipa, Peru. Participants
      were taking no medications and had most permanent teeth present. Participants were asked to
      avoid fish or tea for 12 h before the study and not use fluoridated toothpaste 4 h prior. The
      Comité de Etica de Investigacion, Universidad Católica de Santa María approved the study and
      informed consent was obtained.

      Study Agent DSF [Ag(NH3)2F, CAS RN 33040-28-7, Saforide, Toyo Seiyaku Kasei Co. Ltd. Osaka,
      Japan] was used. DSF is clear and colorless, with a weak odor of ammonia. The 38% solution
      contains between 24.4-28.8% (w/v) silver (Ag), and 5.0-5.9% fluoride (F). Concentrations of
      Ag and F in the DSF used for study were 24.9% and 5.5%, respectively, and the specific
      gravity was 1.253 at 25°C (Certificate of Analysis Lot #155060). Diammine silver fluoride is
      also referred to in the literature as silver diammine fluoride, silver diamine fluoride or
      silver fluoride.

      Procedures Three maxillary teeth (cuspid and premolars) in the same quadrant were treated.
      Three teeth were treated because this was the average number of sensitive teeth reported in
      an epidemiological study of adults [5]. The teeth were free of restorations or cavitation.
      The facial surface was dried with cotton gauze and then DSF was applied to the cervical area
      of each tooth. The teeth were isolated in order to avoid wetting the brush with saliva. Each
      tooth was coated using a separate microbrush, which was dipped in the DSF and weighed before
      and after the application (Model S2000, Kern & Sohn GmbH, Balingen, Germany). The sum of the
      differences in weight of material applied was calculated for each participant.

      Blood Collection Immediately before application of DSF and then again at approximately 30
      min, 1, 2,3 and 4 h after application, blood was obtained from a peripheral vein. The time
      periods were picked based on a previous pharmacokinetics study of fluoride varnish [6]. The
      blood was transferred to a fluoride-free plastic Vacutainer tube with clot activator (BD
      Vacutainer Plus Plastic Serum Tube, BD Diagnostics, New Jersey). The tubes were spun at 1100
      g for 10 min and serum transferred by plastic pipette to plastic cryogenic tubes. The samples
      were frozen and transported to the University of Washington for analysis.

      Analysis Fluoride. Samples were thawed and analyzed in duplicate or triplicate using the
      diffusion and detection method with hexamethyldisiloxane (HMDS) and Orion F-sensitive
      electrode [7-8]. To improve detection, the pre-diffusion step was omitted [9], and seals were
      completed before mixing 1 mL serum with 1 mL HMDS, saturated 3 M H2SO4 [10]. The trapping
      solution was 20 µmoles 1 M NaOH. It was dried down to eliminate variability of evaporation
      and residual HMDS before adding 100 µL 3 M acetic acid to a final pH 3.4. Standards were 10
      and 100 µM fluoride in the same final concentration of NaOH and acetic acid corresponding to
      1 and 10 µM F in the samples. ORION F and sleeve type reference electrodes were coupled with
      an Accumet Basic 0.1 mV meter (Thermo Fisher Scientific, Milwaukee WI).

      Vacutainers were tested for fluoride by adding bovine plasma to 2 tubes and letting them sit
      overnight before combining the 2 samples and comparing them to plasma directly placed in
      diffusion dishes. Average concentrations (μM±SD) were 0.34±0.014 and 0.32±0.012,
      respectively. Therefore, less than 2% F in the serum samples came from the Vacutainers and
      clot enhancer. Recovery (mean±SD) of 1 nmole/ml F added to bovine serum was 101±4%. Fluoride
      levels were averaged and are expressed as µM.

      Silver. Samples were thawed and analyzed using Inductively Coupled Plasma-Mass Spectrometry
      (EPA 6020a Rev.1 2007). Serum (0.5 mL) was brought to 2 mL with dilute acid (final
      concentration: 2% HCl, 1% HNO3; trace metal grade, Fisher, Fairlawn, NJ). Samples were
      centrifuged (1290 g) for 10 min and filtered (0.45 µm, PTFE; all polypropylene syringe) into
      15 mL polypropylene centrifuge tubes. The samples were analyzed on an Agilent 7500CE (Santa
      Clara, CA) inductively-coupled mass spectrometer (ICPMS) with He as the collision cell gas
      and an ASX-510 autosampler (CETAC, Omaha, NB). A Micromist nebulizer (Glass Expansion,
      Pocasset, MA) was used (1.15 L/min carrier gas; no makeup gas). Spray chamber temperature was
      2°C. RF power was 1500 W. Ag was quantified using Y as internal standard. Calibrants were
      diluted with 2% HCl and 1% HNO3 prepared from commercial ICPMS grade solutions (Ultra
      Scientific, N. Kingstown, RI; BDH Aristar, VWR, Radnor, PA) and ranged from 0.5-50 ng/mL.
      Calibration was by weighted (1/cps) linear regression. Data was corrected by procedure blank.
      Values of silver less than 2 ng/mL were recoded to 2. Silver concentrations are expressed as
      ng/mL.

      Soft Tissue Assessment The gingiva adjacent to the application of DSF and the mucosa
      generally were observed at baseline and 24 h after treatment. Erythema, bleeding, white
      changes, ulceration and pigmentation were assessed using methods developed for an earlier
      study [1].
allocation: N/A
intervention model: Single Group Assignment
primary purpose: Treatment
masking: None (Open Label)
measure: Level of Fluoride in Plasma
time frame: baseline, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours
description: Samples were thawed and analyzed in duplicate or triplicate using the diffusion and detection method with hexamethyldisiloxane (HMDS) and Orion F-sensitive electrode
measure: Level of Silver in Plasma
time frame: baseline, 30 minutes, 1 hour, 2 hours, 3 hours, 4 hours
description: Samples were thawed and analyzed using Inductively Coupled Plasma-Mass Spectrometry
measure: Gingival alterations
time frame: baseline, immediately after treatment, 24 hours after treatment
description: The gingiva adjacent to the application of DSF and the mucosa generally were observed at baseline and 24 h after treatment. Erythema, bleeding, white changes, ulceration and pigmentation were assessed using methods developed for an earlier study
arm group label: Levels of SIlver and Fluoride in Plasma
arm group type: Experimental
intervention type: Device
intervention name: DIammine SIlver Fluoride
description: Concentrations of Silver and Fluoride in the DSF used for study were 24.9% and 5.5%, respectively Average total weight (calculated volume) of Diammine Silver Fluoride applied was 7.57 mg (6.04 µL), corresponding to a mean application of 0.33 mg Fluoride based on the lot analysis. The mean total amount of Silver applied to the 3 teeth was 1.50 mg.
arm group label: Levels of SIlver and Fluoride in Plasma
criteria: 
gender: All
minimum age: 10 Years
maximum age: 60 Years
healthy volunteers: Accepts Healthy Volunteers
last name: Peter Milgrom, DDS
role: Principal Investigator
affiliation: University of Washington
last name: Elsa Vasquez, DDS
role: Study Director
affiliation: Universidad Catolica Santa Maria Arequipa
last name: Jorge Castillo, DDS MSD PhD
role: Study Chair
affiliation: Universidad Peruana Cayetano Heredia
facility: 
country: Peru
responsible party type: Sponsor
mesh term: Erythema
mesh term: Fluorides
submitted: May 20, 2013
returned: August 9, 2013
submitted: December 10, 2015
returned: January 14, 2016
submitted: December 3, 2021
returned: February 17, 2022