xml: <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/NCT06020092</url>
    </required_header>
    <id_info>
      <org_study_id>2158-61-46-22-109</org_study_id>
      <nct_id>NCT06020092</nct_id>
    </id_info>
    <brief_title>Histological Analysis of Osteogenic Potential of Biphasic Calcium Phosphate</brief_title>
    <official_title>Histological Analysis of Osteogenic Potential of Biphasic Calcium Phosphate in Alveolar Ridge Augmentation</official_title>
    <sponsors>
      <lead_sponsor>
        <agency>Josip Juraj Strossmayer University of Osijek</agency>
        <agency_class>Other</agency_class>
      </lead_sponsor>
    </sponsors>
    <source>Josip Juraj Strossmayer University of Osijek</source>
    <oversight_info>
      <has_dmc>Yes</has_dmc>
      <is_fda_regulated_drug>No</is_fda_regulated_drug>
      <is_fda_regulated_device>No</is_fda_regulated_device>
      <is_us_export>No</is_us_export>
    </oversight_info>
    <brief_summary>
      <textblock>
        Bone healing after tooth extraction may be affected by defects in the buccal wall of the&#xD;
        alveolus, such as fenestrations and dehiscences. Therefore, it is advisable to perform a&#xD;
        surgical procedure such as guided bone regeneration after tooth extractions. Different&#xD;
        biomaterials are used for this purpose. The aim of this study is to investigate the&#xD;
        qualitative and quantitative histological changes in human biopsies taken after 6 months of&#xD;
        healing of extraction sockets with buccal wall defects. For this purpose, the defects of 36&#xD;
        patients (18 per group) will be augmented with injectable biphasic calcium phosphate (I-BCP)&#xD;
        or bovine xenograft (BX) after extraction. After six months of healing, bone biopsies will be&#xD;
        taken and processed to the qualitative and quantitative histological analysis.&#xD;
      </textblock>
    </brief_summary>
    <detailed_description>
      <textblock>
        The study was designed as a randomized controlled clinical trial. To participate in the&#xD;
        study, patients requires to have at least one tooth predisposed to extraction and to have a&#xD;
        dental implant placed after completion of the healing period. The indications for tooth&#xD;
        extraction are: chronic periapical process, chronic periodontal disease, or fracture of the&#xD;
        vertical bone. The inclusion criteria are: age between 18 and 60 years, intact physical and&#xD;
        mental health, patient must understand the study protocol, patient must sign an informed&#xD;
        consent. Patients will be excluded from the study if they had at least one of the absolute&#xD;
        contraindications to implant prosthetic therapy described by Wang and Hwang. Other exclusion&#xD;
        criteria included the following systemic diseases: uncontrolled diabetes, osteoporosis and&#xD;
        osteopenia, and vitamin D deficiency, bisphosphonate therapy, glucocorticoid therapy,&#xD;
        hypothyroidism, uncontrolled cardiovascular disease (hypertension, coronary artery disease,&#xD;
        congestive heart failure), and the following local factors: use of tobacco products (up to 10&#xD;
        cigarettes per day) and poor oral hygiene. Pregnant and lactating women will not included in&#xD;
        the study.&#xD;
  &#xD;
        In addition to the general health criteria mentioned above, the patient has to meet at least&#xD;
        one intraoral criterion, namely the presence of a buccal bone defect such as fenestration or&#xD;
        dehiscence. Patients who met the inclusion criteria and were willing to participate, are&#xD;
        randomly assigned via a web interface (https://www.randomizer.org/).&#xD;
  &#xD;
        Participants and materials Participants will be recruited from the oral surgery office. A&#xD;
        total of 36 healthy patients will be included in the study. In analyzing the validity of the&#xD;
        test, the approach of equivalence of the tested methods and noninferiority of the tested&#xD;
        method compared with the existing method was used. With a significance level α of 0.05, a&#xD;
        test power of 80%, an expected standard deviation of the predicted result of 10, and an&#xD;
        equivalence limit of d = 10, it was necessary to include at least 36 subjects in the study,&#xD;
        18 subjects per group.&#xD;
  &#xD;
        A total of 18 patients in the test group will receive I-BCP (Maxresorb® inject, Botiss GmbH,&#xD;
        Berlin, Germany). This biomaterial is an alloplastic bone substitute material in the form of&#xD;
        a paste that is placed into the defect with a plastic syringe. The material consists of an&#xD;
        aqueous gel containing granules of biphasic calcium phosphate in the composition of 60% HA&#xD;
        and 40% β-TCP (particle size between 15-50 nm). Patients in the control group will receive BX&#xD;
        (cerabone®, Botiss GmbH, Berlin, Germany). The material is completely anorganic and consists&#xD;
        of 100% HA. For this study, the material will be used in granular form (the size of the&#xD;
        granules is between 0.5 and 1 mm), mixed with physiological solution prior to placement into&#xD;
        the defect.&#xD;
  &#xD;
        First surgical phase and healing period Before the procedure, the patient will be&#xD;
        administered a 3% chlorhexidine solution to rinse the oral cavity for one minute. Local&#xD;
        anesthesia (2% Lidocaine®, Belupo, Koprivnica, Croatia) will be then administered and the&#xD;
        surgical field will be covered with sterile drapes. A full-thickness mucoperiosteal flap will&#xD;
        be elevated at the extraction site. To perform the tooth extraction atraumatically, the roots&#xD;
        of the multirooted teeth will be separated with a drill before extraction. The extraction&#xD;
        will be performed with atraumatic instruments, with as little trauma to the alveolus as&#xD;
        possible. The alveolus is then carefully examined and curetted to remove granulations and&#xD;
        stimulate fresh bleeding in the alveoli. The alveolus will be filled to the edge with I-BCP&#xD;
        in the test group and BX in the control group. In the end bone grafting biomaterial is&#xD;
        covered with a resorbable native collagen membrane obtained from porcine pericardium (Jason®&#xD;
        membrane, Botiss GmbH, Berlin, Germany) in both groups. The mucoperiosteal flap will be then&#xD;
        adjusted and closed so that the collagen membrane was completely covered with soft tissue&#xD;
        (5.0 monofilament). The patient will be then prescribed an analgesic (400 mg ibuprofen) and&#xD;
        instructed to take antibiotics (amoxicillin with clavulanic acid or clindamycin) for the next&#xD;
        7 days. On the 10th day after surgery, the sutures are removed.&#xD;
  &#xD;
        A 6-month period of regeneration of the bone defect will follow. Based on the radiographic&#xD;
        examination, the selection and positioning of the appropriate implant will be performed.&#xD;
  &#xD;
        Second surgical phase Six months after extraction, patients come to the office for placement&#xD;
        of a dental implant (Ankylos®, Dentsply Sirona, Charlotte, USA) in the augmented area. The&#xD;
        preparation of the patient (rinsing of the oral cavity with an antiseptic solution, local&#xD;
        anesthesia, sterile covering of the working area) is the same as for the first surgical&#xD;
        procedure. Subsequently, a full-thickness flap is elevated, and a biopsy of the augmented&#xD;
        area is taken for histological analysis using a trephine drill (2.5 mm inner diameter;&#xD;
        Ustomed® intrumente, Tuttlingen, Germany). It is important to emphasize that the inner&#xD;
        diameter of the trephine drill is smaller than that of the final drill used to prepare the&#xD;
        implant bed to avoid unnecessary removal of the patient's bone. In addition, each biopsy will&#xD;
        be taken from the central part of the augmented area to reduce the possibility of an&#xD;
        incorrect biopsy (e.g., bi-opsy of only the native bone that was not augmented with&#xD;
        biomaterial).&#xD;
  &#xD;
        Histological analysis The trephine drill will be placed in a hermetically sealed plastic tube&#xD;
        containing a 4% formaldehyde solution and sent to the histology laboratory for further&#xD;
        analysis. The tissue will be fixed in 4% formaldehyde solution for two weeks. Subsequently,&#xD;
        the biopsies will be stored in 25% ethylenediaminetetraacetic acid (EDTA) (Decalcifier soft®,&#xD;
        Solvagreen®, Karlsruhe, Austria), which is used for gentle decalcification of bone. After&#xD;
        decalcification, the tissue will be subjected to a standard protocol in a tissue processor&#xD;
        (MTP, SLEE medical GmbH, Mainz, Germany). This will be followed by the formation of paraffin&#xD;
        blocks (MPS/P, SLEE medical GmbH, Mainz, Germany). Six consecutive sections of 5 μm thickness&#xD;
        will be prepared from each biopsy using a rotary microtome (CUT 4062, SLEE medical GmbH,&#xD;
        Mainz, Germany). The specimens will be then stained with Masson trichrome stain and Movat&#xD;
        pentachrome stain. Microscopy of the preparations will be performed using a light microscope&#xD;
        (Leica DMRB, Leica Microsystems GmbH, Wetzlar, Germany) in conjunction with a video camera&#xD;
        (Axio Imager M2, Zeiss, Oberkochen, Germany). Pathohistological analysis will evaluate the&#xD;
        tissue response to the implanted biomaterial, i.e., the presence of fibroblasts, blood&#xD;
        vessels, neutrophils, monocytes/macrophages, and multinucleated giant cells (MGC).&#xD;
  &#xD;
        Histomorphometric processing of the specimens will be performed using the free ImageJ&#xD;
        software (https://imagej.nih.gov/ij/download.html). The surface of the new bone, the sur-face&#xD;
        of the biomaterial, and the surface of the soft tissue will be marked on the specimens. The&#xD;
        surface of the specimen will be then measured, and the following will be calculated from the&#xD;
        data obtained: percentage of new bone formed, percentage of biomaterial remaining, and&#xD;
        percentage of soft tissue.&#xD;
  &#xD;
        Statistical analysis Categorical variables will be reported with absolute and relative&#xD;
        frequencies. Differences between categorical variables were tested with the&#xD;
        Fisher-Freeman-Halton test.&#xD;
  &#xD;
        Normality of distribution will be tested with the Shapiro-Wilk test. Continuous data will be&#xD;
        described by the arithmetic mean and standard deviation. Differences in continuous vari-ables&#xD;
        will be tested with the Mann-Whitney U test (with corresponding difference and 95% confidence&#xD;
        interval of difference).&#xD;
  &#xD;
        All P values will be two-sided. The significance level was set at alpha = 0.05. IBM SPSS&#xD;
        Statistical software, version 27.0.1, will be used for the analysis.&#xD;
      </textblock>
    </detailed_description>
    <overall_status>Completed</overall_status>
    <start_date type="Actual">July 31, 2021</start_date>
    <completion_date type="Actual">July 31, 2023</completion_date>
    <primary_completion_date type="Actual">July 31, 2023</primary_completion_date>
    <phase>Phase 4</phase>
    <study_type>Interventional</study_type>
    <has_expanded_access>No</has_expanded_access>
    <study_design_info>
      <allocation>Randomized</allocation>
      <intervention_model>Parallel Assignment</intervention_model>
      <primary_purpose>Treatment</primary_purpose>
      <masking>None (Open Label)</masking>
    </study_design_info>
    <primary_outcome>
      <measure>Percentage of the newly formed bone, soft tissue and residual biomaterial in bone biopsies harvested 6 months after bone augmentation</measure>
      <time_frame>Up to 8 months</time_frame>
      <description>Histomorphometric processing of the specimens will be performed using the free ImageJ software (https://imagej.nih.gov/ij/download.html). The surface of the new bone, the surface of the biomaterial, and the surface of the soft tissue will be marked on the specimens. Marked surfaces of the specimen will be then measured, and the following will be calculated from the data obtained: percentage of new bone formed, percentage of biomaterial remaining, and percentage of soft tissue</description>
    </primary_outcome>
    <other_outcome>
      <measure>Qualitative histological assessment</measure>
      <time_frame>Up to 8 months</time_frame>
      <description>Tissue response to implanted biomaterial is assessed by descriptive histologic evaluation, i.e., the presence of fibroblasts, blood vessels, neutrophils, monocytes/macrophages, and multinucleated giant cells (MGC).</description>
    </other_outcome>
    <number_of_arms>2</number_of_arms>
    <enrollment type="Actual">36</enrollment>
    <condition>Alveolar Ridge Enlargement</condition>
    <condition>Fenestration</condition>
    <condition>Dehiscence</condition>
    <arm_group>
      <arm_group_label>Injectable biphasic calcium phosphate (maxresorb inject, botiss GmbH)</arm_group_label>
      <arm_group_type>Experimental</arm_group_type>
      <description>This biomaterial is an alloplastic bone substitute material in the form of a paste that is placed into the defect with a plastic syringe. The material consists of an aqueous gel containing granules of biphasic calcium phosphate in the composition of 60% HA and 40% β-TCP (particle size between 15-50 nm).</description>
    </arm_group>
    <arm_group>
      <arm_group_label>Bovine xenograft (cerabone, botiss GmbH)</arm_group_label>
      <arm_group_type>Active Comparator</arm_group_type>
      <description>The material is completely anorganic and consists of 100% HA. For this study, the material will be used in granular form (the size of the granules is between 0.5 and 1 mm), mixed with physiological solution prior to placement into the defect.</description>
    </arm_group>
    <intervention>
      <intervention_type>Procedure</intervention_type>
      <intervention_name>Bone augmentation with I-BCP</intervention_name>
      <description>The extraction socket will be filled after tooth extraction with I-BCP, in order to achieve bone regeneration and prevent volume loss after tooth extraction.</description>
      <arm_group_label>Injectable biphasic calcium phosphate (maxresorb inject, botiss GmbH)</arm_group_label>
    </intervention>
    <intervention>
      <intervention_type>Procedure</intervention_type>
      <intervention_name>Bone augmentation with BX</intervention_name>
      <description>The extraction socket will be filled after tooth extraction with BX, in order to achieve bone regeneration and prevent volume loss after tooth extraction.</description>
      <arm_group_label>Bovine xenograft (cerabone, botiss GmbH)</arm_group_label>
    </intervention>
    <eligibility>
      <criteria>
        <textblock>
          Inclusion Criteria:&#xD;
  &#xD;
            -  age between 18 and 60 years&#xD;
  &#xD;
            -  patients requires to have at least one tooth predisposed to extraction&#xD;
  &#xD;
            -  intact physical and mental health&#xD;
  &#xD;
            -  patient must understand the study protocol&#xD;
  &#xD;
            -  patient must sign an informed consent.&#xD;
  &#xD;
            -  the presence of a buccal bone defect such as fenestration or dehiscence.&#xD;
  &#xD;
          Exclusion Criteria:&#xD;
  &#xD;
            -  patients will be excluded from the study if they had at least one of the absolute&#xD;
               contraindications to implant prosthetic therapy&#xD;
  &#xD;
            -  following systemic diseases: uncontrolled diabetes, osteoporosis and osteopenia, and&#xD;
               vitamin D deficiency&#xD;
  &#xD;
            -  bisphosphonate therapy&#xD;
  &#xD;
            -  glucocorticoid therapy&#xD;
  &#xD;
            -  hypothyroidism&#xD;
  &#xD;
            -  uncontrolled cardiovascular disease (hypertension, coronary artery disease, congestive&#xD;
               heart failure)&#xD;
  &#xD;
            -  pregnant or lactating women&#xD;
  &#xD;
            -  following local factors: use of tobacco products (up to 10 cigarettes per day) and&#xD;
               poor oral hygiene&#xD;
        </textblock>
      </criteria>
      <gender>All</gender>
      <minimum_age>18 Years</minimum_age>
      <maximum_age>60 Years</maximum_age>
      <healthy_volunteers>Accepts Healthy Volunteers</healthy_volunteers>
    </eligibility>
    <overall_official>
      <last_name>Marija Candrlic, PhD</last_name>
      <role>Principal Investigator</role>
      <affiliation>Faculty of Medicine Osijek</affiliation>
    </overall_official>
    <location>
      <facility>
        <name>Faculty of Medicine Osijek</name>
        <address>
          <city>Osijek</city>
          <state>Osijek Baranja</state>
          <zip>31000</zip>
          <country>Croatia</country>
        </address>
      </facility>
    </location>
    <location_countries>
      <country>Croatia</country>
    </location_countries>
    <reference>
      <citation>Candrlic M, Peric Kacarevic Z, Ivanisevic Z, Tomas M, Vcev A, Faj D, Matijevic M. Histological and Radiological Features of a Four-Phase Injectable Synthetic Bone Graft in Guided Bone Regeneration: A Case Report. Int J Environ Res Public Health. 2020 Dec 29;18(1):206. doi: 10.3390/ijerph18010206.</citation>
      <PMID>33383971</PMID>
    </reference>
    <reference>
      <citation>Tomas M, Candrlic M, Juzbasic M, Ivanisevic Z, Matijevic N, Vcev A, Cvijanovic Peloza O, Matijevic M, Peric Kacarevic Z. Synthetic Injectable Biomaterials for Alveolar Bone Regeneration in Animal and Human Studies. Materials (Basel). 2021 May 26;14(11):2858. doi: 10.3390/ma14112858.</citation>
      <PMID>34073551</PMID>
    </reference>
    <verification_date>August 2023</verification_date>
    <study_first_submitted>August 10, 2023</study_first_submitted>
    <study_first_submitted_qc>August 27, 2023</study_first_submitted_qc>
    <study_first_posted type="Actual">August 31, 2023</study_first_posted>
    <last_update_submitted>August 27, 2023</last_update_submitted>
    <last_update_submitted_qc>August 27, 2023</last_update_submitted_qc>
    <last_update_posted type="Actual">August 31, 2023</last_update_posted>
    <responsible_party>
      <responsible_party_type>Principal Investigator</responsible_party_type>
      <investigator_affiliation>Josip Juraj Strossmayer University of Osijek</investigator_affiliation>
      <investigator_full_name>Marija Čandrlić</investigator_full_name>
      <investigator_title>Postdoctoral researcher at Department of Dental Medicine</investigator_title>
    </responsible_party>
    <keyword>bone regeneration</keyword>
    <keyword>alloplast</keyword>
    <keyword>bovine xenograft</keyword>
    <keyword>guided bone regeneration</keyword>
    <condition_browse>
      <!-- CAUTION:  The following MeSH terms are assigned with an imperfect algorithm            -->
      <mesh_term>Hypertrophy</mesh_term>
    </condition_browse>
    <patient_data>
      <sharing_ipd>No</sharing_ipd>
      <ipd_description>I do not plan to make IPD available to other researchers. IPDs will be administrated and stored by one person.</ipd_description>
    </patient_data>
    <!-- Results have not yet been posted for this study                                          -->
  </clinical_study>
  
  
text: 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/NCT06020092
  org study id: 2158-61-46-22-109
  nct id: NCT06020092
  lead sponsor: 
  has dmc: Yes
  is fda regulated drug: No
  is fda regulated device: No
  is us export: No
  Bone healing after tooth extraction may be affected by defects in the buccal wall of the
        alveolus, such as fenestrations and dehiscences. Therefore, it is advisable to perform a
        surgical procedure such as guided bone regeneration after tooth extractions. Different
        biomaterials are used for this purpose. The aim of this study is to investigate the
        qualitative and quantitative histological changes in human biopsies taken after 6 months of
        healing of extraction sockets with buccal wall defects. For this purpose, the defects of 36
        patients (18 per group) will be augmented with injectable biphasic calcium phosphate (I-BCP)
        or bovine xenograft (BX) after extraction. After six months of healing, bone biopsies will be
        taken and processed to the qualitative and quantitative histological analysis.
  The study was designed as a randomized controlled clinical trial. To participate in the
        study, patients requires to have at least one tooth predisposed to extraction and to have a
        dental implant placed after completion of the healing period. The indications for tooth
        extraction are: chronic periapical process, chronic periodontal disease, or fracture of the
        vertical bone. The inclusion criteria are: age between 18 and 60 years, intact physical and
        mental health, patient must understand the study protocol, patient must sign an informed
        consent. Patients will be excluded from the study if they had at least one of the absolute
        contraindications to implant prosthetic therapy described by Wang and Hwang. Other exclusion
        criteria included the following systemic diseases: uncontrolled diabetes, osteoporosis and
        osteopenia, and vitamin D deficiency, bisphosphonate therapy, glucocorticoid therapy,
        hypothyroidism, uncontrolled cardiovascular disease (hypertension, coronary artery disease,
        congestive heart failure), and the following local factors: use of tobacco products (up to 10
        cigarettes per day) and poor oral hygiene. Pregnant and lactating women will not included in
        the study.
  
        In addition to the general health criteria mentioned above, the patient has to meet at least
        one intraoral criterion, namely the presence of a buccal bone defect such as fenestration or
        dehiscence. Patients who met the inclusion criteria and were willing to participate, are
        randomly assigned via a web interface (https://www.randomizer.org/).
  
        Participants and materials Participants will be recruited from the oral surgery office. A
        total of 36 healthy patients will be included in the study. In analyzing the validity of the
        test, the approach of equivalence of the tested methods and noninferiority of the tested
        method compared with the existing method was used. With a significance level α of 0.05, a
        test power of 80%, an expected standard deviation of the predicted result of 10, and an
        equivalence limit of d = 10, it was necessary to include at least 36 subjects in the study,
        18 subjects per group.
  
        A total of 18 patients in the test group will receive I-BCP (Maxresorb® inject, Botiss GmbH,
        Berlin, Germany). This biomaterial is an alloplastic bone substitute material in the form of
        a paste that is placed into the defect with a plastic syringe. The material consists of an
        aqueous gel containing granules of biphasic calcium phosphate in the composition of 60% HA
        and 40% β-TCP (particle size between 15-50 nm). Patients in the control group will receive BX
        (cerabone®, Botiss GmbH, Berlin, Germany). The material is completely anorganic and consists
        of 100% HA. For this study, the material will be used in granular form (the size of the
        granules is between 0.5 and 1 mm), mixed with physiological solution prior to placement into
        the defect.
  
        First surgical phase and healing period Before the procedure, the patient will be
        administered a 3% chlorhexidine solution to rinse the oral cavity for one minute. Local
        anesthesia (2% Lidocaine®, Belupo, Koprivnica, Croatia) will be then administered and the
        surgical field will be covered with sterile drapes. A full-thickness mucoperiosteal flap will
        be elevated at the extraction site. To perform the tooth extraction atraumatically, the roots
        of the multirooted teeth will be separated with a drill before extraction. The extraction
        will be performed with atraumatic instruments, with as little trauma to the alveolus as
        possible. The alveolus is then carefully examined and curetted to remove granulations and
        stimulate fresh bleeding in the alveoli. The alveolus will be filled to the edge with I-BCP
        in the test group and BX in the control group. In the end bone grafting biomaterial is
        covered with a resorbable native collagen membrane obtained from porcine pericardium (Jason®
        membrane, Botiss GmbH, Berlin, Germany) in both groups. The mucoperiosteal flap will be then
        adjusted and closed so that the collagen membrane was completely covered with soft tissue
        (5.0 monofilament). The patient will be then prescribed an analgesic (400 mg ibuprofen) and
        instructed to take antibiotics (amoxicillin with clavulanic acid or clindamycin) for the next
        7 days. On the 10th day after surgery, the sutures are removed.
  
        A 6-month period of regeneration of the bone defect will follow. Based on the radiographic
        examination, the selection and positioning of the appropriate implant will be performed.
  
        Second surgical phase Six months after extraction, patients come to the office for placement
        of a dental implant (Ankylos®, Dentsply Sirona, Charlotte, USA) in the augmented area. The
        preparation of the patient (rinsing of the oral cavity with an antiseptic solution, local
        anesthesia, sterile covering of the working area) is the same as for the first surgical
        procedure. Subsequently, a full-thickness flap is elevated, and a biopsy of the augmented
        area is taken for histological analysis using a trephine drill (2.5 mm inner diameter;
        Ustomed® intrumente, Tuttlingen, Germany). It is important to emphasize that the inner
        diameter of the trephine drill is smaller than that of the final drill used to prepare the
        implant bed to avoid unnecessary removal of the patient's bone. In addition, each biopsy will
        be taken from the central part of the augmented area to reduce the possibility of an
        incorrect biopsy (e.g., bi-opsy of only the native bone that was not augmented with
        biomaterial).
  
        Histological analysis The trephine drill will be placed in a hermetically sealed plastic tube
        containing a 4% formaldehyde solution and sent to the histology laboratory for further
        analysis. The tissue will be fixed in 4% formaldehyde solution for two weeks. Subsequently,
        the biopsies will be stored in 25% ethylenediaminetetraacetic acid (EDTA) (Decalcifier soft®,
        Solvagreen®, Karlsruhe, Austria), which is used for gentle decalcification of bone. After
        decalcification, the tissue will be subjected to a standard protocol in a tissue processor
        (MTP, SLEE medical GmbH, Mainz, Germany). This will be followed by the formation of paraffin
        blocks (MPS/P, SLEE medical GmbH, Mainz, Germany). Six consecutive sections of 5 μm thickness
        will be prepared from each biopsy using a rotary microtome (CUT 4062, SLEE medical GmbH,
        Mainz, Germany). The specimens will be then stained with Masson trichrome stain and Movat
        pentachrome stain. Microscopy of the preparations will be performed using a light microscope
        (Leica DMRB, Leica Microsystems GmbH, Wetzlar, Germany) in conjunction with a video camera
        (Axio Imager M2, Zeiss, Oberkochen, Germany). Pathohistological analysis will evaluate the
        tissue response to the implanted biomaterial, i.e., the presence of fibroblasts, blood
        vessels, neutrophils, monocytes/macrophages, and multinucleated giant cells (MGC).
  
        Histomorphometric processing of the specimens will be performed using the free ImageJ
        software (https://imagej.nih.gov/ij/download.html). The surface of the new bone, the sur-face
        of the biomaterial, and the surface of the soft tissue will be marked on the specimens. The
        surface of the specimen will be then measured, and the following will be calculated from the
        data obtained: percentage of new bone formed, percentage of biomaterial remaining, and
        percentage of soft tissue.
  
        Statistical analysis Categorical variables will be reported with absolute and relative
        frequencies. Differences between categorical variables were tested with the
        Fisher-Freeman-Halton test.
  
        Normality of distribution will be tested with the Shapiro-Wilk test. Continuous data will be
        described by the arithmetic mean and standard deviation. Differences in continuous vari-ables
        will be tested with the Mann-Whitney U test (with corresponding difference and 95% confidence
        interval of difference).
  
        All P values will be two-sided. The significance level was set at alpha = 0.05. IBM SPSS
        Statistical software, version 27.0.1, will be used for the analysis.
  allocation: Randomized
  intervention model: Parallel Assignment
  primary purpose: Treatment
  masking: None (Open Label)
  measure: Percentage of the newly formed bone, soft tissue and residual biomaterial in bone biopsies harvested 6 months after bone augmentation
  time frame: Up to 8 months
  description: Histomorphometric processing of the specimens will be performed using the free ImageJ software (https://imagej.nih.gov/ij/download.html). The surface of the new bone, the surface of the biomaterial, and the surface of the soft tissue will be marked on the specimens. Marked surfaces of the specimen will be then measured, and the following will be calculated from the data obtained: percentage of new bone formed, percentage of biomaterial remaining, and percentage of soft tissue
  measure: Qualitative histological assessment
  time frame: Up to 8 months
  description: Tissue response to implanted biomaterial is assessed by descriptive histologic evaluation, i.e., the presence of fibroblasts, blood vessels, neutrophils, monocytes/macrophages, and multinucleated giant cells (MGC).
  arm group label: Injectable biphasic calcium phosphate (maxresorb inject, botiss GmbH)
  arm group type: Experimental
  description: This biomaterial is an alloplastic bone substitute material in the form of a paste that is placed into the defect with a plastic syringe. The material consists of an aqueous gel containing granules of biphasic calcium phosphate in the composition of 60% HA and 40% β-TCP (particle size between 15-50 nm).
  arm group label: Bovine xenograft (cerabone, botiss GmbH)
  arm group type: Active Comparator
  description: The material is completely anorganic and consists of 100% HA. For this study, the material will be used in granular form (the size of the granules is between 0.5 and 1 mm), mixed with physiological solution prior to placement into the defect.
  intervention type: Procedure
  intervention name: Bone augmentation with I-BCP
  description: The extraction socket will be filled after tooth extraction with I-BCP, in order to achieve bone regeneration and prevent volume loss after tooth extraction.
  arm group label: Injectable biphasic calcium phosphate (maxresorb inject, botiss GmbH)
  intervention type: Procedure
  intervention name: Bone augmentation with BX
  description: The extraction socket will be filled after tooth extraction with BX, in order to achieve bone regeneration and prevent volume loss after tooth extraction.
  arm group label: Bovine xenograft (cerabone, botiss GmbH)
  criteria: 
  gender: All
  minimum age: 18 Years
  maximum age: 60 Years
  healthy volunteers: Accepts Healthy Volunteers
  last name: Marija Candrlic, PhD
  role: Principal Investigator
  affiliation: Faculty of Medicine Osijek
  facility: 
  country: Croatia
  citation: Candrlic M, Peric Kacarevic Z, Ivanisevic Z, Tomas M, Vcev A, Faj D, Matijevic M. Histological and Radiological Features of a Four-Phase Injectable Synthetic Bone Graft in Guided Bone Regeneration: A Case Report. Int J Environ Res Public Health. 2020 Dec 29;18(1):206. doi: 10.3390/ijerph18010206.
  PMID: 33383971
  citation: Tomas M, Candrlic M, Juzbasic M, Ivanisevic Z, Matijevic N, Vcev A, Cvijanovic Peloza O, Matijevic M, Peric Kacarevic Z. Synthetic Injectable Biomaterials for Alveolar Bone Regeneration in Animal and Human Studies. Materials (Basel). 2021 May 26;14(11):2858. doi: 10.3390/ma14112858.
  PMID: 34073551
  responsible party type: Principal Investigator
  investigator affiliation: Josip Juraj Strossmayer University of Osijek
  investigator full name: Marija Čandrlić
  investigator title: Postdoctoral researcher at Department of Dental Medicine
  mesh term: Hypertrophy
  sharing ipd: No
  ipd description: I do not plan to make IPD available to other researchers. IPDs will be administrated and stored by one person.