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<article dtd-version="3.0" article-type="Methods Article - JoVE Produced Video">
  <front>
    <journal-meta>
      <journal-id journal-id-type="publisher">jove</journal-id>
      <journal-id journal-id-type="email">editorial-office@jove.com</journal-id>
      <journal-title-group>
        <journal-title>Journal of Visualized Experiments</journal-title>
      </journal-title-group>
      <issn pub-type="ppub">1940-087X</issn>
      <issn pub-type="epub"></issn>
    </journal-meta>
    <article-meta>
      <article-id pub-id-type="publisher-id">JoVE57294</article-id>
      <article-id pub-id-type="manuscript">JoVE57294</article-id>
      <article-categories>
        <subj-group subj-group-type="Article Type">
          <subject>Methods Article - JoVE Produced Video</subject>
        </subj-group>
        <subj-group subj-group-type="Category">
          <subject>JoVE Immunology and Infection</subject>
        </subj-group>
        <subj-group subj-group-type="Classification">
          <subject>Skeleton</subject>
          <subject>Bacterial Infections</subject>
          <subject>Bone Diseases, Infectious</subject>
          <subject>Bacterial Infections and Mycoses</subject>
        </subj-group>
      </article-categories>
      <title-group>
        <article-title>Treatment with Vancomycin Loaded Calcium Sulphate and Autogenous Bone in an Improved Rabbit Model of Bone Infection</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <role content-type="1" />
          <name>
            <surname>Zhang</surname>
            <given-names>Yang</given-names>
          </name>
          <email>zhangyang0310@163.com</email>
          <xref ref-type="aff" rid="aff1" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="2" />
          <name>
            <surname>Shen</surname>
            <given-names>Lifeng</given-names>
          </name>
          <email>hzshenlf@163.com</email>
          <xref ref-type="aff" rid="aff2" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="3" />
          <name>
            <surname>Wang</surname>
            <given-names>Ping'er</given-names>
          </name>
          <email>apple63209321@126.com</email>
          <xref ref-type="aff" rid="aff3" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="4" />
          <name>
            <surname>Xi</surname>
            <given-names>Weixing</given-names>
          </name>
          <email>877845932@qq.com</email>
          <xref ref-type="aff" rid="aff4" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="5" />
          <name>
            <surname>Yu</surname>
            <given-names>Zhongming</given-names>
          </name>
          <email>mm98239@sohu.com</email>
          <xref ref-type="aff" rid="aff5" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="6" />
          <name>
            <surname>Huang</surname>
            <given-names>Xiaowen</given-names>
          </name>
          <email>bowen8883@126.com</email>
          <xref ref-type="aff" rid="aff6" />
        </contrib>
        <contrib contrib-type="author">
          <role content-type="7" />
          <name>
            <surname>Wang</surname>
            <given-names>Xuping</given-names>
          </name>
          <email>1050979506@qq.com</email>
          <xref ref-type="aff" rid="aff7" />
        </contrib>
        <contrib contrib-type="author" corresp="yes">
          <role content-type="8" />
          <name>
            <surname>Shou</surname>
            <given-names>Dan</given-names>
            <prefix>Dr.</prefix>
          </name>
          <email>shoudanok@163.com</email>
          <xref ref-type="aff" rid="aff8" />
          <xref ref-type="aff" rid="orcid29" />
        </contrib>
        <contrib contrib-type="editor">
          <name>
            <surname>Navani</surname>
            <given-names>Dipesh</given-names>
          </name>
          <degrees>Ph.D.</degrees>
          <email>dipesh.n@jove.com</email>
          <xref ref-type="aff" rid="aff9" />
        </contrib>
        <aff id="aff1">
          <institution>Zhejaing Academy of Traditional Chinese Medicine</institution>
          <institution content-type="dept">Department of Medicine</institution>
          <addr-line content-type="addrline1">No. 132 Tianmushan Road</addr-line>
          <country>CHINA</country>
          <addr-line content-type="city">Hangzhou</addr-line>
          <addr-line content-type="state">Zhejiang</addr-line>
          <addr-line content-type="zipcode">310007</addr-line>
          <institution>Zhejiang Academy of traditional Chinese Medicine</institution>
        </aff>
        <aff id="aff2">
          <institution>Tongde Hospital Of Zhejiang Province</institution>
          <institution content-type="dept">Department of Orthopaedic Surgery</institution>
          <addr-line content-type="addrline1">No. 234 Gucui Road</addr-line>
          <country>CHINA</country>
          <addr-line content-type="city">Hangzhou</addr-line>
          <addr-line content-type="zipcode">310017</addr-line>
          <institution>Tongde Hospital of Zhejiang Province</institution>
        </aff>
        <aff id="aff3">
          <institution>The First Affiliated Hospital of Zhejiang Chinese Medical University</institution>
          <institution content-type="dept">Institute of Orthopaedics and Traumatology</institution>
          <addr-line content-type="addrline1">No. 548 Binwen Road</addr-line>
          <country>CHINA</country>
          <addr-line content-type="city">Hangzhou</addr-line>
          <addr-line content-type="zipcode">310053</addr-line>
          <institution>The First Affiliated Hospital of Zhejiang Chinese Medical University</institution>
        </aff>
        <aff id="aff4">
          <institution>Tongde Hospital Of Zhejiang Province</institution>
          <institution content-type="dept">Department of Clinical Laboratory</institution>
          <addr-line content-type="addrline1">No. 234 Gucui Road</addr-line>
          <country>CHINA</country>
          <addr-line content-type="city">Hangzhou</addr-line>
          <addr-line content-type="zipcode">310017</addr-line>
          <institution>Tongde Hospital of Zhejiang Province</institution>
        </aff>
        <aff id="aff5">
          <institution>Zheajiang Academy of Traditional Chinese Medicine</institution>
          <institution content-type="dept">Department of Medicine</institution>
          <addr-line content-type="addrline1">No. 132 Tianmushan Road</addr-line>
          <country>CHINA</country>
          <addr-line content-type="city">Hangzhou</addr-line>
          <addr-line content-type="state">Zhejiang</addr-line>
          <addr-line content-type="zipcode">310007</addr-line>
          <institution>Zhejiang Academy of Traditional Chinese Medicine</institution>
        </aff>
        <aff id="aff6">
          <institution>Zhejiang Police Vocational Academy</institution>
          <institution content-type="dept">Department of Medicine</institution>
          <addr-line content-type="addrline1">No. 132 Tianmushan Road</addr-line>
          <country>CHINA</country>
          <addr-line content-type="city">Hangzhou</addr-line>
          <addr-line content-type="state">Zhejiang</addr-line>
          <addr-line content-type="zipcode">310007</addr-line>
          <institution>Zhejiang Academy of Traditional Chinese Medicine</institution>
        </aff>
        <aff id="aff7">
          <institution>Zhejiang Academy of Traditional Chinese Medicine</institution>
          <addr-line content-type="addrline1">No. 132 Tianmushan Road</addr-line>
          <country>CHINA</country>
          <addr-line content-type="city">Hangzhou</addr-line>
          <addr-line content-type="zipcode">310007</addr-line>
        </aff>
        <aff id="aff8">
          <institution>Zhejiang Academy of Traditional Chinese Medicine</institution>
          <institution content-type="position">Director</institution>
          <addr-line content-type="addrline1">No. 132 Tianmushan Road</addr-line>
          <country>CHINA</country>
          <addr-line content-type="city">Hangzhou</addr-line>
          <addr-line content-type="state">Zhejiang</addr-line>
          <addr-line content-type="zipcode">310007</addr-line>
          <phone content-type="primary">+8613606803860</phone>
          <institution>Zhejiang Academy of Traditional Chinese Medicine</institution>
        </aff>
        <aff id="orcid29">0000-0003-3465-4072</aff>
        <aff id="aff9">
          <institution>JoVE</institution>
          <addr-line>1 Alewife Center</addr-line>
          <addr-line>Suite 200</addr-line>
          <addr-line>Cambridge, MA  02140</addr-line>
          <country>UNITED STATES</country>
        </aff>
      </contrib-group>
      <history>
        <date date-type="received">
          <day>10</day>
          <month>09</month>
          <year>2017</year>
        </date>
        <date date-type="rev-recd">
          <day>31</day>
          <month>10</month>
          <year>2018</year>
        </date>
        <date date-type="rev-recd">
          <day>16</day>
          <month>10</month>
          <year>2018</year>
        </date>
        <date date-type="rev-recd">
          <day>10</day>
          <month>09</month>
          <year>2018</year>
        </date>
        <date date-type="accepted">
          <day>31</day>
          <month>10</month>
          <year>2018</year>
        </date>
      </history>
      <abstract>
        <p>Bone infection results from bacterial invasion, which is extremely difficult to treat in clinical, orthopaedic, and traumatic surgery. The bone infection may result in sustained inflammation, osteomyelitis, and eventual bone non-union. Establishment of a feasible, reproducible animal model is important to bone infection research and antibiotic treatment. As an in vivo model, the rabbit model is widely used in bone infection research. However, previous studies on rabbit bone infection models showed that the infection status was inconsistent, as the amount of bacteria was variable. This study presents an improved surgical method for inducing bone infection on a rabbit, by blocking the bacteria in the bone marrow. Then, multi-level evaluations can be carried out to verify the modelling method.

In general, debriding necrotic tissue and implantation of vancomycin-loaded calcium sulphate (VCS) are predominant in antibiotic treatment. Although calcium sulphate in VCS benefits osteocyte crawling and new bone growth, massive bone defects occur after debriding. Autogenous bone (AB) is an appealing strategy to overcome bone defects for the treatment of massive bone defects after debriding necrotic bone.

In this study, we used the tail bone as an autogenous bone implanted in the bone defect.Bone repair was measured  using micro-computed-tomography (micro-CT) and histological analysis after animal sacrifice. As a result, in the VCS group, bone non-union was consistently obtained. In contrast, the bone defect areas in the VCS-AB group were decreased significantly. The present modeling method described a reproducible, feasible, stable method to prepare a bone infection model. The VCS-AB treatment resulted in lower bone non-union rates after antibiotic treatment. The improved bone infection model and the combination treatment of VCS and autogenous bone could be helpful in studying the underlying mechanisms in bone infection and bone regeneration pertinent to traumatology orthopaedics applications.</p>
      </abstract>
      <kwd-group>
        <kwd>Bone infection</kwd>
        <kwd>rabbit model</kwd>
        <kwd>Staphylococcus aureus</kwd>
        <kwd>Tibia</kwd>
        <kwd>vancomycin loaded calcium sulfate</kwd>
        <kwd>autogenous bone</kwd>
      </kwd-group>
      <funding-group />
      <counts>
        <fig-count count="0" />
      </counts>
      <custom-meta-group>
        <custom-meta>
          <meta-name>Please indicate whether this article will be Standard Access or Open Access. </meta-name>
          <meta-value>Standard Access (US$2,400);</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Please indicate the &lt;b&gt; city, state/province, and country&lt;/b&gt; where this article will be &lt;b&gt;filmed&lt;/b&gt;. Please do not use abbreviations.</meta-name>
          <meta-value>No. 132 Tianmushan Road, Hangzhou, Zhejiang, China</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Black and White Image Count</meta-name>
          <meta-value>0</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Color Image Count</meta-name>
          <meta-value>0</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Production Notes</meta-name>
          <meta-value>Request Medicine:  Rabbit model for bone infection resulting from bacteria invasion following clinical orthopedic and traumatic surgery</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Acquisitions Editor</meta-name>
          <meta-value>Nandita Singh</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>Assistant who entered lead</meta-name>
          <meta-value>Jennifer Carroll</meta-value>
        </custom-meta>
        <custom-meta>
          <meta-name>ALA Access Type</meta-name>
          <meta-value>JPV Standard Access ($2400)</meta-value>
        </custom-meta>
      </custom-meta-group>
    </article-meta>
  </front>
  <body>
    <fig>
      <label>Figure 1</label>
      <graphic xlink:href="Fig 1.tif" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 3</label>
      <graphic xlink:href="Fig 3.tif" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <table-wrap>
      <label>Table of Materials/Equipment</label>
      <graphic xlink:href="JoVE_Materials.xls" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
    <fig>
      <label>Figure 2</label>
      <graphic xlink:href="Fig 2.tif" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 4</label>
      <graphic xlink:href="renamed_6a89e.tif" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 4</label>
      <graphic xlink:href="Fig 4.tif" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <fig>
      <label>Figure 5</label>
      <graphic xlink:href="Fig 5.tif" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </fig>
    <table-wrap>
      <label>Table of Materials</label>
      <graphic xlink:href="JoVE_Materials revised.xls" xmlns:xlink="http://www.w3.org/1999/xlink" />
    </table-wrap>
  </body>
</article>