Name: treatment with vancomycin loaded calcium sulphate and autogenous bone in an improved rabbit model of bone infection
Uploaded: 2019-03-14T12:30:00
Description: Watch this Scientific Journal Video about Treatment with Vancomycin Loaded Calcium Sulphate and Autogenous Bone in an Improved Rabbit Model of Bone Infection at JoVE.com

这项工作得到了国家自然科学基金 (81803808, 81803808)、浙江省医疗卫生科学技术基金 (2017ky271) 和浙江省科技项目 (2017KY271) 的支持。

本研究中使用的兔子按照《实验动物护理和使用指南》进行处理。所有的实验程序都遵循了浙江中医药研究院生物伦理委员会的规则。
1. 细菌悬浮液的制备
<ol><li>溶解0.5 毫克金黄色葡萄球菌冷冻干燥粉 (atcc 6538) 与0.3 毫升的卢里亚-伯塔尼培养基。将悬架完全混合。</li>
	<li>将细菌悬浮液流到色氨酸大豆琼脂板上, 在37°c 孵育细菌菌落16小时。</li>
	<li>选择单个菌落形成...

<ol>
	<li>Malizos, K. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Global+Forum:+The+Burden+of+Bone+and+Joint+Infection:+A+Growing+Demand+for+More+Resources.">Global Forum: The Burden of Bone and Joint Infection: A Growing Demand for More Resources.</a> Journal of Bone and Joint Surgery-American Volume. 99, 20 (2017).</li><li>Peeters, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Teicoplanin+-+based+antimicrobial+therapy+in+Staphylococcus+aureus+bone+and+joint+infection:+tolerance,+efficacy+and+experience+with+subcutaneous+administration.">Teicoplanin - based antimicrobial therapy in Staphylococcus aureus bone and joint infection: tolerance, efficacy and experience with subcutaneous administration.</a> BMC Infectious Diseases. 16, 622 (2016).</li><li>Sugaya, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Percutaneous+autologous+concentrated+bone+marrow+grafting+in+the+treatment+for+nonunion.">Percutaneous autologous concentrated bone marrow grafting in the treatment for nonunion.</a> European Journal of Orthopeadic Surgery and Traumatology. 24, 671-678 (2014).</li><li>Birt, M. C., Anderson, D. W., Bruce, T. E., Wang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osteomyelitis:+Recent+advances+in+pathophysiology+and+therapeutic+strategies.">Osteomyelitis: Recent advances in pathophysiology and therapeutic strategies.</a> Journal of Orthopeadics. 14, 45-52 (2017).</li><li>Walter, G., Kemmerer, M., Kappler, C., Hoffmann, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+algorithms+for+chronic+osteomyelitis.">Treatment algorithms for chronic osteomyelitis.</a> Deutsches Arzteblatt International. 109, 257-264 (2012).</li><li>Henriksen, K., Neutzsky-Wulff, A. V., Bonewald, L. F., Karsdal, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+communication+on+and+within+bone+controls+bone+remodeling.">Local communication on and within bone controls bone remodeling.</a> Bone. 44, 1026-1033 (2009).</li><li>Mendoza, M. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+vancomycin+powder+on+bone+healing+in+a+rat+spinal+rhBMP-2+model.">The effect of vancomycin powder on bone healing in a rat spinal rhBMP-2 model.</a> Journal of Neurosurgery Spine. 25, 147-153 (2016).</li><li>Cohn Yakubovich, D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Computed+Tomography+and+Optical+Imaging+of+Osteogenesis-angiogenesis+Coupling+to+Assess+Integration+of+Cranial+Bone+Autografts+and+Allografts.">Computed Tomography and Optical Imaging of Osteogenesis-angiogenesis Coupling to Assess Integration of Cranial Bone Autografts and Allografts.</a> Journal of Visualized Experiments. (106), e53459 (2015).</li><li>Brecevich, A. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+Comparison+of+Accell+Evo3+and+Grafton+Demineralized+Bone+Matrix+Putties+against+Autologous+Bone+in+a+Rat+Posterolateral+Spine+Fusion+Model.">Efficacy Comparison of Accell Evo3 and Grafton Demineralized Bone Matrix Putties against Autologous Bone in a Rat Posterolateral Spine Fusion Model.</a> Spine Journal. 17, 855-862 (2017).</li><li>Jensen, L. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Novel+porcine+model+of+implant-associated+osteomyelitis:+A+comprehensive+analysis+of+local,+regional,+and+systemic+response.">Novel porcine model of implant-associated osteomyelitis: A comprehensive analysis of local, regional, and systemic response.</a> Journal of Orthopeadic Research. 35, 2211-2221 (2016).</li><li>de Mesy Bentley, K. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence+of+Staphylococcus+Aureus+Deformation,+Proliferation,+and+Migration+in+Canaliculi+of+Live+Cortical+Bone+in+Murine+Models+of+Osteomyelitis.">Evidence of Staphylococcus Aureus Deformation, Proliferation, and Migration in Canaliculi of Live Cortical Bone in Murine Models of Osteomyelitis.</a> Journal of Bone and Mineral Research. 32, 985-990 (2017).</li><li>Norden, C. W., Kennedy, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+osteomyelitis.+I:+A+description+of+the+model.">Experimental osteomyelitis. I: A description of the model.</a> Journal of Infectious Diseases. 122, 410-418 (1970).</li><li>Mistry, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel,+multi-barrier,+drug+eluting+calcium+sulfate/biphasic+calcium+phosphate+biodegradable+composite+bone+cement+for+treatment+of+experimental+MRSA+osteomyelitis+in+rabbit+model.">A novel, multi-barrier, drug eluting calcium sulfate/biphasic calcium phosphate biodegradable composite bone cement for treatment of experimental MRSA osteomyelitis in rabbit model.</a> Journal of Controlled Release. 239, 169-181 (2016).</li><li>Bernthal, N. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combined+In+vivo+Optical+and+&#181;CT+Imaging+to+Monitor+Infection,+Inflammation,+and+Bone+Anatomy+in+an+Orthopaedic+Implant+Infection+in+Mice.">Combined In vivo Optical and &#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice.</a> Journal of Visualized Experiments. (92), e51612 (2014).</li><li>Koeth, L. M., DiFranco-Fisher, J. M., McCurdy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Reference+Broth+Microdilution+Method+for+Dalbavancin+In+Vitro+Susceptibility+Testing+of+Bacteria+that+Grow+Aerobically.">A Reference Broth Microdilution Method for Dalbavancin In Vitro Susceptibility Testing of Bacteria that Grow Aerobically.</a> Journal of Visualized Experiments. (103), e53028 (2015).</li><li>Uttra, A. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ephedra+gerardiana+aqueous+ethanolic+extract+and+fractions+attenuate+Freund+Complete+Adjuvant+induced+arthritis+in+Sprague+Dawley+rats+by+downregulating+PGE2,+COX2,+IL-1&#946;,+IL-6,+TNF-&#945;,+NF-kB+and+upregulating+IL-4+and+IL-10.">Ephedra gerardiana aqueous ethanolic extract and fractions attenuate Freund Complete Adjuvant induced arthritis in Sprague Dawley rats by downregulating PGE2, COX2, IL-1&#946;, IL-6, TNF-&#945;, NF-kB and upregulating IL-4 and IL-10.</a> Journal of Ethnopharmacology. 224, 482-496 (2018).</li><li>Harrasser, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+model+of+implant-related+osteomyelitis+in+the+metaphysis+of+rat+tibiae.">A new model of implant-related osteomyelitis in the metaphysis of rat tibiae.</a> BMC Musculoskeletal Disorders. 17, 152 (2016).</li><li>Abedon, S. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Commentary:+Phage+Therapy+of+Staphylococcal+Chronic+Osteomyelitis+in+Experimental+Animal+Model.">Commentary: Phage Therapy of Staphylococcal Chronic Osteomyelitis in Experimental Animal Model.</a> Frontiers in Microbiology. 7, 1251 (2016).</li><li>Tan, H. L., Ao, H. Y., Ma, R., Lin, W. T., Tang, T. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+effect+of+quaternized+chitosan-loaded+polymethylmethacrylate+bone+cement+on+methicillin-resistant+Staphylococcus+epidermidis+infection+of+the+tibial+metaphysis+in+a+rabbit+model.">In vivo effect of quaternized chitosan-loaded polymethylmethacrylate bone cement on methicillin-resistant Staphylococcus epidermidis infection of the tibial metaphysis in a rabbit model.</a> Antimicrobial Agents and Chemotherapy. 58, 6016-6023 (2014).</li><li>Chiara, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detection+of+Osteomyelitis+in+the+Diabetic+Foot+by+Imaging+Techniques:+A+Systematic+Review+and+Meta-analysis+Comparing+MRI,+White+Blood+Cell+Scintigraphy,+and+FDG-PET.">Detection of Osteomyelitis in the Diabetic Foot by Imaging Techniques: A Systematic Review and Meta-analysis Comparing MRI, White Blood Cell Scintigraphy, and FDG-PET.</a> Diabetes Care. 40, 1111-1120 (2017).</li><li>Khalid, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Raman+Spectroscopy+detects+changes+in+Bone+Mineral+Quality+and+Collagen+Cross-linkage+in+Staphylococcus+Infected+Human+Bone.">Raman Spectroscopy detects changes in Bone Mineral Quality and Collagen Cross-linkage in Staphylococcus Infected Human Bone.</a> Scientific Reports. 8, 9417 (2018).</li><li>Putters, T. F., Schortinghuis, J., Vissink, A., Raghoebar, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+prospective+study+on+the+morbidity+resulting+from+calvarial+bone+harvesting+for+intraoral+reconstruction.">A prospective study on the morbidity resulting from calvarial bone harvesting for intraoral reconstruction.</a> International Journal of Oral and Maxillofacial Surgery. 44, 513-517 (2015).</li><li>Yin, J., Jiang, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Completely+resorption+of+autologous+skull+flap+after+orthotopic+transplantation:+a+case+report.">Completely resorption of autologous skull flap after orthotopic transplantation: a case report.</a> International Journal of Clinical and Experimental Medicine. 7, 1169-1171 (2014).</li><li>Takehiko, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preliminary+results+of+managing+large+medial+tibial+defects+in+primary+total+arthroplasty:+autogenous+morcellised+bone+graft.">Preliminary results of managing large medial tibial defects in primary total arthroplasty: autogenous morcellised bone graft.</a> International Orthopaedics. 41, 931-937 (2017).</li></ol>

在以往的研究中, 构建了各种动物模型, 研究急性和慢性骨感染;然而, 对理想模型的寻找仍然持续到17,18。此外, 理想的骨感染模型有望模拟临床环境下骨感染的病理特征, 而建模期间, 成本仍然较低, 易于实施。目前, 兔骨感染模型是炎症性骨病研究中最常见的模型, 因为兔子是可用的、可行的、廉价的。在我们之前的研究中, 我们比较了不同体重的兔子的...

骨感染通常是由创伤、骨折或其他骨骼疾病后的细菌或其他微生物入侵引起的。骨感染可能导致高水平的炎症和骨组织破坏。在临床上,金黄色葡萄球菌(金黄色葡萄球菌) 是骨感染的主要病原体 2,3.骨感染是痛苦的, 使人虚弱的, 往往是慢性的, 是极难治疗的4。目前, 清创坏死组织和植入万古霉素负载钙 (vcs) 珠已被证实为控制局部感染5,6的有效策略。然而, 10% 至15% 的患者经历了长期的骨修复过程, 延迟愈合, 或抗感染治疗后不愈合 7。骨缺损的大部分是整形外科医生最困难的问题。自体骨移植被认为是骨不结合治疗8,9的最佳骨置换方法。
到目前为止, 关于骨感染和自体骨植入的研究大多是在各种动物模型中进行的, 如老鼠、兔子、狗、猪和羊10、11.兔子模型最常用于骨感染研究, 这是诺登和肯尼迪在 1970年12,13首次进行的。在我们之前的研究中, 我们采用了 nnoren 方法的兔子模型, 我们发现注入骨髓的金黄色葡萄球菌的数量无法准确量化, 因为从骨髓中漏出的血液导致细菌溶液溢出。
本文提出了一种改良的家兔骨感染的手术方法。在手术结束时, 进行血液生化检查、细菌学检查和组织病理学检查, 以验证骨感染模型。然后, 植入 vcs 抑制感染, 自体骨植入, 促进骨再生。

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			<td height="21" style="height:21px;width:275px;">absorbable surgical suture</td>
			<td style="width:264px;">Jinghuan</td>
			<td style="width:119px;">18S0604A</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">asepsis injector</td>
			<td style="width:264px;">Jinglong</td>
			<td style="width:119px;">20170501</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">bone wax</td>
			<td style="width:264px;">ETHICON</td>
			<td style="width:119px;">JH5CQLM</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">CCD camera</td>
			<td style="width:264px;">Olympus</td>
			<td style="width:119px;">DP72</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">EDTA-K2 anticoagulant blood vessel</td>
			<td style="width:264px;">XINGE</td>
			<td style="width:119px;">20170802</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">Electric bone drill unit</td>
			<td style="width:264px;">Bao Kang</td>
			<td style="width:119px;">BKZ-1</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:275px;">Electric shaver</td>
			<td style="width:264px;">Codos</td>
			<td style="width:119px;">3800</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:275px;">flexible silica gel mold&#160;</td>
			<td style="width:264px;">WRIGHT</td>
			<td style="width:119px;">1527745</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">Hematoxylin and Eosin Staining Kit</td>
			<td style="width:264px;">Beyotime</td>
			<td style="width:119px;">20170523</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">Luria-Bertani culture medium</td>
			<td style="width:264px;">Baisi Biothchnology</td>
			<td style="width:119px;">20170306</td>
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			<td height="21" style="height:21px;width:275px;">Medical-grade calcium sulphate</td>
			<td style="width:264px;">WRIGHT</td>
			<td style="width:119px;">1527745</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">microcomputed tomography (micro-CT)</td>
			<td style="width:264px;">Bruker</td>
			<td style="width:119px;">SkyScan 1172&#160;</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:275px;">Microscope</td>
			<td style="width:264px;">Olympus</td>
			<td style="width:119px;">CX41</td>
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			<td height="21" style="height:21px;width:275px;">New Zealand white rabbits</td>
			<td style="width:264px;">Zhejiang Experimental Animal Center&#160;</td>
			<td style="width:119px;">SCXK 2014-0047</td>
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			<td height="25" style="height:25px;width:275px;">No. 11 scalpel&#160;</td>
			<td style="width:264px;">Yuanlikang</td>
			<td style="width:119px;">20170604</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:275px;">normal saline</td>
			<td style="width:264px;">Mingsheng</td>
			<td style="width:119px;">20170903</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">PBS</td>
			<td style="width:264px;">TBD(Jingyi)</td>
			<td style="width:119px;">20170703-0592</td>
			<td></td>
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			<td height="21" style="height:21px;width:275px;">pentobarbital sodium</td>
			<td style="width:264px;">Merk</td>
			<td style="width:119px;">2070124</td>
			<td></td>
		</tr>
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			<td height="21" style="height:21px;width:275px;">povidone-iodinesolution</td>
			<td style="width:264px;">Lierkang</td>
			<td style="width:119px;">20170114</td>
			<td></td>
		</tr>
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			<td height="42" style="height:42px;width:275px;">S. aureus freeze drying powder</td>
			<td style="width:264px;">China General Microbiological Culture Collection Center</td>
			<td style="width:119px;">ATCC 6538</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">sheep blood agar</td>
			<td style="width:264px;">HuanKai Microbial</td>
			<td style="width:119px;">3103210</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:275px;">tryptic soy agar plates</td>
			<td style="width:264px;">HuanKai Microbial</td>
			<td style="width:119px;">3105697</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">tryptic soy broth tubes</td>
			<td style="width:264px;">HuanKai Microbial</td>
			<td style="width:119px;">3104260</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:275px;">Vancomycin</td>
			<td style="width:264px;">Lilly</td>
			<td style="width:119px;">C599180</td>
			<td></td>
		</tr>
	</tbody>
</table>

treatment with vancomycin loaded calcium sulphate and autogenous bone in an improved rabbit model of bone infection

骨感染是由细菌入侵引起的, 在临床、整形外科和创伤性手术中, 细菌感染是极其困难的。骨感染可能导致持续炎症、骨髓炎和最终骨不愈合。建立可行、可重复的动物模型对骨感染研究和抗生素治疗具有重要意义。作为体内模型, 兔模型在骨感染研究中得到了广泛的应用。然而, 此前对兔骨感染模型的研究表明, 由于细菌数量是可变的, 感染状态不一致。本研究通过阻断骨髓中的细菌, 提出了一种诱导兔骨感染的改进手术方法。然后, 进行多层次的评价, 验证建模方法。
一般情况下, 脱骨坏死组织和植入香康辛负载硫酸钙 (vcs) 是抗生素治疗的主要优势。尽管 vcs 中的硫酸钙有利于骨细胞的爬行和新的骨生长, 但大量的骨缺损发生后, 脱骨。自体骨 (ab) 是一种治疗坏死骨后骨缺损的一种吸引人的方法。
在这项研究中, 我们使用尾骨作为自体骨植入骨缺损。采用显微断层扫描 (微 ct) 和动物牺牲后的组织学分析测量骨修复。因此, 在 vcs 组中, 持续获得骨不愈合。相比之下, vcs-ab 组的骨缺损区明显减少。目前的建模方法描述了一种可重复、可行、稳定的骨感染模型制备方法。vcs-ab 治疗导致抗生素治疗后骨不愈合率降低。改进的骨感染模型以及 vcs 和自体骨的联合治疗有助于研究与创伤骨科应用相关的骨感染和骨再生的潜在机制。

骨感染模型的评价 兔感染金黄色葡萄球菌后, 其病理表现与临床上慢性骨髓炎的典型症状相似。在我们的研究中, 30只家兔被感染, 并作为模型组进行了研究, 10只兔子被作为对照动物。所有模型兔都感染了胫骨局部部位的鼻窦, 从鼻窦流出白色和黄色脓液 (图 3a)。h & e 染色结果表明, 模型组细菌聚集物位于死骨周围, ?...

Watch this Scientific Journal Video about Treatment with Vancomycin Loaded Calcium Sulphate and Autogenous Bone in an Improved Rabbit Model of Bone Infection at JoVE.com

Treatment with Vancomycin Loaded Calcium Sulphate and Autogenous Bone in an Improved Rabbit Model of Bone Infection

本研究通过阻断骨髓中相同数量的细菌, 提出了一种感染金黄色葡萄球菌的改良兔模型。万古霉素负载硫酸钙和自体骨用于抗生素和骨修复治疗。该方案可为骨感染和再生的研究提供参考。

万古霉素滴入硫酸钙和自体骨治疗兔骨感染的改进模型

Bone infection results from bacterial invasion, which is extremely difficult to treat in clinical, orthopedic, and traumatic surgery. The bone infection ...

可生产动物模型对骨感染研究非常重要。然而，目前没有兔子模型的感染状态和细菌的数量是一致的。我们的骨感染兔子模型使用一致的细菌，并由万科霉素加载硫酸钙和自体片段处理。该技术可推广到骨感染和骨非联合治疗的诊断，并用于了解骨感染的发病机制。这种方法可以深入了解大鼠、小鼠和猪的其他骨骼感染模型。将相同体积的细菌悬浮液加载到骨髓中是很困难的。视觉演示可以帮助研究人员轻松准确地执行关键步骤。要准备细菌悬浮液，首先在0.3毫升LB培养基中溶解0.5毫克S.aureus冷冻干燥粉末。将悬浮液彻底混合，将细菌条纹到37摄氏度的试菌大豆加糖盘上，进行16小时的孵育。第二天早上，在37摄氏度下，在试制大豆汤管中挑选一个细菌菌落形成装置，进行24小时培养。当细菌处于中对数生长阶段时，将1毫升细菌转移到1.5毫升管中，通过离心收集细菌。然后，每次洗涤用100微升PBS洗三次细菌细胞颗粒，在最后洗涤后将细菌重新用3毫升新鲜PBS中重新产生。为了建立骨感染模具，确认在三个月大的三公斤雄性新西兰白兔中，爪子捏紧缺乏反应，并用电剃须刀将头发从近侧肉质区域去除，以对抗头发的生长方向。用波维酮碘溶液对裸露的皮肤进行消毒，并使用笔和尺子标记头骨的上端和 S.Aureus 注射的钻孔位置，注意钻孔位置位于蒂比亚尔高原的横向中间。接下来，使用数字11手术刀在皮肤上切口，随后在皮内酮中切口1厘米。使用电骨钻单元，在头骨上打一个直径两毫米的孔，用两毫米直径的两毫米长的骨蜡圆柱按孔。沿蒂比亚高原的水平平面去除任何多余的骨蜡，并确认孔内布满了骨蜡。然后，使用可吸收的手术缝合缝制的月牙和皮肤在垂直床垫缝合，以防止动物咀嚼缝线。并使用一毫升的 asepsis 喷油器将每毫升 S.Aureus 溶液的 8 个菌落形成单位缓慢地注入钻孔中 10 倍。注射后的第7天、第14天、21日、28日，将受感染的兔子放入兔子的抑制器中，将头部和耳朵放在限制器外，从动脉静脉采集血液样本，用于白血球和C反应蛋白分析。麻醉后，使用数字11手术刀做厘米的三分皮肤和内皮切口，并清洁骨蜡。接下来，使用电骨钻单元打两个相邻的直径4.8毫米的孔来脱粒坏死骨，并使用骨勺去除骨坏死骨髓和造粒组织。然后，刮擦和清洁两个洞之间的骨组织，并传播一毫升的脱毛骨髓到羊血红板，在37摄氏度下过夜孵育。第二天早上，选择有30到300个菌落的板块，并计算每个受感染动物形成菌落的单位数量。感染后第28天，在9.5克医用级硫酸钙中加入一克万科霉素盐酸粉，用铲子用三毫升正常盐水搅拌混合物30至45秒。将混合产品加入柔性硅胶模具中，使产品在室温下干燥 15 分钟。然后，弯曲模具以去除装有硫酸钙的万康霉素或VCS珠。对于自体骨的抗生素治疗和植入，首先，在剃掉尾部区域之前重新暴露受伤的头骨，用波维酮碘溶液对暴露的尾皮肤进行消毒。使用手术剪刀切割尾巴，并使用可吸收的手术缝合线关闭垂直床垫缝合的皮肤，以防止动物咀嚼缝线。使用数字 11 手术刀切割尾皮以露出尾骨。去除任何肌肉、软组织和胸膜，并分离每个关节的尾骨。将骨片转移到含有无菌盐水的100毫米塑料盘中。接下来，使用弯曲的剪刀将四块VCS珠植入骨髓腔，并使用弯曲钳子用八块自体尾骨碎片填充骨骼缺陷。然后，用可吸收的手术缝合以床垫缝合的方式关闭周膜和皮肤切口。骨感染模型组的C反应蛋白和白细胞水平高于对照动物。VCS或VCS自生骨治疗后两、四、六、八周，C反应蛋白和白细胞水平显著降低。二维重建图像显示，使用VCS或VCS和自体骨治疗后12周内骨量逐渐增加，而未经治疗的骨骼感染模型组骨质流失显著。为了分析骨骼再生定量指标，可以使用位图数据重建3D模型图像，并在每个骨骼中选择一个直径为9.6毫米的椭圆形椭圆形区域。未经治疗的骨感染模型组中骨体积与组织体积的比例明显低于VCS和VCS自生骨组。VCS自体骨组的皮眼数和厚度分数明显高于未经治疗的骨感染模型和VCS动物。此外，VCS自生骨组的眼皮分离分数明显低于未经治疗的骨感染模型和VCS组。注意选择体重合适的兔子。用骨蜡阻断金黄色葡萄球菌溶液。彻底去皮化坏死骨，打两个相邻的直径4.8毫米的孔。按照该程序，可以执行其他诊断测量方法，如超声波、放射学和计算机断层扫描，以监测骨感染和修复的病理过程。这种改进的骨感染模型以及VCS和自生骨治疗的组合有助于研究骨感染和骨再生的下划线机制。由于金黄色葡萄球菌冻干粉是危险的，细菌悬浮液应始终在隔离的细菌实验室中准备。

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Anatomy and Physiology

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Unit 1

Bone Tissue and the Skeletal System

SCIENTISTS IN ACTION

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

采用实时定量PCR检测供体细胞植入一个具有竞争力的小鼠骨髓移植模型

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

科研

医学

Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia

酪氨酸激酶抑制剂的临床前评价急性白血病的治疗

Receptor tyrosine kinases have been implicated in the development and progression of many cancers, including both leukemia and solid tumors, and are ...

Combined In vivo Optical and &#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Combined In vivo Optical and Ãƒâ€šÃ‚ÂµCT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

结合<em&gt;在体内</em&gt;光学及μCT成像监视感染，炎症和骨解剖在骨科植入物感染的小鼠

Multimodality imaging has emerged as a common technological approach used in both preclinical and clinical research. Advanced techniques that combine in ...

An Improved Method for Rapid Intubation of the Trachea in Mice

在小鼠气管插管快速的改进方法

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Using a Knee Arthrometer to Evaluate Tissue-specific Contributions to Knee Flexion Contracture in the Rat

用膝关节关节仪评价大鼠膝关节屈曲收缩的特定组织贡献

Normal knee range of motion (ROM) is critical to well-being and allows one to perform basic activities such as walking, climbing stairs and sitting. Lost ...

The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing

小鼠闭合性股骨骨折的形成: 骨愈合模型的研究

Bone fractures impose a tremendous socio-economic burden on patients, in addition to significantly affecting their quality of life. Therapeutic strategies ...

Flow Cytometry Analysis of Immune Cell Subsets within the Murine Spleen, Bone Marrow, Lymph Nodes and Synovial Tissue in an Osteoarthritis Model

骨关节炎模型中骨髓、骨髓、淋巴结和合成组织内免疫细胞子集的流动细胞测量分析

Osteoarthritis (OA) is one of the most prevalent musculoskeletal diseases, affecting patients suffering from pain and physical limitations. Recent ...

Evaluation of Amino Acid Consumption in Cultured Bone Cells and Isolated Bone Shafts

Bone development and homeostasis is dependent upon the differentiation and activity of bone forming osteoblasts. Osteoblast differentiation is ...

Improved Rodent Model of Myocardial Ischemia and Reperfusion Injury

Myocardial ischemia and reperfusion injury (MIRI), induced by coronary heart disease (CHD), causes damage to the cardiomyocytes. Furthermore, evidence ...

3D Reconstruction for the Diagnosis and Treatment of Pulmonary Nodules

A 3D Digital Model for the Diagnosis and Treatment of Pulmonary Nodules

A 3D Digital Model for the Diagnosis and Treatment of Pulmonary Nodules (Video) | JoVE 

用于肺结节诊断和治疗的三维数字模型

The three-dimensional (3D) reconstruction of pulmonary nodules using medical images has introduced new technical approaches for diagnosing and treating ...