Microbiome and Innate Immunity with Percutaneous Osseointegrated Prostheses

经皮骨整合假体的微生物组和先天免疫

• 批准号: 9354204
• 负责人: James Peter Beck
• 金额: --
• 依托单位: VA SALT LAKE CITY HEALTHCARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-10-01 至 2019-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9354204
• 关键词: Amputation Stumps; Amputees; Animal Experimentation; Antibiotic Resistance; Antibiotic Therapy; Antibiotics; Bacteria; Bacterial DNA; Bilateral; Biological Markers; Biology; Blood; Canis familiaris; Caring; Chronic; Cities; Clinical; Clinical Trials; Collection; Conflict (Psychology); Contralateral; Custom; DNA; Data; Detection; Devices; Disinfectants; Docking; Down-Regulation; Environment; European; Event; Exudate; Funding; Future; Genes; Goals; Grant; Hygiene; Immune; Immune system; Immunoassay; Implant; Incidence; Infection; Infection prevention; Inflammatory; Intervention; Ipsilateral; Joints; Laboratory Research; Lead; Life; Life Style; Limb Prosthesis; Limb structure; Measurable; Measurement; Measures; Messenger RNA; Methods; Microarray Analysis; Microfluidics; Military Personnel; Molecular; Monitor; Morbidity - disease rate; Natural Immunity; Operative Surgical Procedures; Osseointegration; Outcome Study; Patients; Pattern; Population; Probiotics; Problem Solving; Procedures; Process; Prosthesis; Prosthesis Design; Proteins; Protocols documentation; Residual state; Sampling; Serum; Site; Skin; Soaps; Sodium Chloride; Specimen; Statistical Data Interpretation; Stomas; Suspensions; Swab; Symbiosis; System; Techniques; Technology; Testing; Thigh structure; Time; Tissues; Translating; United States; Utah; Venous; Venous blood sampling; Veterans; Walking; Weight-Bearing state; Wound Healing; animal data; bone; chemokine; chronic wound; commensal microbes; cytokine; design; early onset; effective intervention; healing; human data; implantation; improved; microbiome; microbiota; novel therapeutics; pathogenic bacteria; pilot trial; prevent; probiotic therapy; programs; public health relevance; rRNA Genes; response; skeletal; skin microbiota; success; synthetic polymer Bioplex; virtual; wound; wound closure

DESCRIPTION (provided by applicant): Percutaneous osseointegrated prosthetic attachment or the direct skeletal attachment of artificial limbs, across the amputation stump skin into the residual bone, is a rapidly evolving technology that is now ready to be introduced into the United States, following greater than a decade of European trials that largely involved transfemoral amputees. Translational animal research carried out over the past seven years at the Bone and Joint Research Laboratory, VAH, Salt Lake City, Utah, has clarified principles of osseointegration, and bone and skin interface responses to chronic weight bearing of the residual bone containing a percutaneous implant. This information has produced new prosthetic designs that make the technique safer and more likely to succeed over the long-term. This translational animal data, gathered parallel to the European human data, has made possible an Early Feasibility Device Exemption (IDE) Pilot Program under the direction of the FDA. This trial is anticipated to begin in May 2014, at the VAH in Salt Lake City, Utah, and is funded by VA RR&D grant (RX001208-01). Ten transfemoral amputees, selected from the veteran and active military populations will receive the device. The critical limiting factor to the prior U.S. introduction of this technology has been an up to 30% infection rate at the implant/skin interface in European patients. Evolving designs and surgical techniques have lowered this incidence, but it remains remarkable that 70% to perhaps 95% of European patients now remain infection free with simply mild soap and shower hygiene at the stoma, and avoid the use of antibiotics. In fact antibiotics and disinfectants are often counter productive, and lead to antibiotic resistance. It seems increasingly evident that the key to this improbable success is the biology and mutualistic-commensal microbiota at the skin/implant interface: the bacterial "guard dogs at the gate." This pending 10 patient Pilot Tria offers a unique, 1 year long, opportunity to study the evolving microbiota (Aim 1), and the wound healing and inflammatory cytokine environment of the skin/implant interface and surrounding skin as well as the systemic immune protein response to this percutaneous device (Aim 2); all determined at the same time points. Hopefully, this study will give measurable understanding as to why stomas do or don't become infected and to anticipate the onset of infection (Primary Goal). Serial skin swab bacterial DNA samples will be collected beginning with the Stage 1 surgery (device implantation with wound closure and primary wound healing) and 6 weeks later the Stage 2 surgery (establishment of the stoma; a chronic wound). Amplification of the bacterial 16S rRNA genes, obtained from this DNA, will be followed by sequencing on the Illumina MiSeq platform and will determine the resident microbiota at the stoma and adjacent thigh skin throughout the processes of primary wound healing and chronic stomal maturation. Simultaneous to the microbiome collection, venous blood and stomal exudate specimens will be obtained to compare systemic versus local inflammatory protein responses. The serial venous blood samples will be used to determine the instantaneous status of the host immune system and will be subjected to microarray analysis of immune system proteins using custom TaqMan microfluidic array cards (192 genes). This will measure the "up" and "down" regulation of host mRNA that translates the synthesis of immune system proteins i.e. cytokines and chemokines. Analysis of the stomal-exudate, using the BioPlex multiplex immunoassay and MagPix platform, will measure the levels of 27 wound healing and inflammatory cytokines in the local wound environment. Statistical analyses will compare each patient against himself/herself over time, and against the others (commonality of microbiota and infection vs. a stable, and uninfected stoma).

描述（由申请人提供）： 经皮骨整合假体附着或假肢的直接骨骼附着，穿过截肢残端皮肤进入残余骨，是一种快速发展的技术，现在准备引入美国，在欧洲试验十多年后，主要涉及经股截肢者。过去7年在VAH（盐湖，犹他州）骨与关节研究实验室进行的转化动物研究阐明了骨整合的原理，以及骨和皮肤界面对含经皮植入物的残余骨长期承重的反应。这些信息产生了新的假体设计，使该技术更安全，更有可能长期成功。与欧洲人体数据平行收集的这些转化动物数据使得在FDA指导下的早期可行性器械豁免（IDE）试点项目成为可能。这项试验预计将于2014年5月在犹他湖城的VAH开始开始，由VA RR&D赠款（RX 001208 -01）资助。从退伍军人和现役军人中选出的10名经股截肢者将接受该装置。美国之前引进这项技术的关键限制因素是 在欧洲患者中，植入物/皮肤界面的感染率高达30%。不断发展的设计和手术技术降低了这种发病率，但值得注意的是，70%至95%的欧洲患者现在仍然保持无感染，只需在造口处使用温和的肥皂和淋浴卫生，并避免使用抗生素。事实上，抗生素和消毒剂往往适得其反，并导致抗生素耐药性。越来越明显的是， 这种不太可能的成功的关键是皮肤/植入物界面的生物学和互利共生的微生物群：细菌“看门狗"。“这个悬而未决的10名患者试点试验提供了一个独特的，为期一年的机会，研究不断变化的微生物群（目标1），皮肤/植入物界面和周围皮肤的伤口愈合和炎症细胞因子环境，以及对这种经皮设备的全身免疫蛋白反应（目标2）;所有这些都在同一时间点确定。希望这项研究将提供可衡量的理解，为什么造口会或不会被感染，并预测感染的发生（主要目标）。从第1阶段手术（器械植入，伤口闭合和一期伤口愈合）开始，6周后从第2阶段手术（建立造口;慢性伤口）开始，采集连续皮肤拭子细菌DNA样本。从该DNA获得的细菌16 S rRNA基因的扩增将随后在Illumina MiSeq平台上测序，并将在原发性伤口愈合和慢性造口成熟的整个过程中确定造口和邻近大腿皮肤处的驻留微生物群。在微生物组采集的同时，将采集静脉血和造口渗出液标本，以比较全身与局部炎症蛋白反应。连续静脉血样将用于确定宿主免疫系统的瞬时状态，并将使用定制TaqMan微流控阵列卡（192个基因）进行免疫系统蛋白质的微阵列分析。这将测量翻译免疫系统蛋白质（即细胞因子和趋化因子）合成的宿主mRNA的“上调”和“下调”调节。使用BioFlavor多重免疫测定和MagPix平台分析造口渗出物，将测量局部伤口环境中27种伤口愈合和炎性细胞因子的水平。统计分析将随着时间的推移将每个患者与他/她自己以及与其他患者进行比较（微生物群和感染的共性与稳定和未感染的造口）。