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: 9133179
• 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 2018-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9133179
• 关键词: Accounting; 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; Healed; 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; Primary Infection; Probiotics; Problem Solving; Procedures; Process; Prosthesis; Prosthesis Design; Proteins; Protocols documentation; Residual state; Sampling; Serum; Site; Skin; Soaps; Sodium Chloride; Specimen; Staging; Stomas; Suspension substance; Suspensions; Swab; 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; success; synthetic polymer Bioplex; wound

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）： 经皮骨整合假肢附着或假肢直接骨骼附着，穿过截肢残端皮肤进入残余骨骼，是一项快速发展的技术，经过十多年的欧洲试验，目前已准备好引入美国。涉及经股截肢者。过去七年在犹他州盐湖城 VAH 骨与关节研究实验室进行的转化动物研究阐明了骨整合原理，以及骨和皮肤界面对含有经皮植入物的残余骨的长期负重的反应。这些信息产生了新的假肢设计，使该技术更安全，并且更有可能长期成功。这些与欧洲人类数据平行收集的转化动物数据使 FDA 指导下的早期可行性设备豁免 (IDE) 试点计划成为可能。该试验预计于 2014 年 5 月在犹他州盐湖城的 VAH 开始，由 VA RR&D 拨款 (RX001208-01) 资助。从退伍军人和现役军人中选出的十名股骨截肢者将收到该装置。美国之前引进这项技术的关键限制因素是 欧洲患者植入物/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生率，但值得注意的是，70% 至 95% 的欧洲患者现在只需在造口处使用温和的肥皂和淋浴卫生即可保持无感染，并避免使用抗生素。事实上，抗生素和消毒剂常常适得其反，并导致抗生素耐药性。似乎越来越明显的是 这一看似不可能的成功的关键在于皮肤/植入物界面处的生物学和互利共生微生物群：细菌的“门口看门狗”。 这项待定的 10 名患者试点试验提供了一个为期 1 年的独特机会，用于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全身免疫蛋白反应该经皮装置（目标 2）；全部在同一时间点确定。希望这项研究能够对造口为什么会被感染或不被感染以及预测感染的发生提供可衡量的了解（主要目标）。将从第一阶段手术（装置植入、伤口闭合和初级伤口愈合）开始收集连续皮肤拭子细菌 DNA 样本，并在 6 周后进行第二阶段手术（建立造口；慢性伤口）。从该 DNA 中获得的细菌 16S rRNA 基因的扩增，随后将在 Illumina MiSeq 平台上进行测序，并将在整个初级伤口愈合和慢性造口成熟过程中确定造口和邻近大腿皮肤的常驻微生物群。在收集微生物组的同时，将获得静脉血和造口渗出液样本，以比较全身与局部炎症蛋白反应。系列静脉血样本将用于确定宿主免疫系统的瞬时状态，并将使用定制的 TaqMan 微流体阵列卡（192 个基因）对免疫系统蛋白进行微阵列分析。这将测量宿主 mRNA 的“上”和“下”调节，从而翻译免疫系统蛋白（即细胞因子和趋化因子）的合成。使用 BioPlex 多重免疫分析和 MagPix 平台对造口渗出物进行分析，将测量局部伤口环境中 27 种伤口愈合和炎症细胞因子的水平。统计分析将随着时间的推移将每位患者与自己以及其他患者进行比较（微生物群和感染的共性与稳定且未感染的造口）。