Clinical Engineering (UKCTE), The Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, United Kingdom.
Biomaterials. 2012 Sep;33(27):6380-92. doi: 10.1016/j.biomaterials.2012.05.058. Epub 2012 Jun 26.
Clinical performance of a biomaterial is decided early after implantation as leukocytes interrogate the graft throughout acute inflammation. High degrees of leukocyte activation lead to poor material/patient compliance, accelerated degeneration and graft rejection. A number reactive oxygen species (ROS) are released by leukocytes throughout their interaction with a material, which can be used as a sensitive measure of leukocyte activation. The aim of this study was to compare leukocyte activation by commercially available biologic surgical materials and define the extent manufacturing variables influence down-stream ROS response. Chemiluminescence assays were performed using modifications to a commercially available kit (Knight Scientific, UK). Whole blood was obtained from 4 healthy human adults at 7 day intervals for 4 weeks, combined with Adjuvant K, Pholasin (a highly sensitive ROS excitable photoprotein) and biomaterial, and incubated for 60 min with continuous chemiluminescent measurements. Leukocyte ROS inducers fMLP and PMA were added as controls. Xeno- and allogeneic dermal and small intestinal submucosal (SIS) derived biomaterials were produced commercially (Surgisis Biodesign™, Alloderm(®), Strattice(®)Firm & Pliable & Permacol™) or fabricated in house to induce variations in decellularisation and cross-linking. Statistics were performed using Waller-Duncan post hoc ranking. Materials demonstrated significant differences in leukocyte activation as a function of decellularisation reagent and tissue origin. The data demonstrated SIS was significantly more pro-inflammatory than dermis. Additionally it was deduced that SDS during decellularisation induced pro-inflammatory changes to dermal materials. Furthermore, it was possible to conclude inter-patient variation in leukocyte response. The in vitro findings were validated in vivo which confirmed the chemiluminescence observations, highlighting the potential for translation of this technique as a routine component of pre-surgical evaluation to maximise foreign body compliance.
生物材料的临床性能在植入后早期就已决定，因为白细胞在急性炎症期间会全面检查移植物。白细胞高度激活会导致材料/患者顺应性差、加速退变和移植物排斥。白细胞在与材料相互作用过程中会释放大量活性氧（ROS），这些 ROS 可作为白细胞激活的敏感测量指标。本研究旨在比较市售生物外科材料对白细胞的激活作用，并确定制造变量对下游 ROS 反应的影响程度。使用市售试剂盒（英国 Knight Scientific）进行化学发光测定。在 4 周内每隔 7 天从 4 位健康成年人获得全血，与 Adjuvant K、Pholasin（一种对 ROS 高度敏感的可激发发光蛋白）和生物材料混合，并在 60 分钟内孵育，同时进行连续化学发光测量。添加白细胞 ROS 诱导剂 fMLP 和 PMA 作为对照。商业生产的异种和同种真皮和小肠黏膜下层（SIS）衍生生物材料（Surgisis Biodesign™、Alloderm(®)、Strattice(®)Firm & Pliable & Permacol™）或在实验室中制造，以诱导脱细胞和交联的变化。使用 Waller-Duncan 事后排序进行统计。结果表明，不同材料对白细胞的激活程度与脱细胞试剂和组织来源有关。数据表明，SIS 比真皮更具炎症性。此外，可以推断出 SDS 在脱细胞过程中会导致真皮材料发生促炎变化。此外，还可以得出白细胞反应的个体间差异。体外研究结果在体内得到验证，这证实了化学发光观察结果，突出了将该技术作为术前评估常规组成部分的潜力，以最大限度地提高异物顺应性。
