Thrombogenicity Testing of Medical Devices in a Minimally Heparinized Ovine Blood Loop

Abstract

ISO 10993-4 in vivo thrombogenicity testing is frequently performed for regulatory approval of many blood-contacting medical devices and is often a key part of submission packages. Given the current state of in vivo thrombogenicity assays, a more robust and reproducible assay design, including in vitro models, is needed. This study describes an in vitro assay that integrates freshly harvested ovine blood containing minimal heparin in a closed pumped loop. To confirm the reproducibility of this assay, control materials were identified that elicited either a positive or a negative thrombogenic response. These controls demonstrated reproducibility in the resulting thrombogenicity scores with median scores of 5 and 0 for the positive and negative controls, respectively, which also demonstrated a significant difference (p < 0.0001). For a direct comparison of the in vitro blood loop assay to the traditional in vivo nonanticoagulated venous implant (NAVI) assay, seven sheep were used as blood donors for the loop and then as subjects for an NAVI assay. In each assay—loop or NAVI—three study articles were used: the positive and negative controls and a marketed, approved catheter. The resulting thrombogenicity scores were similar when comparing the loop to the NAVI results. For each study article, the median thrombogenicity scores were the same in these two different assays, being 0, 1, and 5 for the negative control, the marketed catheter, and the positive control, respectively. These data suggest that the in vitro assay performs similarly to the in vivo NAVI assay. This in vitro blood loop method has the potential to predict a materials' in vivo thrombogenicity, can substantially de-risk the materials or coating selection process, and may eventually be able to replace the in vivo models currently in use.