Fig. 5.

Neutrophil Activation and Attachment to CD47 Modified PVC. Chandler Loop studies were utilized to evaluate immune cell interactions with modified polymeric biomaterials. Whole blood was loaded into unmodified PVC, CD47L, CD47L EtO sterilized, CD47B, or CD47B EtO sterilized blood conduits. Blood from the same donor that was collected immediately prior to the initialization of the study, but not exposed to the blood conduits, was utilized as a control. CD62L, a marker of activated neutrophils, was utilized as a cell surface marker to evaluate the activation state of the neutrophils circulating in the whole blood (A) or attached to the PVC surface (C). Whole blood was analyzed by staining with an anti-CD62L FITC-conjugated antibody and measured using flow cytometry. (B) Attached cells were gently removed from the luminal blood conduit surface as detailed in the materials and methods section and stained using an anti-CD62L FITC-conjugated antibody and measured using flow cytometry. Number of attached neutrophils was obtained by counting the total number of anti-CD62L FITC positive cells using the flow cytometer. * indicates P < 0.05; ** indicates P < 0.01; and *** indicates P < 0.001 as compared to the control. Data are the mean ± standard deviation from N = 5 separate experiments with different donors.