A Recombinant Fusion Construct between Human Serum Albumin and NTPDase CD39 Allows Anti-Inflammatory and Anti-Thrombotic Coating of Medical Devices.

Meike-Kristin Abraham,Hans Peter Wendel,Elena Jost,Karlheinz Peter,Viktoria Bongcaron,Yuyang Song,Amy Kate Searle,Xiaowei Wang,Jan David Hohmann,Stefanie Krajewski

doi:10.3390/pharmaceutics13091504

Abstract

Medical devices directly exposed to blood are commonly used to treat cardiovascular diseases. However, these devices are associated with inflammatory reactions leading to delayed healing, rejection of foreign material or device-associated thrombus formation. We developed a novel recombinant fusion protein as a new biocompatible coating strategy for medical devices with direct blood contact. We genetically fused human serum albumin (HSA) with ectonucleoside triphosphate diphosphohydrolase-1 (CD39), a promising anti-thrombotic and anti-inflammatory drug candidate. The HSA-CD39 fusion protein is highly functional in degrading ATP and ADP, major pro-inflammatory reagents and platelet agonists. Their enzymatic properties result in the generation of AMP, which is further degraded by CD73 to adenosine, an anti-inflammatory and anti-platelet reagent. HSA-CD39 is functional after lyophilisation, coating and storage of coated materials for up to 8 weeks. HSA-CD39 coating shows promising and stable functionality even after sterilisation and does not hinder endothelialisation of primary human endothelial cells. It shows a high level of haemocompatibility and diminished blood cell adhesion when coated on nitinol stents or polyvinylchloride tubes. In conclusion, we developed a new recombinant fusion protein combining HSA and CD39, and demonstrated that it has potential to reduce thrombotic and inflammatory complications often associated with medical devices directly exposed to blood.

Highlights

Cardiovascular diseases such as ischemic heart disease and stroke are the world’s leading causes of death
For the generation of our recombinant fusion protein consisting of human serum albumin (HSA) and CD39, the DNA sequence of HSA was inserted into a previously described pSectag2A vector containing the CD39 sequence [39]
Confirmation of successful molecular biology was made by colony screening of clones via polymerase chain reaction (PCR) sequencing, where positive clones resulted in a 2149 bp

Summary

Introduction

Cardiovascular diseases such as ischemic heart disease and stroke are the world’s leading causes of death. Treatment of patients with cardiovascular problems often includes the invasive application of medical devices. Often these medical devices will be directly exposed to blood, e.g., vascular grafts, stents, permanently implantable biosensors. The biomaterials used for blood-contacting devices represent foreign surfaces to human blood and have the potential to induce specific inflammatory and pro-thrombotic reactions that can lead to clinical complications. The underlying pathological mechanisms of these complications are surface-induced reactions of plasma proteins, platelets and leukocytes. Uncoated medical devices often adsorb blood plasma proteins, such as fibrinogen, on their surfaces, thereby inducing an inflammatory process, platelet adhesion and activation of the coagulation [2,3,4,5,6]. The activation of platelet aggregation and the coagulation cascade may lead to severe and life-threatening thrombosis on the surfaces of biomaterials [3,7]

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Discussion

Conclusion