Noviani Maria, Jamiolkowski Ryan M, Grenet Justin E, Lin Qiuyu, Carlon Tim A, Qi Le, Jantzen Alexandra E, Milano Carmelo A, Truskey George A, Achneck Hardean E
From the *Department of Surgery, Duke University Medical Center, Durham, North Carolina; †Cardiovascular & Metabolic Disorders Program, Duke - National University of Singapore Graduate Medical School, Singapore; ‡Department of Biomedical Engineering, Duke University, Durham, North Carolina; §Department of Pathology, Duke University Medical Center, Durham, North Carolina; ¶Hemostemix Inc., Toronto, Ontario, Canada; and ‖Hemostemix Inc., Ness Ziona, Israel.
ASAIO J. 2016 Jul-Aug;62(4):447-53. doi: 10.1097/MAT.0000000000000351.
The most promising alternatives to heart transplantation are left ventricular assist devices and artificial hearts; however, their use has been limited by thrombotic complications. To reduce these, sintered titanium (Ti) surfaces were developed, but thrombosis still occurs in approximately 7.5% of patients. We have invented a rapid-seeding technology to minimize the risk of thrombosis by rapid endothelialization of sintered Ti with human cord blood-derived endothelial cells (hCB-ECs). Human cord blood-derived endothelial cells were seeded within minutes onto sintered Ti and exposed to thrombosis-prone low fluid flow shear stresses. The hCB-ECs adhered and formed a confluent endothelial monolayer on sintered Ti. The exposure of sintered Ti to 4.4 dynes/cm for 20 hr immediately after rapid seeding resulted in approximately 70% cell adherence. The cell adherence was not significantly increased by additional ex vivo static culture of rapid-seeded sintered Ti before flow exposure. In addition, adherent hCB-ECs remained functional on sintered Ti, as indicated by flow-induced increase in nitric oxide secretion and reduction in platelet adhesion. After 15 day ex vivo static culture, the adherent hCB-ECs remained metabolically active, expressed endothelial cell functional marker thrombomodulin, and reduced platelet adhesion. In conclusion, our results demonstrate the feasibility of rapid-seeding sintered Ti with blood-derived hCB-ECs to generate a living antithrombotic surface.
心脏移植最有前景的替代方案是左心室辅助装置和人工心脏；然而，它们的使用受到血栓形成并发症的限制。为了减少这些并发症，人们开发了烧结钛（Ti）表面，但仍有大约7.5%的患者会发生血栓形成。我们发明了一种快速接种技术，通过用人脐血来源的内皮细胞（hCB-ECs）使烧结钛快速内皮化，以将血栓形成风险降至最低。人脐血来源的内皮细胞在数分钟内接种到烧结钛上，并暴露于易发生血栓形成的低流体流动剪切应力下。hCB-ECs在烧结钛上黏附并形成融合的内皮单层。快速接种后立即将烧结钛暴露于4.4达因/平方厘米的剪切应力下20小时，导致约70%的细胞黏附。在流动暴露前，对快速接种的烧结钛进行额外的体外静态培养，细胞黏附并未显著增加。此外，如流动诱导的一氧化氮分泌增加和血小板黏附减少所示，黏附在烧结钛上的hCB-ECs仍保持功能。在体外静态培养15天后，黏附的hCB-ECs仍保持代谢活性，表达内皮细胞功能标志物血栓调节蛋白，并减少血小板黏附。总之，我们的结果证明了用血液来源的hCB-ECs快速接种烧结钛以生成具有抗血栓形成功能的活性表面的可行性。
