Engineered aprotinin for improved stability of fibrin biomaterials
Engineered aprotinin for improved stability of fibrin biomaterials
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DOI：
10.1016/j.biomaterials.2010.08.109
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发表时间：
2011-01-01
期刊：
影响因子：
14
通讯作者：
Hubbell, Jeffrey A.
中科院分区：
文献类型：
--
作者：
Lorentz, Kristen M.;Kontos, Stephan;Hubbell, Jeffrey A.
纤维蛋白在临床上长期用于止血和密封，但由于其体内吸收相对较快，即使添加抑肽酶或其他蛋白酶抑制剂，其在其他应用中的扩展也受到限制。我们报告了一种工程化的抑肽酶变体，可以固定在纤维蛋白内，从而延长寿命。当重组融合到来自 α(2)-纤溶酶抑制剂 (α 2PI1-8) 的转谷氨酰胺酶底物结构域时，所得变体在正常凝血酶/因子 XIIIa 介导的聚合过程中，抑肽酶-α 2PI1-8 共价交联成纤维蛋白基质。用生理纤溶酶浓度进行的挑战显示，含有抑肽酶-α 2PI1-8 的基质在 3 周后保留了 78% 的质量，而含有野生型 (WT) 抑肽酶的基质在 1 周内完全降解；商业密封剂 Omrixil 和 Tisseel 的纤溶酶挑战补充有抑肽酶-α 2PI1-8 或 WT 抑肽酶也显示出更长的寿命 当用人真皮成纤维细胞接种时，补充有抑肽酶-α 2PI1-8 的基质支持细胞生长的时间比含有 WT 抑肽酶的基质长至少 33%。 WT 抑肽酶 我们的结论是，我们的工程重组抑肽酶变体在体外和体内比 WT 抑肽酶更有效地延长纤维蛋白基质的寿命 (C) 2010 Elsevier Ltd 版权所有
Fibrin has been long used clinically for hemostasis and sealing yet extension of use in other applications has been limited due to its relatively rapid resorption in vivo even with addition of aprotinin or other protease inhibitors We report an engineered aprotinin variant that can be immobilized within fibrin and thus provide extended longevity When recombinantly fused to a transglutaminase substrate domain from alpha(2)-plasmin inhibitor (alpha 2PI1-8) the resulting variant aprotinin-alpha 2PI1-8 was covalently crosslinked into fibrin matrices during normal thrombin/factor XIIIa-mediated polymerization Challenge with physiological plasmin concentrations revealed that aprotinin-alpha 2PI1-8-containing matrices retained 78% of their mass after 3 wk whereas matrices containing wild type (WT) aprotinin degraded completely within 1 wk Plasmin challenge of commercial sealants Omrixil and Tisseel supplemented with aprotinin-alpha 2PI1-8 or WT aprotinin showed extended longevity as well When seeded with human dermal fibroblasts aprotinin-alpha 2PI1-8-supplemented matrices supported cell growth for at least 33% longer than those containing WT aprotinin Subcutaneously Implanted matrices containing aprotinin-alpha 2PI1-8 were detectable in mice for more than twice as long as those containing WT aprotinin We conclude that our engineered recombinant aprotinin variant can confer extended longevity to fibrin matrices more effectively than WT aprotinin in vitro and in vivo (C) 2010 Elsevier Ltd All rights reserved