Dimitrievska Sashka, Cai Chao, Weyers Amanda, Balestrini Jenna L, Lin Tylee, Sundaram Sumati, Hatachi Go, Spiegel David A, Kyriakides Themis R, Miao Jianjun, Li Guoyun, Niklason Laura E, Linhardt Robert J
Department of Biomedical Engineering, Yale University, New Haven, CT 06510, USA.
Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY, USA.
Acta Biomater. 2015 Feb;13:177-87. doi: 10.1016/j.actbio.2014.11.015. Epub 2014 Nov 20.
A novel method enabling the engineering of a dense and appropriately oriented heparin-containing layer on decellularized aortas has been developed. Amino groups of decellularized aortas were first modified to azido groups using 3-azidobenzoic acid. Azide-clickable dendrons were attached onto the azido groups through "alkyne-azide" click chemistry, affording a tenfold amplification of adhesions sites. Dendron end groups were finally decorated with end-on modified heparin chains. Heparin chains were oriented like heparan sulfate groups on native endothelial cells surface. X-ray photoelectron spectroscopy, nuclear magnetic resonance imaging, mass spectrometry and Fourier transform infrared FTIR spectroscopy were used to characterize the synthesis steps, building the final heparin layered coatings. The continuity of the heparin coating was verified using fluorescent microscopy and histological analysis. The efficacy of heparin linkage was demonstrated with factor Xa anti-thrombogenic assay and platelet adhesion studies. The results suggest that oriented heparin immobilization to decellularized aortas may improve the in vivo blood compatibility of decellularized aortas and vessels.
已开发出一种新方法,可在脱细胞主动脉上构建致密且取向合适的含肝素层。首先使用3-叠氮基苯甲酸将脱细胞主动脉的氨基修饰为叠氮基。通过“炔烃-叠氮化物”点击化学将可点击叠氮基的树枝状分子连接到叠氮基上,使粘附位点增加了十倍。树枝状分子的末端基团最终用端基修饰的肝素链进行修饰。肝素链的取向类似于天然内皮细胞表面的硫酸乙酰肝素基团。使用X射线光电子能谱、核磁共振成像、质谱和傅里叶变换红外(FTIR)光谱对合成步骤进行表征,构建最终的肝素分层涂层。使用荧光显微镜和组织学分析验证肝素涂层的连续性。通过因子Xa抗血栓形成试验和血小板粘附研究证明了肝素连接的有效性。结果表明,将定向肝素固定在脱细胞主动脉上可能会改善脱细胞主动脉和血管在体内的血液相容性。