Aubin Hug, Mas-Moruno Carlos, Iijima Makoto, Schütterle Nicolas, Steinbrink Meike, Assmann Alexander, Gil Francesc Javier, Lichtenberg Artur, Pegueroles Marta, Akhyari Payam
1 Department of Cardiovascular Surgery, Heinrich-Heine University Düsseldorf , Düsseldorf, Germany .
2 Research Group for Experimental Surgery, Department of Cardiovascular Surgery, Medical Faculty, Heinrich-Heine University Düsseldorf , Düsseldorf, Germany .
Tissue Eng Part C Methods. 2016 May;22(5):496-508. doi: 10.1089/ten.TEC.2015.0556. Epub 2016 Apr 25.
Interface biofunctionalization strategies try to enhance and control the interaction between implants and host organism. Decellularized extracellular matrix (dECM) is widely used as a platform for bioengineering of medical implants, having shown its suitability in a variety of preclinical as well as clinical models. In this study, specifically designed, custom-made synthetic peptides were used to functionalize dECM with different cell adhesive sequences (RGD, REDV, and YIGSR). Effects on in vitro endothelial cell adhesion and in vivo endothelialization were evaluated in standardized models using decellularized ovine pulmonary heart valve cusps (dPVCs) and decellularized aortic grafts (dAoGs), respectively. Contact angle measurements and fluorescent labeling of custom-made peptides showed successful functionalization of dPVCs and dAoGs. The functionalization of dPVCs with a combination of bioactive sequences significantly increased in vitro human umbilical vein endothelial cell adhesion compared to nonfunctionalized controls. In a functional rodent aortic transplantation model, fluorescent-labeled peptides on dAoGs were persistent up to 10 days in vivo under exposure to systemic circulation. Although there was a trend toward enhanced in vivo endothelialization of functionalized grafts compared to nonfunctionalized controls, there was no statistical significance and a large biological variability in both groups. Despite failing to show a clear biological effect in the used in vivo model system, our initial findings do suggest that endothelialization onto dECM may be modulated by customized interface biofunctionalization using the presented method. Since bioactive sequences within the dECM-synthetic peptide platform are easily interchangeable and combinable, further control of host cell proliferation, function, and differentiation seems to be feasible, possibly paving the way to a new generation of multifunctional dECM scaffolds for regenerative medicine.
界面生物功能化策略旨在增强和控制植入物与宿主生物体之间的相互作用。脱细胞细胞外基质（dECM）被广泛用作医用植入物生物工程的平台，已在各种临床前和临床模型中显示出其适用性。在本研究中，使用专门设计的定制合成肽对dECM进行功能化，使其具有不同的细胞粘附序列（RGD、REDV和YIGSR）。分别使用脱细胞羊肺动脉瓣叶（dPVCs）和脱细胞主动脉移植物（dAoGs），在标准化模型中评估对体外内皮细胞粘附和体内内皮化的影响。接触角测量和定制肽的荧光标记显示dPVCs和dAoGs功能化成功。与未功能化的对照相比，用生物活性序列组合对dPVCs进行功能化显著增加了体外人脐静脉内皮细胞的粘附。在功能性啮齿动物主动脉移植模型中，dAoGs上的荧光标记肽在暴露于体循环的情况下在体内持续长达10天。尽管与未功能化的对照相比，功能化移植物的体内内皮化有增强的趋势，但两组均无统计学意义且生物变异性较大。尽管在所用的体内模型系统中未能显示出明显的生物学效应，但我们的初步研究结果确实表明，使用所提出的方法通过定制界面生物功能化可能会调节dECM上的内皮化。由于dECM - 合成肽平台内的生物活性序列易于互换和组合，进一步控制宿主细胞的增殖、功能和分化似乎是可行的，这可能为再生医学的新一代多功能dECM支架铺平道路。
