Department of Biologic and Materials Sciences, University of Michigan School of Dentistry, K1030, 1011 N. University Ave., Ann Arbor, MI 48109, USA.
Biomaterials. 2011 Mar;32(7):1809-15. doi: 10.1016/j.biomaterials.2010.10.046. Epub 2010 Dec 22.
Spatiotemporal control of gene delivery, particularly signaling gradients, via biomaterials poses significant challenges because of the lack of efficient delivery systems for therapeutic proteins and genes. This challenge was addressed by using chemical vapor deposition (CVD) polymerization in a counterflow set-up to deposit copolymers bearing two reactive chemical gradients. FTIR spectroscopy verified the formation of compositional gradients. Adenovirus expressing a reporter gene was biotinylated and immobilized using the VBABM method (virus-biotin-avidin-biotin-materials). Sandwich ELISA confirmed selective attachment of biotinylated adenovirus onto copolymer gradients. When cultured on the adenovirus gradients, human gingival fibroblasts exhibited asymmetric transduction with full confluency. Importantly, gradient transduction occurred in both lateral directions, thus enabling more advanced delivery studies that involve gradients of multiple therapeutic genes.
通过生物材料实现基因传递的时空控制，特别是信号梯度控制，具有重大挑战，因为缺乏有效的治疗性蛋白质和基因的传递系统。通过在逆流装置中使用化学气相沉积（CVD）聚合来沉积带有两个反应性化学梯度的共聚物，解决了这一挑战。傅里叶变换红外光谱（FTIR）证实了组成梯度的形成。表达报告基因的腺病毒被生物素化，并使用 VBABM 方法（病毒-生物素-亲和素-生物素-材料）固定化。夹心 ELISA 证实了生物素化腺病毒选择性地附着在共聚物梯度上。当在腺病毒梯度上培养时，人牙龈成纤维细胞表现出完全汇合的不对称转导。重要的是，转导发生在两个横向方向，从而能够进行更先进的输送研究，包括多个治疗性基因的梯度。
