Low modulus biomimetic microgel particles with high loading of hemoglobin.
Low modulus biomimetic microgel particles with high loading of hemoglobin.
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DOI：
10.1021/bm3007242
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发表时间：
2012-09-10
影响因子：
6.2
通讯作者：
DeSimone, Joseph M.
中科院分区：
文献类型：
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作者：
Chen, Kai;Merkel, Timothy J.;Pandya, Ashish;Napier, Mary E.;Luft, J. Christopher;Daniel, Will;Sheiko, Sergei;DeSimone, Joseph M.
我们合成了极易变形的红细胞样微凝胶颗粒，并将其与牛血红蛋白（Hb）一起装载以增强氧气运输。具有与红细胞（RBC）相似的形状和大小的颗粒使用PRINT®（非润湿模板中的颗粒复制）技术制造。水凝胶的低交联导致这些颗粒的网格密度非常低，允许血红蛋白在整个颗粒中被动扩散。通过EDC/NHS偶联将Hb的赖氨酸基团共价缀合至颗粒中的羧基，将Hb固定在颗粒中。与荧光染料标记的Hb结合的颗粒的共聚焦显微镜证实了Hb在整个颗粒内部的均匀分布，而不是仅表面缀合。获得了高负载比，高达5倍的量的血红蛋白聚合物的重量，没有显着的影响，颗粒的稳定性，形状，虽然颗粒直径略有下降与血红蛋白共轭。通过圆二色性（CD）光谱的蛋白质的分析表明，血红蛋白的二级结构是不受干扰的共轭粒子。通过紫外可见光谱研究，颗粒中的高铁血红蛋白可以保持在低水平，并且负载的Hb仍然可以结合氧。具有中等负载率的Hb负载的颗粒在微流体装置中表现出优异的变形性，容易变形以穿过颗粒直径一半宽的受限孔。与Hb浓度为5.2 g/dL的浓缩颗粒的悬浮液显示出与小鼠血液的粘度相当的粘度，并且即使在恒定的高速率（1,000 1/s）剪切10 min后，颗粒仍保持完整。我们将讨论人工血液的影响，这些颗粒具有控制大小、形状、变形性和Hb加载到RBC模拟物中的能力。
We synthesized extremely deformable red blood cell-like microgel particles and loaded them with bovine hemoglobin (Hb) to potentiate oxygen transport. With similar shape and size as red blood cells (RBCs), the particles were fabricated using the PRINT® (Particle Replication In Non-wetting Templates) technique. Low crosslinking of the hydrogel resulted in very low mesh density for these particles, allowing passive diffusion of hemoglobin throughout the particles. Hb was secured in the particles through covalent conjugation of the lysine groups of Hb to carboxyl groups in the particles via EDC/NHS coupling. Confocal microscopy of particles bound to fluorescent dye-labeled Hb confirmed the uniform distribution of Hb throughout the particle interior, as opposed to the surface conjugation only. High loading ratios, up to 5 times the amount of Hb to polymer by weight, were obtained, without a significant effect on particle stability, shape, though particle diameter decreased slightly with Hb conjugation. Analysis of the protein by circular dichroism (CD) spectroscopy showed that the secondary structure of Hb was unperturbed by conjugation to the particles. Methemoglobin in the particles could be maintained at a low level and the loaded Hb could still bind oxygen as studied by UV-vis spectroscopy. Hb-loaded particles with moderate loading ratios demonstrated excellent deformability in microfluidic devices, easily deforming to pass through restricted pores half as wide as the diameter of the particles. The suspension of concentrated particles with Hb concentration of 5.2 g/dL showed comparable viscosity to that of mouse blood, and the particles remained intact even after being sheared at a constant high rate (1,000 1/s) for 10 min. Armed with the ability to control size, shape, deformability, and loading of Hb into RBC mimics, we will discuss the implications for artificial blood.
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影响因子：
4.9
作者：
Alexis F;Pridgen E;Molnar LK;Farokhzad OC
通讯作者：
Farokhzad OC
影响因子：
15
作者：
Gershevitz, O;Sukenik, CN
通讯作者：
Sukenik, CN
影响因子：
6.2
作者：
Patton, JN;Palmer, AF
通讯作者：
Palmer, AF
影响因子：
4.9
作者：
Li, Shyh-Dar;Huang, Leaf
通讯作者：
Huang, Leaf
DOI：
10.1073/pnas.80.12.3568
发表时间：
1983-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
BUSTAMANTE, C;TINOCO, I;MAESTRE, MF
通讯作者：
MAESTRE, MF