Institute of Biomaterials and Biomedical Engineering, University of Toronto, Canada; Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Canada.
Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Canada.
Biomaterials. 2017 Feb;118:74-83. doi: 10.1016/j.biomaterials.2016.11.036. Epub 2016 Nov 25.
Polymer beads made of 45% methacrylic acid co methyl methacrylate (MAA beads) promote vascular regenerative responses in contrast to control materials without methacrylic acid (here polymethyl methacrylate beads, PMMA). In vitro and in vivo studies suggest that MAA copolymers induce differences in macrophage phenotype and polarization and inflammatory responses, presumably due to protein adsorption differences between the beads. To explore differences in protein adsorption in an unbiased manner, we used high resolution shotgun mass spectrometry to identify and compare proteins that adsorb from human plasma or serum onto MAA and PMMA beads. From plasma, MAA beads adsorbed many complement proteins, such as C1q, C4-related proteins and the complement inhibitor factor H, while PMMA adsorbed proteins, such as albumin, C3 and apolipoproteins. Because of the differences in complement protein adsorption, follow-up studies focused on using ELISA to assess complement activation. When incubated in serum, MAA beads generated significantly lower levels of soluble C5b9 and C3a/C3a in comparison to PMMA beads, indicating a decrease in complement activation with MAA beads. The differences in adsorbed protein on the two materials likely alter subsequent cell-material interactions that ultimately result in different host responses and local vascularization.
由 45%甲基丙烯酸甲酯共甲基丙烯酸(MAA 珠)制成的聚合物珠与不含甲基丙烯酸的对照材料(这里是聚甲基丙烯酸甲酯珠,PMMA)相比,促进血管再生反应。体外和体内研究表明,MAA 共聚物诱导巨噬细胞表型和极化以及炎症反应的差异,推测是由于珠之间的蛋白质吸附差异所致。为了以无偏倚的方式探索蛋白质吸附的差异,我们使用高分辨率鸟枪法质谱鉴定和比较从人血浆或血清中吸附到 MAA 和 PMMA 珠上的蛋白质。从血浆中,MAA 珠吸附了许多补体蛋白,如 C1q、C4 相关蛋白和补体抑制剂因子 H,而 PMMA 珠吸附了白蛋白、C3 和载脂蛋白等蛋白。由于补体蛋白吸附的差异,后续研究集中于使用 ELISA 评估补体激活。当在血清中孵育时,与 PMMA 珠相比,MAA 珠产生的可溶性 C5b9 和 C3a/C3a 的水平明显降低,表明 MAA 珠的补体激活减少。这两种材料上吸附的蛋白质的差异可能改变随后的细胞-材料相互作用,最终导致不同的宿主反应和局部血管化。