Allémann E, Gravel P, Leroux J C, Balant L, Gurny R
School of Pharmacy, University of Geneva, Switzerland.
J Biomed Mater Res. 1997 Nov;37(2):229-34. doi: 10.1002/(sici)1097-4636(199711)37:2<229::aid-jbm12>3.0.co;2-9.
After intravenous administration, nanoparticles suffer a major drawback in that they are rapidly and massively taken up by the cells of the mononuclear phagocyte system. The mechanisms involved in the opsonization, adhesion, and internalization of biodegradable nanoparticles by the mononuclear phagocyte system are still poorly understood. In this work, the kinetics of blood protein adsorption onto nanoparticles of poly(D,L-lactic acid) prepared by the salting-out technique was investigated. Nanoparticles of 312 nm were incubated for variable periods of time (5-60 min) in human serum and citrated plasma. After incubation, the particles were washed and the proteins detached from them, denatured, and analyzed by two-dimensional polyacrylamide gel electrophoresis. In plasma, the predominant protein was immunoglobulin G (IgG), and the amount adsorbed was not dependent on incubation time. Albumin amounts were high for short incubation periods but decreased as a function of time, whereas apolipoprotein E levels increased significantly as a function of the incubation period. Owing to the possible complement cascade inactivation by addition of citrate to plasma, the kinetics of adsorption was also evaluated in serum. In this medium, adsorption of complement C3 components onto the surface of the nanoparticles was clearly evidenced by spots of increasing intensity and area, reaching levels comparable to those of the omnipresent IgG. This result confirms the important role of complement components in the opsonization process of poly(D,L-lactic acid) particles.
静脉给药后，纳米颗粒存在一个主要缺点，即它们会被单核吞噬细胞系统的细胞迅速大量摄取。单核吞噬细胞系统对可生物降解纳米颗粒进行调理、黏附及内化的相关机制仍了解甚少。在这项研究中，我们研究了通过盐析技术制备的聚（D，L-乳酸）纳米颗粒上血液蛋白吸附的动力学。将312 nm的纳米颗粒在人血清和枸橼酸盐血浆中孵育不同时间（5 - 60分钟）。孵育后，洗涤颗粒，将从颗粒上分离的蛋白质变性，并通过二维聚丙烯酰胺凝胶电泳进行分析。在血浆中，主要蛋白质是免疫球蛋白G（IgG），吸附量不依赖于孵育时间。孵育时间短时白蛋白量较高，但随时间减少，而载脂蛋白E水平随孵育时间显著增加。由于向血浆中添加枸橼酸盐可能使补体级联失活，因此也在血清中评估了吸附动力学。在这种介质中，纳米颗粒表面补体C3成分的吸附通过强度和面积不断增加的斑点得到明确证实，达到了与普遍存在的IgG相当的水平。这一结果证实了补体成分在聚（D，L-乳酸）颗粒调理过程中的重要作用。
