The Function Of Macrophage Influenced By PS Microspheres With Different Surface Roughness And Its Mechanism

Objective: To investigate the IL-1? secretion of macrophage influenced by PS microspheres with different surface roughness and its mechanism.Methods: 1.Microspheres were prepared by microfluidic technique and emulsion solvent evaporation method, and its surface morphology was characterized by scanning electron microscopy(SEM) and transmission electron microscopy(TEM).2.The phagocytosis of microspheres with different roughness were observed with confocal microscope, and the phagocytic rate of microspheres with different roughness were detected by flow cytometry. 3.ELISA were used to detect the release of IL-1? in Ana-1 macrophages stimulated by microspheres with different roughness.4.Microspheres with different roughness were injected into mouse peritoneal to detect the inflammatory response.5.PCR was used to verify the inflammatory pathway of NLRP3 induced by microspheres in macrophage.Results:1. By the microfluidic technique and emulsion solvent evaporation method,we obtained the uniform PS microspheres with different surface roughness; 2. In confocal microscopy and flow cytometry, we concluded that different PS microspheres were swallowed by Ana-1 cells in different periods. The longer time incubated, the more microspheres were phagocyted, and more Ana-1 cells were recruited around the microspheres with rough surface; 3.ELISA experiments showed that the cells of experimental group released more IL-1? than control group, and the amount of IL-1? in coarse group were more than that in smooth group. 4. In vivo study showed that more neutrophils were recruited after injection of rough spheres compared those of smooth spheres or PBS(p<0.05); 5. PCR analysis showed different microspheres could up-regulate inflammasome m RNA, such as NLRP3, ASC,caspase-1, and their counts were different among different microparticle groups.Conclusions: Polymer microsphere with controllable surface roughness prepared by methods mentioned above. And we validated that the roughness of polymer microsphere could influence cell biological functions(such as IL-1? secretion).Based on the results, it will provide a new idea for designing a biocompatibility adjuvant or therapeutic microspheres in the future.