We provide a facile and low-cost method (F-L) to fabricate a two-dimensional positive master using a milling technique for polydimethylsiloxane (PDMS)-based microchannel molding. This method comprises the following steps: (1) a positive microscale master of the geometry is milled on to an acrylic block; (2) pre-cured PDMS is used to mold the microscale positive master; (3) the PDMS plate is peeled off from the master and punctured with a blunt needle; and (4) the PDMS plate is O₂ plasma bonded to a glass slide. Using this technique, we can fabricate microchannels with very simple protocols quickly and inexpensively. This method also avoids breakage of the end mill (ϕ = 0.4 mm) of the computerized numerical control (CNC) system when fabricating the narrow channels (width < 50 µm). The prominent surface roughness of the milled bottom-layer could be overcomed by pre-cured PDMS with size trade-off in design. Finally, emulsion formation successfully demonstrates the validity of the proposed fabrication protocol. This work represents an important step toward the use of a milling technique for PDMS-based microfabrication.

Introduction: The integrity of the erythrocyte membrane cytoskeletal network controls the morphology, specific surface area, material exchange, and state of erythrocytes in the blood circulation. The antioxidant properties of resveratrol have been reported, but studies on the effect of resveratrol on the hypoxia-induced mechanical properties of erythrocytes are rare.Methods: In this study, the effects of different concentrations of resveratrol on the protection of red blood cell mor-phology and changes in intracellular redox levels were examined to select an appropriate concentration for further study. The Young’s modulus and surface roughness of the red blood cells and blood viscosity were measured via atomic force microsco-py and a blood rheometer, respectively. Flow cytometry, free hemoglobin levels, and membrane lipid peroxidation levels were used to characterize cell membrane damage in the presence and absence of resveratrol after hypoxia. The effects of oxida-tive stress on the erythrocyte membrane proteins band 3 and spectrin were further investigated by immunofluorescent label-ing and Western blotting.Results and discussion: Resveratrol changed the surface roughness and Young’s modulus of the erythrocyte mem-brane, reduced the rate of eryptosis in erythrocytes after hypoxia, and stabilized the intracellular redox level. Further data showed that resveratrol protected the erythrocyte membrane proteins band 3 and spectrin. Moreover, resistance to band 3 pro-tein tyrosine phosphorylation and sulfhydryl oxidation can protect the stability of the erythrocyte membrane skeleton net-work, thereby protecting erythrocyte deformability under hypoxia. The results of the present study may provide new insights into the roles of resveratrol in the prevention of hypoxia and as an antioxidant.

Activated carbon (AC) is an effective adsorbent for creatinine removal in hemoperfusion. However, the hemocompatibility and adsorption capacity of AC was required to be improved further. Heparin has different anticoagulant mechanisms due to its different molecular weights. Thus, it was necessary to study the surface modification with unfractionated heparin (UFH) or low-molecular-weight heparin (LMWH) on improvement of hemocompatibility and adsorption. In this study, UFH and LMWH were, respectively, grafted on AC through polyethyleneimine as an intermediate layer. The modification of AC regarding morphology, mechanical strength, and pore structure was characterized by X-ray photoelectron spectroscopy (XPS), scanning electron microscope (SEM), texture analyzer (TA), and surface area analyzer. It was found that, compared with AC, the morphology and mechanical strength of AC-UFH and AC-LMWH could be well maintained, but the specific surface area was decreased due to the grafting of macromolecules. Furthermore, AC-UFH and AC-LMWH showed better hemocompatibility on protein adsorption, clotting time, and platelet activation compared with AC, in which AC-LMWH had lower fibrinogen adsorption and longer clotting time than AC-UFH. In addition, it was found that AC, AC-UFH, and AC-LMWH had no significant effect on blood cell composition. Finally, the adsorption capacity of adsorbents for creatinine was evaluated. Although there was no significant difference between AC-UFH and AC-LMWH, it was found that heparin could be interacted with creatinine to enhance the adsorption capacity when compared with polyethyleneimine-modified AC. This study deepened the understanding of anticoagulation of heparinized surface and provided a theoretical basis for adsorption in hemoperfusion.

Activated carbon (AC) is an effective adsorbent for creatinine removal in hemoperfusion. However, the hemocompatibility and adsorption capacity of AC was required to be improved furthermore. Heparin, a widely used anticoagulant biomacromolecule, could improve the hemocompatibility of the absorbents in different ways based on its molecular weight. Thus, it was necessary to study the surface modification with unfractionated heparin (UFH) or low molecular weight heparin (LMWH) on improvement of hemocompatibility and adsorption. In this study, UFH and LMWH were respectively grafted on AC through polyethyleneimine as an intermediate layer. The modification of AC about morphology, mechanical strength and pore structure was characterized by XPS, SEM, TA and BET. It was found that compared with AC, the morphology and mechanical strength AC-UFH and AC-LMWH could be well maintained, but the BET surface was decreased due to the grafting of macromolecules. Furthermore, AC-UFH and AC-LMWH showed the better hemocompatibility on protein adsorption, APTT and platelet activation compared with AC, in which AC-LMWH had lower fibrinogen adsorption and longer APTT than AC-UFH. Besides, it was found that AC, AC-UFH and AC-LMWH had no significant effect on blood cell composition. Finally, the adsorption capacity of adsorbents for creatinine was evaluated. Although there was no significant difference between AC-UFH and AC-LMWH, it was found that heparin could be interacted with creatinine to enhance the adsorption capacity when compared with AC-PEI. This study deepened the understanding of anticoagulation of heparinized surface and provided a theoretical basis for adsorbents in hemoperfusion.