微流控芯片的微米级结构使流体在通道中显示和宏观尺度不同的特殊性能，同时反应速度快、消耗试剂少等特点提供了实用性优势，在生物医学领域有巨大的发展前景。近几年来，新兴的可集成不同反应室的微流控芯片已成为合成核壳纳米颗粒的强大工具。由于微流体动力学增强了微通道中液体的混合，可制备粒径均一的微纳米颗粒，且不同批次之间差异小。然而，利用微流控芯片制备细胞膜包覆的微纳米颗粒研究较少。在已有利用微流控技术研究中，微流体难以在微通道充分使细胞膜与纳米颗粒混合，限制了微流控对制备细胞膜包覆纳米颗粒的应用。挤压制备方法破坏细胞膜包覆的核壳结构，导致细胞膜包覆率较低，过程耗时较长。针对这一问题，本研究提出了外部场力与微流控技术的结合，设计并制备基于超声辅助的软管微流控芯片，用于快速混合制备微纳米颗粒，提高细胞膜包覆率。

设计并制备不同形状软管支撑模板，进一步组装成软管微流控芯片。根据流量、流速比和超声功率的荧光实验分析，不同形状芯片对于荧光素混合效率没有显著影响。其中，混合单元数为3的直通道软管微流控芯片的混合效率最高。在流速比为1：5时，混合效率最高为78.83 %；在同等流量条件下，当超声功率达到90 %时，混合效率可提高19.6 %。从而说明超声辅助的软管微流控芯片可以提高荧光素等小分子混合效率。进一步探究在软管微流控芯片上，超声功率对细胞膜包覆荧光微球的包覆率影响。

利用荧光微球模拟纳米探针进行癌细胞膜包覆实验，探究流量、流速比和超声功率参数对包覆率影响。综合流量和超声功率参数，可得出包覆率最高的参数是流量为100 μL/min，超声功率为90 %，其包覆率是对照组的2.32倍。流速比越高，超声功率对于包覆率的影响越大；注入细胞膜与荧光微球的流速比Rf为3时，超声功率为90 %，包覆率是对照组的1.83倍；最佳细胞膜和纳米探针的材料比为3:1从以上实验结果表明，超声功率改变对细胞膜包覆率有显著增强的作用。通过扫描电子显微镜进一步证明了细胞膜成功包覆到荧光微球上，证明了超声辅助软管微流控芯片可制备细胞膜包覆微米级荧光微球。

基于超声辅助的软管微流控芯片制备AuNS@SiO₂@CCMs-EC109、AuNS@SiO₂@CCMs-4T1和AuNS@SiO₂@RBCM三种纳米探针 。对比分析AuNS@SiO₂纳米探针的电镜、粒径后发现，细胞膜被成功包覆在AuNS@SiO₂纳米探针表面。在细胞水平上，0 ~ 50 սg/mL浓度 的纳米探针的细胞活性均大于90 %，证明了纳米探针具备良好生物相容性。光热治疗结果表明，当AuNS@SiO₂@CCMs-EC109纳米探针的浓度是50 μg/mL对EC-109食管癌细胞杀伤效果可将细胞存活率降低至17.18%。由此可见，细胞膜包覆后的纳米探针具备更优秀的光热治疗效果。相比于相同浓度的AuNS@SiO₂@RBCM纳米探针对EC-109细胞存活率为23.92%，说明同源靶向性强，有利于癌症治疗。

本研究在实现小分子物质快速混合基础上，制备癌细胞膜包覆微米级荧光微球和多种细胞膜包覆纳米级探针。超声辅助的软管微流控芯片提高了细胞膜对于纳米探针的包覆率。在保证细胞膜功能完好的同时，进一步降低制备细胞膜包覆纳米颗粒的材料成本和时间成本。因此，超声辅助的软管微流控芯片在制备细胞膜包覆微纳米颗粒具有广泛的生物医学应用及其在临床转化的商业价值。

The micron-level structure of the microfluidic chip enables the fluid to display different special properties in the channel and at the macro scale. At the same time, the characteristics of fast reaction speed and less consumption of reagents provide practical advantages, and it has a great development prospect in the field of biomedical science. In recent years, emerging microfluidic chips that can integrate different reaction chambers have become a powerful tool for the synthesis of core-shell nanoparticles. Because microfluidic dynamics enhances the mixing of liquids in microchannels, micro-nanoparticles with uniform particle sizes can be prepared with little difference between batches. However, the use of microfluidic chips to prepare membrane-coated micro and nano particles is rarely studied. The external force caused by the traditional extrusion preparation method can destroy the core-shell structure of the membrane coating, resulting in a low membrane coating rate, and

the process takes a long time. The extrusion method destroys the core-shell structure of the membrane coating, resulting in a low coating rate and a long process time. In view of this problem, this study proposed the combination of external field force and microfluidic technology, designed and prepared the ultrasonic-assisted soft microfluidic chip, In view of this problem, this study proposed the combination of external field force and microfluidic technology, designed and prepared the ultrasonic-assisted soft microfluidic chip, which is used for the rapid mixing and preparation of micro and nano particles and improving the coating rate of cell membrane.

 

The supporting templates of different shapes of hoses were prepared and further assembled into hose-microfluidic chips. According to the fluorescence experiment results of different quantity, flow rate ratio and ultrasonic power, different shapes of chips have no significant effect on the luciferin mixing efficiency. Among them, the microfluidic chip with the number of mixing units of 3 has the highest mixing efficiency. When the flow rate ratio was 1:5, the mixing efficiency reached the highest of 78.83%. Under the same flow condition, when the ultrasonic power reaches 90%, the mixing efficiency can be increased by 19.6%.

 

Combined with the flow rate and ultrasonic power parameters, the highest coating rate was obtained when the flow rate was 100 μL/min and the ultrasonic power was 90 %. The coating rate was 2.32 times that of the control group.The higher the velocity ratio, the greater the influence of ultrasonic power on the coating rate.When the flow rate ratio of injected cell membrane to fluorescent microspheres was 35, the ultrasonic power was 90%, and the coating rate was 1.83 times that of the control group.The optimal material ratio between the cell membrane and the nanoprobe is 3:1. From the above experimental results, it is indicated that the change of ultrasonic power can significantly enhance the cell membrane coating rate.Through scanning electron microscopy, it was further proved that the cell membrane was successfully coated on the fluorescent microspheres, and it was proved that the ultrasonic-assisted flexible tube microfluidic chip could prepare the micron-sized fluorescent microspheres coated on the cell membrane.

 

A flexible tube microfluidic chip was used to fabricate the AuNS@SiO₂ @CCMS-EC109、AuNS@SiO₂@CCMS-4T1 and AuNS@SiO₂@RBCM nanoprobe. By comparing the electron microscopy and particle size of the AuNS@SiO₂ nano-probe, it was found that the cancer cell membrane was successfully coated on the surface of the nano-probe. At the cellular level, the cell activity of the nanoprobes at different concentrations was more than 90%, which proved that the nanoprobes had good biocompatibility. The results of photothermal therapy showed that the killing effect of AuNS@SiO₂@CCMS-EC109 nano-probe at 50 μg/mL on EC-109 esophageal cancer cells could reduce the cell survival rate to 17.18%. In conclusion, the membrane coated nanoprobe has a better photothermal therapeutic effect. Compared with the same AuNS@SiO₂@RBCM nanoprobe,the cell survival rate was 23.92%,indicating strong homologous targeting, which is beneficial to cancer treatment.

 

In this study, on the basis of fast mixing of small molecules, micron-scale fluorescent microspheres coated with cancer cell membrane and nanoscale probes coated with various cell membranes were prepared. Meanwhile, the material cost and time cost of preparing membrane-coated nanoparticles can be further reduced. Therefore, ultrasound-assisted soft microfluidic chips have a wide range of biomedical applications and commercial value in clinical transformation in the preparation of membrane-coated micro and nano particles.

The micron-level structure of the microfluidic chip enables the fluid to display different special properties in the channel and at the macro scale. At the same time, the characteristics of fast reaction speed and less consumption of reagents provide practical advantages, and it has a great development prospect in the field of biomedical science. In recent years, emerging microfluidic chips that can integrate different reaction chambers have become a powerful tool for the synthesis of core-shell nanoparticles. Because microfluidic dynamics enhances the mixing of liquids in microchannels, micro-nanoparticles with uniform particle sizes can be prepared with little difference between batches. However, the use of microfluidic chips to prepare membrane-coated micro and nano particles is rarely studied. The external force caused by the traditional extrusion preparation method can destroy the core-shell structure of the membrane coating, resulting in a low membrane coating rate, and
the process takes a long time. The extrusion method destroys the core-shell structure of the membrane coating, resulting in a low coating rate and a long process time. In view of this problem, this study proposed the combination of external field force and microfluidic technology, designed and prepared the ultrasonic-assisted soft microfluidic chip, In view of this problem, this study proposed the combination of external field force and microfluidic technology, designed and prepared the ultrasonic-assisted soft microfluidic chip, which is used for the rapid mixing and preparation of micro and nano particles and improving the coating rate of cell membrane.

The supporting templates of different shapes of hoses were prepared and further assembled into hose-microfluidic chips. According to the fluorescence experiment results of different quantity, flow rate ratio and ultrasonic power, different shapes of chips have no significant effect on the luciferin mixing efficiency. Among them, the microfluidic chip with the number of mixing units of 3 has the highest mixing efficiency. When the flow rate ratio was 1:5, the mixing efficiency reached the highest of 78.83%. Under the same flow condition, when the ultrasonic power reaches 90%, the mixing efficiency can be increased by 19.6%.

Combined with the flow rate and ultrasonic power parameters, the highest coating rate was obtained when the flow rate was 100 μL/min and the ultrasonic power was 90 %. The coating rate was 2.32 times that of the control group.The higher the velocity ratio, the greater the influence of ultrasonic power on the coating rate.When the flow rate ratio of injected cell membrane to fluorescent microspheres was 35, the ultrasonic power was 90%, and the coating rate was 1.83 times that of the control group.The optimal material ratio between the cell membrane and the nanoprobe is 3:1. From the above experimental results, it is indicated that the change of ultrasonic power can significantly enhance the cell membrane coating rate.Through scanning electron microscopy, it was further proved that the cell membrane was successfully coated on the fluorescent microspheres, and it was proved that the ultrasonic-assisted flexible tube microfluidic chip could prepare the micron-sized fluorescent microspheres coated on the cell membrane.

A flexible tube microfluidic chip was used to fabricate the AuNS@SiO2 @CCMS-EC109、AuNS@SiO2@CCMS-4T1 and AuNS@SiO2@RBCM nanoprobe. By comparing the electron microscopy and particle size of the AuNS@SiO2 nano-probe, it was found that the cancer cell membrane was successfully coated on the surface of the nano-probe. At the cellular level, the cell activity of the nanoprobes at different concentrations was more than 90%, which proved that the nanoprobes had good biocompatibility. The results of photothermal therapy showed that the killing effect of AuNS@SiO2@CCMS-EC109 nano-probe at 50 μg/mL on EC-109 esophageal cancer cells could reduce the cell survival rate to 17.18%. In conclusion, the membrane coated nanoprobe has a better photothermal therapeutic effect. Compared with the same AuNS@SiO2@RBCM nanoprobe,the cell survival rate was 23.92%,indicating strong homologous targeting, which is beneficial to cancer treatment.

In this study, on the basis of fast mixing of small molecules, micron-scale fluorescent microspheres coated with cancer cell membrane and nanoscale probes coated with various cell membranes were prepared. Meanwhile, the material cost and time cost of preparing membrane-coated nanoparticles can be further reduced. Therefore, ultrasound-assisted soft microfluidic chips have a wide range of biomedical applications and commercial value in clinical transformation in the preparation of membrane-coated micro and nano particles.