实验技术与管理2025，Vol.42Issue(3)：72-79,8.DOI:10.16791/j.cnki.sjg.2025.03.009
基于微流控技术的PLGA多孔载细胞微球制备、表征与性能评价
Preparation,characterization,and performance evaluation of PLGA porous cell-loaded microspheres based on microfluidic technology
摘要
Abstract
[Objective]A poly(lactic-co-glycolic)acid(PLGA)copolymer scaffold system has good biocompatibility and can effectively support cell adhesion and proliferation,which makes it a potential material for three-dimensional cell culture.The preparation of porous microspheres can effectively increase the specific surface area and thus facilitate cell adhesion,nutrient delivery,and metabolic waste removal.However,the traditional emulsion method has many disadvantages,such as uneven particle size and incomplete pore expansion,which greatly reduce the effective utilization rate of PLGA porous microspheres.Moreover,the method of using alkali corrosion for pore expansion weakens the mechanical properties of microspheres and potentially increases cytotoxicity.[Methods]In view of the shortcomings of traditional preparation,we propose a method for the rapid preparation of PLGA porous microspheres using microfluidic technology.In this technology,dichloromethane is used to dissolve PLGA to obtain the oil phase.Then,a gelatin solution acting as the porogen is emulsified with the PLGA solution to acquire the injection oil phase.The polyvinyl alcohol solution serves as the water phase and collection phase.Further,emulsion drops are obtained through phase shearing on a microfluidic platform.After solvent volatilization and gelatin elution,PLGA porous microspheres are finally prepared.Compared with the traditional method,the combination of the emulsion method and microfluidic technology regulates the particle size of PLGA microspheres and facilitates pore formation in these microspheres simultaneously,which simplifies the preparation process.Subsequently,in-depth research is conducted to optimize the parameters for preparing PLGA porous microspheres using this method,such as the flow rate ratio and pore-forming agent concentration.Besides,the biocompatibility and cell adhesiveness of microspheres are tested.[Results]In this article,microspheres were successfully prepared at an oil flow rate of 1 mL/min.The average particle size distributions at 1∶10,1∶20,and 1∶30 oil-to-water flow ratios were 932.36±65.68,88.76±8.14,and 72.92±6.26 μm,respectively.The pore size of microspheres decreased with increasing gelatin concentration.The above results demonstrated the controllability of porous microsphere preparation.Furthermore,cytotoxicity and live-dead staining experiments showed that microspheres had good biocompatibility.In addition,in the coculture experiment,cells could effectively adhere to porous microspheres.These results indicated that PLGA porous microspheres prepared by the proposed method had the potential for three-dimensional cell culture and tissue engineering.[Conclusions]This study proposes that a combination of the traditional emulsion method and microfluidic technology can be used to prepare a variety of PLGA porous microspheres,which is expected to improve the application prospects of these porous microspheres.This entire study provides physical,chemical,and biological related experimental knowledge,such as that related to microfluidic systems,liquid-liquid phase transitions,cell compatibility,and cell adhesion;it also covers relative characterization techniques and data analysis points,including scanning electron microscopy,enzyme-linked immunosorbent assay,and inverted fluorescence microscopy.As a laboratory technology design,it can effectively cultivate students'comprehensive experimental abilities from problem analysis,experimental design,and implementation to data processing and analysis.关键词
多孔微球/微流控技术/PLGA/细胞培养支架Key words
porous microspheres/microfluidic technology/PLGA/cell culture scaffold分类
社会科学引用本文复制引用
周嘉安,江庆泉,宋文婧..基于微流控技术的PLGA多孔载细胞微球制备、表征与性能评价[J].实验技术与管理,2025,42(3):72-79,8.基金项目
国家自然科学基金项目(32071333) （32071333）
华南理工大学校级教研教改项目青年重点项目(x2clc249110) （x2clc249110）
