静电喷雾法制备聚乳酸-羟基乙酸共聚物/羟基磷灰石微载体的工艺及其效果评价

目的：探讨静电喷雾法制备聚乳酸-羟基乙酸共聚物/羟基磷灰石（PLGA/HA）微载体的工艺过程，阐明PLGA/HA微载体的优势表征。方法：采用HA （20 nm，99.9%）和PLGA （LA/GA=50/50，Mw30k），利用静电喷雾法制备PLGA/HA微载体，观察不同HA质量浓度（1%、3%、5%）、不同电压（4.0、4.5、>5.0kV）对微载体形貌的影响，获得最佳制球参数。以PLGA微载体为PLGA组，PLGA+1% HA为1% PLGA/HA组，PLGA+3% HA为3% PLGA/HA组，PLGA+5% HA为5% PLGA/HA组，单纯细胞作为空白对照组。采用扫描电镜（SEM）、细胞增殖实验、细胞荧光染色实验和傅里叶红外光谱（FTIR）等检测PLGA/HA微载体表征。结果：SEM观察，PLGA/HA微载体颗粒均匀，均为椭圆形或圆形，无异常形态球，表面光滑，无锐利边缘，球体之间无粘连、无聚集，不同浓度配比微载体之间无明显差别。细胞增殖实验检测，黏附细胞数量5% PLGA/HA组 > 3% PLGA/HA组 > 1% PLGA/HA组 > PLGA组 > 空白对照组（P<0.05），细胞数量随HA浓度增加而升高，5% PLGA/HA组微载体对细胞亲和性最佳，微载体无细胞毒性。细胞荧光染色实验检测，MC3T3-E1细胞在微载体上黏附状态良好。FTIR检测显示HA特征吸收峰，表明复合微载体中含有PLGA与HA。结论：成功利用静电喷雾技术建立PLGA/HA微载体的制备工艺，该方法操作简单便捷，制球效果优良，在骨组织工程方面有广泛应用前景。

Preparation process of polylactic acid-glycolic acid copolymer/hydroxyapatite microcarrier by electrostatic spraying method and its effect evaluation

Objective: To investigate the preparation process of polylactic acid-glycolic acid copolymer/hydroxyapatite (PLGA/HA) microcarriers by electrostatic spraying method, and to elucidate the superiority of PLGA/HA microcarriers.Methods: The PLGA/HA microcarriers were prepared by electrostatic spraying method using nano-HA (20 nm, 99.9%) and PLGA (LA/GA=50/50, Mw30k),and the influence of different concentrations (1%, 3%, 5%) and different voltages (4.0, 4.5, > 5.0 kV) of HA in the morphology of the microcarriers was investigated and the best ball making parameters were obtained.The PLGA microcarriers were used as PLGA group, PLGA+1%HA as 1% PLGA/HA group, PLGA+3% HA as 3% PLGA/HA group, PLGA+5%HA as 5%PLGA/HA group, and the simple cells were used as blank control group.The characteristics of PLGA/HA microcarriers were detected by scanning electron microscope (SEM), cell proliferation test, cell fluorescence staining experiment, and Fourier transform infrared spectroscopy (FTIR).Results: The SEM results showed that the microcarrier particles were uniform, all of them were elliptical or circular, without abnormal shape spheres, with smooth surface, without sharp edges, adhesion between the spheres and aggregation, and there were no significant differences between the different concentrations of microcarriers.The cell proliferation test results showed that the order of adhesion cells was 5%PLGA/HA group > 3%PLGA/HA group > 1% PLGA/HA group > PLGA group > blank control group (P<0.05); the number of cells was increased with the increasing of HA concentration; the microcarriers in 5%PLGA/HA group had the best cell affinity and the microcarriers had no cytotoxity.The cell fluorescence staining experiment showed that the MC3T3-E1 cells adhered well on the microcarriers.The FTIR analysis results showed that HA characteristic absorption peak was observed, indicating that the composite microcarrier contained PLGA and HA.Conclusion: The preparation process of PLGA/HA microcarriers is successfully established by electrostatic spraying method.The method is simple and convenient to operate, and has excellent ball-making effect.It has broadly application prospects in bone tissue engineering.