【摘要】 目的 探究负载基质细胞衍生因子-1α(stromal cell-derived factor-1 alpha, SDF-1α)的同轴静电喷雾微球对人牙髓干细胞(human dental pulp stem cells, hDPSCs)迁移与分化的作用。方法 采用静电喷雾法制备SDF-1α/PLGA微球，扫描电镜(phosphate buffer solution, SEM)与透射电镜(transmission electron microscope, TEM)观察形貌与结构，ImageJ软件分析粒径，ELISA测定包封率及体外释放实验。CCK-8检测微球对hDPSCs增殖的影响，Transwell实验评估微球促进细胞迁移的能力，实时荧光定量PCR(quantitative real time PCR, RT-qPCR)检测碱性磷酸酶(Alkaline Phosphatase, ALP)、牙本质基质蛋白-1(dentin matrix protein-1,DMP-1)、牙本质涎磷蛋白(dentin sialophosphoprotein, DSPP)、血管生成素-1(angiopoietin-1,Ang-1)等基因的表达情况。结果 SDF-1α/PLGA微球均匀分散，呈核壳结构，平均粒径约(0.79±0.13)μm,包封率为(73.8±0.30)%。微球在24 h内出现初始突释，随后释放逐渐平缓，30 d累计释放达71.04%。SDF-1α/PLGA微球在第1天微球组与对照组OD值分别为(0.45±0.04)和(0.43±0.04),差异无统计学意义(P>0.05);第3天微球组显著高于对照组，分别为(1.25±0.06)和(0.89±0.05)(P<0.05);第5天微球组OD值为(1.69±0.08),显著高于对照组的(1.38±0.06)(P<0.05)。第4周微球缓释液诱导的迁移细胞数量为(106±9)个/视野，与对照组的33±7个/视野相比，差异有统计学意义(P<0.05)。与对照组相比，微球组的ALP、DMP-1、DSPP、Ang-1、CD31相对表达量分别提高了2.06倍、1.81倍、1.80倍、2.12倍、2.00倍，差异均具有统计学意义(均P<0.05)。结论 负载SDF-1α的同轴静电喷雾微球可有效促进hDPSCs迁移和成牙本质及成血管向分化。更多还原

【Abstract】 Objective This study investigates the effects of SDF-1α-loaded coaxial electrosprayed microspheres on the migration and differentiation of human dental pulp stem cells(hDPSCs).Methods SDF-1α/PLGA microspheres were fabricated by electrostatic spraying. Their morphology and structure were characterized by scanning electron microscopy(SEM) and transmission electron microscopy(TEM), and particle size was analyzed using ImageJ. Encapsulation efficiency and in vitro release were measured by ELISA. The effects of microspheres on hDPSCs proliferation were assessed by CCK-8 assay, cell migration by Transwell assay, and gene expression(ALP, DMP-1, DSPP, Ang-1, CD31) by real-time quantitative PCR(RT-qPCR).Results SDF-1α/PLGA microspheres were uniformly dispersed with a core-shell structure, an average diameter of approximately(0.79±0.13) μm, and an encapsulation efficiency of(73.86±0.30)%. An initial burst release occurred within 24 hours, followed by a sustained release reaching 71.04% over 30 days. On day 1, OD values in the SDF-1α/PLGA microsphere and control groups were(0.45±0.04) and(0.43±0.04), respectively, with no significant difference(P>0.05). By day 3, the microsphere group(1.25±0.06) was significantly higher than the control(0.89±0.05)(P<0.05), and this trend continued on day 5(1.69±0.08 vs. 1.38±0.06, P<0.05). At week 4, microsphere-conditioned medium induced(106±9) migrated cells/field, significantly more than the control(33±7/field, P<0.05). Expression levels of ALP, DMP-1, DSPP, Ang-1, and CD31 were significantly elevated in the microsphere group by 2.06-, 1.81-, 1.80-, 2.12-, and 2.00-fold, respectively(P<0.05).Conclusion The SDF-1α-loaded electrosprayed coaxial microspheres significantly promoted hDPSCs migration and differentiation toward odontogenic and angiogenic lineages.更多还原