功能化载药黑磷纳米片的制备表征及抗骨肉瘤活性评价

目的：制备可稳定分散的载药纳米黑磷，并将其用于光热/化疗协同的纳米制剂。方法：采用液相剥离法，以N-甲基吡咯烷酮（NMP）为剥离剂，聚乙二醇二胺（H₂N-PEG-NH₂）、壳聚糖（CS）及A型明胶（Gel）为修饰剂，筛选并载带化疗药物盐酸阿霉素（DOX）制备出分散性良好的载药黑磷纳米片。通过透射电镜、傅立叶变换红外光谱仪及流式细胞仪等仪器，进行形貌结构表征及体外细胞学评价。结果：剥离方法筛选试验表明BPNSs最佳制备工艺为：水浴超声300 W，10 h，探头超声180 W，6 h，温度<30℃，表现出外观形貌均一；修饰剂筛选试验表明H₂N-PEG-NH₂/BPNSs质量比为10：1，BPNSs/DOX质量比为1：1，表现出良好的稳定性及负载率；体外释放试验表明，DOX在弱酸性条件下结合近红外光照（NIR）释放迅速，展现出光控释药特性；体外细胞学试验表明PEG-BPNSs-DOX具有良好的抗骨肉瘤活性。结论：PEG-BPNSs-DOX可稳定分散，制备简单，具有NIR光控释药特性及良好的光热性能，有利于光热/化疗协同治疗骨肉瘤，在纳米递送系统中具有广阔应用前景。

Preparation and characterization of functionalized drug-loaded black phosphorus nanoparticles and evaluations of anti-osteosarcoma activity

OBJECTIVE To prepare drug-loaded black phosphorus nanomaterials (BPNSs) stably dispersed and apply in nano-preparations with synergistic effects of photothermal therapy and chemotherapy.METHODS With liquid phase exfoliation method, using N-methylpyrrolidone (NMP) as a liquid phase stripping agent, polyethylene glycol diamine (H₂N-PEG-NH₂), chitosan (CS) and A-type gelatin (Gel) as modifiers, chemotherapeutic agent doxorubicin hydrochloride (DOX) was screened and loaded for preparing drug-loaded black phosphorus nanoparticles with an excellent dispersion. The morphology, structure and in vitro cytology were characterized by transmission electron microscope, Fourier transform infrared spectrometer (FTIR) and flow cytometry.RESULTS The optimal preparation technology of BPNSs was as follows:water bath ultrasonic 300 W, 10 h, probe ultrasonic 180 W, 6 h, peeling temperature <30℃. Exfoliation product showed uniform appearance; modifier screening experiment indicated that mass ratio of H₂N-PEG-NH₂/BPNSs was 10 and mass ratio of BPNS 1, showing decent stability and loading rate; release experiment in vitro showed that DOX released rapidly in weak acid condition plus near infrared light (NIR), showing the characteristics of NIR photocontrolled release; and cytological experiments in vitro showed that PEG-BPNSs-DOX had excellent anti-osteosarcoma activity.CONCLUSION PEG-BPNSs-DOX may be stably dispersed. With a simple preparation method, it has the characteristics of NIR photocontrolled release and excellent photothermal properties, benefitting the photothermal/chemotherapy synergistic treatment of osteosarcoma. It has a broad prospect in the application of a nano-delivery system.