| Objective:To study the inhibitory effect of 20(S)-protopanaxadiol(PPD)-loaded polycaprolactone(PCL)nanofiber membranes prepared by electrospinning on laryngeal malignant tumors in vitro and in vivo,and to investigate whether electrospun nanofiber membranes are feasible as anti-tumor carriers for PPD.Methods:PCL nanofiber membranes loaded with PPD were prepared by electrospinning with hexafluoroisopropanol as solvent.The changes of fiber diameter and morphology for nonafibers with different concentrations of PPD were observed by scanning electron microscopy.The thermal stability and mechanical properties of the fiber membranes were tested using thermal analyzer and universal capability tester.Qualitative analysis of the drug-loaded nanofiber membranes were performed using Fourier infrared spectrometers;the in vitro degradation test was used to investigate whether the drugloaded nanofiber has biodegradability;the high performance liquid chromatograph was used to study the drug release characteristics of the drug-loaded nanofiber membrane;MTT assay was used to detect the cytotoxicity of drug-loaded nanofiber membranes on human laryngeal cancer cells(Hep-2);Annexin V-EGFP/PI double staining method was used to detect the apoptosis of Hep-2 cells in vitro.BALB/c nude mouse Hep-2 subcutaneous tumor model was prepared to study the in vivo antitumor effect of the nanofiber membrane.Results:(1)Scanning electron microscopy showed that the surface of drug-loaded nanofibers was smooth,and the fiber diameter increased with the increase of drug concentration.(2)Thermogravimetric analysis showed that the weight loss of all nanofiber membranes was between 300-350°C;Mechanical tests showed that when the loading content of PPD was lower than 10% of total polymers,the fracture stress of nanofibers increased gradually,and when the loading content exceeded 10%,the fracture stress decreased;Fourier infrared spectroscopy showed that the spectrum of the drug-loaded nanofiber membrane contained both functional peaks of PPD and PCL,and no new functional group absorption peak appeared.(3)In vitro degradation experiments revealed that when the nanofibers were immersed in phosphate buffered saline(PBS)for 15 days,the nanofibers were deformed and fractured.(4)The results of in vitro drug release experiments showed that the drug was rapidly released from the drug-loaded nanofiber membrane in the first 12 h,and then the drug released slowly.(5)The results of MTT assay showed that the survival rate of tumor cells decreased with the increase of drug concentration.When the drug concentration reached 30?g/ml,the cytotoxicity of the PPD group was better than the drug-loaded nanofiber group after 1 day.Statistical analysis showed a significant difference(p<0.01).Then the cell survival rate of the drug-loaded nanofiber membrane group was lower than the PPD group after 3 day(p<0.01).The results of apoptosis experiments showed that the apoptotic rates of blank group,PPD group,and drug-loaded nanofiber membrane group were 2.6±0.4,16.8±2.1,and 29.9±2.4,respectively.There were significant statistical differences between the groups(p<0.01).(6)Animal experiments showed that there was no significant difference in tumour volume and weight between the blank group and the PCL nanofiber membrane group after 9 days(p>0.05).However,the tumor volume and weight of the PPD group and the drug-loaded nanofiber membrane group were significantly lower than those of the first two groups(p<0.01).Conclusion:(1)The drug-loaded nanofibers equipped with good thermal stability and mechanical properties were successfully prepared.No new substances were generated during the synthesis.It is speculated that this method does not affect the structure and efficacy of PPD.(2)Drug-loaded nanofiber membranes could be degraded,and PPD from drugloaded nanofiber could be released in a prolonged mode.(3)Cellular and animal experiments showed that the nanofiber membrane has good antitumor effect in vitro and in vivo,indicating that the PCL nanofiber membrane could be used as a carrier for PPD. |
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