Preparation Of Composite Nanofibers Containing Combination Chemotherapy Drugs And Its Killing Effect On Oral Squamous Cell Carcinoma Cell Line

OSCC is a common malignant tumor of the head and neck.Chemotherapy,as one of the important clinical therapy of OSCC,has puzzling problems as the clinical side effects and hardly maintained the effective drug concentrations.Nanofiber membranes,prepared by a one-step method called electrospinning technology,can be directly implanted into the lesion as a carrier for chemotherapy drugs,so that the drugs can be released locally to ensure the therapeutic effect and reduce the pain caused by toxic and side effects.According to the previous research of oral squamous carcinoma?OSCC?by our study group,the autophagy inhibitor chloroquine phosphate?CQ?down-regulates autophagy of oral squamous carcinoma cells,which can enhance the inhibitory effect of chemotherapy drug cisplatin?DDP?on OSCC and improve the therapeutic effect.In addition,the combination of methylseleninic acid?MSA?and cisplatin?DDP?can effectively enhance the killing effect on OSCC cells.Therefore,in this study,polyvinyl alcohol?PVA?and poly?lactic acid??PLA?,polymers with good biocompatibility and degradability,were selected as carrier materials to prepare one kind of three-layer composite nanofiber membranes containing MSA,DDP and CQ.This article mainly includes two parts:?1?Preparation and characterization of PLA nanofiber membranes with monolayer structure loaded with MSA and DDP,and to study the inhibitory effect of MSA/DDP-containing membranes on OSCC cells;?2?Preparation and characterization of a three-layer PVA/PLA/PVA composite fibrous membranes containing CQ,MSA,and DDP,and to study the inhibitory effect of CQ/MSA/DDP-containing membranes on OSCC cells.1.Preparation and characterization of MSA and DDP-loaded PLA nanofibers and observing its killing effect on CAL-27 cell line.Results:?1?MTS results showed that both MSA and DDP significantly inhibited the proliferation of CAL-27 cell line,and the inhibiton was concentration-dependent;the half-inhibitory concentration(IC₅₀)of MSA against CAL-27 cells was 0.3?M,and IC₅₀ of DDP against CAL-27 cells was 10?M.When 0.15?M MSA and 7.5?M DDP were combined,the inhibitory effect on CAL-27 cells was stronger.The concentration of the two drugs was relatively low,which was the optimal combination concentration.?2?SEM showed that the 8%wt PLA nanofibers were even,the nanofiber diameter was distributed between 0.6-1.7?m.Based on this,MSA,DDP and MSA+DDP-loaded PLA nanofibers were prepared,and the fibers were smooth with no lumps,indicating that the drugs were well wrapped in the nanofibers.?3?Blank PLA membranes had good biocompatibility.?4?The degradation time of the drug-loaded PLA electrospinning nanofibers is about 17 days.?5?PLA nanofiber membranes loaded with DDP and MSA has a strong killing effect on CAL-27 cells.MSA enhances the sensitivity of CAL-27 cells to DDP.2.Preparation and characterization of a three-layer PVA/PLA/PVA composite fibrous membranes containing CQ,MSA,and DDP,and to study the inhibitory effect of CQ/MSA/DDP-containing membranes on CAL-27 cells.Results:?1?SEM showed that the 7%wt PVA nanofibers had ideal morphology,and the diameter is distributed between 31-204nm.On this basis,PVA+SA nanofibers,CQ/?PVA+SA?nanofibers,PVA+SA/PLA/PVA+SA nanofibers and CQ ?PVA+SA?/DDP+MSA?PLA?/CQ?PVA+SA?composite nanofibers were prepared,the nanofibers were smooth without lumps,indicating that the drugs were well wrapped in the nanofiber.?2?The blank PVA+SA membranes had good biocompatibility.?3?The CQ?PVA+SA?/DDP+MSA?PLA?/CQ?PVA+SA?-composite nanofibers can produce sustained and strong killing effect on CAL-27 cells.Conclusion:CQ?PVA+SA?/DDP+MSA?PLA?/CQ?PVA+SA?composite nanofibers were prepared,which had a slow-release effect,and enhances the killing effect of DDP on CAL-27 cell line.