Preparation And Application Of Cell-Penetrating Peptide-modified Albumin Nanoparticles In Glioma Tumor

Objective: Glioblastoma is the major brain tumor.In the last 20 years,although the traditional treatments such as surgery,radiotherapy and chemotherapy have been greatly improved,the survival of patients with glioma has not been significantly prolonged accordingly,still less than two years.Chemotherapy is the first line of treatment for glioblastoma.The use of chemotherapeutics is hampered by the presence of the blood-brain barrier.As a result,the concentration of drugs is hard to reach a sufficient level in the tumor sites,while normal organs are exposed to high level of chemotherapy drugs.Combination chemotherapy has been widely used in the clinic to solve this drug resistance problem.In addition,the combination of several chemotherapeutics is a promising strategy for eliminating toxic and side effects from medications.Therefore,this study is designed to construct a new albumin nanoparticles delivery system with the combination of two medications and evaluate the treatment efficacy of glioblastoma.Method: Preparation of cell-penetration peptide modified albumin nanoparticles.Main methods: firstly,adopt the method of chemical coupling through crosslinking agent will cell-penetration peptide modified albumin.It was characterized by polyacrylamide gel electrophoresis(SDS-PAGE),further confirmed the final product of BSA-LMWP.Secondly,using of high concentrations of urea solution with sodium borohydride reduction conditions to degenerate of albumin.During the dilution period in the aqueous solution to hydrophobic drug paclitaxel and fenretinide loaded together,formation of drug-loading albumin nanoparticles.It was purified by the Sephadex G50 column.The albumin nanoparticles were characterized by zeta potential and size.Secondly,the albumin nanoparticles were subjected to in vitro studies,including cytotoxicity,cellular uptake in U87 glioma cells by qualitative and quantitative determination.We verified the ability to cross the blood-brain barrier and evaluate the penetrating ability of albumin nanoparticles by tumor spheroids.Thirdly,in vivo imaging and anti-glioma effect of the albumin nanoparticles were investigated using two kinds of animal models which were U87 xenograft and orthotopic tumors.Finally,this study was also tested the biocompatibility safety assessment after treatment.Results: The 150 nm albumin nanoparticles was successfully built with good form,uniform dispersion.The drug delivery system of the nanoparticles could not only be stored up efficiently in the tumor tissue by EPR effect,but also make the tumor tissue for higher level of expression of SPARC and gp60 as well as combine with albumin nanoparticles and transfer it.Therefore,we developed a novel albumin nanoparticles system which transported drugs across the BBB and penetrated brain tumors and then played a powerful role in inhibiting the growth of glioma.Conclusion: We developed a new and green nontoxic preparation method of albumin nanoparticles,which overcomes the defects from the traditional methods,such as complicated preparation facility,the low loading capacity and lower safety of prepared albumin nanoparticles.Modified albumin nanoparticles with cell-penetration peptide could effectively accumulate and permeation of albumin nanoparticles in the tumor site.In vitro results indicated the cell-penetration peptide modified albumin nanoparticles is a good treatment for glioblastoma with higher safety,which provides a new method for treatment of glioblastoma.This is a great development potential of tumor treatment.