Novel Delivery of Mitoxantrone with Hydrophobically Modified Pullulan Nanoparticles to Inhibit Bladder Cancer Cell and the Effect of Nano-drug Size on Inhibition Efficiency

Abstract

Reducing the dosage of chemotherapeutic drugs via enhancing the delivery efficiency using novel nanoparticles has great potential for cancer treatment. Here, we focused on improving mitoxantrone delivery by using cholesterol-substituted pullulan polymers (CHPs) and selected a suitable nano-drug size to inhibit the growth of bladder cancer cells. We synthesized three kinds of CHPs, named CHP-1, CHP-2, CHP-3. Their chemical structures were identified by NMR, and the degree of cholesterol substitution was 6.82%, 5.78%, and 2.74%, respectively. Their diameters were 86.4, 162.30, and 222.28 nm. We tested the release rate of mitoxantrone in phosphate-buffered saline for 48 h: the release rate was 38.73%, 42.35%, and 58.89% for the three CHPs. The hydrophobic substitution degree in the polymer was associated with the self-assembly process of the nanoparticles, which affected their size and therefore drug release rate. The release of the three drug-loaded nanoparticles was significantly accelerated in acid release media. The larger the nanoparticle, the greater the drug release velocity. At 24 h, the IC50 value was 0.25 M, for the best inhibition of mitoxantrone on bladder cancer cells.3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) experiments demonstrated that drug-loaded CHP-3 nanoparticles with the largest size were the most toxic to bladder cancer cells. Immunofluorescence and flow cytometry revealed that drug-loaded CHP-3 nanoparticles with the largest size had the strongest effect on promoting apoptosis of bladder cancer cells. Also, the three drug-loaded nanoparticles could all inhibit the migration of MB49 cells, with large-size CHP-3 nanoparticles having the most powerful inhibition.

Preparation of Cholesteryl-Modified Aminated Pullulan Nanoparticles to Evaluate Nanoparticle of Hydrophobic Degree on Drug Release and Cytotoxicity

“…In a previous study [23,24], the drug loading of CHP NPs depended on the degree of cholesterol substitution in the polymer, and after reaching the critical concentration of aggregation to form NPs, the drug loading increased with increasing degree of substitution. Amino NPs had higher inhibitory activity in cancer cells than the two other kinds Journal of Nanomaterials of NPs by grafting cholesterol without modification and the carboxyl group [35].…”

“…A small amount of solid succinic anhydride cholesterol ester, amino pullulan polysaccharide, and different CHPNs was dissolved in deuterium DMSO and analyzed by NMR. The degree of substitution of CHS was determined on the basis of the α-1,4 and α-1,6 glycosidic bonds and the area under the methylene peak [24]: Drug release was measured by the dialysis method. Briefly, 5 mg drug-loaded NPs were dispersed in 10 mL phosphate buffer (pH 7.4, temperature 37.5°C) and loaded into a dialysis bag (M W = 8-12 kDa) and separated at 25 mL, pH 7.4.…”

“…Previous study of cholesterol-modified pullulan polysaccharide showed that it has high loading capacity and a sustained-release effect [23]. The hydrophobicity of NPs can be changed by changing the substitution degree of cholesterol, which changes the size and drug loading and release of NPs [24]. However, because of the large surface area and high surface activity, NPs can easily agglomerate, which will have adverse effects [10,25].…”

Interaction of Mitoxantrone-Loaded Cholesterol Modified Pullulan Nanoparticles with Human Serum Albumin and Effect on Drug Release

“…Transform Infrared (FT-IR) Spectroscopy. The synthesis of CHP conjugate was previously reported [23]. The FT-IR spectra of pullulan, cholesterol succinate (CHS), and CHP were obtained as KBr pellets for FT-IR spectroscopy (Nicolet Nexus 470-ESP, USA) at room temperature.…”

Butylglyceryl Pectin Nanoparticles: Synthesis, Formulation and Characterization

“…In addition to their low cost, biocompatibility and biodegradability, the ease of chemical modification with various conjugates make polysaccharides very promising materials for the design of various carriers for drug delivery applications [1,2,3]. Among the common types of chemical modifications applied to polysaccharides, the introduction of alkyl groups leads to the adjustment of the hydrophilic lipophilic balance of the macromolecule, which is being used in turn for tuning its interactions with living cells and ultimately for controlling its bioactivity.…”