Glucose cryoprotectant affects glutathione-responsive antitumor drug release from polysaccharide nanoparticles.
The aim of this work was to prepare polysaccharide-based nanoparticles (NPs) sensitive to glutathione (GSH), and to elucidate the effect of the concentration of glucose used as cryoprotectant during freeze-drying on the GSH-responsiveness. NPs were obtained via ionic interaction between negatively charged polysaccharides, chondroitin sulfate and dermatan sulfate, and the positively charged thiolated chitosan (CSSH), and crosslinking of CSSH before or after the nanoparticles formation with a disulfide-bond containing crosslinker, N,N'-bis(acryloyl)cystamine (BAC). NPs were freeze-dried with glucose at two different concentrations (0.5 and 5.0%w/w) and then characterized as methotrexate delivery systems, studying the effect of GSH concentration on drug release, efficacy against tumor cells and cellular internalization. Non-loaded NPs were highly compatible with murine fibroblasts and showed a suitable size for being used in anticancer therapy. When methotrexate-loaded NPs were freeze-dried with the highest glucose concentration, they lost their responsiveness to GSH concentration in vitro. Drug-loaded NPs were shown to inhibit the growth of tumor cells (HeLa and CHO-K1) with greater efficiency than free methotrexate, disregarding the concentration of glucose used for freeze-drying. Nevertheless, confocal microscopy studies revealed that cellular internalization of NPs freeze-dried with 5.0% glucose is more difficult than for NPs freeze-dried with lower glucose concentration. Thus, concentration of glucose cryoprotectant should be taken into account during development of NPs intended to release the drug as a function of GSH levels, due to the specific interactions of glucose with GSH.
University of Calabria, Department of Pharmaceutical Sciences, Edificio Polifunzionale, Arcavacata di Rende (CS) 87036, Italy; Universidad de Santiago de Compostela, Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Farmacia, 15782 Santiago de Compostela, Spain. Electronic address: manuela.curcio@unical.it.
Manuela
M
Curcio
Universidad de Santiago de Compostela, Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Farmacia, 15782 Santiago de Compostela, Spain.
Bárbara
B
Blanco-Fernández
Universidad de Santiago de Compostela, Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Farmacia, 15782 Santiago de Compostela, Spain.
Alejandro
A
Costoya
Universidad de Santiago de Compostela, Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Farmacia, 15782 Santiago de Compostela, Spain.
Angel
A
Concheiro
University of Calabria, Department of Pharmaceutical Sciences, Edificio Polifunzionale, Arcavacata di Rende (CS) 87036, Italy.
Francesco
F
Puoci
Universidad de Santiago de Compostela, Departamento de Farmacia y Tecnología Farmacéutica, Facultad de Farmacia, 15782 Santiago de Compostela, Spain. Electronic address: carmen.alvarez.lorenzo@usc.es.
Carmen
C
Alvarez-Lorenzo
D000818:Animals; D000964:Antimetabolites, Antineoplastic / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D041702:BALB 3T3 Cells; D016466:CHO Cells; D002470:Cell Survival / Q000187:drug effects; D002626:Chemistry, Pharmaceutical; D048271:Chitosan / Q000031:analogs & derivatives / Q000737:chemistry; D002809:Chondroitin Sulfates / Q000737:chemistry; D003412:Cricetulus; D003432:Cross-Linking Reagents / Q000737:chemistry; D003451:Cryoprotective Agents / Q000737:chemistry / Q000494:pharmacology*; D003871:Dermatan Sulfate / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D004337:Drug Carriers; D005260:Female; D005612:Freeze Drying; D005947:Glucose / Q000737:chemistry / Q000494:pharmacology*; D005978:Glutathione / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008727:Methotrexate / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology*; D051379:Mice; D018613:Microscopy, Confocal; D050997:Nanomedicine; D053758:Nanoparticles; D011109:Polymethacrylic Acids / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry*; D012995:Solubility; D013678:Technology, Pharmaceutical / Q000379:methods; D002583:Uterine Cervical Neoplasms / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology
D000964:Antimetabolites, Antineoplastic; D003432:Cross-Linking Reagents; D003451:Cryoprotective Agents; D004337:Drug Carriers; D011109:Polymethacrylic Acids; D011134:Polysaccharides; D003871:Dermatan Sulfate; D002809:Chondroitin Sulfates; D048271:Chitosan; D005978:Glutathione; D005947:Glucose; D008727:Methotrexate
Cell interaction; Chitosan; Chondroitin sulfate; Dermatan sulfate; Glucose cryoprotectant; Glutathione-responsiveness; Methotrexate