bromochloroacetic-acid and Hemolysis

Rapid endothelialization and regulation of smooth muscle cell proliferation are crucial for small-diameter vascular grafts to address poor compliance, thromboembolism, and intimal hyperplasia, and achieve revascularization. As a gaseous signaling molecule, nitric oxide (NO) regulates cardiovascular homeostasis, inhibits blood clotting and intimal hyperplasia, and promotes the growth of endothelial cells. Due to the instability and burst release of small molecular NO donors, a novel biomacromolecular donor has generated increasing interest. In the study, a low toxic NO donor of S-nitrosated keratin (KSNO) was first synthesized and then coelectrospun with poly(ε-caprolactone) to afford NO-releasing small-diameter vascular graft. PCL/KSNO graft was capable to generate NO under the catalysis of ascorbic acid (Asc), so the graft selectively elevated adhesion and growth of human umbilical vein endothelial cells (HUVECs), while inhibited the proliferation of human aortic smooth muscle cells (HASMCs) in the presence of Asc. In addition, the graft displayed significant antibacterial properties and good blood compatibility. Animal experiments showed that the biocomposite graft could inhibit thrombus formation and preserve normal blood flow via single rabbit carotid artery replacement for 1 month. More importantly, a complete endothelium was observed on the lumen surface. Taken together, PCL/KSNO small-diameter vascular graft has potential applications in vascular tissue engineering with rapid endothelialization and vascular remolding.

Topics: Animals; Aorta; Biocompatible Materials; Blood Vessel Prosthesis; Cell Adhesion; Cell Death; Cell Line; Cell Proliferation; Cell Survival; Escherichia coli; Hemolysis; Human Umbilical Vein Endothelial Cells; Humans; Keratins; Mice; Microbial Sensitivity Tests; Myocytes, Smooth Muscle; Nitric Oxide; Nitrosation; Platelet Adhesiveness; Polyesters; Rabbits; Spectroscopy, Fourier Transform Infrared; Tissue Scaffolds

Development of pH/GSH/enzyme triple stimuli-responsive drug delivery system is promising for tumor therapy due to more acidic, higher glutathione (GSH) level, and overexpressed trypsin under tumor microenvironment. Herein, keratin/doxorubicin (DOX) complex nanoparticles (KDNPs) were for the first time prepared using a drug-induced ionic gelation technique without cross-linker, organic solvent and surfactant. The resultant KDNPs had high drug loading efficacy and performed considerably stable in aqueous solution. Drug delivery curves showed that KDNPs exhibited triple-responsive characters (pH, GSH, and enzyme). Under tumor microenvironments (acid and high GSH level), KDNPs performed surface charge conversion of negative-to-positive and enhanced permeation retention effect (EPR), which both benefited the drug accumulation. Furthermore, the overexpressed trypsin would cleave the peptide bonds within KDNPs and enhance the DOX release. KDNPs were demonstrated to be internalized by A549 cells through endocytosis by cellular uptake assay. Cytotoxicity assay indicated that KDNPs could inhibit the proliferation of tumor cells efficiently. In vivo cytotoxicity and hemolysis tests suggested that KDNPs exhibited excellent biocompatibility as well as good blood compatibility. In vivo antitumor efficacy demonstrated that KDNPs had a strong antitumor effect similar to that of free DOX, but with nearly no side effects. Intriguingly, KDNPs were able to catalyze endogenous NO donor in blood to release NO in tumor tissue, resulting in the prolonged blood circulation time and improved therapeutic activity of drug. In conclusion, keratin-based drug carriers are potential for cancer therapy in clinical medicine.

Topics: A549 Cells; Animals; Antineoplastic Agents; Cell Death; Doxorubicin; Drug Carriers; Drug Delivery Systems; Drug Liberation; Glutathione; Hemolysis; Humans; Hydrogen-Ion Concentration; Keratins; Kinetics; Mice, Nude; Nanoparticles; Nitric Oxide; Particle Size; Rabbits; Spectrophotometry, Ultraviolet; Static Electricity; Trypsin

Smart drug carriers are the current need of the hour in controlled drug delivery applications. In this work, pH and redox dual responsive keratin based drug-loaded nanoparticles (KDNPs) were fabricated through two-step strategies. Keratin nanoparticles were first prepared by desolvation method and chemical crosslinking, followed by electrostatic adsorbing doxorubicin (DOX) to afford drug loaded keratin nanoparticles (KDNPs). The size, size distribution, and morphology of the KDNPs were characterized with dynamic light scattering (DLS) and Scan electronic microscope (SEM). Drug delivery profiles showed that KDNPs exhibited pH and glutathione (GSH) dual-responsive characters. Under tumor tissue/cell microenvironments (more acidic and high GSH level), KDNPs tended to accumulate at the tumor region through a potential enhanced permeability and retention (EPR) effect and perform surface negative-to-positive charge conversion. Hemolysis assay indicated that KDNPs had good blood compatibility. Cellular uptake assay demonstrated that KDNPs could be internalized by A 549 cells through endocytosis. Intriguingly, KDNPs were capable of promoting nitric oxide (NO) release from endogenous donor of S-nitrosoglutathione in the presence of GSH. All of these results demonstrated that keratin based drug carriers had potential for drug/NO delivery and cancer therapy in clinical medicine.

Topics: A549 Cells; Animals; Cell Death; Cell Survival; Doxorubicin; Drug Delivery Systems; Drug Liberation; Dynamic Light Scattering; Endocytosis; Glutathione; Hemolysis; Humans; Hydrogen-Ion Concentration; Keratins; Kinetics; Mice; Nanoparticles; NIH 3T3 Cells; Nitric Oxide; Particle Size; Rabbits; Static Electricity

Effect of pyrrolidone derivatives on lipid membrane and protein conformation has been assessed to obtain fundamental information about a mechanism of transdermal penetration enhancer. Their effects on a release of 6-carboxyfluorescein from liposome were examined. The lipophilic derivatives enhanced a release of dye and the enhancing effect showed a concentration dependency. Especially 1-lauryl-2-pyrrolidone showed the highest effect at the lowest concentration. The pyrrolidone derivatives also increased a hemolysis of rat erythrocytes. The derivatives slightly liberated SH group of keratin but did not change the electrophoresis pattern of keratin. 1-Methyl-2-pyrrolidone increased and retained a hydration of rat skin although 1-hexyl- and 1-lauryl-2-pyrrolidone showed no increase. These results suggest that the high enhancing effect of II, III and IX, as shown in the previous study, may be predominantly due to their interaction with skin lipid and their penetration behavior into the lipid.

Topics: Animals; Drug Carriers; Electrophoresis, Polyacrylamide Gel; Fluoresceins; Galvanic Skin Response; Hemolysis; Keratins; Liposomes; Male; Membrane Lipids; Molecular Structure; Protein Conformation; Pyrrolidinones; Rats; Rats, Inbred Strains; Skin Absorption; Sulfhydryl Compounds; Water

Topics: Animals; Blood Platelets; Dogs; Erythrocytes; Feathers; Hematocrit; Hemolysis; Infusions, Intravenous; Infusions, Parenteral; Keratins; Leukocytes; Pharmacology; Plasma Substitutes; Protein Hydrolysates; Research