Nitric Oxide-Releasing Porous Coating with Antibacterial Activity and Blood Compatibility.

Nitric oxide (NO)-releasing coating is promising to enhance the biocompatibility of medical devices. In this study, polyurethane (PU) and S-nitrosated keratin (KSNO) were dissolved with dimethyl sulfoxide (DMSO) and tetrahydrofuran (THF) to prepare a coating solution. This solution is facile to form a porous coating on various substrates based on solvent-evaporation-induced phase separation (SEIPS). The coating could continuously release NO up to 200 h in the presence of ascorbic acid (Asc). In addition, the coating could accelerate endothelialization by promoting the viability of human umbilical vein endothelial cells (HUVECs) while inhibiting the proliferation of human umbilical artery smooth muscle cells (HUASMCs). Furthermore, the coating had good antibacterial activity and blood compatibility. Taken together, this universal coating provides wider potential applications in the field of cardiovascular implants.

Endothelium-Mimicking Bilayer Vascular Grafts with Dual-Releasing of NO/H2S for Anti-Inflammation and Anticalcification.

Vascular complications caused by diabetes impair the activities of endothelial nitric oxide synthase (eNOS) and cystathionine γ-lyase (CSE), resulting in decreased physiological levels of nitric oxide (NO) and hydrogen sulfide (H2S). The low bioavailability of NO and H2S hinders the endothelialization of vascular grafts. In this study, endothelium-mimicking bilayer vascular grafts were designed with spatiotemporally controlled dual releases of NO and H2S for in situ endothelialization and angiogenesis. Keratin-based H2S donor was synthesized and electrospun with poly(l-lactide-co-ε-caprolactone) (PLCL) as the outer layer of the graft to release H2S. Hyaluronic acid, one of the major glycosaminoglycans in endothelial glycocalyx, was complexed with Cu ions as the inner layer to mimic glutathione peroxidase (GPx) and maintain long-term physiological NO flux. The synergistic effects of NO and H2S of bilayer grafts selectively promoted the regeneration and migration of human umbilical vascular endothelial cells (HUVECs), while inhibiting the overproliferation of human umbilical artery smooth muscle cells (HUASMCs). Bilayer grafts could effectively prevent vascular calcification, reduce inflammation, and alleviate endothelial dysfunction. The in vivo study in a rat abdominal aorta replacement model for 1 month showed that the graft had a good patency rate and had potential for vascular remodeling in situ.

Recent advances in keratin for biomedical applications.

The development of keratin-based biomaterials provides an approach to addressing related environmental pollutants and turns waste into wealth. Keratin possesses various merits, such as biocompatibility, biodegradability, hemostasis, non-immunogenicity, antibacterial activity, antioxidation, multi-responsiveness, and abundance in nature. Additionally, keratin biomaterials have been extensively employed in various biomedical applications such as drug delivery, wound healing, and tissue engineering. This review focuses on the properties and biomedical applications of keratin biomaterials. It is anticipated to provide valuable insights for the research and development of keratin biomaterials.

Hydrogen sulfide releasing poly(γ-glutamic acid) biocomposite hydrogel with monitoring, antioxidant, and antibacterial properties for diabetic wound healing.

Adverse factors such as high levels of glucose, oxidative stress, inflammation, and bacterial infection impede diabetic wound healing and even worsen wounds. Owing to its outstanding anti-inflammatory and antioxidant properties as well as the potential to promote cell migration and proliferation, hydrogen sulfide(H2S) gas therapy is promising for chronic diabetic wound recovery. In this work, a multifunctional poly(γ-glutamic acid)(PGA) hydrogel encapsulated with keratin-based H2S donor(KTC), ciprofloxacin(Cip), and anthocyanins(Ant) was developed. The resultant hydrogel was capable of releasing H2S, thereby promoting cell proliferation and enhancing anti-inflammation and antioxidant activity. The release of antibiotic Cip was accelerated under a diabetic wound microenvironment, thereby enhancing the antibacterial activity of the hydrogel. The encapsulated Ant could serve as a pH monitor, sensitively indicating wound pH conditions in situ and indirectly reflecting wound infection. In vivo results in diabetic wound healing suggested that PGA/Ant/KTC/Cip hydrogel reduced inflammation and promoted angiogenesis and collagen deposition, thereby accelerating wound healing.

Chlorhexidine-loaded polysulfobetaine/keratin hydrogels with antioxidant and antibacterial activity for infected wound healing.

Multifunctional hydrogel dressings are promising for wound healing. In the study, chlorhexidine(CHX) loaded double network hydrogels were prepared by free radical polymerization of sulfobetaine and oxidative self-crosslinking of reduced keratin. The introduced keratin and CHX endowed hydrogels with cytocompatibility, antioxidant capability as well as enhanced antibacterial activity due to the antifouling property of polysulfobetaine. These hydrogels exhibited acidity, glutathione(GSH), and trypsin triple-responsive release behaviors, resulting in the accelerated release of CHX under wound microenvironments. Intriguingly, the freeze-drying hydrogels could be ground to powders and sprinkled on the irregular wound bed, followed by absorbing wound fluid to reform hydrogel in situ. These xerogel powders were more convenient for sterilization, formulation, and storage. Further, these xerogel powders could be rejected after being mixed with an appropriate amount of water. In vivo infected wound healing confirmed that the xerogel powder dressing significantly promoted collagen deposition and reduced inflammation, thereby accelerating the closure and regeneration of skin wounds. Taken together, these degradable xerogel powders have great potential applications for wound healing.

Multiresponsive Keratin-Polysulfobetaine Conjugate-Based Micelles as Drug Carriers with a Prolonged Circulation Time.

A protein-polymer conjugate combines the chemical properties of a synthetic polymer chain with the biological properties of a protein. In this study, the initiator terminated with furan-protected maleimide was first synthesized through three steps. Then, a series of zwitterionic poly[3-dimethyl(methacryloyloxyethyl)ammonium propanesulfonate] (PDMAPS) was synthesized via atom transfer radical polymerization (ATRP) and optimized. Subsequently, well-controlled PDMAPS was conjugated with keratin via thiol-maleimide Michael addition. The keratin-PDMAPS conjugate (KP) could self-assemble in an aqueous solution to form micelles with low critical micelle concentration (CMC) values and good blood compatibility. The drug-loaded micelles exhibited triple responsiveness to pH, glutathione (GSH), and trypsin under tumor microenvironments. In addition, these micelles showed high toxicity against A549 cells while low toxicity on normal cells. Furthermore, these micelles performed prolonged blood circulation.

Bilayer dressing based on aerogel/electrospun mats with self-catalytic hydrogen sulfide generation and enhanced antioxidant ability.

Hydrogen sulfide (H2S) releasing wound dressings have attracted much attention for their ability to promote cell proliferation, stimulate angiogenesis, and resist inflammation. Mimicking the skin structure, a bilayer wound dressing based on aerogel/mats with H2S release capability was designed and fabricated. A bio-macromolecular H2S donor based on a keratin-TA conjugate (KTC) was first synthesized through a thiol-disulfide exchange reaction. As an inner layer, KTC was then loaded into a gelatin hydrogel with large pores to absorb the wound exudates and generate H2S self-catalytically. Subsequently, polyurethane was electrospun with glutathione (GSH) to be used as an outer layer with small pores, which provided mechanical support, supplied GSH, and prevented bacterial invasion. The bilayer dressing was capable of generating H2S self-catalytically, achieving a controlled and sustained release. The dressing could also promote cell proliferation and migration. In addition, the dress possessed enhanced antioxidant ability and reactive oxygen species (ROS) scavenging capability. The bilayer dressing on promoting wound healing was investigated in a full-thickness excisional cutaneous wound model in rats. The results demonstrated that it could reduce inflammation, promote vascularization, and facilitate hair follicle regeneration, thereby accelerating wound healing. Overall, the bilayer dressing has great potential applications in the field of the wound dressing.

Tannic Acid-Assisted Immobilization of Copper(II), Carboxybetaine, and Argatroban on Poly(ethylene terephthalate) Mats for Synergistic Improvement of Blood Compatibility and Endothelialization.

Due to thrombosis and intimal hyperplasia, small-diameter vascular grafts have poor long-term patency. A combination strategy based on nitric oxide (NO) and anticoagulants has the potential to address those issues. In this study, poly(ethylene terephthalate) (PET) mats were prepared by electrospinning and coated with tannic acid (TA)/copper ion complexes. The chelated copper ions endowed the mats with sustained NO generation by catalytic decomposition of endogenous S-nitrosothiol. Subsequently, zwitterionic carboxybetaine acrylate (CBA) and argatroban (AG) were immobilized on the mats. The introduced AG and CBA had synergistic effects on the improvement of blood compatibility, resulting in reduced platelet adhesion and prolonged blood clotting time. The biocomposite mats selectively promoted the proliferation and migration of human umbilical vein endothelial cells while inhibiting the proliferation and migration of human umbilical arterial smooth muscle cells under physiological conditions. In addition, the prepared mats exhibited antibacterial activity against Escherichia coli and Staphylococcus aureus. Collectively, the prepared mats hold great promise as artificial small-diameter vascular grafts.

Preparation of MSNs@Keratin as pH/GSH dual responsive drug delivery system.

Designing a drug delivery system that is responsive in a tumor microenvironment is important to potentiate the efficacy and reduce the side effects of antitumor drugs. In this study, the surface of mesoporous silica nanoparticles (MSNs) were aminated with 3-aminopropyl triethoxysilane (APTES) and then coupled with keratin, as a gatekeeper, to afford MSNs-NH2@Keratin. The average sizes and morphologies of MSNs and MSNs-NH2@Keratin were characterized with dynamic light scattering and transmission electron microscopy, respectively. The loading content and encapsulation efficiency of doxorubicin (DOX) were calculated to be 17.1 ± 1.7% and 71.3 ± 2.1%. Drug-loaded MSNs-NH2@Keratin exhibited pH and glutathione (GSH) dual responsiveness under tumor microenvironment. The nanoparticles could be uptaken by tumor cells to effectively inhibit tumor cell growth. Moreover, the sizes of nanoparticle were stable in the serum. Collectively, our findings demonstrated the potential of DOX-loaded MSNs-NH2@Keratin in the treatment of cancer.