Injectable zein gel with in situ self-assembly as hemostatic material.

Zein is a biocompatible and biodegradable corn protein with promising properties for biomedical applications. It is hydrophobic with the ability to self-assemble in an aqueous medium. It can also form a gel in hydroalcoholic solvents at higher concentrations. Few studies have investigated the biomedical significance of zein gels. Herein, we exploited the injectability and water-responsive increase in stiffness of zein gel to achieve hemostasis by physical blockage of the wound and clot formation. The release of components from the gel further aided blood clotting and gave a higher clot strength than a natural clot, which can prevent rebleeding. Rabbit aortic injury and swine femoral artery injury models were used to evaluate the hemostatic efficacy of the zein gel. Zein gel was effective in both hemostatic models without applying external compression due to an in situ increase in stiffness, while the control (Celox™ Gauze) required external compression at the wound site. The zein gel was easily removed after hemostasis due to hydrophobic self-assembly. Overall, zein gel is proposed as an effective hemostatic product for any wound shape owing to its good shape adaptability and rapid in situ blood-responsive stiffness increase.

Improvement of mechanical properties of zein porous scaffold by quenching/electrospun fiber reinforcement.

As a novel bone substitute material, zein-based scaffolds (ZS) should have suitable mechanical properties and porosity. ZS has shown good compressive properties matching cancellous bone, but there is still a demand to improve its mechanical properties, especially tensile and bending properties without adding plasticizers. The present study explored two simple and environment-friendly factors for this purpose: fiber reinforcement and quenching. Addition of electrospun zein fibers enhanced all mechanical properties significantly including compressive, tensile, and bending moduli; compressive and bending strengths of ZS with both higher (70-80%) and lower (50-60%) porosities, no matter whether heating treated or not treated. Especially, all these parameters were further enhanced significantly by addition of heating treated fibers. AFM provided evidence that high temperature modification could significantly alter the micro-elastic properties of zein electrospun fibers, i.e., increased stiffness of fibers. Quenching treatment also enhanced compressive, tensile, and bending strengths significantly. Finally, quenching treated ZS were implanted into critical-sized bone defects (15 mm) of the rabbit model to compare the repair efficacy with a commercial ß-tricalcium phosphate product. The results demonstrated that there were no remarkable differences in bone reconstructions between these two materials.

Effects of the micro/nanostructure of electrospun zein fibres on cells in simulated blood flow environment.

In order to prevent thrombosis, reduce intima hyperplasia, and to maintain long-term patency after implantation of an artificial blood vessel, the formation of intact endothelial cells layer on an inner surface of graft is desirable. The present study aimed to improve endothelial cell adhesion by regulating the morphology of the inner surface of artificial blood vessels. Zein fibre membranes with three fibre diameters (small, ~100 nm; medium, ~500 nm; and large, ~1000 nm) were constructed by electrospinning. A flow chamber device was designed to simulate the blood flow environment. The morphology and adhesion of human umbilical vein fusion cells (EA.hy926) on the surface of the fibre membranes were studied under a shear stress of approximately 15 dynes/cm2. The results showed that oriented electrospun zein fibre surfaces with both medium- and large-diameter fibres can regulate the morphology of endothelial cells (EA.hy926), which are aligned by the fibre direction. The three fibre membranes improved the adhesion of endothelial cells significantly compared to that on the flat membrane. When the fibre direction was fixed parallel to the fluid direction, the medium-diameter oriented-fibre membrane could significantly improve the ability endothelial cells to resist shear stress, and there was a significant difference at 1, 2 and 4 h time points compared with the shear stress resistance on the small-diameter and large-diameter oriented-fibre membranes. When the fibre direction was perpendicular to the fluid direction, again the medium-diameter oriented-fibre membrane improved the ability of endothelial cells to resist shear stress significantly at 1 and 2 h time points. It was concluded that by changing the diameter and arrangement of electrospun fibres, cell morphology control and shear stress resistance can be achieved.

Enhancement of mechanical strength of TCP-alginate based bioprinted constructs.

To overcome the mechanical drawback of bioink, we proposed a supporter model to enhance the mechanical strength of bioprinted 3D constructs, in which a unit-assembly idea was involved. Based on Computed Tomography images of critical-sized rabbit bone defect, the 3D re-construction was accomplished by a sequenced process using Mimics 17.0, BioCAM and BioCAD software. 3D constructs were bioprinted using polycaprolactone (PCL) ink for the outer supporter under extrusion mode, and cell-laden tricalcium phosphate (TCP)/alginate bioink for the inner filler under air pressure dispensing mode. The relationship of viscosity of bioinks, 3D bioprinting pressure, TCP/alginate ratio and cell survival were investigated by the shear viscosities analysis, live/dead cell test and cell-counting kit 8 measurement. The viscosity of bioinks at 1.0 s-1-shear rate could be adjusted within the range of 1.75 ±â€¯0.29 Pa·s to 155.65 ±â€¯10.86 Pa·s by changing alginate concentration, corresponding to 10 kPa-130 kPa of printing pressure. This design with PCL supporter could significantly enhance the compressive strength and compressive modulus of standardized 3D mechanical testing specimens up to 2.15 ±â€¯0.14 MPa to 2.58 ±â€¯0.09 MPa, and 42.83 ±â€¯4.75 MPa to 53.12 ±â€¯1.19 MPa, respectively. Cells could maintain the high viability (over 80%) under the given printing pressure but cell viability declined with the increase of TCP content. Cell survival after experiencing 7 days of cell culture could be achieved when the ratio of TCP/alginate was 1 : 4. All data supported the feasibility of the supporter and unit-assembly model to enhance mechanical properties of bioprinted 3D constructs.

Formulation of zein based compression coated floating tablets for enhanced gastric retention and tunable drug release.

The present study aimed to investigate the potential of zein (a protein obtained from corn) for development of gastroretentive floating tablets for the first time. A compression coated tablet design with outer floating layer and inner drug containing layer was followed to achieve floating over gastric fluid with sustained release of drug. Captopril was used as a model drug for this purpose. Eight formulations were developed and the influence of different components on drug release and floating behavior was evaluated. The drug in coating layer was found to be released at faster rate while sustained release behavior was observed from core layer. In vivo pharmacokinetic studies on rabbits showed significant increase in bioavailability and mean residence time (MRT). Moreover, radiographic study exhibited gastric retention of prepared tablets >12 h. In conclusion, zein can be used for development of gastroretentive floating tablets and by adjusting amount of different formulation factors, desired drug release rate can be achieved.