Helan Xu

Novel 3D Electrospun Scaffolds with Fibers Oriented Randomly and Evenly in Three Dimensions to Closely Mimic the Unique Architectures of Extracellular Matrices in Soft Tissues: Fabrication and Mechanism Study

In this work, novel electrospun scaffolds with fibers oriented randomly and evenly in three dimensions (3D) including in the thickness direction were developed based on the principle of electrostatic repulsion. This unique structure is different from most electrospun scaffolds with fibers oriented mainly in one direction. The structure of novel 3D scaffolds could more closely mimic the 3D randomly oriented fibrous architectures in many native extracellular matrices (ECMs). The cell culture results of this study indicated that, instead of becoming flattened cells when cultured in conventional electrospun scaffolds, the cells cultured on novel 3D scaffolds could develop into stereoscopic topographies, which highly simulated in vivo 3D cellular morphologies and are believed to be of vital importance for cells to function and differentiate appropriately. Also, due to the randomly oriented fibrous structure, improvement of nearly 5 times in cell proliferation could be observed when comparing our 3D scaffolds with 2D counterparts after 7 days of cell culture, while most currently reported 3D scaffolds only showed 1.5- to 2.5-fold improvement for the similar comparison. One mechanism of this fabrication process has also been proposed and showed that the rapid delivery of electrons on the fibers was the crucial factor for formation of 3D architectures.

Hollow nanoparticles from zein for potential medical applications

Hollow nanoparticles from corn storage protein zein, with average diameters as small as 65 nm and capable of loading a large amount of drug and penetrating into the cell cytoplasm, have been developed for potential drug delivery applications. As an important protein co-product of corn-based ethanol, zein is biocompatible and has been proved to be useful for medical applications through in vitro and in vivo evaluations. Zein can overcome the limitations of inorganic or metal nanoparticles that tend to accumulate in the organs and tissues and is therefore preferable for drug delivery applications. However, it has been observed that only small proteins and peptides are able to penetrate into cells and zein with a molecular weight of 14–44 kDa may not be able to enter into the cells. In this research, hollow zein nanoparticles have been developed and the potential of the hollow zein nanoparticles to load drugs and enter the cell cytoplasm was investigated. Hollow zein nanoparticles developed in this research were capable of loading as high as 369 mg g−1 of the drug metformin at an equilibrium concentration of 3 g L−1. Metformin in hollow zein nanoparticles showed a more sustained and controlled release profile than that in solid zein nanoparticles. Hollow zein nanoparticles were found to be able to enter the fibroblast cells 1 hour after incubation. The biocompatibility, nano-scale diameters, potential for loading a large amount of drugs and the ability to penetrate into cells make hollow zein nanoparticles ideal candidates for carrying various payloads for intracellular drug delivery.

Water-Stable Three-Dimensional Ultrafine Fibrous Scaffolds from Keratin for Cartilage Tissue Engineering

Intrinsically water-stable scaffolds composed of ultrafine keratin fibers oriented randomly and evenly in three dimensions were electrospun for cartilage tissue engineering. Keratin has been recognized as a biomaterial that could substantially support the growth and development of multiple cell lines. Besides, three-dimensional (3D) ultrafine fibrous structures were preferred in tissue engineering due to their structural similarity to native extracellular matrices in soft tissues. Recently, we have developed a nontraditional approach to developing 3D fibrous scaffolds from alcohol-soluble corn protein, zein, and verified their structural advantages in tissue engineering. However, keratin with highly cross-linked molecular structures could not be readily dissolved in common solvents for fiber spinning, which required the remarkable drawability of solution. So far, 3D fibrous scaffolds from pure keratin for biomedical applications have not been reported. In this research, the highly cross-linked keratin from chicken feathers was de-cross-linked and disentangled into linear and aligned molecules with preserved molecular weights, forming highly stretchable spinning dope. The solution was readily electrospun into scaffolds with ultrafine keratin fibers oriented randomly in three dimensions. Due to the highly cross-linked molecular structures, keratin scaffolds showed intrinsic water stability. Adipose-derived mesenchymal stem cells could penetrate much deeper, proliferate, and chondrogenically differentiate remarkably better on the 3D keratin scaffolds than on 2D PLA fibrous scaffolds, 3D soy protein fibrous scaffolds, or 3D commercial nonfibrous scaffolds. In summary, the electrospun 3D ultrafine fibrous scaffolds from keratin could be promising candidates for cartilage tissue engineering.

Biodegradable hollow zein nanoparticles for removal of reactive dyes from wastewater

In this study, biodegradable hollow zein nanoparticles with diameters less than 100 nm were developed to remove reactive dyes from simulated post-dyeing wastewater with remarkably high efficiency. Reactive dyes are widely used to color cellulosic materials, such as cotton and rayon. Wastewater from reactive dyeing process contains up to 50% dye and electrolytes with concentrations up to 100 g L(-1). Current methods to remove reactive dyes from wastewater are suffering from low adsorption capacities or low biodegradability of the sorbents. In this research, biodegradable zein nanoparticles showed high adsorption capacities for dyes. Hollow zein nanoparticles showed higher adsorption for Reactive Blue 19 than solid structures, and the adsorption amount increased as temperature decreased, pH decreased or initial dye concentration increased. At pH 6.5 and pH 9.0, increasing electrolyte concentration could improve dye adsorption significantly. Under simulated post-dyeing condition with 50.0 g L(-1) salt and pH 9.0, maximum adsorption of 1016.0 mg dye per gram zein nanoparticles could be obtained. The adsorption capacity was much higher than that of various biodegradable adsorbents developed to remove reactive dye. It is suggested that the hollow zein nanoparticles are good candidates to remove reactive dye immediately after dyeing process.

Intrinsically water-stable electrospun three-dimensional ultrafine fibrous soy protein scaffolds for soft tissue engineering using adipose derived mesenchymal stem cells

Soy protein, the plant protein from soybean, was electrospun into intrinsically water-stable scaffolds with large volume and ultrafine fibers oriented randomly and evenly in three dimensions (3D) to simulate native extracellular matrices of soft tissues. The 3D ultrafine fibrous scaffolds from proteins could be favored in soft tissue engineering. However, protein-based biomaterials usually suffered from poor water stability, while the highly crosslinked proteins which had water stability were usually difficult to be fabricated into fibers. Soy protein was a typical protein with intrinsic water stability, attributed to its 1.2% cysteine content. Soy protein has been developed into 3D non-fibrous structures, coarse fibers and films for tissue engineering applications, but not ultrafine fibrous structures. In this research, the disulfide crosslinks in soy protein were cleaved to facilitate its dissolution in an aqueous solvent system. The obtained solution was electrospun into bulky scaffolds composed of ultrafine fibers oriented randomly in three dimensions. Without external crosslinking, the fibrous soy protein scaffolds demonstrated long-term water stability, and maintained their fibrous structures after incubated in PBS for up to 28 days. In vitro study showed that the 3D soy protein scaffolds well supported uniform distribution and adipogenic differentiation of adipose derived mesenchymal stem cells. In summary, the 3D ultrafine fibrous soy protein structures could be good candidates as scaffolds in soft tissue engineering.

Intrinsically water-stable keratin nanoparticles and their in vivo biodistribution for targeted delivery.

Highly water-stable nanoparticles of around 70 nm and capable of distributing with high uptake in certain organs of mice were developed from feather keratin. Nanoparticles could provide novel veterinary diagnostics and therapeutics to boost efficiency in identification and treatment of livestock diseases to improve protein supply and ensure safety and quality of food. Nanoparticles could penetrate easily into cells and small capillaries, surpass detection of the immune system, and reach targeted organs because of their nanoscale sizes. Proteins with positive and negative charges and hydrophobic domains enable loading of various types of drugs and, hence, are advantageous over synthetic polymers and carbohydrates for drug delivery. In this research, the highly cross-linked keratin was processed into nanoparticles with diameters of 70 nm under mild conditions. Keratin nanoparticles were found supportive to cell growth via an in vitro study and highly stable after stored in physiological environments for up to 7 days. At 4 days after injection, up to 18% of the cells in kidneys and 4% of the cells in liver of mice were penetrated by the keratin nanoparticles.

Cellulose nanocrystal-reinforced keratin bioadsorbent for effective removal of dyes from aqueous solution

High-efficiency and recyclable three-dimensional bioadsorbents were prepared by incorporating cellulose nanocrystal (CNC) as reinforcements in keratin sponge matrix to remove dyes from aqueous solution. Adsorption performance of dyes by CNC-reinforced keratin bioadsorbent was improved significantly as a result of adding CNC as filler. Batch adsorption results showed that the adsorption capacities for Reactive Black 5 and Direct Red 80 by the bioadsorbent were 1201 and 1070mgg-1, respectively. The isotherms and kinetics for adsorption of both dyes on bioadsorbent followed the Langmuir isotherm model and pseudo-second order model, respectively. Desorption and regeneration experiments showed that the removal efficiencies of the bioadsorbent for both dyes could remain above 80% at the fifth recycling cycles. Moreover, the bioadsorbent possessed excellent packed-bed column operation performance. Those results suggested that the adsorbent could be considered as a high-performance and promising candidate for dye wastewater treatment.

Electrospun ultrafine fibrous wheat glutenin scaffolds with three-dimensionally random organization and water stability for soft tissue engineering

Wheat glutenin, the highly crosslinked protein from wheat, was electrospun into scaffolds with ultrafine fibers oriented randomly and evenly in three dimensions to simulate native extracellular matrices of soft tissues. The scaffolds were intrinsically water-stable without using any external crosslinkers and could support proliferation and differentiation of adipose-derived mesenchymal stem cells for soft tissue engineering. Regeneration of soft tissue favored water-stable fibrous protein scaffolds with three-dimensional arrangement and large volumes, which could be difficult to obtain via electrospinning. Wheat glutenin is an intrinsically water-stable protein due to the 2% cysteine in its amino acid composition. In this research, the disulfide crosslinks in wheat glutenin were cleaved while the backbones were preserved. The treated wheat glutenin was dissolved in aqueous solvent with an anionic surfactant and then electrospun into bulky scaffolds composed of ultrafine fibers oriented randomly in three dimensions. The scaffolds could maintain their fibrous structures after incubated in PBS for up to 35 days. In vitro study indicated that the three-dimensional wheat glutenin scaffolds well supported uniform distribution and adipogenic differentiation of adipose derived mesenchymal stem cells.

Cytocompatible and water-stable camelina protein films for tissue engineering

In this research, films with compressive strength and aqueous stability were developed from camelina protein (CP) for tissue engineering. Protein based scaffolds have poor mechanical properties and aqueous stability and generally require chemical or physical modifications to make them applicable for medical applications. However, these modifications such as crosslinking could reduce biocompatibility and/or degradability of the scaffolds. Using proteins that are inherently water-stable could avoid modifications and provide scaffolds with the desired properties. CP with a high degree of disulfide cross-linkage has the potential to provide water-stable biomaterials, but it is difficult to dissolve CP and develop scaffolds. In this study, a new method of dissolving highly cross-linked proteins that results in limited hydrolysis and preserves the protein backbone was developed to produce water-stable films from CP without any modification. Only 12 % weight loss of camelina films was observed after 7 days in phosphate buffer saline (PBS) at 37°C. NIH 3T3 fibroblasts could attach and proliferate better on camelina films than on citric acid cross-linked collagen films. Therefore, CP films have the potential to be used for tissue engineering, and this extraction-dissolution method can be used for developing biomedical materials from various water-stable plant proteins.

A Sustainable Slashing Industry Using Biodegradable Sizes from Modified Soy Protein To Replace Petro-Based Poly(Vinyl Alcohol)

Biodegradable sizing agents from triethanolamine (TEA) modified soy protein could substitute poly(vinyl alcohol)(PVA) sizes for high-speed weaving of polyester and polyester/cotton yarns to substantially decrease environmental pollution and impel sustainability of textile industry. Nonbiodegradable PVA sizes are widely used and mainly contribute to high chemical oxygen demand (COD) in textile effluents. It has not been possible to effectively degrade, reuse or replace PVA sizes so far. Soy protein with good biodegradability showed potential as warp sizes in our previous studies. However, soy protein sizes lacked film flexibility and adhesion for required high-speed weaving. Additives with multiple hydroxyl groups, nonlinear molecule, and electric charge could physically modify secondary structure of soy protein and lead to about 23.6% and 43.3% improvement in size adhesion and ability of hair coverage comparing to unmodified soy protein. Industrial weaving results showed TEA-soy protein had relative weaving efficiency 3% and 10% higher than PVA and chemically modified starch sizes on polyester/cotton fabrics, and had relative weaving efficiency similar to PVA on polyester fabrics, although with 3- 6% lower add-on. In addition, TEA-soy sizes had a BOD5/COD ratio of 0.44, much higher than 0.03 for PVA, indicating that TEA-soy sizes were easily biodegradable in activated sludge.