Ya-Jing Ye

Silk fibroin/chitosan scaffold with tunable properties and low inflammatory response assists the differentiation of bone marrow mesenchymal stem cells

The physical and chemical properties of the scaffold are known to play important roles in three-dimensional (3D) cell culture, which always determine the cellular fate or the results of implantation. To control these properties becomes necessary for meeting the requirements of a variety of tissue engineering applications. In this study, a series of silk fibroin/chitosan (SF/CS) scaffolds with tunable properties were prepared using freeze-drying method, and the rat bone marrow-derived mesenchymal stem cells (BM-MSCs) were seeded in these scaffolds to evaluate their availability of use in tissue engineering. The 3D structure, mechanical properties and degradation ability of SF/CS scaffold can be tuned by changing the total concentration of the precursor solution and the blending ratio between SF and CS. BM-MSCs cultured in the SF/CS scaffold exhibited excellent proliferation and multiple morphologies. The induction of osteogenic and adipogenic differentiation of BM-MSCs were successful in this scaffold when cultured in vitro. Subcutaneous implantation of the SF/CS scaffolds did not cause any inflammatory response within four weeks, which revealed good compatibility. Moreover, the implanted scaffold allowed host cells to invade, adhere, grow and form new blood vessels. With these excellent performance, SF/CS scaffold has great potential in preparing implants for tissue engineering applications.

Advances in Fe-based biodegradable metallic materials

In recent years, biodegradable metallic materials, such as Mg-, Fe-, Zn- and W-based materials, have been the focus of many studies. As one of the two most studied types of biodegradable metallic materials, Fe-based materials have aroused a great deal of interest because of their outstanding mechanical properties, which are similar to stainless steel. The processing methods can directly affect the microstructure of the material and influence the mechanical and degradation properties of the material. Furthermore, biocompatibility is directly affected by the degradation properties. Therefore, the processing methods, mechanical properties, degradability and biocompatibility are several of the main concerns in the study of biodegradable Fe-based materials. Here, we systematically summarize recent studies on Fe-based materials and discuss these findings in terms of their processing methods, degradability and biocompatibility.

Interferometry measurement of protein concentration evolution during crystallization and dissolution with improved reliability and versatility

Interferometry is a widely used technique for real-time monitoring of concentration variation during processes in a solution like crystal growth and dissolution. This paper reports a protein concentration measurement system using a conventional Mach–Zehnder interferometer that allows monitoring the time course evolution of the protein concentration distribution during the processes of crystallization or dissolution. The demonstrated method showed improved reliability and versatility in that it (1) enables measurements under versatile temperature conditions and (2) eliminates the unpredictable errors caused by the environmental disturbances. The measurement system described in this paper can also be applied in measurements in other solution systems.

A protein crystallisation screening kit designed using polyethylene glycol as major precipitant

Crystallisation of proteins is usually achieved with the help of chemical agents. Because there are few general guidelines in determining what agents will help to crystallise a specific protein, suitable crystallisation agents are often found via exhaustive trial-and-error tests by mixing many chemical agents (the collection of which is called a crystallisation screening kit) one-by-one with the protein. Currently, many commercially available crystallisation screening kits have been developed and utilised in practical crystallisation screen experiments. However, information regarding the design of new screening kits has yet to be expanded using a large amount of experimental data. Here, we show the step-by-step design processes of a polyethylene glycol-based screening kit. It was found that the screening performance could be improved by modifying the crystallisation screening kits according to the accumulated data (such as those in the Biological Macromolecule Crystallisation Database (BMCD)), the screening test results and existing knowledge. The screening kit designed in this paper can be used for practical protein crystallisation screen experiments and the method can be used in the design of other crystallisation screening kits.

A new method to realize high-throughput protein crystallization in a superconducting magnet

We present a new method for the realization of high-throughput protein crystallization screening using an array of 96 capillaries aligned in a circle. In this method, each capillary represents a single crystallization condition, and all capillaries experience identical magnetic field conditions. After crystallization, the crystals in the capillary can be directly diffracted without harvesting. This method proved easy to perform and is applicable for use in magnetic fields and may be further extended for use in other circumstances, for example, under space microgravity conditions.

A novel layer-structured scaffold with large pore sizes suitable for 3D cell culture prepared by near-field electrospinning

Electrospinning is a powerful method for preparing porous materials that can be applied as biomedical materials for implantation or tissue engineering or as scaffolds for 3D cell culture experiments. However, this technique is limited in practical applications because the pore size of 3D scaffolds directly prepared by conventional electrospinning is usually less than several tens of micrometres, which may not be suitable for 3D cell culture and tissue growth. To allow for satisfactory 3D cell culture and tissue engineering, the pore size of the scaffold should be controllable according to the requirement of the specific cells to be cultured. Here, we show that layer-structured scaffolds with pore sizes larger than 100μm can be obtained by stacking meshes prepared by direct-writing using the near-field electrospinning (NFES) technique. In the study, we prepared composite scaffolds made of polycaprolactone (PCL) and hydroxyapatite (HAp) via the above-mentioned method and tested the effectiveness of the novel scaffold in cell culture using mouse pre-osteoblast cells (MC3T3-E1). The pore size and the degradability of the PCL/HAp scaffolds were characterized. The results showed that the average pore size of the scaffolds was 167μm, which was controllable based on the required application; the degradation rate was controllable depending on the ratio of PCL to HAp. The biocompatibility of the scaffolds in vitro was studied, and it was found that the scaffolds showed no toxicity and that the cells could effectively attach, proliferate, and differentiate in the 3D skeleton of the scaffolds. Our studies showed that a simple modification of the preparation procedure can lead to a new way to fabricate novel layer-structured 3D scaffolds with controllable structures and pore sizes suitable for practical applications in implantation, tissue engineering and 3D cell culture.

From 2D to 3D: The morphology, proliferation and differentiation of MC3T3-E1 on silk fibroin/chitosan matrices

It has been widely accepted that cell culture in two-dimensional (2D) conditions may not be able to represent growth in three-dimensional (3D) conditions. Systematic comparisons between 2D and 3D cell cultures are needed to appropriately use the existing 2D results. In this work, we conducted a comparative study between 2D and 3D cell cultures of MC3T3-E1 using the same type of material (a mixture of silk fibroin (SF) and chitosan (CS)). Our results showed 3D SF/CS scaffold exhibited different effects on cell culture compared with the 2D cases. 1) The cells grown in 3D scaffold showed multiple morphologies. 2) The proliferation of cells in 3D scaffold was long-term and sustainable. 3) Cell differentiation occurred throughout the entire 3D scaffold. The results showed that cell culture in 3D SF/CS scaffold exhibited different features than 2D cases and 3D SF/CS scaffold could be a promising material for 3D cell culture.

Correlation Between Protein Sequence Similarity and X-Ray Diffraction Quality in the Protein Data Bank

As the most widely utilized technique to determine the 3-dimensional structure of protein molecules, X-ray crystallography can provide structure of the highest resolution among the developed techniques. The resolution obtained via X-ray crystallography is known to be influenced by many factors, such as the crystal quality, diffraction techniques, and X-ray sources, etc. In this paper, the authors found that the protein sequence could also be one of the factors. We extracted information of the resolution and the sequence of proteins from the Protein Data Bank (PDB), classified the proteins into different clusters according to the sequence similarity, and statistically analyzed the relationship between the sequence similarity and the best resolution obtained. The results showed that there was a pronounced correlation between the sequence similarity and the obtained resolution. These results indicate that protein structure itself is one variable that may affect resolution when X-ray crystallography is used.

Silk fibroin/chitosan thin film promotes osteogenic and adipogenic differentiation of rat bone marrow-derived mesenchymal stem cells:

As a biodegradable polymer thin film, silk fibroin/chitosan composite film overcomes the defects of pure silk fibroin and chitosan films, respectively, and shows remarkable biocompatibility, appropriate hydrophilicity and mechanical properties. Silk fibroin/chitosan thin film can be used not only as metal implant coating for bone injury repair, but also as tissue engineering scaffold for skin, cornea, adipose, and other soft tissue injury repair. However, the biocompatibility of silk fibroin/chitosan thin film for mesenchymal stem cells, a kind of important seed cell of tissue engineering and regenerative medicine, is rarely reported. In this study, silk fibroin/chitosan film was prepared by solvent casting method, and the rat bone marrow-derived mesenchymal stem cells were cultured on the silk fibroin/chitosan thin film. Osteogenic and adipogenic differentiation of rat bone marrow-derived mesenchymal stem cells were induced, respectively. The proliferation ability, osteogenic and adipogenic differentiation abilities of rat bone marrow-derived mesenchymal stem cells were systematically compared between silk fibroin/chitosan thin film and polystyrene tissue culture plates. The results showed that silk fibroin/chitosan thin film not only provided a comparable environment for the growth and proliferation of rat bone marrow-derived mesenchymal stem cells but also promoted their osteogenic and adipogenic differentiation. This work provided information of rat bone marrow-derived mesenchymal stem cells behavior on silk fibroin/chitosan thin film and extended the application of silk fibroin/chitosan thin film. Based on the results, we suggested that the silk fibroin/chitosan thin film could be a promising material for tissue engineering of bone, cartilage, adipose, and skin.

A periodic magnetic field as a special environment for scientific research created by rotating permanent magnet pairs

A magnetic field is an often-encountered physical environment that can affect many processes, including chemical, physical, and biochemical processes. Utilization of magnetic fields is thus very helpful in a wide variety of applications, such as scientific research in various disciplines, materials processing (e.g., crystal growth and separation) in industry, and nuclear fusion. There are many different types of magnetic fields generated by different magnets, such as superconducting magnets, electromagnets, hybrid magnets, pulsed magnets, and permanent magnets. In this paper, we introduce a newly designed periodic magnetic field generated by rotating permanent magnet pairs. Preliminary tests showed that the periodic magnetic field is valuable in water evaporation, silver deposition, and protein crystallization. Apparently, in such a new environment that can generate a periodic magnetic field, a periodic force field will also be simultaneously generated on the sample. Further work shall be carried out to explore the potential applications of this magnetic field.