Da-Wei Li

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.

A novel porous Fe/Fe-W alloy scaffold with a double-layer structured skeleton: Preparation, in vitro degradability and biocompatibility

A novel porous Fe/Fe-W alloy scaffold with a double-layer structured skeleton was prepared for the first time by electrodeposition. The microstructure of the scaffold was analysed by X-ray diffraction (XRD), scanning electron microscopy (SEM), energy-dispersive X-ray spectroscopy (EDS) and mercury porosimetry. Mechanical property, in vitro degradability and biocompatibility were tested by tensile test, immersion and a cytotoxicity test. The results showed that the scaffolds exhibited a cellular structure that is similar to that of cancellous bone and had a considerably large specific surface area. The skeleton of the scaffolds showed a double-layer structure that was composed of a hollow Fe skeleton wrapped in a thin layer of Fe-W alloy. The tensile strength and the apparent density are close to that of cancellous bone. It was also found that the different surface microstructures showed different effects on in vitro degradability and biocompatibility. In the immersion test, the corrosion rate decreased gradually as the immersion time increased. In the cytotoxicity test, the extraction medium of the pure Fe scaffold showed the lowest cell viability, followed by that of 1.5FeW as a close second. The extraction media of FeW, Fe1.5W and Fe2W were similar, and their cell viability was far above that of the Fe and 1.5FeW scaffolds. The structural style of the scaffolds presented in this paper is potentially useful and applicable to developing degradable scaffolds with a tailored corrosion rate.

An ignored variable: solution preparation temperature in protein crystallization

Protein crystallization is affected by many parameters, among which certain parameters have not been well controlled. The temperature at which the protein and precipitant solutions are mixed (i.e., the ambient temperature during mixing) is such a parameter that is typically not well controlled and is often ignored. In this paper, we show that this temperature can influence protein crystallization. The experimental results showed that both higher and lower mixing temperatures can enhance the success of crystallization, which follows a parabolic curve with an increasing ambient temperature. This work illustrates that the crystallization solution preparation temperature is also an important parameter for protein crystallization. Uncontrolled or poorly controlled room temperature may yield poor reproducibility in protein crystallization.

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.

An investigation of the effects of varying pH on protein crystallization screening

Protein crystallization occurs when the solution conditions are suitable for nucleation and growth of the crystals. pH, as one of the most important parameters that governs the protein crystallization process, can affect the conformation, activity, electrostatic interactions and solubility of protein in the solution. Hence, manipulating the pH is an important path to success in crystallizing proteins. In this paper, we show that varying pH levels over the incubation time during the crystallization process can help increase the chance of obtaining protein crystals. The process of varying the pH can be considered as one way to automatically search for a suitable pH for successful crystallization. This discovery can guide us to develop a new screening strategy in which varying the pH against the incubation time is the major process used during the crystallization screening. Our screening experiments verified that varying the pH is indeed a promising method to achieve efficient crystallization screening of a protein.

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.

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.

Effect of the weather conditions during solution preparation on lysozyme crystallization

Protein crystallization is a delicate process that is always sensitive to environmental factors. When the environmental factors are not well controlled or not controlled at all, identical crystallization droplets from the same mother liquid may yield different crystallization results. One environmental factor, the weather conditions during crystallization solution preparation, is not usually considered as a parameter for protein crystallization. In this paper, it is shown that the weather parameters during preparation of the crystallization experiment, including the ambient temperature, humidity, pressure and particulate matter in the air, can all affect the reproducibility of lysozyme crystallization. An identical lysozyme crystallization experiment was repeated for an entire year, and the weather conditions when each crystallization experiment was set up were recorded along with the crystallization results. Among the parameters recorded, the humidity during the experiment setup showed the strongest effect on lysozyme crystallization. On the basis of these results, it is suggested that the weather conditions during crystallization solution preparation should be considered as a potential factor that can influence protein crystallization.