Porous nanofibrous dressing enables mesenchymal stem cell spheroid formation and delivery to promote diabetic wound healing

Delayed and nonhealing of diabetic wounds imposes substantial economic burdens and physical pain on patients.Mesenchymal stem cells(MSCs)promote diabetic wound healing.Particularly when MSCs aggregate into multicellular spheroids,their therapeutic effect is enhanced.However,traditional culture platforms are inadequate for the efficient preparation and delivery of MSC spheroids,resulting in inefficiencies and inconveniences in MSC spheroid therapy.In this study,a three-dimensional porous nanofibrous dressing(NFD)is prepared using a combination of electrospinning and homogeneous freeze-drying.Using thermal crosslinking,the NFD not only achieves satisfactory elasticity but also maintains notable cytocompatibility.Through the design of its structure and chemical composition,the NFD allows MSCs to spontaneously form MSC spheroids with controllable sizes,serving as MSC spheroid delivery systems for diabetic wound sites.Most importantly,MSC spheroids cultured on the NFD exhibit improved secretion of vascular endothelial growth factor,basic fibroblast growth factor,and hepatocyte growth factor,thereby accelerating diabetic wound healing.The NFD provides a competitive strategy for MSC spheroid formation and delivery to promote diabetic wound healing.

Spheroid construction strategies and application in 3D bioprinting

Tissue engineering has been striving toward designing and producing natural and functional human tissues.Cells are the fundamental building blocks of tissues.Compared with traditional two-dimensional cultured cells,cell spheres are threedimensional(3D)structures that can naturally form complex cell–cell and cell–matrix interactions.This structure is close to the natural environment of cells in living organisms.In addition to being used in disease modeling and drug screening,spheroids have significant potential in tissue regeneration.The 3D bioprinting is an advanced biofabrication technique.It accurately deposits bioinks into predesigned 3D shapes to create complex tissue structures.Although 3D bioprinting is efficient,the time required for cells to develop into complex tissue structures can be lengthy.The 3D bioprinting of spheroids significantly reduces the time required for their development into large tissues/organs during later cultivation stages by printing them with high cell density.Combining spheroid fabrication and bioprinting technology should provide a new solution to many problems in regenerative medicine.This paper systematically elaborates and analyzes the spheroid fabrication methods and 3D bioprinting strategies by introducing spheroids as building blocks.Finally,we present the primary challenges faced by spheroid fabrication and 3D bioprinting with future requirements and some recommendations.

Dynamic spheroidization kinetics behavior of Ti-6.5Al-2Zr-1Mo-1V alloy with lamellar microstructure

Abstract： The dynamic spheroidization kinetics behavior of Ti-6.5Al-2Zr-1Mo-1V alloy with a lamellar initial microstructure was studied by isothermal hot compression tests in the temperature range of 750-950℃ and strain rates of 0.001-10 s^-1. The results show that the spheroidized fraction of alpha lamellae increases with the increase of temperature and the decrease of strain rate. The spheroidization kinetics curves predicted by JMAK equation agree well with experimental ones. The corresponding SEM and TEM observations indicate that the dynamic spheroidization process can be divided into two stages. The primary stage is boundary splitting formed by two competing mechanisms which are dynamic recrystallization and mechanical twin. In the second stage, the penetration of beta phase into the alpha/alpha grain boundaries is dominant, which is controlled in nature by diffusion of the chemical elements such as Al, Mo and V.

3D （Three-Dimensional） Caco-2 Spheroids： Optimized in vitro Protocols to Favor Their Differentiation Process and to Analyze Their Cell Growth Behavior

3D （Three-dimensional） Caco-2 spheroids closely recapitulating in vivo physiological organization of intestinal epithelial cells, provide an excellent in vitro model system to study their pathophysiology and their response to stressful stimuli. The objective of this technical note is to provide optimized in vitro experimental protocols for culturing 3D Caco-2 spheroids and for analyzing their cell growth features. An optimized 3D Caco-2 spheroid culturing technique based on a new configuration of the culture medium is provided A methodological approach to determine the distribution of the cell cycle phases in disaggregated Caco-2 spheroids by using cytofluorimetric analysis is also described. The optimized culturing protocol favors 3D Caco-2 spheroid differentiation process, as evaluated by the number of well-differentiated spheroids with a single hollow lumen. The cytofluorimetric analysis allows rapid collection of cell cycle phase data from high numbers of spheroid samples, thus, permitting to estimate their growth dynamics in a relatively short time. The optimized technical approaches described here can be applied in systematic manner to a variety of research activities utilizing 3D Caco-2 spheroids. Ease of use, time and economic saving advantages deriving from these protocols further highlight their potential.

Carbide refinement in M42 high speed steel by rare earth metals and spheroidizing treatment

The influence of rare earth metals and heat treatment on the microstructure and performance of M42 steel has been investigated by means of an optical microscope OM scanning electron microscope SEM energy dispersive spectroscopy EDS transmission electron microscope TEM electron back-scatter diffraction EBSD and X-ray diffraction XRD . The results show that M2 C is the prevailing type of eutectic carbides in M42 steel. After modification with rare earth metals M2 C eutectic carbides change from the ordered lamellar structure into a circular structure.Despite different morphologies the two carbides present the same characteristics of microstructure and growth orientation.Compared with lamellar carbides M2 C carbides with the circular structure are much easier to decompose and spheroidize after heating which remarkably refines the carbide dimensions.The refined carbides improve the supersaturation of alloying elements in martensite and increase the hardness of M42 steel by 1.5 HRC.

Primary porcine hepatocytes with portal vein serum cultured on microcarriers or in spheroidal aggregates

AIM To develop a culture mode providingdurable biomaterials with high yields andactivities used in bioartificial liver.METHODS Hepatocytes were isolated from awhole pig liver by Seglen’s method of orthotopicperfusion with collagenase.In culture onmicrocarriers,primary porcine hepatocyteswere inoculated at a concentration of 5×10~7/mLinto the static culture systems containing 2 g/LCytodex-3,then supplemented with 100 mL/Lfetal calf serum(FCS)or 100 mL/L porcineportal vein serum(PPVS)respectively.Inspheroidal aggregate culture hepatocytes wereinoculated into 100 mL siliconized flasks at aconcentration of 5.0×10~6/mL.RESULTS In culture on microcarriershepatocytes tended to aggregate on Cytodex-3obviously after being inoculated.Typical multi-cellular aggregated spheroids could be found inthe two systems 24 h-48 h after hepatocyteswere cultured.The morphological charact-eristics and synthetic functions were maintainedfor 5 wk in FCS culture system and 8 wk in PPVSculture system.In spheroidal aggregate cultureabout 80%-90% isolated hepatocytes becameaggregated spheroids 24h after cultured insuspension and mean diameter of the spheroidswas 100μm.The relationship among thehepatocytes resembled that in the liver in vivo.Synthetic functions of albumin and urea of the spheroids were twice those of hepatocytescultured on monolayers.CONCLUSION As high-yields and high-activitymodes of culture on microcarriers or inspheroidal aggregate culture with portal veinserum are promising to provide biomaterials forbioartificial liver(BAL)efficiently.

Effect of Plasma Spheroidization Process on the Microstructure and Crystallographic Phases of Silica, Alumina and Nickel Particles

During the plasma spheroidization process powders undergo different changes in their microstructures and crystal phases. In this paper, simple calculation of heat transfer between the plasma and a suspended particle was performed based on three hypotheses for the purpose of guiding experiments. Experimental investigation of the crystal phases and microstructural changes during the plasma processing was made using silica, alumina and nickel powders as starting materials. It has been revealed from the experimental results that these materials undergo different changes in crystal phases and microstructures, and these changes are essentially determined by the structures, properties and aggregate states of the starting materials.

Spheroidization and Ostwald Growth of Carbide in Isothermal Process of Hot-Deformed High Carbon Chromium Cast Steel

In isothermal spheroidizing process,the spheroidization and growth of the carbide formed in hot-deformed high-carbon chromium cast steel at high temperature were investigated.The results showed that the spheroidizing growth of carbide proceeds in such a way that the bigger carbide particles swallow the smaller ones,and the short rhabdoid carbides dissolve and are spheroidized by itself.When the samples were held at 720℃ for more than 3 h,the spheroidization is not obvious.The feature of the process is the size increment and the amount decrement of carbide particles.The empirical equation for growth rate of carbides was obtained.The volume fraction of carbides keeps constant.The growth process agrees well with Ostwald Ripening Law.

New Spheroidizing Technique of Ultra-High Carbon Steel With Aluminum Addition

A new spheroidizing process of ultra-high carbon steel （UHCS） containing C 1.55%, Cr 1.45%, and Al 1.5% in mass percent has been proposed. The effect of processing parameters on the microstructure was analyzed. The UHCS produced by this new process has a microstructure with recrystallized ferrite matrix and fine and uniform carbide particles. After this spheroidizing, the UHCS exhibits good mechanical properties at ambient temperature, for example σb= 1 100 MPa, σs =915 MPa, δ=8% and high ratio of σs/σb.

Numerical Simulation of Shrinkage Cavity Formation in Spheroidal Graphite Iron Castings Based on Micro Modeling

A micro-modeling method (MM) for the quantitative prediction of the shrinkage cavity formation in SGiron castings is proposed. The mathematical models describing the volume changes during the solidification ofspheroidal graphite cast iron are established based on the models of solidification kinetics. The shrinkage cavityformation of T-shaped SG iron castings is calculated with MM method. The calculated results are compared with theexperimental results. It is shown that the predicted size, shape and distribution of shrinkage cavity by MM methodare in good agreement with the measured results.

Characterizing the Spheroidization Grade and Strength of 15CrMo Steel through Determining Fractal Dimension

The fractal dimension（FD） of surfaces has been widely used to characterize the properties of materials.However,most of the previous researches were concentrated on the correlation between the FD of surfaces and mechanical properties of materials,such as impact energy and fracture toughness,etc.The aim of this paper is to characterize the spheroidization grade and strength of 15CrMo steel through determination of FD of cementite phase on the basis of two-dimension microstructural image.Two methods,namely slit-island method（SIM） and box-counting method（BCM）,are used to determine the value of FD.It is found that the FD value evaluated by using BCM is generally higher than that evaluated by SIM.This phenomenon may be due to the difference in the principles used in different methods.Whether SIM or BCM is used,the spheroidization grade in 15CrMo steel linearly increases with decreasing the value of FD.The relationship between the FD value,D,and spheroidization grade,Sg,can be approximately expressed as D≌-0.11Sg＋A,where A is a constant value which is depended on the evaluation method.Both the ultimate strength and the yielding strength of 15CrMo steel increase with increasing FD of cementite phase.There may be a common relationship between the FD of cementite phase and strength of 15CrMo steel.When the FD of cementite phase in 15CrMo steel is determined,the strength of this steel can be evaluated.The present paper can provide a novel method to evaluate the strength and spheroidization grade of carbon steel through determination of fractal dimension（FD） of cementite phase.

Spheroidization of silica powders by radio frequency inductively coupled plasma with Ar–H2 and Ar–N2 as the sheath gases at atmospheric pressure

Amorphous spherical silica powders were prepared by inductively coupled thermal plasma treatment at a radio frequency of 36.2 MHz. The effects of the added content of hydrogen and nitrogen into argon(serving as the sheath gas), as well as the carrier gas flow rate, on the spheroidization rate of silica powders, were investigated. The prepared silica powders before and after plasma treatment were examined by scanning electron microscopy, X-ray diffraction, and laser granulometric analysis. Results indicated that the average size of the silica particles increased, and the transformation of crystals into the amorphous state occurred after plasma treatment. Discharge image processing was employed to analyze the effect of the plasma temperature field on the spheroidization rate. The spheroidization rate of the silica powder increased with the increase of the hydrogen content in the sheath gas. On the other hand, the spheroidization rate of the silica power first increased and then decreased with the increase of the nitrogen content in the sheath gas. Moreover, the amorphous content increased with the increase of the spheroidization rate of the silica powder.

Growth Pattern and Its Indication of Spheroidal Nano-Micro Crystal Aggregates of Pyrite in the Baiyunpu Pb-Zn Polymetallic Deposit, Central Hunan

The Baiyunpu deposit lies in the southwest plunging Dachengshan anticline in central Hunan, which is a large Pb-Zn polymetallic deposit. The orebodies were surrounded by the Qiziqiao Formation limestone in the Middle Devonian, and its geological occurrence is consistent with the wall rocks. A large number of spheroidal pyrite aggregates are found unevenly distributed in the ores. The spheroidal aggregates are made up of kernels and concentric rings. The kernels are composed of approximately epigranular pyrite nanocrystals, while the rings are composed of accumulated pyrite microcrystals growing along the radial direction. The spheroidal pyrite aggregate and its outer zones can be divided into five areas（A–E）. The results of electron probe micro analysis（EPMA） show that from the zone A1 to B, Co/Ni 〈1, the sum of Co and Ni is 0.08%–0.26%, S/Fe increases from 2.06 to 2.15. While from the zone C to E, Ni cannot be detected and S/Fe decreases from 2.22 to 2.08. Powder X-ray diffraction（XRD） analysis in the micro zone shows obvious crystalline characteristics in the aggregates. Moving from the inside outwards, the maximum diffraction peak intensity of the（111） and（220） crystal planes of pyrite increases, and the crystallinity improves. The degree of change in the（111） plane is the most prominent. Considering the theory of crystal growth along with the geologic features of the depositional environment where the spheroidal pyrite aggregates developed, we confirm that the spheroidal aggregates are the result of nano-micro crystalline gathering and growth occurring by the following sequence of processes：nano-crystalline nucleation and growth, gathering into a ball, oriented growth of microcrystals, continuous accumulation, and adjustment of grain boundaries. The formation of the spheroidal pyrite aggregates in the late Qiziqiao Formation of the Middle Devonian occurred in a neutral to weak alkaline and reductive sedimentary environment in the normal oxygen-rich shallowwater carbonate platform edge. The variations in the S/Fe ratio and crystallisation characteristics indicate that during pyrite crystal growth, the sulphur fugacity was high locally and rose constantly, the degree of supersaturation decreased locally and the growth environment was stable relatively.

Study on the RF inductively coupled plasma spheroidization of refractory W and W-Ta alloy powders

Spherical powders with good flowability and high stacking density are mandatory for powder bed additive manufacturing. Nevertheless, the preparation of spherical refractory tungsten and tungsten alloy powders is a formidable task. In this paper, spherical refractory metal powders processed by high-energy stir ball milling and RF inductively coupled plasma were investigated. By utilizing the technical route, pure spherical tungsten powders were prepared successfully, the flowability increased from 10.7 s/50 g to 5.5 s/50 g and apparent density increased from 6.916 g cm-3 to 11.041 g cm-3. Alloying element tantalum can reduce the tendency to micro- crack during tungsten laser melting and rapid solidification process. Spherical W-6Ta （%wt） powders were prepared in this way, homogeneous dispersion of tantalum in a tungsten matrix occurred but a small amount of flake-like shape particles appeared after high-energy stir ball milling. The flake-like shape particles can hardly be spheroidized in subsequent RF inductively coupled plasma process, might result from the unique suspended state of flaky particles under complex electric and magnetic fields as well as plasma-particle heat exchange was different under various turbulence models. As a result, the flake-like shape particles cannot pass through the high-temperature area of thermal plasma torch and cannot be spheroidized properly.

DETERMINATION OF EFFECTIVE MODULI FOR FOAM PLASTICS BASED ON THREE PHASE SPHEROIDAL MODEL

The relations of bulk modulus, shear modulus, Young's modulus and the Poisson's ratio with porosity of foam plastics are determined by a three phase spheroidal model commonly used in Composite Mechanics. The results are compared with those using differential scheme. It is shown that the material properties derived from the present model normally are larger than those obtained by differential scheme for foam plastics with identical porosity. The differences in shear moduli and Young's moduli obtained by the two methods are small but they are larger for bulk moduli of incompressible matrix and Poisson's ratios. The Young's moduli of high density foam plastics derived by the present model agree better with experimental ones.

Spatiotemporal pharmacometabolomics based on ambient mass spectrometry imaging to evaluate the metabolism and hepatotoxicity of amiodarone in HepG2 spheroids

Three-dimensional(3D)cell spheroid models combined with mass spectrometry imaging(MSI)enables innovative investigation of in vivo-like biological processes under different physiological and pathological conditions.Herein,airflow-assisted desorption electrospray ionization-MSI(AFADESI-MSI)was coupled with 3D HepG2 spheroids to assess the metabolism and hepatotoxicity of amiodarone(AMI).High-coverage imaging of>1100 endogenous metabolites in hepatocyte spheroids was achieved using AFADESI-MSI.Following AMI treatment at different times,15 metabolites of AMI involved in Ndesethylation,hydroxylation,deiodination,and desaturation metabolic reactions were identified,and according to their spatiotemporal dynamics features,the metabolic pathways of AMI were proposed.Subsequently,the temporal and spatial changes in metabolic disturbance within spheroids caused by drug exposure were obtained via metabolomic analysis.The main dysregulated metabolic pathways included arachidonic acid and glycerophospholipid metabolism,providing considerable evidence for the mechanism of AMI hepatotoxicity.In addition,a biomarker group of eight fatty acids was selected that provided improved indication of cell viability and could characterize the hepatotoxicity of AMI.The combination of AFADESI-MSI and HepG2 spheroids can simultaneously obtain spatiotemporal information for drugs,drug metabolites,and endogenous metabolites after AMI treatment,providing an effective tool for in vitro drug hepatotoxicity evaluation.

Eutectic solidification mode of spheroidal graphite cast iron and graphitization

The shrinkage and chilling tendency of spheroidal graphite (abbreviated SG) cast iron is much greater than that of the flake graphite cast iron in spite of its higher amount of C and Si contents. Why? The main reason should be the difference in their graphitization during the eutectic solidification. In this paper, we discuss the difference in the solidification mechanism of both cast irons for solving these problems using unidirectional solidification and the cooling curves of the spheroidal graphite cast iron. The eutectic solidification rate of the SG cast iron is controlled by the diffusion of carbon through the austenite shell, and the final thickness is 1.4 times the radius of the SG, therefore, the reduction of the SG size, namely, the increase in the number, is the main solution of these problems.

Formation mechanism of spheroidal carbide in ultra-low carbon ductile cast iron

The formation mechanism of the spheroidal carbide in the ultra-low carbon ductile cast iron fabricated by the metal mold casting technique was systematically investigated. The results demonstrated that the spheroidal carbide belonged to eutectic carbide and crystallized in the isolated eutectic liquid phase area. The formation process of the spheroidal carbide was related to the contact and the intersection between the primary dendrite and the secondary dendrite of austenite. The oxides of magnesium, rare earths and other elements can act as heterogeneous nucleation sites for the spheroidal carbide. It was also found that the amount of the spheroidal carbide would increase with an increase in carbon content. The cooling rate has an important influence on the spheroidal carbide under the same chemical composition condition.

Solving ground eigenvalue and eigenfunction of spheroidal wave equation at low frequency by supersymmetric quantum mechanics method

The spheroidal wave functions are found to have extensive applications in many branches of physics and mathematics. We use the perturbation method in supersymmetric quantum mechanics to obtain the analytic ground eigenvalue and the ground eigenfunction of the angular spheroidal wave equation at low frequency in a series form. Using this approach, the numerical determinations of the ground eigenvalue and the ground eigenfunction for small complex frequencies are also obtained.

USING HIGH Mg-CONTENT CORED-WIRE TO SPHEROIDIZE AND DESULPHURIZE DUCTILE IRON MELT IN INDUSTRIAL EXPERIMENTS

Mg-content cored-wire was used to desulphurize and spheroidize ductile iron melt in industrial experiments. The optimal feeding speed and suitable treatment temperature were determined in the experiments. And cored-wire method and pouring method were compared. Conclusions have been drawn that, under these conditions in the experiments, the optimal feeding speed is 15m/min, treatment temperature should be as low as possible, 1400-1450℃ generally; and cored-wire method can act more effective in ductile iron melt desulphurization and spheroid than pouring method.