A thixotropic nanocomposite gel for three-dimensional cell culture

Thixotropic materials, which become less viscous under stress and return to their original state when stress is removed, have been used to deliver gel-cell constructs and therapeutic agents. Here we show that a polymer-silica nanocomposite thixotropic gel can be used as a three-dimensional cell culture material. The gel liquefies when vortexed--allowing cells and biological components to be added--and resolidifies to trap the components when the shear force from spinning is removed. Good permeability of nutrients and gases through the gel allows various cell types to proliferate and be viable for up to three weeks. Human mesenchymal stem cells cultured in stiffer gels developed bone-like behaviour, showing that the rheological properties of the gel can control cell differentiation. No enzymatic, chemical, or photo-crosslinking, changes in ionic strength or temperature are required to form or liquefy the gel, offering a way to sub-culture cells without using trypsin-a protease commonly used in traditional cell culture techniques.     

Human-like collagen/nano-hydroxyapatite scaffolds for the culture of chondrocytes

Three dimensional (3D) biodegradable porous scaffolds play a key role in cartilage tissue repair. Freeze-drying and cross-linking techniques were used to fabricate a 3D composite scaffold that combined the excellent biological characteristics of human-like collagen (HLC) and the outstanding mechanical properties of nano-hydroxyapatite (nHA). The scaffolds were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD) and compression tests, using Relive® Artificial Bone (RAB) scaffolds as a control. HLC/nHA scaffolds displayed homogeneous interconnected macroporous structure and could withstand a compression stress of 2.67 ± 0.37 MPa, which was higher than that of the control group. Rabbit chondrocytes were seeded on the composite porous scaffolds and cultured for 21 days. Cell/scaffold constructs were examined using SEM, histological procedures, and biochemical assays for cell proliferation and the production of glycosaminoglycans (GAGs). The results indicated that HLC/nHA porous scaffolds were capable of encouraging cell adhesion, homogeneous distribution and abundant GAG synthesis, and maintaining natural chondrocyte morphology compared to RAB scaffolds. In conclusion, the presented data warrants the further exploration of HLC/nHA scaffolds as a potential biomimetic platform for chondrocytes in cartilage tissue engineering.     

Medical application of 3-D polymer gel dosemeter evaluated by nuclear magnetic resonance

A polymer-gel dosemeter, which can be evaluated by nuclear magnetic resonance (NMR), was prepared and then a few samples were homogeneously irradiated by a Leksell gamma knife using an 18 mm collimator (60Co gamma photons) to obtain a calibration curve (NMR 1/T2 response to absorbed dose). To measure dose distribution from the Leksell gamma knife, a testing flask tilled with the gel was fixed in the head phantom and then irradiated based on a calculated treatment plan. Evaluation of dosemeters was performed on a Siemens EXPERT 1T NMR scanner. Dose profiles in X, Y and Z coordinates through the ellipsoidal shape of the dose distribution were obtained to compare experimental results from the irradiated phantom with the treatment planning system calculations. The use of a polymer-gel dosemeter for a verification of stereotactic procedures has some unique advantages which can be summarised as follows: (1) the dosemeter itself is tissue equivalent, (2) three-dimensional dose distributions can be measured, (3) the dosemeter allows patient's procedures to be simulated without any limitations.     

Fibrillized peptide microgels for cell encapsulation and 3D cell culture

One of the advantages of materials produced by self-assembly is that in principle they can be formed in any given container to produce materials of predetermined shapes and sizes. Here, we developed a method for triggering peptide self-assembly within the aqueous phase of water-in-oil emulsions to produce spherical microgels composed of fibrillized peptides. Size control over the microgels was achieved by specification of blade type, speed, and additional shear steps in the emulsion process. Microgels constructed in this way could then be embedded within other self-assembled peptide matrices by mixing pre-formed microgels with un-assembled peptides and inducing gelation of the entire composite, offering a route towards multi-peptide materials with micron-scale domains of different peptide formulations. The gels themselves were cytocompatible, as was the microgel fabrication procedure, enabling the encapsulation of NIH 3T3 fibroblasts and C3H10T-1/2 mouse pluripotent stem cells with good viability.     

具有三维聚焦之微型流式细胞/颗粒计数仪

本研究提出一种新式的微型细胞颗粒计数器,此微型细胞颗粒计数器结合二维水力聚焦及微文件流结构(micro-weir structure)于微管道中,可进行高精确度及均一性的细胞或颗粒计数,本研究是利用简单的等向性(isotropie)的湿式蚀刻技术来制作微文件流结构于玻璃基材上,而此微型细胞颗粒计数器的主要构成的组件包含二维边鞘流的聚焦结构,其主要功能为将细胞或颗粒聚焦于x-Y平面上,而微文件流结构的目的在于将细胞或颗粒在z方向的筛选,最后利用雷射诱导荧光系统将细胞或颗粒侦测出.在实验与数值分析的结果,显示此微型细胞颗粒计数器确实可进行高精确度及均一性的细胞或颗粒的计数,并且可以提供一个微型化的生物分析系统.     

Sparse 2-D arrays for 3-D phased array imaging--design methods

One of the most promising techniques for limiting complexity for real-time 3-D ultrasound systems is to use sparse 2-D layouts. For a given number of channels, optimization of performance is desirable to ensure high quality volume images. To find optimal layouts, several approaches have been followed with varying success. The most promising designs proposed are Vernier arrays, but also these suffer from high peaks in the sidelobe region compared with a dense array. In this work, we propose new methods based on the principles of suppression of grating lobes to form symmetric and non-symmetric regular sparse periodic and radially periodic designs. The proposed methods extend the concept of sparse periodic layouts by exploiting either an increased number of symmetry axes or radial symmetry. We also introduce two new strategies to form designs with nonoverlapping elements. The performance of the new layouts range from the performance of Vernier arrays to almost that of dense arrays. Our designs have simplicity in construction, flexibility in the number of active elements, and the possibility of trade off sidelobe peaks against sidelobe energy.     

2-D array for 3-D ultrasound imaging using synthetic aperture techniques

A two-dimensional (2-D) array of 256 X 256 = 65,536 elements, with total area 4 X 4 = 16 cm2, serves as a flexible platform for developing acquisition schemes for 3-D rectilinear ultrasound imaging at 10 MHz using synthetic aperture techniques. This innovative system combines a simplified interconnect scheme and synthetic aperture techniques with a 2-D array for 3-D imaging. A row-column addressing scheme is used to access different elements for different transmit events. This addressing scheme is achieved through a simple interconnect, consisting of one top, one bottom single-layer, flex circuits that, compared to multilayer flex circuits, are simpler to design, cheaper to manufacture, and thinner so their effect on the acoustic response is minimized. We present three designs that prioritize different design objectives: volume acquisiton time, resolution, and sensitivity, while maintaining acceptable figures for the other design objectives. For example, one design overlooks time-acquisition requirements, assumes good noise conditions, and optimizes for resolution, achieving -6 dB and -20 dB beamwidths of less than 0.2 and 0.5 mm, respectively, for an F/2 aperture. Another design can acquire an entire volume in 256 transmit events, with -6 dB and -20 dB beamwidths in the order of 0.4 and 0.8 mm, respectively.     

3-D culture of human umbilical vein endothelial cells with reversible thermosensitive hydroxybutyl chitosan hydrogel

The aim of this study was to present a non-trypsin 3D cell culture method with a reversible thermosensitive HBCS hydrogel. In this study, hydroxybutyl chitosan (HBCS) was synthesized by grafting hydroxybutyl groups on chitosan molecule chains. The prepared HBCS was water-soluble, and the reversible phase transformation temperature was 26 °C. Scanning electron microscope images illuminated the 3-D network of hydrogel formed irregular porous structure which ranged from 50–250 μm. Cell viability assay indicated that HBCS solution could promote the proliferation of human umbilical vein endothelial cells (HUVECs), and the boost of proliferation was enhanced with the increase of HBCS concentration. HBCS had no harm to the nitric oxide (NO) synthesis functionality of HUVECs. HUVECs could grow and reproduce inside the hydrogel, and showed good vitality after 14-days culture. Meanwhile, cells cultured inside the hydrogel could be passaged successively through the reversible phase transformation process of HBCS. The results revealed that HBCS have the potential to be used for 3-D cell culture without the use of trypsin.     

三维有限元模拟中的网格重划

详细论述了网格重划技术的判断准则、新旧网格之间场量传递等,提出了两种进行包含测试的新方法交点判断法和矢量判断法,并成功地应用于新旧网格之间三维场量的传递.实例应用验证了该方法的正确性和有效性.     

3-D biomodelling technology for maxillofacial reconstruction

Biomodel is a product of rapid prototyping process that represents a new approach for surgical planning and simulation. Biomodels were found to be particularly effective to locate an exact defect with good measurement accuracy and without any risk to the patients' health. These models could provide surgeons a realistic impression of complex structures before surgical intervention, allowing a define diagnosis and a precise preoperative simulation of skeleton modifying interventions.This work reports on the application of innovative 3-D biomodelling technology in the maxillofacial reconstruction. This technology allows for the calculation of the exact contours, angulations, length and general morphology of iliac crest and fibula flaps for maxillofacial reconstruction in a total of 28 clinical cases. Clinical results have shown that by using this technique, much better facial symmetry and improved functionality were achieved after surgery, which resulted in enormous benefits for treated patients.     

A 2-D model that accounts for 3-D fringing in MEMS devices

MEMS devices such as comb drives and rotary drives are geometrically simple in that each of the components may be represented as a ‘sweep’ of a 2-D cross-section through a given height. This simplicity leads to simpler CAD requirements, geometric robustness, faster visualization, etc. Further, 3-D electrostatic simulation may be simplified to a 2-D problem over the cross-section if one neglects 3-D fringing. Such 2-D simulations provide a quick feedback to the designer on various parameters such as capacitance and electrostatic forces.However, as is well known, 3-D simulations cannot be avoided if fringing is significant, or when these devices need to be fully optimized. Such 3-D simulations unfortunately involve constructing the full 3-D geometry, volume/surface mesh, etc.In this paper, we demonstrate that one can pose and solve a 2-D problem that accounts for 3-D fringing. The proposed technique does not require the construction of the 3-D CAD model or surface/volume mesh. Instead, the 3-D electrostatics problem is collapsed to 2-D via a novel dimensional reduction method. Once the 2-D problem is solved, the full 3-D field and associated charges/forces can be recovered, as a post-processing step. The simplicity and computational efficiency of the technique lends itself well to parametric study and design optimization.     

A miniaturized catheter 2-D array for real-time, 3-D intracardiac echocardiography

The design, fabrication, and characterization of a 112 channel, 5 MHz, two-dimensional (2-D) array transducer constructed on a six layer flexible polyimide interconnect circuit is described. The transducer was mounted in a 7 Fr (2.33 mm outside diameter) catheter for use in real-time intracardiac volumetric imaging. Two transducers were constructed: one with a single silver epoxy matching layer and the other without a matching layer. The center frequency and -6 dB fractional bandwidth of the transducer with a matching layer were 4.9 MHz and 31%, respectively. The 50 omega pitch-catch insertion loss was 80 dB, and the typical interelement crosstalk was -30 dB. The final element yield was greater than 97% for both transducers. The transducers were used to acquire real-time, 3-D images in an in vivo sheep model. We present in vivo images of cardiac anatomy obtained from within the coronary sinus, including the left and right atria, aorta, coronary arteries, and pulmonary veins. We also present images showing the manipulation of a separate electrophysiological catheter into the coronary sinus.     

2-D multiplane finite element technique for solving a class of 3-D problems

A finite element technique, for efficient solution of a class of 3-D elasticity problems, is presented. In this method, standard 2-D finite elements are used along with a ‘connector’ element. An element, previously used to model material interfaces, is shown to provide the properties for use as a ‘connector’ element, if input variables are redefined. The accuracy of the technique is illustrated with a sample solution.     

A class of data structures for 2-D and 3-D adaptive mesh refinement

A ‘family’ of tree data structures for adaptive mesh refinement is described and details concerning the associated logic are provided. The data structures encompass triangular elements and quadrilateral elements in two dimensions and quadrilateral bricks in three dimensions. Furthermore, both linear (bilinear) and quadratic (biquadratic) element types, respectively, are developed. Representative refinement results are given for the bilinear, trilinear and biquadratic types and associated performance studies made for the refinement procedure.     

Finite element-computation of the electromechanical J-Integral for 2-D and 3-D crack analysis

Piezoelectric ceramics find an application in many fields of technology. They may serve as sensors or actuators, mostly beeing exposed to high electric and mechanical loads. Therefore, fracture mechanics of piezoelectrics is an important field preserving strength and reliability under different conditions of application. This paper deals with the calculation of electromechanical energy release rates for arbitrary cracks in spatial piezoelectric structures applying a generalized J-integral. The crack problem is solved using a commercial FEM-code obtaining electric and mechanical field variables in nodes and integration points. These results serve as input data for the numerical computation of the electromechanical J-integral. The results are compared to findings from analytical and alternative numerical methods.     

Finite element simulation and modeling of 2-D arrays for 3-D ultrasonic imaging

Issues of modeling and design of 2-D arrays in three dimensions with finite element code are discussed. These ultrasonic arrays are used for real time dynamic imaging of the heart. Topics include optimization, sensitivity, and performance and methods to speed up the run times required for computer simulations of large three-dimensional models. Empirical results from 45×45 2-D arrays are also presented     

Sol–gel process for vegetal cell encapsulation

A sol–gel process was carried out at room temperature to obtain porous silica matrixes for entrapment of vegetal cells. The process uses sodium silicate as a precursor rather than alkoxides to avoid generation of by-products that is detrimental to Chlorella vulgaris cells. The influence of several factors influencing the gelification was explored to optimize the encapsulation process. The technique produces a transparent matrix and is then very suitable for vegetal cell entrapment since they require light for their photosynthetic activity. Activity of the algal cells was determined by measurement of the intensity of fluorescence emission at 682 nm. This measurement also allowed to evaluate the efficiency of the immobilization technique and assess the long-term activity of the encapsulated whole cells.     

Numerical modeling of ductile crack growth in 3-D using computational cell elements

This study describes a 3-D computational framework to model stable extension of a macroscopic crack under mode I conditions in ductile metals. The Gurson-Tvergaard dilatant plasticity model for voided materials describes the degradation of material stress capacity. Fixed-size, computational cell elements defined over a thin layer at the crack plane provide an explicit length scale for the continuum damage process. Outside this layer, the material remains undamaged by void growth, consistent with metallurgical observations. An element vanish procedure removes highly voided cells from further consideration in the analysis, thereby creating new tractionfree surfaces which extend the macroscopic crack. The key micro-mechanics parameters are D, the thickness of the computational cell layer, and f ₀ , the initial cell porosity. Calibration of these parameters proceeds through analyses of ductile tearing to match R-curves obtained from testing of deep-notch, through-crack bend specimens. The resulting computational model, coupled with refined 3-D meshes, enables the detailed study of non-uniform growth along the crack front and predictions of specimen size, geometry and loading mode effects on tearing resistance, here described by J-a curves. Computational and experimental studies are described for shallow and deep-notch SE(B) specimens having side grooves and for a conventional C(T) specimen without side grooves. The computational models prove capable of predicting the measured R-curves, post-test measured crack profiles, and measured load-displacement records.