Functionally heterogeneous porous scaffold design for tissue engineering

Porous scaffolds with interconnected and continuous pores have recently been considered as one of the most successful tissue engineering strategies. In the literature, it has been concluded that properly interconnected and continuous pores with their spatial distribution could contribute to perform diverse mechanical, biological and chemical functions of a scaffold. Thus, there has been a need for reproducible and fabricatable scaffold design with controllable and functional gradient porosity. Improvements in Additive Manufacturing (AM) processes for tissue engineering and their design methodologies have enabled the development of controlled and interconnected scaffold structures. However homogeneous scaffolds with uniform porosity do not capture the intricate spatial internal micro architecture of the replaced tissue and thus are not capable of capturing the design. In this work, a novel heterogeneous scaffold modeling is proposed for layered-based additive manufacturing processes. First, layers are generated along the optimum build direction considering the heterogeneous micro structure of tissue. Each layer is divided into functional regions based on the spatial homogeneity factor. An area weight based method is developed to generate the spatial porosity function that determines the deposition pattern for the desired gradient porosity. To design a multi-functional scaffold, an optimum deposition angle is determined at each layer by minimizing the heterogeneity along the deposition path. The proposed methodology is implemented and illustrative examples are also provided. The effective porosity is compared between the proposed design and the conventional uniform porous scaffold design. Sample designed structures have also been fabricated with a novel micro-nozzle biomaterial deposition system. The result has shown that the proposed methodology generates scaffolds with functionally gradient porosity.     

Computer-aided characterization for effective mechanical properties of porous tissue scaffolds

Performance of various functions of the tissue structure depends on porous scaffold microstructures with specific porosity characteristics that influence the behavior of the incorporated or ingrown cells. Understanding the mechanical properties of porous tissue scaffold is important for its biological and biomechanical tissue engineering application. This paper presents a computer aided characterization approach to evaluate the effective mechanical properties of porous tissue scaffold. An outline of a computer-aided tissue engineering approach for design and fabrication of porous tissue scaffold, procedure of computer-aided characterization and its interface with design model, development of a computational algorithm for finite element implementation and numerical solution of asymptotic homogenization theory is presented. Application of the algorithm to characterize the effective mechanical properties of porous poly-ε-caprolactone scaffold manufactured by precision extruding freeform deposition will also be presented, along with a parametric study of the process and design parameter to the structural properties of tissue scaffold.     

Creation of a unit block library of architectures for use in assembled scaffold engineering

Guided tissue regeneration is gaining importance in the field of orthopaedic tissue engineering as need and technology permits the development of site-specific engineering approaches. Computer Aided Design (CAD) and Finite Element Analysis (FEA) hybridized with manufacturing techniques such as Solid Freeform Fabrication (SFF), is hypothesized to allow for virtual design, characterization, and production of scaffolds optimized for tissue replacement. However, a design scope this broad is not often realized due to limitations in preparing scaffolds both for biological functionality and mechanical longevity. To aid scientists in fabrication of a successful scaffold, we propose characterization and documentation of a library of micro-architectures, capable of being seamlessly merged according to the mechanical properties (stiffness, strength), flow perfusion characteristics, and porosity, determined by the scientist based on application and anatomic location. The methodology is discussed in the sphere of bone regeneration, and examples of catalogued shapes are presented. Similar principles may apply for other organs as well.     

Internal architecture design and freeform fabrication of tissue replacement structures

Modeling, design and fabrication of tissue scaffolds with intricate architecture, porosity and pore size for desired tissue properties presents a challenge in tissue engineering. This paper will present the details of our development in the design and fabrication of the interior architecture of scaffolds using a novel design approach. The interior architecture design (IAD) approach seeks to generate layered scaffold freeform fabrication tool path without forming complicated 3D CAD scaffold models. This involves: applying the principle of layered manufacturing to determine the scaffold individual layered process planes and layered contours; defining the 2D characteristic patterns of the scaffold building blocks (unit cells) to form the Interior Scaffold Pattern; and the generating the process tool path for freeform fabrication of these scaffolds with the specified interior architecture. Feasibility studies applying the IAD algorithm to example models with multi-interior architecture and the generation of fabrication planning instructions will also be presented.     

Bayesian computer-aided experimental design of heterogeneous scaffolds for tissue engineering

This paper presents a Bayesian methodology for computer-aided experimental design of heterogeneous scaffolds for tissue engineering applications. These heterogeneous scaffolds have spatial distributions of growth factors designed to induce and direct the growth of new tissue as the scaffolds degrade. While early scaffold designs have been essentially homogenous, new solid freeform fabrication (SFF) processes enable the fabrication of more complex, biologically inspired heterogeneous designs with controlled spatial distributions of growth factors and scaffold microstructures. SFF processes dramatically expand the number of design possibilities and significantly increase the experimental burden placed on tissue engineers in terms of time and cost. Therefore, we use a multi-stage Bayesian surrogate modeling methodology (MBSM) to build surrogate models that describe the relationship between the design parameters and the therapeutic response. This methodology is well suited for the early stages of the design process because we do not have accurate models of tissue growth, yet the success of our design depends on understanding the effect of the spatial distribution of growth factors on tissue growth. The MBSM process can guide experimental design more efficiently than traditional factorial methods. Using a simulated computer model of bone tissue regeneration, we demonstrate the advantages of Bayesian versus factorial methods for designing heterogeneous fibrin scaffolds with spatial distributions of growth factors enabled by a new SFF process.     

The optimization of hybrid scaffold fabrication process in precision deposition system using design of experiments

In recent tissue engineering field, it is being reported that the fabrication of three-dimensional (3D) scaffolds having high porous and controlled internal/external architectures can give potential contributions in cell adhesion, proliferation and differentiation. To fabricate these scaffolds, various rapid prototyping technologies are being applied to. The rapid prototyping technology has made it possible to fabricate solid free-form 3D microstructures in layer-by-layer process. In this research, we introduce the development of precision deposition system, which is one of rapid prototyping technologies, and the fabrication result of scaffold using design of experiments (DOE) to optimize the deposition process. The precision deposition system required the combination of several technologies, including motion control, thermal control, pneumatic control, and CAD/CAM software. Through the organization of experimental approach using DOE, the fabrication process of hybrid scaffold, which is composed of blended poly-caprolactone, poly-lactic-co-glycolic acid and tricalcium phosphate, is established to get a uniform line width, line height and porosity efficiently.     

Optimal design of a 3D-printed scaffold using intelligent evolutionary algorithms

Fabrication of three-dimensional structures has gained increasing importance in the bone tissue engineering (BTE) field. Mechanical properties and permeability are two important requirement for BTE scaffolds. The mechanical properties of the scaffolds are highly dependent on the processing parameters. Layer thickness, delay time between spreading each powder layer, and printing orientation are the major factors that determine the porosity and compression strength of the 3D printed scaffold.In this study, the aggregated artificial neural network (AANN) was used to investigate the simultaneous effects of layer thickness, delay time between spreading each layer, and print orientation of porous structures on the compressive strength and porosity of scaffolds. Two optimization methods were applied to obtain the optimal 3D parameter settings for printing tiny porous structures as a real BTE problem. First, particle swarm optimization algorithm was implemented to obtain the optimum topology of the AANN. Then, Pareto front optimization was used to determine the optimal setting parameters for the fabrication of the scaffolds with required compressive strength and porosity. The results indicate the acceptable potential of the evolutionary strategies for the controlling and optimization of the 3DP process as a complicated engineering problem.     

A control approach for pore size distribution in the bone scaffold based on the hexahedral mesh refinement

Tissue engineering is the application of that knowledge to the building or repairing of tissues. Generally, engineered tissue is a combination of living cells and a support structure called scaffolds. Modeling, design and fabrication of tissue scaffold with intricate architecture, porosity and pore size for desired tissue properties presents a challenge in tissue engineering. In this paper, a control approach for pore size distribution in the bone scaffold based on the hexahedral mesh refinement is presented. Firstly, the bone scaffold modeling approach based on the shape function in the finite element method is provided. The resulting various macroporous morphologies can be obtained. Then conformal refinement algorithm for all-hexahedral element mesh is illustrated. Finally, a modeling approach for constructing tissue engineering (TE) bone scaffold with defined pore size distribution is presented. Before the conformal refinement of all-hexahedral element mesh, a 3D mesh with various hexahedral elements must be provided. If all the pores in the bone scaffold need to be reduced, that means that the whole hexahedral mesh needs to be refined. Then the solid entity can be re-divided with altered subdivision parameters. If the pores in the local regions of bone need to be reduced, that means that 3D hexahedral mesh in the local regions needs to be refined. Based on SEM images, the pore size distribution in the normal bone can be obtained. Then, according to the conformal refinement of all-hexahedral element meshes, defined hexahedral size distribution can be gained, which leads to generate defined pore size distribution in the bone scaffold, for the pore morphology and size are controlled by various subdivided hexahedral elements. Compared to other methods such as varying processing parameters in supercritical fluid processing and multi-interior architecture design, the method proposed in this paper enjoys easy-controllability and higher accuracy.     

Biodegradable polylactic acid microstructures for scaffold applications

In this research, we present a simple and cost effective soft lithographic process to fabricate polylactic acid (PLA) scaffolds for tissue engineering. In which, the negative photoresist JSR THB-120N was spun on a glass subtract followed by conventional UV lithographic processes to fabricate the master to cast the PDMS elastomeric mold. A thin poly(vinyl alcohol) (PVA) layer was used as a mode release such that the PLA scaffold can be easily peeled off. The PLA precursor solution was then cast onto the PDMS mold to form the PLA microstructures. After evaporating the solvent, the PLA microstructures can be easily peeled off from the PDMS mold. Experimental results show that the desired microvessels scaffold can be successfully transferred to the biodegradable polymer PLA. Encouraging progress in bovine endothelial cells seeding was observed.     

A tracer metric numerical model for predicting tortuosity factors in three-dimensional porous tissue scaffolds

One of the critical functions of a tissue-engineered construct is to be able to provide adequate nutrient and oxygen supply into the interior of the construct. An insufficient supply will lead to slower cellular proliferation rates and eventual apoptosis. The supply of the nutrients is largely governed by the transport properties of the construct which in turn is dependent on the porosity, tortuosity and surface chemistry of the tissue construct. The design and fabrication of scaffolds with tailored properties is thus a crucial step in the growth of tissue within their host environment. This paper discusses the development of a numerical characterization technique to measure the three-dimensional tortuosity factors for any given interconnected porous design. Tortuosity factors are obtained in the three orthogonal principal directions for several candidate unit cell architectures. The proposed numerical technique has been validated with models of known tortuosity. The developed technique will provide a basis for the study of transport properties of the designed scaffold and its effect on cellular function and response through the development of dynamic culture bioreactors.     

Characterization of the influence of fabrication methods on microstructure failure

Single-crystal silicon microstructures, of identical design, exhibit different failure rates (following fabrication or mechanical shock testing) due to various processes. The microstructures fabricated with a boron diffusion and subsequent removal of the boron-diffused layer have a higher survival rate to the fabrication process and to mechanical shock. The survival rate (a survivor has an intact proof mass and beam) through the process is increased by 26.5%. At a 3680g shock, the boron-diffused devices have a 2.3% lower failure rate but the difference is not statistically significant. These results have been developed with wafer-level shock testing, which permits bulk testing of many samples in a cost-effective manner.     

基于多模态成像技术监测基因修饰支架中血管生成及修复临界性骨缺损的研究

基因修饰促血管化的支架是促进骨再生的有效方法之一,它可以将目的基因转移到内源性细 胞中实现生长因子原位、持续表达,诱导内源性细胞的增殖、迁移和分化,从而促进骨组织再生。该 文以慢病毒介导基因修饰多孔 PLGA/nHAp 复合支架诱导血管生成为研究对象,采用静电吸附和低温冷冻方法制备基因修饰多孔 PLGA/nHAp 复合支架;以小鼠顶骨临界性骨缺损为模型,利用多模态双光子及光声显微成像技术在体、实时、连续监测,研究骨修复过程中基因修饰多孔支架诱导血管化的动态过程。体外实验结果显示,慢病毒颗粒从支架上的持续释放长达 5 天,缓释出的病毒颗粒可以有效转染 293T 细胞并表达 PDGF-BB 因子。体内实验结果表明,慢病毒介导基因修饰多孔 PLGA/nHAp 复合支架,可以实现 PDGF-BB 因子原位表达,促进体内局部及系统性干细胞等细胞迁移,加快血管诱导生成并提高骨缺损部位骨组织的再生能力。同时,在该研究中,成功使用并比较了多模态双光子及光 声显微成像技术在体、实时、连续监测 3D 骨组织支架内血管形成的动态变化过程,并验证了基因修饰对于提高 3D 打印支架的生物学反应性的作用。该研究为研究不同支架对血管生成作用的监测与鉴定提供了新的技术手段。     

Microassembly Fabrication of Tissue Engineering Scaffolds With Customized Design

This paper presents a novel technique to fabricate scaffold/cell constructs for tissue engineering by robotic assembly of microscopic building blocks (of volume 0.5$,times,$0.5$,times,$0.2 ${hbox{mm}}^{3}$ and 60 $mu {hbox{m}}$ thickness). In this way, it becomes possible to build scaffolds with freedom in the design of architecture, surface morphology, and chemistry. Biocompatible microparts with complex 3-D shapes were first designed and mass produced using MEMS techniques. Semi-automatic assembly was then realized using a robotic workstation with four degrees of freedom integrating a dedicated microgripper and two optical microscopes. Coarse movement of the gripper is determined by pattern matching in the microscopes images, while the operator controls fine positioning and accurate insertion of the microparts. Successful microassembly was demonstrated using SU-8 and acrylic resin microparts. Taking advantage of parts distortion and adhesion forces, which dominate at micro-level, the parts cleave together after assembly. In contrast to many current scaffold fabrication techniques, no heat, pressure, electrical effect, or toxic chemical reaction is involved, a critical condition for creating scaffolds with biological agents.      

用软刻蚀制备组织工程3D聚合物微结构

软刻蚀制备组织工程3D可生物降解、生物相容性好的几何图案聚合物微结构,方法简单,使用聚二甲基硅氧烷(PDMS)作为弹性铸膜.高分辨率透明掩膜图案用CorelDraw软件绘制并用高分辨率行式打印在塑料片上.制备微结构的聚合物材料为摩尔比85/15的聚乳酸/乙醇酸(PLGA)共聚物,对其进行预处理使之具有适当的硬度和柔韧性.用铸造、旋涂两种成模方法制备形体尺寸为10～100 μm的PLGA肢手架.对影响肢手架结构完整及横向分辨率的因素进行了讨论和优化,并对两种成型方法的横向、纵向分辨率进行了比较.作为脚手架在组织工程上应用的概念验证,用热层压方法制造用于细胞培养的多层聚合物结构.实验结果表明,软刻蚀技术提供了一种实现组织工程微尺度3D聚合物结构的有效工具.     

An approach to integrating shape and biomedical attributes in vascular models

Computational models have been used widely in tissue engineering research and have proven to be powerful tools for bio-mechanical analysis (i.e., blood flow, growth models, drug delivery, etc). This paper focuses on developing higher-fidelity models for vascular structures and blood vessels that integrate computational shape representations with biomedical properties and features. Previous work in computer-aided vascular modeling comes from two communities. For those in biomedical imaging, the goal of past research has been to develop image understanding techniques for the interpretation of x-ray, magnetic resonance imaging (MRI), or other radiological data. These representations are predominantly discrete shape models that are not tied to physiological properties. The other corpus of existing work comes from those interested in developing physiological models for vascular growth and behavior based on bio-medical attributes. These models usually either have a highly simplified shape representation, or lack one entirely. Further, neither of these representations are suitable for the kind of interactive modeling required by tissue engineering applications.This paper aims to bridge these two approaches and develop a set of mathematical tools and algorithms for feature-based representation and computer-aided modeling of vascular trees for use in computer-aided tissue engineering applications. The paper offers a multi-scale representation based on swept volumes and a feature-based representation that can attribute the geometric representation with information about blood flow, pressure, and other biomedical properties. The paper shows how the resulting representation can be used as part of an overall approach for designing and visualizing vascular scaffolds. As a real-world example, we show how this computational model can be used to develop a tissue scaffold for liver tissue engineering. Such scaffolds may prove useful in a number of biomedical applications, including the growth of replacement tissue grafts and in vitro study of the pharmacological affects of new drugs on tissue cultures.     

Microstructure interpolation for macroscopic design

We present a method for multiple length scale structural optimisation. We first optimise isotropic microstructures for maximum bulk modulus at five solid fractions. Shape interpolation between these optimised microstructures produces a continuous set that smoothly varies in both geometry and mechanical properties. This smooth set is used for macroscopic optimisation via the material distribution method. The approach is computationally efficient and the geometric smoothness makes it clear how the microstructures can be transitioned between neighbouring elements. Performance comparisons are made to traditional structural optimisation for some example compliance optimisation problems. The interpolated microstructure designs are most advantageous for two dimensional problems involving multiple loading cases. In these cases, intermediate densities are utilised to more effectively distribute the load. In three dimensions, the method would be useful for a number of applications where specific microstructural requirements, such as a connected pore space, are needed within a multiple-scale design.     

Measurement of Young’s modulus and residual stress of thin SiC layers for MEMS high temperature applications

Silicon carbide (SiC) is a promising material for applications in harsh environments. Standard silicon (Si) microelectromechanical systems (MEMS) are limited in operating temperature to temperatures below 130°C for electronic devices and below 600°C for mechanical devices. Due to its large bandgap SiC enables MEMS with significantly higher operating temperatures. Furthermore, SiC exhibits high chemical stability and thermal conductivity. Young’s modulus and residual stress are important mechanical properties for the design of sophisticated SiC-based MEMS devices. In particular, residual stresses are strongly dependent on the deposition conditions. Literature values for Young’s modulus range from 100 to?400?GPa, and residual stresses range from 98 to?486?MPa. In this paper we present our work on investigating Young’s modulus and residual stress of SiC films deposited on single crystal bulk silicon using bulge testing. This method is based on measurement of pressure-dependent membrane deflection. Polycrystalline as well as single crystal cubic silicon carbide samples are studied. For the samples tested, average Young’s modulus and residual stress measured are 417?GPa and 89?MPa for polycrystalline samples. For single crystal samples, the according values are 388?GPa and 217?MPa. These results compare well with literature values.     

基于落锤装置的体积弹性模量测定方法

体积弹性模量是液压工作系统中传压介质一个重要的物理参数,为求解超高压状态下液压介质体积弹性模量随压力的变化,论证了超高压状态下液压介质采用常量体积弹性模量的不适用性,构建了体积弹性模量修正方法,提出了一套用落锤液压发生装置测定液压介质体积弹性模量与体积相对压缩量之间关系的试验方案并通过了试验验证。验证结果表明,体积弹性模量修正后,压力预测准确度在2.0%以内,在压力外推2.5%的范围内,预报准确度在3.5%以内。测量结果可满足工程应用的要求,对于在超高压状态下或一些特定场合提高压力测量的精度具有工程指导意义。     

Computer-aided design of porous artifacts

Heterogeneous structures represent an important new frontier for 21st century engineering. Human tissues, composites, ‘smart’ and multi-material objects are all physically manifest in the world as three-dimensional (3D) objects with varying surface, internal and volumetric properties and geometries. For instance, a tissue engineered structure, such as bone scaffold for guided tissue regeneration, can be described as a heterogeneous structure consisting of 3D extra-cellular matrices (made from biodegradable material) and seeded donor cells and/or growth factors.The design and fabrication of such heterogeneous structures requires new techniques for solid models to represent 3D heterogeneous objects with complex material properties. This paper presents a representation of model density and porosity based on stochastic geometry. While density has been previously studied in the solid modeling literature, porosity is a relatively new problem. Modeling porosity of bio-materials is critical for developing replacement bone tissues. The paper uses this representation to develop an approach to modeling of porous, heterogeneous materials and provides experimental data to validate the approach. The authors believe that their approach introduces ideas from the stochastic geometry literature to a new set of engineering problems. It is hoped that this paper stimulates researchers to find new opportunities that extend these ideas to be more broadly applicable for other computational geometry, graphics and computer-aided design problems.     

Supporting self‐regulated learning in computer‐based learning environments: systematic review of effects of scaffolding in the domain of science education

Despite the widespread assumption that students require scaffolding support for self‐regulated learning (SRL) processes in computer‐based learning environments (CBLEs), there is little clarity as to which types of scaffolds are most effective. This study offers a literature review covering the various scaffolds that support SRL processes in the domain of science education. Effective scaffolds are categorized and discussed according to the different areas and phases of SRL. The results reveal that most studies on scaffolding processes focus on cognition, whereas few focus on the non‐cognitive areas of SRL. In the field of cognition, prompts appear to be the most effective scaffolds, especially for processes during the control phase. This review also shows that studies have paid little attention to scaffold designs, learner characteristics, or various task characteristics, despite the fact that these variables have been found to have a significant influence. We conclude with the implications of our results on future design and research in the field of SRL using CBLEs.