Kerm Sin Chian

In vitro cell infiltration and in vivo cell infiltration and vascularization in a fibrous, highly porous poly(D,L‐lactide) scaffold fabricated by cryogenic electrospinning technique

One of the obstacles limiting the application of electrospun scaffolds for tissue engineering is the nanoscale pores that inhibit cell infiltration. In this article, we describe a technique that uses ice crystals as templates to fabricate cryogenic electrospun scaffolds (CES) with large three-dimensional and interconnected pores using poly(D,L-lactide) (PLA). Manipulating the humidity of the electrospinning environment the pore sizes are controlled. We are able to achieve pore sizes ranging from 900 +/- 100 microm(2) to 5000 +/- 2000 microm(2) depending on the relative humidity used. Our results show that cells infiltrated the CES up to 50 microm in thickness in vitro under static culture conditions whereas cells did not infiltrate the conventional electrospun scaffolds. In vivo studies demonstrated improved cell infiltration and vascularization in the CES compared with conventionally prepared electrospun scaffolds. In gaining control of the pore characteristics, we can then design CES that are optimized for specific tissue engineering applications.

Development of a rigid polyurethane foam from palm oil

The reactions between polymeric diphenyl methane diisocyanate (polymeric MDI) and conventional polyols to produce foamed polyurethane products are well documented and published. Current polyurethane foams are predominantly produced from these reactions whereby the polyol components are usually obtained from petrochemical processes. This article describes a new development in polyurethane foam technology whereby a renewable source of polyol derived from refined–bleached–deodorized (RBD) palm oil is used to produce polyurethane foams. Using very basic foam formulation, rigid polyurethane foams were produced with carbon dioxide as the blowing agent generated from the reaction between excess polymeric MDI with water. The foams produced from this derivatized RBD palm oil have densities in excess of 200 kg/m3 and with compression strengths greater than 1 MPa.

Effect of electrospun poly(D,L-lactide) fibrous scaffold with nanoporous surface on attachment of porcine esophageal epithelial cells and protein adsorption.

Electrospun scaffolds have been increasingly used in tissue engineering applications due to their size-scale similarities with native extracellular matrices. Their inherent fibrous features may be important in promoting cell attachment and proliferation on the scaffolds. In this study, we explore the technique of fabricating electrospun fibers with nano-sized porous surfaces and investigate their effects on the attachment of porcine esophageal epithelial cells (PEECs). Porosity was introduced in electrospun poly(D,L-lactide) fibers by creating vapor-induced phase separation conditions during electrospinning. The nanoporous fiber scaffolds were mechanically weaker than the conventional solid fiber scaffolds and solvent-cast films of the same polymer. However, the nanoporosity of the fibers was found to enhance the levels of adsorbed protein from a dilute solution of fetal bovine serum. The amount of protein adsorbed by nanoporous fiber scaffolds was approximately 80% higher than the solid fiber scaffolds. This corresponds to an estimated 62% increase in surface area of the porous fibers than the solid fibers. By comparison, the solvent-cast films adsorbed low levels of protein from the FBS solution. In addition, the porous fibers were found to be advantageous in enhancing initial cell attachment as compared with the solid fibers and solvent-cast films. It was observed that nanoporous fiber scaffolds seeded with PEECs had significantly greater number of viable cells attached than the solid fiber scaffolds after 10 and 24 h in culture. Hence, our results indicate that nanosized porous surfaces on electrospun fibers enhance both protein adsorption and cell attachment. These findings provide a method to improve cell-matrix interactions of electrospun scaffolds for tissue engineering applications.

Prediction and verification of process induced warpage of electronic packages

Abstract During the manufacturing, testing and service, thermally induced deformations and stresses will occur in IC devices and packages, which may cause various kinds of product failures. FEM techniques are widely used to predict the thermal deformations and stresses and their evolutions. However, due to the complexity of the real engineering problems, various assumptions and simplifications have to be made in conducting FEM modelling. Therefore, the applicability of the predicted results depend strongly on the reliability and accuracy of the developed FEM-based prediction models which should be verified before applications. In this paper, FEM models are developed to predict the thermal deformations of certain electronic packages and naked die samples under packaging and testing loading. For all the package constituents, appropriate material properties and models are used, including temperature-dependent visco-elasticity, anisotropy, and temperature-dependent elasticity and plasticity. To verify the developed FE models, a series of optical metrology tests are performed. A compact 3D interferometry testing system that can measure simultaneously out-of plane and in-plane deformations has been developed. Thermal deformation measurements are performed on samples of both real electronic packages and naked dies attached on a leadframe. Identical deformation patterns were found for the measured fringe patterns in the U -, V -, and W -fields and the simulated ones. Also, quantitatively, the maximum deformation mismatch between the predicted and tested results is within 15%. It is concluded that the thermally induced deformations predicted by the non-linear FEM models match well with measured deformations for both the naked die and the real packages.

Esophageal Tissue Engineering: An In-Depth Review on Scaffold Design

Treatment of esophageal cancer often requires surgical procedures that involve removal. The current approaches to restore esophageal continuity however, are known to have limitations which may not result in full functional recovery. In theory, using a tissue engineered esophagus developed from the patients own cells to replace the removed esophageal segment can be the ideal method of reconstruction. One of the key elements involved in the tissue engineering process is the scaffold which acts as a template for organization of cells and tissue development. While a number of scaffolds range from traditional non‐biodegradable tubing to bioactive decellularized matrix have been proposed to engineer the esophagus in the past decade, results are still not yet favorable with many challenges relating to tissue quality need to be met improvements. The success of new esophageal tissue formation will ultimately depend on the success of the scaffold being able to meet the essential requirements specific to the esophageal tissue. Here, the design of the scaffold and its fabrication approaches are reviewed. In this paper, we review the current state of development in bioengineering the esophagus with particular emphasis on scaffold design. Biotechnol. Bioeng. 2012;109: 1–15.

Fabrication and in vitro and in vivo cell infiltration study of a bilayered cryogenic electrospun poly(D,L-lactide) scaffold

Cryogenic electrospinning has previously been demonstrated for controlling the pore sizes of electrospun scaffolds, which has been impossible with traditional electrospinning processes. This article describes the application of the cryogenic technique to fabricate a bilayered electrospun poly(D,L-lactide) scaffold (BLES) in a single uninterrupted process. The resulting BLES consisted of a traditional electrospun (ES) fibrous layer with a dense pore area of 17 +/- 3 microm(2) adjacent to a cryogenic electrospun layer (CES) with a pore area of 3300 +/- 500 microm(2). The significance of this bilayered scaffold was to mimic the anatomical structure of tissues with dense basement membrane followed by loose and highly porous connective tissue such as skin and blood vessels. Cell infiltration in the BLES was compared in vitro and in vivo. Both studies suggested the CES supported high cell infiltration, whereas the ES could serve as a physical barrier to prevent cell infiltration across the CES-ES boundary because of its size exclusion. The bilayered structure produced by this technique suggests a great potential for engineering tissues with similar architectures.

Effects of CO2 Laser Irradiation on the Surface Properties of Magnesia-Partially Stabilised Zirconia (MgO-PSZ) Bioceramic and the Subsequent Improvements in Human Osteoblast Cell Adhesion

In order to acquire the surface properties favouring osseo-integration at the implant and bone interface, human foetal osteoblast cells (hFOB) were used in an in vitro test to examine changes in cell adhesion on a magnesia-partially stabilised zirconia (MgO-PSZ) bioceramic after CO2 laser treatment. The surface roughness, microstructure, crystal size and surface energy of untreated and CO2 laser-treated MgO-PSZ were fully characterised. The in vitro cell evaluation revealed a more favourable cell response on the CO2 laser-treated MgO-PSZ than on the untreated sample. After 24-h cell incubation, no cell was observed on the MgO-PSZ, whereas a few cells attached on the CO2 laser-treatedMgO-PSZandshowedwellspreadandgood attachment. Moreover, the cell coverage density indicating cell proliferation generally increases with CO2 laser power densities applied in the experiments. The enhancement of the surface energy of the MgO-PSZ, especially its polar component caused by the CO2 laser treatment, was found to play a significant role in the initial cell attaching, thus enhancing the cell growth. Moreover, the change in topography induced by the CO2 laser treatment was identified as one of the factors influencing the hFOB cell response.

Continuous Cell Separation Using Dielectrophoresis through Asymmetric and Periodic Microelectrode Array

The study presents a dielectrophoretic cell separation method via three-dimensional (3D) nonuniform electric fields generated by employing a periodic array of discrete but locally asymmetric triangular bottom microelectrodes and a continuous top electrode. Traversing through the microelectrodes, heterogeneous cells are electrically polarized to experience different strengths of positive dielectrophoretic forces, in response to the 3D nonuniform electric fields. The cells that experience stronger positive dielectrophoresis are streamed further in the perpendicular direction to the fluid flow, leaving the cells that experience weak positive dielectrophoresis, which continue to traverse the microelectrode array essentially along the laminar flow streamlines. The proposed method has achieved 87.3% pure live cells harvesting efficiency from a live/dead NIH-3T3 cells mixture, and separation of MG-63 cells from erythrocytes with a separation efficiency of 82.8%. The demonstrated cell separation shows promising applications of the DEP separator for cell separation in a continuous mode.

Rapid freeze prototyping technique in bio‐plotters for tissue scaffold fabrication

Purpose – The purpose of this paper is to develop a new bio‐plotter using a rapid freeze prototyping (RFP) technique and to investigate its potential applications in fabricating tissue scaffolds.Design/methodology/approach – The development of cryogenic bio‐plotters including design steps of hardware as well as software is addressed. Effects of structural parameters and process parameters on the properties of tissue scaffolds are demonstrated through simulation and experimental results.Findings – The paper finds that the RFP method is suitable to fabricate macro‐ and micro‐porous scaffolds, especially for temperature‐sensitive polymers. In addition, through simulation and experiment results, it also shows that macro‐ and micro‐porous properties could be manipulated by structural parameters and process parameters, respectively.Research limitations/implications – This paper shows that the chamber temperature is an important process parameter that can provide the means to control the micro‐porous structure o...

Dependence of alignment direction on magnitude of strain in esophageal smooth muscle cells

The response of cells in vitro to mechanical forces has been the subject of much research using devices to exert controlled mechanical stimulation on cultured cells or isolated tissue. In this study, esophageal smooth muscle cells were seeded on flexible polyurethane membranes to form a confluent cell layer. The cells were then subjected to uniform cyclic stretch of varying magnitudes at a frequency of approximately five cycles per minute in a custom made mechatronic bioreactor, providing similar strains experienced in the in vivo mechanical environment of the esophagus. The results show that the orientation response is dependent on the magnitude of cyclic stretch applied. Smooth muscle cells showed parallel alignment to the force direction at low cyclic strains (2%) compared to the hill‐valley morphology of static controls. At higher strains (5% and 10% magnitude), the cells exhibited a consistent alignment perpendicular to the strain. To our knowledge, this is the first time that the alignment directions dependence on strain magnitude has been demonstrated. MTS analysis indicated that cell metabolism was reduced when mechanical strain was applied, and proliferation was inhibited by mechanical strain. Protein expression indicates a decrease in smooth muscle α‐actin, indicative of changes in cell phenotype, an increase in vimentin, which is associated with increased cell motility, and an increase in desmin, indicating differentiation in stimulated cells. Biotechnol. Bioeng. 2009;102: 1703–1711.