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Thomas Boland

Publications and source records attributed to Thomas Boland.

15 recordsLinked to original sources

The role of independently variable grafting density and layer thickness of polymer nanolayers on peptide adsorption and cell adhesion.

This contribution demonstrates a simple and reproducible method for fabricating surface-tethered polymer brushes that vary in grafting density and layer thickness for peptide adsorption and cell-adhesion studies. Surface-initiated atom transfer radical polymerization was used together with thiol self-assembly to generate these nanothin polymer brush layers of poly((polyethylene glycol) methacrylate). A kinetic study was done to measure the layer thickness growth rate at room temperature from flat gold substrates presenting different polymerization initiator molecule surface densities. The polymer brush layers transition from mushroom to brush regimes with increasing grafting density. A crossover density of 0.038+/-0.005 chains/nm(2) was determined for the PPEGMA polymer brushes. The results described in this paper show that layer properties such as wettability and dry layer thickness depend strongly on initiator surface density. Ultimately, the adsorbed concentration of an RGD-containing synthetic peptide Gly-Arg-Gly-Asp-Ser and the adhesion and spreading of cells were correlated with surface properties, which continues to be a major research theme in biomedical and biomaterials research.

3T3 Cells↗

Viability and electrophysiology of neural cell structures generated by the inkjet printing method.

Complex cellular patterns and structures were created by automated and direct inkjet printing of primary embryonic hippocampal and cortical neurons. Immunostaining analysis and whole-cell patch-clamp recordings showed that embryonic hippocampal and cortical neurons maintained basic cellular properties and functions, including normal, healthy neuronal phenotypes and electrophysiological characteristics, after being printed through thermal inkjet nozzles. In addition, in this study a new method was developed to create 3D cellular structures: sheets of neural cells were layered on each other (layer-by-layer process) by alternate inkjet printing of NT2 cells and fibrin gels. These results and findings, taken together, show that inkjet printing is rapidly evolving into a digital fabrication method to build functional neural structures that may eventually find applications in neural tissue engineering.

Animals↗

Application of inkjet printing to tissue engineering.

Recent advances in organ printing technology for applications relating to medical interventions and organ replacement are described. Organ printing refers to the placement of various cell types into a soft scaffold fabricated according to a computer-aided design template using a single device. Computer aided scaffold topology design has recently gained attention as a viable option to achieve function and mass transport requirements within tissue engineering scaffolds. An exciting advance pioneered in our laboratory is that of simultaneous printing of cells and biomaterials, which allows precise placement of cells and proteins within 3-D hydrogel structures. This advance raises the possibility of spatially controlling not only the scaffold structure, but also the type of tissue that can be grown within the scaffold and the thickness of the tissue as capillaries and vessels could be constructed within the scaffolds. Here we summarize recent advances in printing cells and materials using the same device.

Animals↗

Inkjet printing of viable mammalian cells.

The purpose of this study was to explore the use of a commercial thermal printer to deposit Chinese Hamster Ovary (CHO) and embryonic motoneuron cells into pre-defined patterns. These experiments were undertaken to verify the biocompatibility of thermal inkjet printing of mammalian cells and the ability to assemble them into viable constructs. Using a modified Hewlett Packard (HP) 550C computer printer and an HP 51626a ink cartridge, CHO cells and rat embryonic motoneurons were suspended separately in a concentrated phosphate buffered saline solution (3 x). The cells were subsequently printed as a kind of "ink" onto several "bio-papers" made from soy agar and collagen gel. The appearance of the CHO cells and motoneurons on the bio-papers indicated an healthy cell morphology. Furthermore, the analyses of the CHO cell viability showed that less than 8% of the cells were lysed during printing. These data indicate that mammalian cells can be effectively delivered by a modified thermal inkjet printer onto biological substrates and that they retain their ability to function. The computer-aided inkjet printing of viable mammalian cells holds potential for creating living tissue analogs, and may eventually lead to the construction of engineered human organs.

Animals↗

Construction of high-density bacterial colony arrays and patterns by the ink-jet method.

We have developed a method for fabricating bacterial colony arrays and complex patterns using commercially available ink-jet printers. Bacterial colony arrays with a density of 100 colonies/cm(2) were obtained by directly ejecting Escherichia coli (E. coli) onto agar-coated substrates at a rapid arraying speed of 880 spots per second. Adjusting the concentration of bacterial suspensions allowed single colonies of viable bacteria to be obtained. In addition, complex patterns of viable bacteria as well as bacteria density gradients were constructed using desktop printers controlled by a simple software program.

Biological Assay↗

Experimental and numerical modeling of variable friction between nanoregions in coventional and crosslinked UHMWPE.

Recently, highly crosslinked UHMWPE components have been promoted for their high abrasive wear resistance over conventional UHMWPE (PE) in total joint replacement (TJR) prostheses to minimize osteolysis and consequent implant loosening. This study was aimed at investigating the role of friction gradients induced by localized coefficients of friction at both crystalline and amorphous nanoregions in PE, and crystalline and crosslinked nanoregions in crosslinked UHMWPE (XPE), in submicron wear debris generation. An abrasive wear study performed on both XPE and PE using atomic force microscopy (AFM) illustrated that the onset of plastic deformation for XPE occurred at a normal load that was approximately 3 times higher when compared to PE. Coefficients of friction (mu d) of 0.2, 0.35, and 0.61, experimentally derived using AFM, were used as representative mu d for crystalline, amorphous, and crosslinked nanoregions, respectively, in a numerical Hertzian model. An increase in mu (0.2 +/- 0.02, 0.35 +/- 0.01 and 0.6 +/- 0.04) was observed with a decrease in crystallinity and storage modulus at 22 degrees C. Using the Hertzian contact model, it was observed that variability in friction between nanoregions contributed to higher magnitude stresses for XPE (0.2 to 0.61; maximum sigma eff = 2.8) compared to PE (0.2 to 0.35; maximum sigma eff = 1.1) over a negligible thickness of the interfacial zone (IZ) between nanoregions. The experimentally observed increase in abrasive wear resistance of XPE could be attributed to an increase in the thickness of the interfacial zone between nanoregions with mu changing gradually from crystalline to crosslinked nanoregions, a situation that may not be observed with PE. This would cause a decrease in the friction gradient and resulting stresses thereby agreeing with the observed experimental higher abrasive wear resistance for XPE. However, in both PE and XPE, the presence of stress concentrations over a period of time could lead to irreversible damage of the material eventually generating submicron wear debris. Hence, semicrystalline, inhomogenous UHMWPE with several nanoregions (amorphous and crystalline) would be at a disadvantage for bearing application in terms of abrasive wear resistance compared to UHMWPE with relatively lower number of nanoregions and crosslinked nanoregions.

Biocompatible Materials↗

A quantitative approach to studying structures and orientation at self-assembled monolayer/fluid interfaces.

Self-assembled monolayers (SAMs) have become a standard tool for exploring surface interactions. Although well characterized, SAMs are known to undergo structural and conformational changes in the presence of solution, yet the ability to quantify these changes remains an obstacle due to limited analytical techniques. In this study, we determine changes in structure and conformation of CH3, OH, and COOH terminated hexadecanethiols on gold in water by means of a new technique known as evanescence reflection spectroscopy. This FTIR application, in conjunction with a semiempirical formalism, is capable of providing both qualitative and quantitative understanding of the molecular structure and orientation at the solid/liquid interface.

Journal Article↗

Cell and organ printing 1: protein and cell printers.

We have developed several devices for positioning organic molecules, molecular aggregates, cells, and single-cell organisms onto solid supports. These printers can create stable, functional protein arrays using an inexpensive technology. The cell printer allows us to create cell libraries as well as cellular assemblies that mimic their respective position in organs. The printers are derived from commercially available ink-jet printers that are modified to dispense protein or cell solutions instead of ink. We describe here the modifications to the print heads, and the printer hardware and software that enabled us to adapt the ink-jet printers for the manufacture of cell and protein arrays. The printers have the advantage of being fully automated and computer controlled, and allow for the high-throughput manufacture of protein and cell arrays.

Animals↗

Cell and organ printing 2: fusion of cell aggregates in three-dimensional gels.

We recently developed a cell printer (Wilson and Boland, 2003) that enables us to place cells in positions that mimic their respective positions in organs. However, this technology was limited to the printing of two-dimensional (2D) tissue constructs. Here we describe the use of thermosensitive gels to generate sequential layers for cell printing. The ability to drop cells on previously printed successive layers provides a real opportunity for the realization of three-dimensional (3D) organ printing. Organ printing will allow us to print complex 3D organs with computer-controlled, exact placing of different cell types, by a process that can be completed in several minutes. To demonstrate the feasibility of this novel technology, we showed that cell aggregates can be placed in the sequential layers of 3D gels close enough for fusion to occur. We estimated the optimum minimal thickness of the gel that can be reproducibly generated by dropping the liquid at room temperature onto a heated substrate. Then we generated cell aggregates with the corresponding (to the minimal thickness of the gel) size to ensure a direct contact between printed cell aggregates during sequential printing cycles. Finally, we demonstrated that these closely-placed cell aggregates could fuse in two types of thermosensitive 3D gels. Taken together, these data strongly support the feasibility of the proposed novel organ-printing technology.

Animals↗

Frictional properties of poly(MPC-co-BMA) phospholipid polymer for catheter applications.

A fundamental understanding of surface properties of the biomaterials at a nanometer scale should be generated in order to understand cellular responses of the tissue to biomaterials thereby minimizing or eliminating tissue trauma at a macrometer scale. In this study poly(2-methacryloyloxyethyl phosphorylcholine-co-n-butyl methacrylate) ([poly(MPC-co-BMA]) was evaluated as a potential coating material for vascular applications to provide smooth catheterization using atomic force microscopy (AFM) techniques.A uniform coating of [poly(MPC-co-BMA] equivalent to a thickness of 2.5 microm on a polyurethane (PU) catheter material was provided using dip casting technique. Using a contact mode AFM, no significant difference in surface roughness (R(a)) and frictional force (f) between uncoated (R(a)=10.2+/-1.9 nm, f=0.907+/-0.02) and coated (R(a)=11.7+/-1.8 nm, f=0.930+/-0.06) surfaces was observed under dry conditions. However, under wet conditions the R(a) of the coated surface (3.4+/-1.0 nm) was significantly lower than uncoated PU surface (9.0+/-1.8 nm). The coating on PU substrate offered the least frictional resistance (f=0.004+/-0.001) illustrating enhanced boundary lubrication capability due to hydration of phosphorylcholine polymer as compared to a significantly higher f for uncoated PU (0.017+/-0.007) surfaces. These tribological and chemical characteristics of the [poly(MPC-co-BMA)] coating could increase the overall efficacy of PU for clinical applications.

Catheterization↗

Organ printing: computer-aided jet-based 3D tissue engineering.

Tissue engineering technology promises to solve the organ transplantation crisis. However, assembly of vascularized 3D soft organs remains a big challenge. Organ printing, which we define as computer-aided, jet-based 3D tissue-engineering of living human organs, offers a possible solution. Organ printing involves three sequential steps: pre-processing or development of "blueprints" for organs; processing or actual organ printing; and postprocessing or organ conditioning and accelerated organ maturation. A cell printer that can print gels, single cells and cell aggregates has been developed. Layer-by-layer sequentially placed and solidified thin layers of a thermo-reversible gel could serve as "printing paper". Combination of an engineering approach with the developmental biology concept of embryonic tissue fluidity enables the creation of a new rapid prototyping 3D organ printing technology, which will dramatically accelerate and optimize tissue and organ assembly.

Artificial Organs↗

In vitro evaluation of phosphonylated low-density polyethylene for vascular applications.

The use of catheters for vascular applications is often complicated by the development of friction between the catheter material and the vessel wall, which leads to endothelial cell removal and intimal lesions. Phosphonylation, a chemical surface treatment, has been proposed as a means of increasing the hydrophilicity of low-density polyethylene (LDPE), a commonly used catheter material, in efforts to impart lubricity to the material and reduce vascular tissue damage. In an in vitro tribological study, phosphonylated LDPE produced a lower coefficient of friction and allowed greater retention of endothelial cells on vessels as compared to untreated LDPE when the materials were reciprocated against normal porcine aorta. Chemical characterizations of the LDPE before and after friction testing involving Fourier transform infrared and energy-dispersive X-ray (EDX) confirmed the phosphorus content on phosphonylated LDPE. Election spectroscopy for chemical analysis (ESCA) and atomic force micrscope (AFM) analyses verified that proteins initially adsorb to both the phosphonylated and untreated LDPE surfaces and that the proteins interfere with water to lubricate the surfaces. However, with repeated friction, proteins are removed from the surface and hydrophilicity, as imparted by phosphonylation, becomes a principal factor in the lubrication process.

Animals↗

Rapid prototyping of tissue-engineering constructs, using photopolymerizable hydrogels and stereolithography.

One of the most important aspects of tissue engineering is the design of the scaffold providing the mechanical strength and access to nutrients for the new tissue. For customized tissue engineering, it is essential to be able to fabricate three-dimensional scaffolds of various geometric shapes, in order to repair defects caused by accidents, surgery, or birth. Rapid prototyping or solid free-form fabrication (SFF) techniques hold great promise for designing three-dimensional customized scaffolds, yet traditional cell-seeding techniques may not provide enough cell mass for larger constructs. This article presents a novel attempt to fabricate three-dimensional scaffolds, using hydrogels combined with cell encapsulation to fabricate high-density tissue constructs. A commercially available stereolithography technique was applied to fabricate scaffolds using poly(ethylene oxide) and poly(ethylene glycol)dimethacrylate photopolymerizable hydrogels. Mechanical characterization shows the constructs to be comparable with soft tissues in terms of elasticity. High cell viability was achieved and high-density constructs fabricated.

Animals↗