Eleftherios Sachlos

Novel collagen scaffolds with predefined internal morphology made by solid freeform fabrication

Novel collagen scaffolds possessing predefined and reproducible internal channels with widths of 135 microm and greater have been produced. The process employed to make the collagen scaffold utilises a sacrificial mould, manufactured using solid freeform fabrication technology, and critical point drying technique. A computer aided design (CAD) file of the mould to be produced is created. This mould is manufactured using a phase change ink-jet printer. A dispersion of collagen is then cast into the mould and frozen. The mould is dissolved away with ethanol and the collagen scaffold is then critical point dried with liquid carbon dioxide. The effect of processing on the tertiary structure of collagen is assessed by monitoring the wavenumber of the N-H stretching vibration peak using Fourier transform infra-red spectroscopy and it is found that processing does not denature the collagen. Ultraviolet-visual spectroscopy was used to detect the presence of any contamination from the sacrificial mould on the collagen. The ability to use computer aided design and manufacture (CAD/CAM) provides a route to optimise scaffold designs using collagen in tissue engineering applications.

The impact of critical point drying with liquid carbon dioxide on collagen–hydroxyapatite composite scaffolds

Collagen-hydroxyapatite composites for bone tissue engineering are usually made by freezing an aqueous dispersion of these components and then freeze-drying. This method creates a foamed matrix which may not be optimum for growing cell colonies larger than a few hundred micrometres due to the limited diffusion of nutrients and oxygen, and the limited removal of waste metabolites. Incorporating a network of microchannels in the interior of the scaffold which may permit the flow of nutrient-rich media has been proposed as a method to overcome these diffusion constraints. A novel three-dimensional printing and critical point drying technique previously used to make collagen scaffolds has been modified to create collagen-hydroxyapatite scaffolds. This study investigates the properties of collagen and collagen-hydroxyapatite scaffolds and whether subjecting collagen and hydroxyapatite to critical point drying with liquid carbon dioxide results in any changes to the individual components. Specifically, the hydroxyapatite component was characterized before and after processing using wavelength-dispersive X-ray spectroscopy, X-ray diffraction and infrared spectroscopy. Critical point drying did not induce elemental, crystallographic or molecular changes in the hydroxyapatite. The quaternary structure of collagen was characterized using transmission electron microscopy and the quarter-staggered array characteristic of native collagen remained after processing. Microstructural characterization of the composites using scanning electron microscopy showed the hydroxyapatite particles were mechanically interlocked in the collagen matrix. The in vitro biological response of MG63 osteogenic cells to the composite scaffolds were characterized using the Alamar Blue, PicoGreen, alkaline phosphate and Live/Dead assays, and revealed that the critical point dried scaffolds were non-cytotoxic.

On the manufacturability of scaffold mould using a 3D printing technology

Purpose – To investigate the effect of operation parameters and printing configuration on the manufacturability of moulds in the manufacture of tissue engineering scaffolds using a 3D printing system.Design/methodology/approach – The scaffold moulds were built using proprietary biocompatible materials using a modified Solidscape T66 ink‐jet printing system. The manufacturability of biological scaffold moulds has been investigated in terms of resolution, accuracy, and minimum and maximum manufacturable features.Findings – The results demonstrated that the 3D system used in this study is able to fabricate structures with high reproducibility and flexibility. It was found that thermal degradation of BioSupport material had an adverse effect on resolution and accuracy of moulds printed for scaffold manufacturing. The maximum features, including maximum length and height, are geometrical dimension and orientation dependent. The system could produce a longer and higher features when the mould was aligned perpen...

A Process to Make Collagen Scaffolds with an Artificial Circulatory System Using Rapid Prototyping

Abstract : Tissue engineering aims to produce biological substitutes to restore or repair damaged human tissues or organs. The principle strategy behind tissue engineering involves seeding relevant cell(s) onto porous 3D biodegradable scaffolds. The scaffold acts as a temporary substrate where the cells can attach and then proliferate and differentiate. Collagen is the major protein constituent of the extracellular matrix in the human body and therefore an attractive scaffold material. Current collagen scaffolds are foams which limit the mass transport of oxygen and nutrients deep into the scaffold, and consequently cannot support the growth of thick-cross sections of tissue (greater than 500 micrometers). We have developed a novel process to make collagen and collagen-hydroxyapatite scaffolds containing an internal artificial circulatory system in the form of branching channels using a sacrificial mould, casting and critical point drying technique. The mould is made using a commercial rapid prototyping system, the Model-Maker II, and is designed to possess a series of connected shafts. The mould is dissolved away and the solvent itself removed by critical point drying with liquid carbon dioxide. Processed hydroxyapatite has been characterised by XRD and FTIR analysis. Tissue engineering with collagen scaffolds possessing controlled internal microarchitecture may be the key to growing thick cross-sections of human tissue.

Characterisation of Collagen Scaffolds Using X-ray Microtomography

Abstract : Collagen scaffolds have been produced that incorporate predefined internal channels. The scaffolds were obtained with the aid of sacrificial moulds that have been manufactured using a rapid prototyping technique. A computer aided design file of the mould was created and then realised using an ink-jet printer. A dispersion of collagen was then cast into the mould and frozen. Ethanol was used to dissolve the mould leaving the collagen which was then freeze dried to produce the final product. The scaffold was then analysed using X-ray microtomography (XMT) to determine whether the desired internal structure was obtained. It was found necessary to add saturated potassium iodide (KI) solution to the scaffold in order to analyses it satisfactorily by XMT. The resultant images indicate that the desired internal structure was obtained.