Nazia Mehrban

Effect of cold-setting calcium- and magnesium phosphate matrices on protein expression in osteoblastic cells

Bone loss due to accidents or tissue diseases requires replacement of the structure by either autografts, allografts, or artificial materials. Reactive cements, which are based on calcium phosphate chemistry, are commonly used in nonload bearing areas such as the craniofacial region. Some of these materials are resorbed by the host under physiological conditions and replaced by bone. The aim of this study was to test different calcium and magnesium cement composites in vitro for their use as bone substitution material. Phase composition of calcium deficient hydroxyapatite (Ca(9) (PO(4) )(5) HPO(4) OH), brushite (CaHPO(4) ·2H(2) O), and struvite (MgNH(4) PO(4) ·6H(2) O) specimens has been determined by means of X-ray diffraction, and compressive strength was measured. Cell growth and activity of osteoblastic cells (MG 63) on the different surfaces was determined, and the expression of bone marker proteins was analyzed by western blotting. Cell activity normalized to cell number revealed higher activity of the osteoblasts on brushite and struvite when compared to hydroxyapatite and also the expression of osteoblastic marker proteins was highest on brushite scaffolds. While brushite sets under acidic conditions, formation of struvite occurs under physiological pH, similar to hydroxyapatite cements, providing the possibility of additional modifications with proteins or other active components.

3D bioprinting for tissue engineering: Stem cells in hydrogels

Surgical limitations require alternative methods of repairing and replacing diseased and damaged tissue. Regenerative medicine is a growing area of research with engineered tissues already being used successfully in patients. However, the demand for such tissues greatly outweighs the supply and a fast and accurate method of production is still required. 3D bioprinting offers precision control as well as the ability to incorporate biological cues and cells directly into the material as it is being fabricated. Having precise control over scaffold morphology and chemistry is a significant step towards controlling cellular behaviour, particularly where undifferentiated cells, i.e., stem cells, are used. This level of control in the early stages of tissue development is crucial in building more complex systems that morphologically and functionally mimic in vivo tissue. Here we review 3D printing hydrogel materials for tissue engineering purposes and the incorporation of cells within them. Hydrogels are ideal materials for cell culture. They are structurally similar to native extracellular matrix, have a high nutrient retention capacity, allow cells to migrate and can be formed under mild conditions. The techniques used to produce these materials, as well as their benefits and limitations, are outlined.

Effect of plasma surface modification on the biocompatibility of UHMWPE.

In this paper active screen plasma nitriding (ASPN) is used to chemically modify the surface of UHMWPE. This is an unexplored and new area of research. ASPN allows the homogeneous treatment of any shape or surface at low temperature; therefore, it was thought that ASPN would be an effective technique to modify organic polymer surfaces. ASPN experiments were carried out at 120 °C using a dc plasma nitriding unit with a 25% N(2) and 75% H(2) atmosphere at 2.5 mbar of pressure. UHMWPE samples treated for different time periods were characterized by nanoindentation, FTIR, XPS, interferometry and SEM. A 3T3 fibroblast cell line was used for in vitro cell culture experiments. Nanoindentation of UHMWPE showed that hardness and elastic modulus increased with ASPN treatment compared to the untreated material. FTIR spectra did not show significant differences between the untreated and treated samples; however, some changes were observed at 30 min of treatment in the range of 1500-1700 cm(-1) associated mainly with the presence of N-H groups. XPS studies showed that nitrogen was present on the surface and its amount increased with treatment time. Interferometry showed that no significant changes were observed on the surfaces after the treatment. Finally, cell culture experiments and SEM showed that fibroblasts attached and proliferated to a greater extent on the plasma-treated surfaces leading to the conclusion that ASPN surface treatment can potentially significantly improve the biocompatibility behaviour of polymeric materials.

Spherical indentation analysis of stress relaxation for thin film viscoelastic materials

The mechanical testing of thin layers of soft materials is an important but difficult task. Spherical indentation provides a convenient method to ascertain material properties whilst minimising damage to the material by allowing testing to take place in situ. However, measurement of the viscoelastic properties of these soft materials is hindered by the absence of a convenient yet accurate model which takes into account the thickness of the material and the effects of the underlying substrate. To this end, the spherical indentation of a thin layer of viscoelastic solid material is analysed. It is assumed that the transient mechanical properties of the material can be described by the generalised standard linear solid model. This model is incorporated into the theory and then solved for the special case of a stress relaxation experiment taking into account the finite ramp time experienced in real experiments. An expression for the force as a function of the viscoelastic properties, layer thickness and indentation depth is given. The theory is then fitted to experimental data for the spherical indentation of poly(dimethyl)siloxane mixed with its curing agent to the ratios of 5:1, 10:1 and 20:1 in order to ascertain its transient shear moduli and relaxation time constants. It is shown that the theory correctly accounts for the effect of the underlying substrate and allows for the accurate measurement of the viscoelastic properties of thin layers of soft materials.

Comparing physicochemical properties of printed and hand cast biocements designed for ligament replacement

Abstract Abstract In order to combat the low regenerative capabilities of ligaments, full ‘bone to bone’ replacements are required, which will integrate with bone while providing a smooth transition to the replacement soft tissue (tissues surrounding organs in the body, not being bone). This study investigated the use of three-dimensional powder printing technology to form calcium phosphate brackets, previously used for forming bespoke scaffold geometries, to 95±0·1% accuracy of their original computer aided design. The surface and internal structures of the printed samples were characterised both chemically and morphologically and compared with hand moulded cements in the dry state and after 3 days of immersion in phosphate buffered saline. X-ray diffraction, Raman spectroscopy and SEM all showed the presence of brushite in the hand moulded samples and brushite and monetite within the printed samples. Furthermore, the printed structures have a higher level of porosity in the dry state in comparison to the hand moulded samples (36±2·2% compared to 24±0·7%) despite exhibiting a compressive strength of almost double the hand cast material. Although the compressive strength of the printed cements decreases after the 3 day immersion, there was no significant difference between the printed and hand moulded cements under the same conditions. Three-dimensional powder printing technology has enabled the manufacture of bespoke calcium phosphate brackets with properties similar to those reported for hand moulded cements.

Structural changes to resorbable calcium phosphate bioceramic aged in vitro.

This work investigates the effect of mammalian cell culture conditions on 3D printed calcium phosphate scaffolds. The purpose of the studies presented was to characterise the changes in scaffold properties in physiologically relevant conditions. Differences in crystal morphologies were observed between foetal bovine serum-supplemented media and their unsupplemented analogues, but not for supplemented media containing tenocytes. Scaffold porosity was found to increase for all conditions studied, except for tenocyte-seeded scaffolds. The presence of tenocytes on the scaffold surface inhibited any increase in scaffold porosity in the presence of extracellular matrix secreted by the tenocytes. For acellular conditions the presence or absence of sera proteins strongly affected the rate of dissolution and the distribution of pore diameters within the scaffold. Exposure to high sera protein concentrations led to the development of significant numbers of sub-micron pores, which was otherwise not observed. The implication of these results for cell culture research employing calcium phosphate scaffolds is discussed.

A Comparative Study of Iota Carrageenan, Kappa Carrageenan and Alginate as Tissue Engineering Scaffolds

The increasingly aging population and lack of donor tissue has pushed tissue regeneration to the forefront of modern biomedical research and biopolymers traditionally found in the food and pharmaceutical industry are increasingly being used in tissue engineering. The development of tissue engineering provides the opportunity to take a sample of cells from a patient which are then cultured in vitro to organise into functional tissue that can then be implanted back into the patient, ultimately, overcoming problems of tissue rejection and the need for donor tissue. To grow replacement tissue requires a suitable substrate or scaffold for cells to attach, proliferate and organise into viable tissue that can be safely implanted into the body. Biopolymers and hydrogel forming biopolymers in particular, have been shown to be a promising scaffold material due having properties that resemble the environment of the mammalian extracellular matrix (ECM). In addition properties such as mild gelation conditions, potential for good mass transport of nutrients and waste molecules, nontoxic nature and biological compatibility provide an advantage over synthetic polymers and ceramic and metallic materials which have also been used as tissue engineering scaffolds.

Silsesquioxane polymer as a potential scaffold for laryngeal reconstruction

Cancer, disease and trauma to the larynx and their treatment can lead to permanent loss of structures critical to voice, breathing and swallowing. Engineered partial or total laryngeal replacements would need to match the ambitious specifications of replicating functionality, outer biocompatibility, and permissiveness for an inner mucosal lining. Here we present porous polyhedral oligomeric silsesquioxane-poly(carbonate urea) urethane (POSS-PCUU) as a potential scaffold for engineering laryngeal tissue. Specifically, we employ a precipitation and porogen leaching technique for manufacturing the polymer. The polymer is chemically consistent across all sample types and produces a foam-like scaffold with two distinct topographies and an internal structure composed of nano- and micro-pores. While the highly porous internal structure of the scaffold contributes to the complex tensile behaviour of the polymer, the surface of the scaffold remains largely non-porous. The low number of pores minimise access for cells, although primary fibroblasts and epithelial cells do attach and proliferate on the polymer surface. Our data show that with a change in manufacturing protocol to produce porous polymer surfaces, POSS-PCUU may be a potential candidate for overcoming some of the limitations associated with laryngeal reconstruction and regeneration.

Facile synthesis of novel hybrid POSS biomolecules via “Click” reactions

A novel alkyne-terminated cubic-octameric POSS was synthesised in high yield (82–90%). The X-ray crystal structure revealed intra- and intermolecular hydrogen bonding between the amide groups of the arms. Hybrid biomaterials were synthesised in nearly quantitative yields via a click reaction with (i) azido-N-Fmoc-norleucine and (ii) 3′-azido-3′-deoxythymidine.

Monitoring biomineralization of biomaterials in vivo

In this chapter we have sought to provide readers with an overview of in vivo techniques currently used for the imaging of biomineralisation materials. To achieve this we have provided a background of what biomineralisation entails and why there is a need for biomaterials. Furthermore, we have briefly reviewed some of the in vitro techniques used for the basic understanding of mineralisation events and the prediction of how the materials may behave in vivo. For the latter we have attempted to guide the reader through how the techniques work and how they are used to monitor the biomineralisation process, with focus on integration of the implanted material with its surroundings. The technologies described here are constantly being updated, guided by the complexity in monitoring needs. We therefore advise the reader to update this list regularly.