A study of the relationship between process conditions and mechanical strength of mineralized red algae in the preparation of a marine-derived bone void filler

Bone void fillers that can enhance biological function to augment skeletal repair have significant therapeutic potential in bone replacement surgery. This work focuses on the development of a unique microporous (0.5-10 mu m) marine-derived calcium phosphate bioceramic granule. It was prepared fro Corallina officinalis, a mineralized red alga, using a novel manufacturing process. This involved thermal processing, followed by a low pressure-temperature chemical synthesis reaction. The study found that the ability to maintain the unique algal morphology was dependent on the thermal processing conditions. This study investigates the effect of thermal heat treatment on the physiochemical properties of the alga. Thermogravimetric analysis was used to monitor its thermal decomposition. The resultant thermograms indicated the presence of a residual organic phase at temperatures below 500 degrees C and an irreversible solid-state phase transition from mg-rich-calcite to calcium oxide at temperatures over 850 degrees C. Algae and synthetic calcite were evaluated following heat treatment in an air-circulating furance at temperatures ranging from 400 to 800 degrees C. The highest levels of mass loss occurred between 400-500 degrees C and 700-800 degrees C, which were attributed to the organic and carbonate decomposition respectively. The changes in mechanical strength were quantified using a simple mechanical test, which measured the bulk compressive strength of the algae. The mechanical test used may provide a useful evaluation of the compressive properties of similar bone void fillers that are in granular form. The study concluded that soak temperatures in the range of 600 to 700 degrees C provided the optimum physiochemical properties as a precursor to conversion to hydroxyapatite (HA). At these temperatures, a partial phase transition to calcium oxide occurred and the original skeletal morphology of the alga remained intact.

Biocompatibility of Calcium Phosphate Bone Cement with Optimised Mechanical Properties

The broad aim of this work was to investigate and optimise the properties of calcium phosphate bone cements (CPCs) for use in vertebroplasty to achieve effective primary fixation of spinal fractures. The incorporation of collagen, both bovine and from a marine sponge (Chondrosia reniformis), into a CPC was investigated. The biological properties of the CPC and collagen-CPC composites were assessed in vitro through the use of human bone marrow stromal cells. Cytotoxicity, proliferation and osteoblastic differentiation were evaluated using lactate dehydrogenase, PicoGreen and alkaline phosphatase activity assays respectively. The addition of both types of collagen resulted in an increase in cytotoxicity, albeit not to a clinically relevant level. Cellular proliferation after 1, 7 and 14 days was unchanged. The osteogenic potential of the CPC was reduced through the addition of bovine collagen but remained unchanged in the case of the marine collagen. These findings, coupled with previous work showing that incorporation of marine collagen in this way can improve the physical properties of CPCs, suggest that such a composite may offer an alternative to CPCs in applications where low setting times and higher mechanical stability are important.

Allogenic Peripheral Blood Derived Mesenchymal Stem Cells (MSCs) Enhance Bone Regeneration in Rabbit Ulna Critical-Sized Bone Defect Model

Mesenchymal stem cells (MSCs) were demonstrated to exist within peripheral blood (PB) of several mammalian species including human, guinea pig, mice, rat, and rabbit. Whether or not the PB derived MSCs (PBMSCs) could enhance the regeneration of large bone defects have not been reported. In this study, rabbit MSCs were obtained from mononuclear cells (MNCs) cultures of both the PB and bone marrow (BM) origin. The number of PBMSCs was relatively lower, with the colony forming efficiency (CFE) ranging from 1.2-13 per million MNCs. Under specific inductive conditions, PBMSCs differentiated into osteoblasts, chondrocytes, and adipocytes, showing multi- differentiation ability similar to BMMSCs. Bilateral 20 mm critical-sized bone defects were created in the ulnae of twelve 6-month old New Zealand white rabbits. The defects were treated with allogenic PBMSCs/Skelite (porous calcium phosphate resorbable substitute), BMMSCs/Skelite, PBMNCs/Skelite, Skelite alone and left empty for 12 weeks. Bone regeneration was evaluated by serial radiography, peripheral quantitative computed tomography (pQCT), and histological examinations. The x-ray scores and the pQCT total bone mineral density in the PBMSCs/Skelite and BMMSCs/Skelite treated groups were significantly greater than those of the PBMNCs/Skelite and Skelite alone groups (p

Investigating Approaches to Three-Dimensional Printing of Hydroxyapatite Scaffolds for Bone Regeneration

This study investigated the feasibility of manufacturing hydroxyapatite (HA)-based scaffolds using 3D printing technology by incorporating different binding additives, such as maltodextrin and polyvinyl alcohol (PVOH), into the powder formulation. Different grades of PVOH were evaluated in terms of their impact on the printing quality. Results showed that scaffolds with high architectural accuracy in terms of the design and excellent green compressive strength were obtained when the PVOH (high viscosity) was used as the binding additive for HA.

Marine Collagen Reinforcement of Calcium Phosphate Bone Cements: A Biological Assessment

Induction of in vivo responses by implanted biomaterials is of great interest in the medical device field. Calcium phosphate bone cements (CPCs) can potentially promote natural bone remodelling and ingrowth in vivo and, as such are becoming more common place in a range of orthopaedic procedures. However, concerns remain regarding their mechanical and handling properties. Compressive modulus and fracture toughness of CPCs can be improved, without compromising injectability and setting time, through the incorporation of bovine collagen fibres1. Incorporation of marine derived collagen fibres has also yielded similar improvements2. It is hypothesised that, due to its role in bone formation and function, that incorporation of collagen in CPCs will also result in biological benefits.
The biological properties of α-TCP-CPC were largely unchanged by the incorporation of marine derived collagen. However, as a result of significant improvements to the mechanical properties, its incorporation may still result in a suitable alternative to some commercially available bone cements.

Investigation of influence of process variables on mechanical strength, size and homogeneity of pharmaceutical granules produced by fluidised hot melt granulation

The overall aim of the project was to study the influence of process variables on the distribution of a model active pharmaceutical ingredient (API) during fluidised melt granulation of pharmaceutical granules with a view of optimising product characteristics. Granules were produced using common pharmaceutical excipients; lactose monohydrate using poly ethylene glycol (PEG1500) as a meltable binder. Methylene blue was used as a model API. Empirical models relating the process variables to the granules properties such as granule mean size, product homogeneity and granule strength were developed using the design of experiment approach. Fluidising air velocity and fluidising air temperature were shown to strongly influence the product properties. Optimisation studies showed that strong granules with homogeneous distribution of the active ingredient can be produced at high fluidising air velocity and at high fluidising air temperatures.

Development of composite tissue scaffolds containing naturally sourced mircoporous hydroxyapatite

The aims of this work were to investigate the conversion of a marine alga into hydroxyapatite (HA), and furthermore to design a composite bone tissue engineering scaffold comprising the synthesised HA within a porous bioresorbable polymer. The marine alga Phymatolithon calcareum, which exhibits a calcium carbonate honeycomb structure, with a natural architecture of interconnecting permeable pores (microporosity 4-11 mu m), provided the initial raw material for this study. The objective was to convert the alga into hydroxyapatite while maintaining its porous morphology using a sequential pyrolysis and chemical synthesis processes. Semi-quantitative XRD analysis of the post-hydrothermal material (pyrolised at 700-750 degrees C), indicated that the calcium phosphate (CaP) ceramic most likely consisted of a calcium carbonate macroporous lattice, with hydroxyapatite crystals on the surface of the macropores. Cell visibility (cytotoxicity) investigations of osteogenic cells were conducted on the CaP ceramic (i.e., the material post-hydrothermal analysis) which was found to be non-cytotoxic and displayed good biocompatibility when seeded with MG63 cells. Furthermore, a hot press scaffold fabrication technique was developed to produce a composite scaffold of CaP (derived from the marine alga) in a polycaprolactone (PCL) matrix. A salt leaching technique was further explored to introduce macroporosity to the structure (50-200 mu m). Analysis indicated that the scaffold contained both micro/macroporosity and mechanical strength, considered necessary for bone tissue engineering applications. (C) 2008 Published by Elsevier B.V.