Correction to: The effect of cationically-modified phosphorylcholine polymers on human osteoblasts in vitro and their effect on bone formation in vivo.

The article "The effect of cationically modified phosphorylcholine polymers on human osteoblasts in vitro and their effect on bone formation in vivo", written by Jonathan M. Lawton, Mariam Habib, Bingkui Ma, Roger A. Brooks, Serena M. Best, Andrew L. Lewis, Neil Rushton and William Bonfield, was originally published Online First without open access. After publication in volume 28, issue 9, page 144 it was noticed that the copyright was wrong in the PDF version of the article. The copyright of the article should read as "

The effect of cationically-modified phosphorylcholine polymers on human osteoblasts in vitro and their effect on bone formation in vivo.

The effect of introducing cationic charge into phosphorylcholine (PC)-based polymers has been investigated in this study with a view to using these materials as coatings to improve bone formation and osseointegration at the bone-implant interface. PC-based polymers, which have been used in a variety of medical devices to improve biocompatibility, are associated with low protein adsorption resulting in reduced complement activation, inflammatory response and cell adhesion. However, in some applications, such as orthopaedics, good integration between the implant and bone is needed to allow the distribution of loading stresses and a bioactive response is required. It has previously been shown that the incorporation of cationic charge into PC-based polymers may increase protein adsorption that stimulates subsequent cell adhesion. In this paper, the effect of cationic charge in PC-based polymers on human osteoblasts (HObs) in vitro and the effect of these polymers on bone formation in the rat tibia was assessed. Increasing PC positive surface charge increased HOb cell adhesion and stimulated increased cell differentiation and the production of calcium phosphate deposits. However, when implanted in bone these materials were at best biotolerant, stimulating the production of fibrous tissue and areas of loosely associated matrix (LAM) around the implant. Their development, as formulated in this study, as bone interfacing implant coatings is therefore not warranted.

Design, construction and performance of a portable handheld electrohydrodynamic multi-needle spray gun for biomedical applications.

Electrohydrodynamic (EHD) processing has attracted substantial interest in the technological and pharmaceutical sectors in recent years. Given the complexity of the process, exploring new ideas for EHD electrospraying and electrospinning delivery is a challenge. In this article, the design, construction and testing of a portable handheld EHD multi-needle device are described to produce multifunctional particles and fibers. Solid and encapsulated polymer particles and fibers were generated in order to study the performance of the device. The intrinsic properties of the feed solution/suspension and the processing conditions were adjusted to ensure robustness of the process and give uniform and reproducible products, with diameters ranging from the sub-micrometer scale to a few micrometers. These products have a broad range of applications in many advanced industrial sectors e.g. drug delivery systems, wound dressing patches, low calorie food products and cosmetics.

Modification of the release characteristics of estradiol encapsulated in PLGA particles via surface coating.

BACKGROUND: Drug-loaded poly(lactide-co-glycolide) particles (100-4500 nm in diameter) were prepared via the electrospraying method. An extensive study was then carried out to determine the parameters affecting the release profile of estradiol (the drug or active pharmaceutical ingredient) in order to facilitate minimum initial burst release of estradiol. RESULTS AND DISCUSSION: The three most important factors affecting estradiol release were identified as: particle size, coating of the particles with chitosan/gelatin and the concentration of the coating agent. It was shown that coating the particles with chitosan significantly reduced the burst and initial release without affecting the subsequent release profile. CONCLUSIONS: This work demonstrates a powerful method of generating drug-loaded polymeric particles with modified release behavior and control over the initial release phase. The surface-modified particles may be useful in controlled therapeutic delivery systems to minimize undesirable side effects.

An electrically driven jetting technique for diverse high-resolution surface structures of nanometre hydroxyapatite crystals.

Nanometre hydroxyapatite (nHA) coated metallic materials have been successfully used for bone tissue implantation for several decades now due to its sound biological and mechanical properties. The microstructure and surface topography of the implant material are well-known to play a crucial role in influencing cellular responses to implants and bone tissue regeneration ultimately. Recently, a novel jet-based patterning technique, template-assisted electrohydrodynamic atomisation (TAEA) spraying, has been devised to prepare depositions with defined surface topography for guiding the cellular response. In this study, an improvement investigation of this patterning process was carried out to precisely control the nHA surface structure in terms of geographies and dimensions via an angular needle jetting during the patterning process. More importantly, the mechanism of such improvement of the TAEA patterning technique was also discussed and uncovered. A range of diverse nHA surface structures with high-resolution was therefore achieved, which paves the way for the research of the new generation implant materials with defined cellular response.

A novel route for processing cobalt-chromium-molybdenum orthopaedic alloys.

Spark plasma sintering has been used for the first time to prepare the ASTM F75 cobalt-chromium-molybdenum (Co-Cr-Mo) orthopaedic alloy composition using nanopowders. In the preliminary work presented in this report, the effect of processing variables on the structural features of the alloy (phases present, grain size and microstructure) has been investigated. Specimens of greater than 99.5 per cent theoretical density were obtained. Carbide phases were not detected in the microstructure but oxides were present. However, harder materials with finer grains were produced, compared with the commonly used cast/wrought processing methods, probably because of the presence of oxides in the microstructure.

Design of a multiphase osteochondral scaffold III: Fabrication of layered scaffolds with continuous interfaces.

There is a need to improve current treatments for articular cartilage injuries. This article is the third in a series describing the design and development of an osteochondral scaffold based on collagen-glycosaminoglycan and calcium phosphate technologies for regenerative repair of articular cartilage defects. The previous articles in this series described methods for producing porous, three-dimensional mineralized collagen-GAG (CGCaP) scaffolds whose composition can be reproducibly varied to mimic the composition of subchondral bone, and pore microstructure and mineral phase can be modified. This article describes a method, "liquid-phase cosynthesis," that enables the production of porous, layered scaffolds that mimic the composition and structure of articular cartilage on one side, subchondral bone on the other side, and the continuous, gradual or "soft" interface between these tissues: the tidemark of articular joints. This design enables the layered scaffolds to be inserted into the subchondral bone at an osteochondral defect site without the need for sutures, glue, or screws, with a highly interconnected porous network throughout the entire osteochondral defect. Moreover, the differential moduli of the osseous and cartilaginous compartments enable these layered scaffolds to exhibit compressive deformation behavior that mimics the behavior observed in natural articular joints.

Design of a multiphase osteochondral scaffold. I. Control of chemical composition.

This is the first in a series of articles that describe the design and development of a family of osteochondral scaffolds based on collagen-glycosaminoglycan (collagen-GAG) and calcium phosphate technologies, engineered for the regenerative repair of defects in articular cartilage. The osteochondral scaffolds consist of two layers: a mineralized type I collagen-GAG scaffold designed to regenerate the underlying subchondral bone and a nonmineralized type II collagen-GAG scaffold designed to regenerate cartilage. The subsequent articles in this series describe the fabrication and properties of a mineralized scaffold as well as a two-layer (one mineralized, the other not) osteochondral scaffold for regeneration of the underlying bone and cartilage, respectively. This article describes a technology through which the chemical composition-particularly the calcium phosphate mass fraction-of triple coprecipitated nanocomposites of collagen, glycosaminoglycan, and calcium phosphate can be accurately and reproducibly varied without the need for titrants or other additives. Here, we describe how the mineral:organic ratio can be altered over a range that includes that for articular cartilage (0 wt % mineral) and for bone (75 wt % mineral). This technology achieves the objective of mimicking the composition of two main tissue types found in articular joints, with particular emphasis on the osseous compartment of an osteochondral scaffold. Exclusion of titrants avoids the formation of potentially harmful contaminant phases during freeze-drying steps crucial for scaffold fabrication, ensuring that the potential for binding growth factors and drugs is maintained.

Design of a multiphase osteochondral scaffold. II. Fabrication of a mineralized collagen-glycosaminoglycan scaffold.

This paper is the second in a series of papers describing the design and development of an osteochondral scaffold using collagen-glycosaminoglycan and calcium phosphate technologies engineered for the regenerative repair of articular cartilage defects. The previous paper described a technology (concurrent mapping) for systematic variation and control of the chemical composition of triple coprecipitated collagen, glycosaminoglycan, and calcium phosphate (CGCaP) nanocomposites without using titrants. This paper describes (1) fabricating porous, three-dimensional scaffolds from the CGCaP suspensions, (2) characterizing the microstructure and mechanical properties of such scaffolds, and (3) modifying the calcium phosphate mineral phase. The methods build on the previously demonstrated ability to vary the composition of a CGCaP suspension (calcium phosphate mass fraction between 0 and 80 wt %) and enable the production of scaffolds whose pore architecture (mean pore size: 50-1000 microm), CaP phase chemistry (brushite, octacalcium phosphate, apatite) and crosslinking density (therefore mechanical properties and degradation rate) can be independently controlled. The scaffolds described in this paper combine the desirable biochemical properties and pore architecture of porous collagen-glycosaminoglycan scaffolds with the strength and direct bone-bonding properties of calcium phosphate biomaterials in a manner that can be tailored to meet the demands of a range of applications in orthopedics and regenerative medicine.

Preparation of novel bioactive nano-calcium phosphate-hydrogel composites.

Nano-sized hydroxyapatite (nHA) and carbonate-substituted hydroxyapatite (nCHA) particles were incorporated into a poly-2-hydroxyethylmethacrylate/polycaprolactone (PHEMA/PCL) hydrogel at a filler content of 10 wt%. Fourier transform infrared absorption, transmission electron microscopy, x-ray diffraction and scanning electron microscopy were used to analyse the physical and chemical characteristics of the calcium phosphate fillers and resultant composites. Nano-sized calcium phosphate particles were produced with a needle-like morphology, average length of 50 nm and an aspect ratio of 3. The nanoparticles were uniformly distributed in the polymer matrix. The addition of both HA and CHA in nano-form enhanced the bioactivity and biocompatibility of the PHEMA/PCL matrix. The carbonate-substitution has allowed for improved bioactivity and biocompatibility of the resultant composite, indicating the potential of this material for use in bone tissue engineering.

A novel jet-based nano-hydroxyapatite patterning technique for osteoblast guidance.

Surface topography is well known to play a crucial role in influencing cellular responses to an implant material and is therefore important in bone tissue regeneration. A novel jet-based patterning technique, template-assisted electrohydrodynamic atomization spraying, was recently devised to control precisely the surface structure as well as its dimensions. In the present study, a detailed investigation of this patterning process was carried out. A range of nano-hydroxyapatite (nHA) line-shaped patterns <20 microm in width were successfully deposited on a commercially pure Ti surface by controlling the flow of an nHA suspension in an electric field. In vitro studies showed that the nHA patterns generated are capable of regulating the human osteoblast cell attachment and orientation.

Osteoclastogenesis on hydroxyapatite ceramics: the effect of carbonate substitution.

Human osteoclasts derived from CD14+ve precursors were cultured on discs of stoichiometric hydroxyapatite (HA) and carbonate-substituted hydroxyapatite (CHA) of varying carbonate contents. The development of osteoclasts was qualitatively different on ceramics compared to dentine, occurring in discrete, confluent subpopulations, which suggests local cell signaling may be important in the process. Resorption was quantified by scanning electron microscopy, surface profilometry, and by calcium release into the culture medium. Cells were characterised by a number of histochemical markers of the osteoclast phenotype. Resorption of the ceramic increased with increasing carbonate content up to 2.35 wt %, when resorption trails and pits characteristic of osteoclast activity were seen. Controlling carbonate content may be one way of controlling the rate of resorption of synthetic HA ceramics.

Spreading of an anchorage-dependent cell on a selectively ligand-coated substrate mediated by receptor-ligand binding.

A continuum model was proposed to simulate a biological cell adhering to a substrate surface domain with selectively coated ligands. Cell adhesion was modeled by a traction-separation law depending on the receptor-ligand distance and their densities. It was found that the obtained spreading patterns are consistent with published experimental data and that adhesion requires a nonuniform distribution of receptors at the spreading front with convex fronts requiring a much higher receptor density than concave fronts. Adhesion strength was also found to be more influential than adhesion energy on the spreading kinetics.

The role of electrosprayed apatite nanocrystals in guiding osteoblast behaviour.

Apatite nanocrystals, which mimic the dimensions of natural bone mineral, were electrosprayed on glass substrates, as a suitable synthetic biomedical material for osteoblast outgrowth was explored. A variety of topographic patterns were deposited and the influence of these designs on osteoblast alignment and cell differentiation was investigated. Patterned cell growth and enhanced cell differentiation were seen. Osteoblasts were also cultured on apatite nanocrystals chemically modified with either carbonate or silicon ions. Enhanced cell proliferation and early formation of mineral nodules were observed on apatite nanocrystals with silicon addition. This work highlights the importance of the combined effects of surface topography and surface chemistry in the guidance of cell behaviour.

Biological control of apatite growth in simulated body fluid and human blood serum.

The surface transformation reactions of bioactive ceramics were studied in vitro in standard K9-SBF solution and in human blood serum (HBS)-containing simulated body fluid (SBF). The calcium phosphate ceramics used for this study were stoichiometric hydroxyapatite (HA), beta-tricalcium phosphate (beta-TCP) and brushite. Immersion of each calcium phosphate tested in this study, in simulated body fluid, led to immediate surface precipitation of apatite. The use of HBS resulted in a delay in the onset of precipitation and a significant inhibition of the dissolution reaction normally observed for brushite in solution. However, apatite formation still occurred. The use of HBS and SBF in this investigation, which has shown the ability to induce similar crystal growth as that observed in vivo, suggests that there is scope for the use of serum proteins in simulated body fluid in order to create a protein-rich surface coating on biomedical substrates.

Development of nano-sized hydroxyapatite reinforced composites for tissue engineering scaffolds.

Nano-sized hydroxyapatite (nanoHA) reinforced composites, mimicking natural bone, were produced. Examination by transmission electron microscopy revealed that the nanoHA particles had a rod-like morphology, 20-30 nm in width and 50-80 nm in length. The phase composition of hydroxyapatite was confirmed by X-ray diffraction. The nanoHA particles were incorporated into poly-2-hydroxyethylmethacrylate (PHEMA)/polycaprolactone (PCL) matrix to make new nanocomposites: nanoHA-PHEMA/PCL. Porous nanocomposite scaffolds were then produced using a porogen leaching method. The interconnectivity of the porous structure of the scaffolds was revealed by non-destructive X-ray microtomography. Porosity of 84% was achieved and pore sizes were approximately around 300-400 microm. An in vitro study found that the nanocomposites were bioactive as indicated by the formation of a bone-like apatite layer after immersion in simulated body fluid. Furthermore, the nanocomposites were able to support the growth and proliferation of primary human osteoblast (HOB) cells. HOB cells developed a well organized actin cytoskeletal protein on the nanocomposite surface. The results demonstrate the potential of the nanocomposite scaffolds for tissue engineering applications for bone repair.

Predicting the tensile strength of compacted multi-component mixtures of pharmaceutical powders.

PURPOSE: Pharmaceutical tablets are generally produced by compacting a mixture of several ingredients, including active drugs and excipients. It is of practical importance if the properties of such tablets can be predicted on the basis of the ones for constituent components. The purpose of this work is to develop a theoretical model which can predict the tensile strength of compacted multi-component pharmaceutical mixtures. METHODS: The model was derived on the basis of the Ryshkewitch-Duckworth equation that was originally proposed for porous materials. The required input parameters for the model are the relative density or solid fraction (ratio of the volume of solid materials to the total volume of the tablets) of the multi-component tablets and parameters associated with the constituent single-component powders, which are readily accessible. The tensile strength of tablets made of various powder blends at different relative density was also measured using diametrical compression. RESULTS: It has been shown that the tensile strength of the multi-component powder compacts is primarily a function of the solid fraction. Excellent agreement between prediction and experimental data for tablets of binary, ternary and four-component blends of some widely used pharmaceutical excipients was obtained. CONCLUSION: It has been demonstrated that the proposed model can well predict the tensile strength of multi-component pharmaceutical tablets. Thus, the model will be a useful design tool for formulation engineers in the pharmaceutical industry.

The structure of the bond between bone and porous silicon-substituted hydroxyapatite bioceramic implants.

The significance of micrometer-sized strut porosity in promoting bone ingrowth into porous hydroxyapatite (HA) scaffolds has only recently been noted. In this study, silicon-substituted HA (0.8 wt % Si-HA) with approximately 8.5% of the total porosity present as microporosity within the struts of the implant was prepared for high-resolution transmission electron microscopy (HR-TEM) via both ultramicrotomy and focused ion beam milling. Between the struts of the porous Si-HA, pores with varying shapes and sizes (1-10 microm in diameter) were characterized. Within the struts, the Si-HA contained features such as grain boundaries and triple-junction grain boundaries. Bone ingrowth and dissolution from a Si-HA implant were studied using HR-TEM after 6 weeks in vivo. Minor local dissolution occurred within several pores within the struts. Organized, mineralized collagen fibrils had grown into the strut porosity at the interface between the porous Si-HA implant and the surface of the surrounding bone. In comparison, deeper within the implant, disorganized and poorly mineralized fibers were observed within the strut porosity. These findings provide valuable insight into the development of bone around porous Si-HA implants.