Applications of coralline hydroxyapatite with bioabsorbable containment and reinforcement as bone graft substitute : An experimental study

The aim of the present experimental study was to find out if the applications of coralline hydroxyapatite (HA) can be improved by using bioabsorbable containment or binding substance with particulate HA in mandibular contour augmentation and by using bioabsorbable fibre-reinforced HA blocks in filling bone defects and in anterior lumbar interbody fusion. The use of a separate curved polyglycolide (PGA) containment alone or together with a fast resorbing polyglycolide/polylactide (PGA/PLA) binding substance were compared to the conventional non-contained method in ridge augmentation in sheep. The contained methods decreased HA migration, but the augmentations did not differ significantly. The use of the containment caused a risk for wound dehiscence and infection. Histologically there was a rapid connective tissue ingrowth into the HA graft and it was more abundant with the PGA containment compared to the non-contained augmentation and even additionally rich when the HA particles were bound with PGA/PLA copolymer. However, the bone ingrowth was best in the non-contained augmentation exceeding 10-12 % of the total graft area at 24 weeks. Negligible or no bone ingrowth was seen in the cases where the polymer composite was added to the HA particles and, related to that, foreign-body type cells were seen at the interface between the HA and host bone. The PGA and poly-dl/l-lactide (PDLLA) fibre-reinforced coralline HA blocks were studied in the metaphyseal and in the diaphyseal defects in rabbits. A rapid bone ingrowth was seen inside the both types of implants. Both PGA and PDLLA fibres induced an inflammatory fibrous reaction around themselves but it did not hinder the bone ingrowth. The bone ingrowth pattern was directed according to the loading conditions so that the load-carrying cortical ends of the implants as well as the implants sited in the diaphyseal defects were the most ossified. The fibre-reinforced coralline HA implants were further studied as stand-alone grafts in the lumbar anterior interbody implantation in pigs. The strength of the HA implants proved not to be adequate, the implants fractured in six weeks and the disc space was gradually lost similarly to that of the discectomized spaces. Histologically, small quantities of bone ingrowth was seen in some of the PGA and PDLLA reinforced coralline implants while no bone formation was identified in any of the PDLLA reinforced synthetic porous HA implants. While fragmented, the inner structure of the implants was lost, the bone ingrowth was minimal, and the disc was replaced by the fibrous connective tissue. When evaluated radiologically the grade of ossification was assessed as better than histologically, and, when related to the histologic findings, CT was more dependable than the plain films to show ossification of the implanted disc space. Local kyphosis was a frequent finding along with anterior bone bridging and ligament ossification as a consequence of instability of the implanted segment.

The nanocomposite scaffold of poly(lactide-co-glycolide) and hydroxyapatite surface-grafted with L-lactic acid oligomer for bone repair

Nanohydroxyapatite (op-HA) surface-modified with L-lactic acid oligomer (LAc oligomer) was prepared by LAc oligomer grafted onto the hydroxyapatite (HA) surface. The nanocomposite of op-HA/PLGA with different op-HA contents of 5, 10, 20 and 40 wt.% in the composite was fabricated into three-dimensional scaffolds by the melt-molding and particulate leaching methods. PLGA and the nanocomposite of HA/PLGA with 10 wt.% of ungrafted hydroxyapatite were used as the controls. The scaffolds were highly porous with evenly distributed and interconnected pore structures, and the porosity was around 90%. Besides the macropores of 100-300 mu m created by the leaching of NaCl particles, the micropores (1-50 mu m) in the pore walls increased with increasing content of op-HA in the composites of op-HA/PLGA. The op-HA particles could disperse more uniformly than those of pure HA in PLGA matrix. The 20 wt.% op-HA/PLGA sample exhibited the maximum mechanical strength, including bending strength (4.14 MPa) and compressive strength (2.31 MPa). The cell viability and the areas of the attached osteoblasts on the films of 10 wt.% op-HA/PLGA and 20 wt.% op-HA/PLGA were evidently higher than those on the other composites.

The Surface Modification of Hydroxyapatite Nanoparticles by the Ring Opening Polymerization of gamma-Benzyl-L-glutamate N-carboxyanhydride

The surface modification of hydroxyapatite (HA) nanoparticles by the ring opening polymerization (ROP) of gamma-benzyl-L-glutamate N-carboxyanhydride (BLG-NCA) was proposed to prepare the poly(gamma-benzyl-L-glutamate) (PBLG)-grafted HA nanoparticles (PBLG-g-HA) for the first time. HA nanoparticles were firstly treated by 3-aminopropylthriethoxysilane (APS) and then the terminal amino groups of the modified HA particles initiated the ROP of BLG-NCA to obtain PBLG-g-HA. The process was monitored by XPS and FT-IR. The surface grafting amounts of PBLG on HA ranging from 12.1 to 43.1% were characterized by thermal gravimetric analysis (TGA). The powder X-ray diffraction (XRD) analysis confirmed that the ROP only underwent on the surface of HA nanoparticles without changing its bulk properties. The SEM measurement showed that the PBLG-g-HA hybrid could form an interpenetrating net structure in the self-assembly process.

Composites of poly(lactide-co-glycolide) and the surface modified carbonated hydroxyapatite nanoparticles

To improve the mechanical properties of the composites of poly(lactide-co-glycolide) (PLGA, LA/GA = 80/20) and the carbonate hydroxyapatite (CHAP) particles, the rice-form or claviform CHAP particles with 30-40 nm in diameter and 100-200 nm in length were prepared by precipitation method. The uncalcined CHAP particles have a coarse surface with a lot of global protuberances, which could be in favor of the interaction of the matrix polymer to the CHAP particles. The nanocomposites of PLGA and surface grafted CHAP particles (g-CHAP) were prepared by solution mixing method. The structure and properties of the composites were subsequently investigated by the emission scanning electron microscopy, the tensile strength testing, and the cell culture. When the contents of g-CHAP were in the range of 2-15 wt %, the PLGA/g-CHAP nanocomposites exhibited an improved elongation at break and tensile strength. At the 2 wt % content of g-CHAP, the fracture strain was increased to 20%) from 4-5% for neat PLGA samples. Especially at g-CHAP content of 15 wt %, the tensile strength of PLGA/g-CHAP composite was about 20% higher than that of neat PLGA materials. The tensile moduli of composites were increased with the increasing of filler contents, so that the g-CHAP particles had both reinforcing and toughening effects on the PLGA composites. The results of biocompatibility test showed that the higher g-CHAP contents in PLGA composite facilitated the adhesion and proliferation properties of osteoblasts on the PLGA/g-CHAP composite film.

Grafting polymerization of L-lactide on the surface of hydroxyapatite nano-crystals

A novel method of grafting ring-opening polymerization of L-lactide (LLA) onto the surface of hydroxyapatite nano-particles (n-HAP) was developed. PLLA was directly connected onto the HAP surface through a chemical linkage. The PLLA-g-HAP particles could be stably dispersed in organic solvent such as chloroform for several weeks. The n-HAP particles still retained the original dimension and shape after the grafting of PLLA. Compared with the P-31 MAS-NMR spectrum of pure HAP powders, there appeared a downfield displacement of 1.2 ppm in the spectrum of PLLA-g-HAP. Fourier transformation infrared (FT-IR) spectra further confirmed the existence of PLLA on the surface of PLLA-g-HAP. The amount of grafted polymer determined by thermal gravimetric analysis (TGA) was about 6% in weight. The tensile strength and elongation at break of the PLLA/PLLA-g-HAP composite containing 8 wt% of PLLA-g-HAP were 55 MPa and about 10-13%, respectively, while those of the PLLA/n-HAP composites were 40 MPa and 3-5%, respectively.

Morphology and compression behaviour of biodegradable scaffolds produced by the sintering process

Porous poly(L-lactic acid) (PLA) scaffolds of 85 per cent and 90 per cent porosity are prepared using polymer sintering and porogen leaching method. Different weight fractions of 10 per cent, 30 per cent, and 50 per cent of hydroxyapatite (HA) are added to the PLA to control the acidity and degradation rate. The three-dimensional (3D) morphology and surface porosity are tested using micro-computer tomography (micro-CT), optical microscopy, and scanning electron microscopy (SEM). Results indicate that the surface porosity does not change on the addition of HA. The micro-CT examinations show a slight decrease in the pore size and increase in the wall thickness accompanied by reduced anisotropy for the scaffolds containing HA. Scanning electron micrographs show detectable interconnected pores for the scaffold with pure PLA. Addition of the HA results in agglomeration of the HA particles and reduced leaching of the porogen. Compression tests of the scaffold identify three stages in the stress-strain curve. The addition of HA results in a reduction in the modulus of the scaffold at the first stage of elastic bending of the wall, but this is reversed for the second and third stages of collapse of the wall and densification in the compression tests. In the scaffolds with 85 per cent porosity, the addition of a high percentage of HA could result in 70 per cent decrease in stiffness in the first stage, 200 per cent increase in stiffness in the second stage, and 20 per cent increase in stiffness in the third stage. The results of these tests are compared with the Gibson cellular material model that is proposed for prediction of the behaviour of cellular material under compression. The pH and molecular weight changes are tracked for the scaffolds within a period of 35 days. The addition of HA keeps the pH in the alkaline region, which results in higher rate of degradation at an early period of observation, followed by a reduced rate of degradation later in the process. The final molecular weight is higher for the scaffolds with HA than for scaffolds of pure PLA. The manufactured scaffolds offer acceptable properties in terms of the pore size range and interconnectivity of the pores and porosity for non-load-bearing bone graft substitute; however, improvement to the mixing of the phases of PLA and HA is required to achieve better integrity of the composite scaffolds. © 2008 IMechE.

PMMA-co-EHA cements for osteoprotheses

The main purpose of this thesis was to produce new formulations of PMMA-co- EHA and study its feasibility as being an alternative to traditional PMMA bone cements. Thus, were originally produced several co-polymers of PMMA-co-EHA and its mechanical properties and in vitro behaviour were evaluated. The copolymers were obtained by radical polymerization and several formulations were produced by partial replacement of MMA (up to about 50%) for EHA. Overall, the results suggest that the partial replacement of MMA by EHA decreased the modulus of the materials and, consequently, increased its flexibility. Then, PMMA commercial beads were added to PMMA-co-EHA formulations (to get bone cement) and the general properties of the resulting bone cements were evaluated. In general, the results revealed that the partial replacement of MMA by EHA led to beneficial changes in curing parameters (there was a reduction of the peak temperature and an increase of curing/setting time), in the in vitro behaviour (the water capacity increased) and in the mechanical properties (the bending strength increased) of new cements. The in vitro cellular response of new formulations of PMMA-co-EHA was compared with that of traditional PMMA bone cement. To this end, we tested the cell adhesion and proliferation of osteoblast-like MG63 cells and human cells from bone marrow. The results revealed that both types of cells were able to attach and proliferate in both formulations. The only exception was observed for the formulation prepared with the highest percentage of EHA, where a few cells that adhere failed to proliferate. Moreover, it was found that increasing the amount of EHA in cement led to an increasing inhibition of cell growth, especially during the first week of culture. This was related to increased water uptake capacity by the new formulations and consequent release of some of its toxic components. Finally, PMMA commercial beads were partially replaced by HA particles and the influence of this substitution on the curing parameters, the mechanical properties and in vitro behaviour of the resulting composites was also evaluated. Incorporation of HA into the bone cements induced a number of significant changes in its final properties: 1) decrease the peak temperature; 2) increase of curing time, 3) increasing the value of elastic modulus accompanied by decrease of the strength/tension. This last finding was related to poor interfacial adhesion between the various components of the bone cements and a heterogeneous distribution (possible agglomeration) of HA particles.

Biological Performance of Chemical Hydroxyapatite Coating Associated With Implant Surface Modification by Laser Beam: Biomechanical Study in Rabbit Tibias

Purpose: Considering the potential of the association between laser ablation and smaller scale hydroxyapatite (HA) coatings to create a stable and bioactive surface on titanium dental implants, the aim of the present study was to determine, by the removal torque test, the effects of a surface treatment created by laser-ablation (Nd:YAG) and, later, thin deposition of HA particles by a chemical process, compared to implants with only laser-ablation and implants with machined surfaces.Materials and Methods: Forty-eight rabbits received I implant by tibia of the following surfaces: machined surface (MS), laser-modified surface (LMS), and biomimetic hydroxiapatite coated surface (HA). After 4, 8, and 12 weeks of healing, the removal torque was measured by a torque gauge. The surfaces studied were analyzed according to their topography, chemical composition, and roughness.Results: Average removal torque in each period was 23.28, 24.0, and 33.85 Ncm to MS, 33.0, 39.87, and 54.57 Ncm to LMS, and 55.42, 63.71 and 64.0 Ncm to HA. The difference was statistically significant (P < .05) between the LMS-MS and HA-MS surfaces in all periods of evaluation, and between LMS-HA to 4 and 8 weeks of healing. The surface characterization showed a deep, rough, and regular topography provided by the laser conditioning, that was followed by the HA coating.Conclusions: Based on these results, it was possible to conclude that the implants with laser surface modification associated with HA biomimetic coating can shorten the implant healing period by the increase of bone implant interaction during the first 2 months after implant placement. (C) 2009 American Association of Oral and Maxillofacial Surgeons J Oral Maxillofac Surg 67:1706-1715, 2009

Osseointegração de implantes de titânio com modificação da superfície por ablação a laser e recobrimento por hidroxiapatita biomimética: estudo biomecânico e histomorfométrico em tíbia de coelhos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The effect of monoalkyl phosphates and fluoride on dissolution of hydroxyapatite, and interactions with saliva

The aims were to investigate the effect of monoalkyl phosphates (MAPs) and fluoride on dissolution rate of native and saliva-coated hydroxyapatite (HA). Fluoride at 300 mg/l (as NaF) inhibited dissolution of native HA by 12%, while potassium and sodium dodecyl phosphates (PDP, SDP), at 0.1% or higher, inhibited dissolution by 26-34%. MAPs, but not fluoride, also showed persistence of action. MAPs at 0.5% and fluoride at 300 mg/l were then tested separately against HA pre-treated with human saliva for 2 or 18 h. Agents were applied with brushing to half the specimens, and without brushing to the other half. In control (water-treated) specimens, pre-treatment of HA with human saliva reduced dissolution rate on average by 41% (2 h) and 63% (18 h). Brushing did not have a statistically significant effect on dissolution rate of saliva-coated specimens. In brushed specimens, fluoride significantly increased the inhibition due to 2- or 18-hour saliva pre-treatment. It is hypothesised that brushing partially removes the salivary film and allows KOH-soluble calcium fluoride formation at the surfaces of HA particles. Inhibition was reduced by PDP in 2-hour/non-brushed specimens and in 18-hour/brushed specimens. PDP did not affect dissolution rates in the remaining groups and SDP did not affect dissolution rate in any group. Possible reasons for these variable results are discussed. The experiments show that pre-treatment with saliva can significantly modify results of tests on potential anti-erosive agents and it is recommended that saliva pre-treatment should be a routine part of testing such agents.

CFD-DEM simulation of propagation of sound waves in fluid particles fluidised medium

In this work, speed of sound in 2 phase mixture has been explored using CFD-DEM (Computational Fluid Dynamcis - Discrete Element Modelling). In this method volume averaged Navier Stokes, continuity and energy equations are solved for fluid. Particles are simulated as individual entities; their behaviour is captured by Newton's laws of motion and classical contact mechanics. Particle-fluid interaction is captured using drag laws given in literature.The speed of sound in a medium depends on physical properties. It has been found experimentally that speed of sound drops significantly in 2 phase mixture of fluidised particles because of its increased density relative to gas while maintaining its compressibility. Due to the high rate of heat transfer within 2 phase medium as given in Roy et al. (1990), it has been assumed that the fluidised gas-particle medium is isothermal.The similar phenomenon has been tried to be captured using CFD-DEM numerical simulation. The disturbance is introduced and fundamental frequency in the medium is noted to measure the speed of sound for e.g. organ pipe. It has been found that speed of sound is in agreement with the relationship given in Roy et al. (1990). Their assumption that the system is isothermal also appears to be valid.

Fabrication and luminescent properties of the core-shell structured YNbO4 : Eu3+/Tb3+@SiO2 spherical particles

The core-shell structured YNbO4:Eu3+/Tb3+@SiO2 particles were realized by coating the YNbO4:Etr(3+)/Tb3+ phosphors onto the surface of spherical silica via a sol-gel process. The obtained materials were characterized by means of X-ray diffraction (XRD), field emission scanning electron microscopy (FE-SEM), transmission electron microscopy (TEM), Fourier transform IR spectroscopy (FT-IR), photoluminescence (PL) spectra, and cathodoluminescence (CL) spectra.

Sonochemical effects on ZnO particles

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

On the penetration of particles through filters and masks.

El principal objetivo de este trabajo es la evaluación de la eficiencia de filtros y máscaras destinados a la protección de los trabajadores contra partículas sólidas respirables. Se ha diseñado un dispositivo que simula los ciclos respiratorios del ser humano de forma automática. Filtros y máscaras se ensayan en el interior de una cámara, en una atmósfera controlada, y se introducen partículas de polvo en suspensión en su interior. Las partículas de polvo pasan a través de los filtros y se toma una muestra en vía húmeda. Se toma una muestra de polvo después de pasar por los filtros y se compara con la inicial, en una forma cualitativa y cuantitativa, mediante gravimetría y difracción laser. Analizados los resultados, se ha observado una variabilidad significativa en la eficiencia de máscaras y filtros pertenecientes a una misma categoría, por lo que se han sugerido recomendaciones a la normativa actual.

Diesel bus emissions of submicrometer particles measured in a tunnel study

The emission factors of a bus fleet consisting of approximately three hundreds diesel powered buses were measured in a tunnel study under well controlled conditions during a two-day monitoring campaign in Brisbane. The number concentration of particles in the size range 0.017-0.7 m was monitored simultaneously by two Scanning Mobility Particle Sizers located at the tunnel’s entrance and exit. The mean value of the number emission factors was found to be (2.44±1.41)×1014 particles km-1. The results are in good agreement with the emission factors determined from steady-state dynamometer testing of 12 buses from the same Brisbane City bus fleet, thus indicating that when carefully designed, both approaches, the dynamometer and on-road studies, can provide comparable results, applicable for the assessment of the effect of traffic emissions on airborne particle pollution.