The effect of particle size on the immunodepressive properties of silica

Five silica preparations were tested in the mouse for their effects on the in vivo humoral immune response to SRBC, the in vitro macrophage phagocytosis of SRBC and the in vitro blastogenic response of non-adherent spleen cells to Con A and LPS¹. One silica preparation was composed exclusively of small particles and produced the greatest depression of all functions tested with the least variability in results. Four of the silica preparations were heterogeneous with respect to particle size, and the heterogeneous preparations containing the highest percentage of small particles had the most depressive effect on the in vivo humoral immune response to SRBC and on in vitro macrophage phagocytosis of SRBC but not on the in vitro blastogenic response to mitogens. All 4 heterogeneous preparations produced great variability in each of the functions tested.Small sized particles from the 4 heterogeneous preparations were prepared by differential sedimentation, and were more depressive in the in vitro assays for macrophage phagocytosis and lymphocyte blastogenesis than the corresponding unfractionated heterogeneous preparation. The large sized particles either had no effect or enhanced both functions. All unfractionated silica preparations were toxic for non-adherent spleen cells, but the homogeneous small particle preparation and the small particles from the heterogeneous preparations were more cytotoxic. These data indicate that silica has a direct depressive effect on both macrophage and lymphocyte functions and that the depressive effects are inversely related to particle size.     

Effect of particle size on phagocytosis of latex particles by guinea-pig polymorphonuclear leucocytes

The effect of the size of foreign particles on phagocytosis by guinea-pig polymorphonuclear leucocytes (PMNs) was investigated using latex particles having approximately the same surface potentials but different sizes. Two types of latexes with different negative surface potentials, those having amino groups (Ami-latexes) and those having carboxylic acid groups (Ca-latexes), were used in this experiment. Since Ca-latexes had different surface potentials for different particle sizes, the carboxylic acid groups were modified by the carbodiimide method to make the surface potentials almost equal. Phagocytosis was estimated by the rate of oxygen consumption. It was found that the phagocytosis reached its maximum when the latex particle size was about 0.5 microns. This finding was plausibly explained by considering the two probabilities that a latex particle comes into contact with a PMN surpassing a potential barrier and the contact takes place at certain receptor areas favourable to phagocytosis on the PMN surface.     

Influence of powder particle size distribution on complex viscosity and other properties of acrylic bone cement for vertebroplasty and kyphoplasty

For use in vertebroplasty and kyphoplasty, an acrylic bone cement should possess many characteristics, such as high radiopacity, low and constant viscosity during its application, low value of the maximum temperature reached during the polymerization process (T(max)), a setting time (t(set)) that is neither too low nor too high, and high compressive strength. The objective of this study was to investigate the influence of the powder particle distribution on various properties of one acrylic bone cement; namely, residual monomer content, T(max), t(set), complex viscosity, storage and loss moduli, injectability, and quasi-static compressive strength and modulus. It was found that the formulations that possessed the most suitable complex viscosity-versus-mixing time characteristics are those in which the ratio of the large poly(methyl methacrylate) beads (of mean diameter 118.4 microm) to the small ones (of mean diameter 69.7 microm) was at least 90% w/w. For these formulations, the values of the other properties determined were acceptable.     

Influence of particle size in the effect of polyethylene on human osteoblastic cells

The influence of two different sizes of polyethylene particles (< 30 and 20-200 microm) on osteoblastic function has been studied in primary human bone cell cultures. Cells were obtained from trabecular bone fragments of patients undergoing knee reconstructive surgery. On reaching confluency, cells were subcultured in three flasks: < 30 microm polyethylene particles were added to the first flask, 20-200 microm particles to the second flask and none to the third flask, which was the control. The resulting subcultures were incubated until confluence. Osteoblastic function was evaluated by assaying the secretion of osteocalcin, alkaline phosphatase, and C-terminal type I procollagen (PICP), with or without 1.25(OH)2D3 stimulation in the cell-conditioned medium. Adding < 30 microm polyethylene particles to these osteoblastic cell cultures increased the levels of osteocalcin secreted after 1,25(OH)2D3 stimulation. Treating stimulated or basal osteoblastic cultures with either polyethylene particle size did not affect alkaline phosphatase secretion. However, the addition of <30 microm polyethylene particles decreased PICP levels in the basal and stimulated cultures. A parallel series of osteoblastic cultures was treated with < 30 microm polyethylene particles and stimulated or not with 1,25(OH)2D3 to determine the effect on osteocalcin mRNA expression using RT-PCR amplification. Polyethylene particle-treated cultures had higher osteocalcin mRNA expression regardless of whether they had been stimulated with 1,25(OH)2D3 or not. We conclude that particle size affects the influence of polyethylene on osteoblastic function markers. Particles with a diameter of less than 30 microm increase osteocalcin expression and secretion.     

An analytical model for the dissolution of different particle size samples of Bioglass in TRIS-buffered solution

We analyzed the early stages of reactivity of three different particle size samples of Bioglass 45S5 and a bulk sample in TRIS-buffered solution at pH 8. Ion release, measured with ion-coupled plasma emission spectroscopy, and pH variations are reported. It was demonstrated that differences in the initial surface area influence the increase in pH, the rate of elemental release, and the rate of calcium phosphate reprecipitation. In particular, a thicker Ca/P layer was obtained on larger particles. The equilibrium value of Si in solution was independent of sample form and amount of sample dissolved, and was always close to the value observed when bulk silica is dissolved at pH 8. An analytical model is proposed for cation release, based on a two-step mechanism. It was found that the early stage of dissolution was nearly diffusion controlled for larger particles and bulk samples. The second stage was similar to a first-order homogeneous dissolution. The influence of sample surface area/solution volume ratio seemed to be more complex than that proposed in the early works presented in the literature. It is suggested that variation of surface area has a significant impact on the course of the dissolution.     

Effect of particle size on the in vitro and in vivo degradation rates of poly(DL-lactide-co-glycolide) microcapsules

Three different sieve size fractions of ergot-containing biodegradable microcapsules were examined both in vitro and in vivo. The sieve sizes and average particle diameter, (micron), were: less than 45-75 (mean = 30); 75-106 (mean = 79); 106-177 (mean = 130). These microcapsules contained ca. 9% drug and were produced from 50:50 poly(DL-lactide-co-glycolide). The objective was to determine the effect of particle size on in vivo and in vitro degradation rates. The microcapsules were injected into rat gastrocnemius muscle and excised and examined at various time points up to 70 days. Initially a minimal tissue response was noted which was characterized by a sharply localized acute inflammatory reaction. Following this, connective tissue and foreign body giant cells engulfed the microcapsules at 20-30 days. Only vestiges of the microcapsules were found surrounded by minimal connective tissue and foreign body giant cells after 60-70 days. The tissue reaction was a minimal, sharply localized foreign body giant cell and connective tissue process for all three size groups of microcapsules. The largest microcapsules (mean = 130 microns) exhibited a slightly greater tendency to undergo in vivo and in vitro degradation relative to the other groups. However, it can be concluded that over the microcapsule size ranges examined minimal differences in the degradation properties of the polymeric matrices and consequently those of the microcapsules were noted.     

Effects of digestion protocols on the isolation and characterization of metal-metal wear particles. I. Analysis of particle size and shape

Isolation of metal wear particles from hip simulator lubricants or tissues surrounding implants is a challenging problem because of small particle size, their tendency to agglomerate, and their potential for chemical degradation by digestion reagents. To provide realistic measurements of size, shape, and composition of metal wear particles, it is important to optimize particle isolation and minimize particle changes due to the effects of the reagents. In this study (Part I of II), transmission electron microscopy (TEM) was used to examine and compare the effects of different isolation protocols, using enzymes or alkaline solutions, on the size and shape of three different types of cobalt-based alloy particles produced from metal-metal bearings. The effect on particle composition was examined in a subsequent study (Part II). Large particles (<1200 nm) were generated by dry abrasion of CoCrMo alloy against itself and small particles (<300 nm) were generated by hip simulator testing of a metal-metal implant pair in the presence of either distilled-deionized water or a 95% bovine serum solution. The reagents changed particle size and to a lesser extent particle shape. For both large particles and small particles generated in water, the changes in size were more extensive after alkaline than after enzymatic protocols and increased with alkaline concentration and time in solution, up to twofold at 2 h and threefold at 48 h. However, when isolating particles from 95% serum, an initial protective effect of serum proteins and/or lipids was observed. Because of this protective effect, there was no significant difference in particle size and shape for both oval and needle-shaped particles after 2 h in 2N KOH and after enzymatic treatments. However, round particles were significantly smaller after 2 h in 2N KOH than after enzymatic treatments. Particle composition may also have been affected by the 2N KOH treatment, as suggested by a difference in particle contrast under TEM, an issue examined in detail in Part II.     

复合磷酸钙颗粒溶解性能测试研究

目的研究复合磷酸钙颗粒(HA∶TCP=3∶7)在TRIS缓冲溶液中的体外溶解特性。方法配制一系列浓度逐级递增的Ca~(2+)标准溶液,绘制Ca~(2+)离子溶液的工作曲线。将复合磷酸钙颗粒(HA∶TCP=3∶7)置于pH值为7.4的TRIS缓冲溶液中浸泡,每小时取样一次(初始阶段每3分钟取样一次),用高速离心技术提取上层清液,采用电感耦合等离子体质谱法测定各时间点提取液的Ca~(2+)浓度,绘制Ca~(2+)浓度对浸泡时间的溶解曲线。结果 Ca~(2+)离子溶液的工作曲线的线性方程为y=4043.9x+23783,相关系数R为0.9999,方法检出限为0.0063 g/m L,定量限为0.021 g/mL。浸泡后最初的0.5 h以内,Ca~(2+)浓度随时间延长急剧升高,0.5 h处有明显转折,0.5 h至20 h,Ca~(2+)浓度仍随时间增加而升高,但是增加的速率较前0.5 h明显下降,20 h以后溶解达到基本平衡。复合磷酸钙颗粒的溶解过程分为3个阶段,首先是Ca~(2+)离子浓度快速升高的溶解过程,第二阶段是Ca~(2+)离子溶解与HA或Ca HPO_4·2H_2O沉积的竞争过程,第三阶段是Ca~(2+)离子浓度不再继续增加的稳定过程。结论复合磷酸钙颗粒(HA∶TCP=3∶7)在TRIS缓冲溶液中的溶解具有一定的规律性,24 h后溶出的Ca~(2+)离子浓度在1200 g/L左右。     

Mechanical properties of glass-ceramic A-W-polyethylene composites: effect of filler content and particle size

Composites which comprise a bioactive filler and ductile polymer matrix are desirable as implant materials since both their biological and mechanical properties can be tailored for a given application. In the present study three-point bending was used to characterise biomedical materials composed of glass-ceramic apatite-wollastonite (A-W) particulate reinforced polyethylene (PE) (denoted as AWPEX). The effects of filler volume fraction, varied from 10 to 50 vol%, and average particle size, 4.4 and 6.7 microm, on the bending strength, yield strength, mode of fracture, Young's modulus and strain to failure were investigated. HAPEX, a commercially used composite of hydroxyapatite and polyethylene, with a 40 vol% filler content, was used for comparison. Increasing the filler content caused an increase in Young's modulus, yield strength and bending strength, and a decreased strain to failure. When filler particle size was increased, the Young's modulus, yield and bending strengths were found to be slightly reduced. A transition in fracture behaviour from ductile to brittle behaviour was observed in samples containing between 30 and 40 vol% filler.     

Influence of motor unit properties on the size of the simulated evoked surface EMG potential

The purpose of the study was to quantify the influence of selected motor unit properties on the simulated amplitude and area of evoked muscle potentials detected at the skin surface. The study was restricted to a motor unit population simulating a hand muscle whose potentials were recorded on the skin over the muscle. Peak-to-peak amplitude and area of the evoked potential were calculated from the summed motor unit potentials and compared across conditions that simulated variation in different motor unit properties. The simulations involved varying the number of activated motor units, muscle fiber conduction velocities, axonal conduction velocities, neuronal activation times, the shape of the intracellular action potential, and recording configurations commonly used over hand muscles. The results obtained for the default condition simulated in this study indicated that ~7% of the motor unit potentials were responsible for 50% of the size of the evoked potential. Variation in the amplitude and area of the evoked muscle potential was directly related to the number of active motor units only when the stimulus activated motor units randomly, and not when activation was based on a parameter such as motor unit size. Independent adjustments in motor unit properties had variable effects on the size of the evoked muscle potential, including when the stimulus activated only a subpopulation of motor units. These results provide reference information that can be used to assist in the interpretation of experimentally observed changes in the size of evoked muscle potentials.     

Rheological properties of acrylic bone cement during curing and the role of the size of the powder particles

A dynamic compressive rheometric technique was used to determine the true or complex viscosity (eta*) of three poly (methyl methacrylate), PMMA-based bone cement formulations (one commercially available and two experimental), as a function of the time that elapsed from commencement of hand mixing of the cement constituents (t). For each cement, two rheological parameters [namely, the time of onset of cure (t(ons)) and the critical cure rate (CCR), which is herein defined as the complex viscosity rate computed at t(ons)] were determined from the eta*-t data. For each cement, particle analysis was used to obtain the powder particle size distribution, from which the following parameters were obtained: (a) the overall mean particle diameter D(m), and (b) the relative amounts of small-sized PMMA beads (mean diameter d between 0 and 40 mum) (alpha) and large-sized PMMA beads (d > 75 mum) (beta). It is seen that the key particle parameter is not D(m) per se but alpha and beta. Thus, the highest values of t(ons) and CCR were obtained from a cement with the highest values of alpha and beta. An explanation for these trends is given, and two areas for further research work in this field are outlined.     

Influence of intra-amoebic and other growth conditions on the surface properties of Legionella pneumophila.

The surface properties of Legionella pneumophila were examined by analyzing outer membrane (OM) proteins, lipopolysaccharides (LPS), and cellular fatty acids after growth within Acanthamoeba polyphaga and in vitro under various nutrient-depleted conditions. Intra-amoeba-grown legionellae were found to differ in several respects from cells grown in vitro; most notably, they contained a 15-kDa OM protein and a monounsaturated straight-chain fatty acid (18:1(9)). These compounds were also found in abundant quantities in the host amoeba. Immunoblot analysis of intra-amoeba-grown legionellae with antiacanthamoebic serum revealed that both the bacterial whole cells and Sarkosyl-extracted OMs contained amoebic antigens. The findings suggest that the 15-kDa OM protein is likely to be of amoebic origin and associates with the OM of the bacterium. It is proposed that disruption of amoebic membranes, as a result of intra-amoebic infection, may liberate macromolecules, including a 15-kDa polypeptide, a major constituent of the amoebic membrane, which adhere to the surface of the legionellae. Growth under specific nutrient depletions also had a significant effect on the surface composition of L. pneumophila. Cells grown under phosphate depletion were markedly sensitive to protease K digestion and contained lower levels of LPS, as observed by silver staining of the digests on polyacrylamide gels. Intra-amoeba-grown cells contained more bands than the in vitro-grown organisms, reflecting further differences in the nature of the LPS. The whole-cell fatty acids of the phosphate-depleted cells were appreciably different from those of cells grown under other nutritional conditions. We found no evidence for expression of iron-regulated OM proteins under iron depletion.     

Influence of the degree of acetylation on the enzymatic degradation and in vitro biological properties of trimethylated chitosans

Chitosan derivatives such as N,N,N-trimethylated chitosan (TMC) are currently being investigated for the delivery of drugs, vaccines and genes. However, the influence of the extent of N-acetylation of these polymers on their enzymatic degradability and biological properties is unknown. In this study, TMCs with a degree of acetylation (DA) ranging from 11 to 55% were synthesized by using a three-step method. First, chitosan was partially re-acetylated using acetic anhydride followed by quantitative dimethylation using formaldehyde and sodium borohydrate. Then, in presence of an excess amount of iodomethane, TMC was synthesized. The TMCs obtained by this method showed neither detectable O-methylation nor loss in acetyl groups (¹H NMR) and a slight increase in molecular weight (GPC) with increasing degree of substitution, implying that no chain scission occurred during synthesis. The extent of lysozyme-catalyzed degradation of TMC, and that of its precursors chitosan and dimethyl chitosan, was highly dependent on the DA and polymers with the highest DA showed the largest decrease in molecular weight. On Caco-2 cells, TMCs with a high DA (～50%), a DQ of around 44% and with or without O-methylated groups, were not able to open tight junctions in the trans-epithelial electrical resistance (TEER) assay, in contrast with TMCs (both O-methylated and O-methyl free; concentration 2.5 mg/ml) with a similar DQ but a lower DA which were able to reduce the TEER with 30 and 70%, respectively. Additionally, TMCs with a high DA (～50%) demonstrated no cell toxicity (MTT, LDH release) up to a concentration of 10 mg/ml.     

The effect of different powder particle size on mechanical properties of sintered alumina, resin- and glass-infused alumina

In this study, the compaction and sintering behavior of fine alumina powders of different particle sizes and the effect of matrix particle size on biaxial strength and fracture toughness of infused matrices were investigated. Three different alumina powders, In-Ceram alumina, A16SG, and RC172 were selected, representing a range of particle size and shape. RC172 and A16SG were dry-pressed. In-Ceram alumina was slip-cast following manufacturer's recommendations. Dry-pressed ceramic blocks were sectioned into disks with a thickness of 1.5-mm. Uninfused disks were sintered at four temperatures between 1250 degrees C and 1400 degrees C. For glass or resin infused specimens, alumina disks were sintered at 1250 degrees C for 2 h and separated into two groups for glass infusion and resin (UDMA/TEGDMA) infusion. Disks were tested for biaxial flexural strength with a universal testing machine (Instron) at 0.5-mm/min crosshead speeds. One-way ANOVA and Duncan's multiple range tests revealed that alumina disks with different smaller particle sizes have significantly higher biaxial strength (p < 0.05). The strength of the alumina matrix was greatly increased by glass and resin infusion. The biaxial strength of resin-infused alumina increased as particle size decreased, whereas strength of glass-infused alumina was constant.     

Influence of packaging conditions on the properties of gamma-irradiated UHMWPE following accelerated ageing and shelf ageing

The ageing behaviour of ultra-high molecular weight polyethylene (UHMWPE) has been studied following gamma irradiation (25 or 40 kGy) in air or in inert atmosphere (vacuum packed). Accelerated ageing procedures used elevated temperature (70 degrees C) and/or pressurised oxygen (5 bar). Shelf ageing for up to six months was also performed. The variation in polymer properties with depth into the polymer was determined using density measurements, infra-red spectroscopy and differential scanning calorimetry. Tensile properties of the irradiated polymer after accelerated ageing were also determined. Accelerated ageing in air for 20 days or pressurised oxygen for 4 days resulted in peaks in polymer density, crystallinity and degree of oxidation at the polymer surface. Accelerated ageing in pressurised oxygen for 8 days resulted in peaks in these properties 500 microns below the polymer surface. Where gamma irradiation was performed in inert atmosphere the amount of polymer degradation following accelerated ageing (in oxidising conditions) was generally lower but still significant. Differences were also observed in tensile properties between material gamma-irradiated in air and in vacuum. This study indicated that performing gamma sterilisation procedures under inert conditions can reduce the level of UHMWPE degradation when exposed to an oxidising environment.     

Influence of process conditions during impulsed electrostatic droplet formation on size distribution of hydrogel beads

In the medical applications of microencapsulation of living cells there are strict requirements concerning the high size uniformity and the optimal diameter, the latter dependent on the kind of therapeutic application, of manufactured gel beads. The possibility of manufacturing small size gel bead samples (diameter 300 microm and below) with a low size dispersion (less than 10%), using an impulsed voltage droplet generator, was examined in this work. The main topic was the investigation of the influence of values of electric parameters (voltage U, impulse time tau and impulse frequency f) on the quality of obtained droplets. It was concluded that, owing to the implementation of the impulse mode and regulation of tau and f values, it is possible to work in a controlled manner in the jet flow regime (U> critical voltage UC). It is also possible to obtain uniform bead samples with the average diameter, deff, significantly lower than the nozzle inner diameter dI (bead diameters 0.12-0.25 mm by dI equal to 0.3 mm, size dispersion 5-7%). Alterations of the physical parameters of the process (polymer solution physico-chemical properties, flow rate, distance between nozzle and gellifying bath) enable one to manufacture uniform gel beads in the wide range of diameters using a single nozzle.     

Phagocytosis and degradation of DNA-anti-DNA complexes by human phagocytes. II. Influence of the size of the complexes

The influence was studied of the size of DNA-anti-DNA complexes on their capacity to induce phagocytosis and degradation by monocytes and neutrophils from human blood. An estimate of the size of the complexes was obtained by precipitation analysis, sucrose-gradient velocity sedimentation and filtration over Nucleopore filters. The results show that only those complexes are processed that precipitate in 15 min at 3000 × g. These complexes have an S value of more than 25 000 and a diameter of more than 0.4 μm. Likewise, tetanus-anti-tetanus complexes also had to be precipitable in 15 min at 3000 × g to be processed by the phagocytes.     

Determination of the size of airborne flour particles

Air sampling was done in a UK bakery and a flour mill with an eight-stage cascade impactor to determine the size of airborne flour particles. The concentration of captured material was determined by a competitive inhibition radioimmunoassay. Comparison of the different sites revealed that 8.79% and 52.14% of the particles in the bakery dough-brake and roll-production areas had a diameter 6.0 μm, respectively, and 19.7% of the particles in the flour mill packing area were of this size. We conclude that in dusty areas up to 20% of the airborne flour particles are of a diameter likely to allow them to be deposited in the bronchial airways and alveoli.