Biomedical subjects

E Maconnachie

Publications and source records attributed to E Maconnachie.

9 recordsLinked to original sources

PubMed1990

The effect of fluoride on the resorption of dentine by osteoclasts in vitro.

Osteoclasts were isolated mechanically from bones of prehatch chicks and cultured on dentine for 24 or 39 h in medium without or with added sodium fluoride at concentrations of 0.15, 1, 15, or 30 mg/liter. Fixed and toluidine blue-stained specimens were examined by light microscopy and osteoclasts were counted, all cells were removed and resorption pits were counted. The dentine was then prepared for scanning electron microscopy (SEM) and pits located on a raster search using backscattered electron imaging. Stereophotogrammetric measurements were made of the depths, plan and true surface areas and volumes of pits, and the volume:area ratio and a three-dimensional form factor calculated for each. The presence or absence of a ring of demineralization around each pit was recorded. Pits with the same plan area were significantly (p less than 0.001) less deep in cultures containing 15 or 30 mg/liter NaF and the incidence of rings around pits was higher (controls 2.7%, 15 mg/liter 31.3%, 30 mg/liter NaF 39.7%). In one experiment, pits were larger in the specimen cultured with added 1 mg/liter NaF, but in a much larger series of 631 (251 in the control and 380 in the 1 mg/liter NaF cultures) pits measured, depths, volumes and volume:plan-area ratios were significantly lower (p less than 0.001), and rings round pits 2.5 times more common, in the fluoride-treated group. A complex interplay of stimulation of some cells in the culture with indirect stimulation of osteoclasts and direct inhibition of osteoclastic resorptive function may be operating at this concentration of fluoride. In vivo, therapeutic doses of sodium fluoride would be expected to act not only to increase the formation of osteoid but also to suppress osteoclastic activity.

Animals↗

PubMed1990

Human cranial bone structure and the healing of cranial bone grafts: a study using backscattered electron imaging and confocal microscopy.

This study explored the microstructure of human cranial bone at different ages, and the survival, remodelling and modelling of cranial bone grafts. A combination of reflection and fluorescence confocal optical microscopy and scanning electron microscopy in the backscattered electron imaging mode was employed to examine highly polished block faces of plastic-embedded bone fragments as harvested for grafting, or recovered after a period in situ as a graft. The methods enabled remarkably detailed information on bone content, maturation and turnover to be gleaned from tiny scraps of bone. Microfractures in the harvested bone were repaired at the graft site, with welding of old and new bone indicating revascularization. Human cranial bone grafts successfully stimulated bone cell differentiation, supported new bone formation on resorbed and unresorbed surfaces, and underwent bone turnover. The type and organization of new bone reflected the growth rate and maturation of the graft rather than the age of the patient.

Adult↗

PubMed1990

SEM study of surface alterations of bioactive glasses and glass--ceramics in a bony implantation bed.

Back scattered electron (BSE) images of glasses and glass-ceramics with different degrees of bone-bonding behaviour have been investigated. After the implantation of probes in the femur of rats changes of the surface material, induced by leaching and corrosion (material response) and of the tissue in the drill hole (host response), were studied. The results demonstrate advantages of using BSE images and give further information for understanding the sequential changes in the implant material which occur mainly in bone-bonding materials. The density of the leached zone was not homogeneous and it therefore might be possible that preferential leaching of the glass matrix contributes to the mineralization of extracellular matrix. These findings could be of use for further development of surface reactive materials.

Animals↗

PubMed1987

The effects of inhibitors of cysteine-proteinases and collagenase on the resorptive activity of isolated osteoclasts.

The effects of specific inhibitors of cysteine-proteinases ((Z-Phe-Ala-CHN2: benzyloxycarbonyl-phenyl-alanyl alanyl diazomethane and E-64: trans-epoxy-succinyl-L-leucylamido(4-guanidino)-butane) and collagenase and collagenase ((Cl-1: N-(3-N-benzyloxycarbonyl amino-1-R-carboxypropyl)-L-leucyl-O-methyl-L-tyrosine N-methylamide) have been tested on the osteoclastic resorption of dentine. Chick osteoclasts were cultured in the presence or absence of 12.5 microM Z-Phe-Ala-CHN2, 40 or 60 microM E-64, or 40 or 100 microM Cl-1 for 1 or 2 days. In addition, osteoclasts were cultured on oyster shell calcitostracum with or without 12.5 microM Z-Phe-Ala-CHN2. Specimens were studied by light microscopy to count cells and resorption features and by scanning electron microscopy (SEM) stereophotogrammetry for the measurement of the depths, plan-areas and volumes of resorption pits. The numbers, depths and volumes (but not the plan-areas) of the resorption pits in dentine were significantly reduced by Z-Phe-Ala-CHN2 and E-64. Thus, for a given plan-area, the volumes and the depths of resorption pits were smaller in these experimental groups compared with control dentine specimens. The overall inhibition of resorption was at least 75%. Cl-1 did not have this inhibitory effect on the numbers or sizes of resorption pits in dentine. When the oyster calcitostracum was used as a substrate for the osteoclasts, Z-Phe-Ala-CHN2 did not reduce the numbers or volumes of pits, but increased the plan-areas and prevented the formation of deeper pits.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1986

Scanning electron microscopy in bone pathology: review of methods, potential and applications.

This article reviews the applications of SEM methods to human bone pathologies referring to studies made at UCL. We consider the methods which may be most suitable; these prove to be not "routine" in the context of most bio-medical applications of SEM. Valuable information can be obtained from a bone sample if its edges are ground flat, before making either a matrix surface preparation by washing away all the cells or a mineralizing front preparation, by also dissolving the osteoid-for which hydrogen peroxide is recommended to produce a robust specimen. BSE contrast from a cut block surface can be used to measure bone phase volume. SE contrasts from natural surfaces (trabeculae, canals and lacunae) can be used to study forming, resting and resorbing surfaces both qualitatively and quantitatively (except in the case of histological osteomalacia, where the existence of osteoid will go undetected and reversal lines will be difficult to distinguish from recently resorbed surfaces). We also recommend the use of PMMA embedded bone blocks, which can be used as obtained from the pathologist, but are better embedded by a more rigorous procedure. BSE image analysis can be used to quantitate bone density fractions opening up a completely new investigative method for the future. Osteoid can be measured automatically using CL if the bone sample is block stained with brilliant sulphaflavine before embedding or if a scintillant is added to the embeddant. We give examples of observations made from a number of bone diseases: vitamin D resistant rickets, osteogenesis imperfecta; osteomalacia; osteoporosis; hyperparathyroidism; fluorosis; Paget's disease; tumour metastasis to bone.

Adolescent↗

PubMed1986

Variation in the sizes of resorption lacunae made in vitro.

The assessment of in vitro osteoclastic activity has, until recently, been dependent on the analysis of organ culture experiments. We have developed a single cell resorption assay so that the resorptive function of individual osteoclasts could be studied. This paper examines the biological variation in the sizes of resorption lacunae produced by bone cell cultures derived from neonate rats and rabbits, and prehatch or hatchling chicks. Cultures were run for 24h for all species; and in addition for 48h for rat, 9 or 12 hours for rabbit and 3-7 hours for chick. The numbers of the nuclei of osteoclasts seeded on to plastic were counted for all three species. SEM stereophotogrammetry was used to measure areas, volumes, and maximum and average depths of the lacunae using specially designed instruments and software. Rat osteoclasts were smallest, and more chick osteoclasts were very large. There was a species difference in the onset of resorption and the sizes of pits produced, the chick osteoclasts being more vigorous resorbers than the rabbit ones, and the rat least so. For a given plan area, chick lacunae were deeper. There was a high correlation between area and volume. The range of maximum depths for a given area was high, however. Thus the mean of a few measurements of depths should not be used to calculate volume from area. At 24 hours, 77% of the rat, 47% of the rabbit and 28% of the chick lacunae were less than 1,000 microns 3 in volume; and 11% of the rat, 17% of the rabbit and 22% of the chick lacunae were between 1,000 and 2,000 microns 3 in volume. The mean values at 24 hours were 981, 2796, and 4582 microns 3 for rat, rabbit and chick lacunae respectively.

Animals↗

PubMed1981

Morphological correlations with dimensional change during SEM specimen preparation.

We have previously reported details of the dimensional changes taking place during the processing of soft tissue specimens for scanning electron microscopy. Mouse embryo limbs were used for many of these measurements and the present paper deals with the associated morphological findings. Effects of fixation, dehydration and drying are considered. Freeze drying and critical point drying of glutaraldehyde and glutaraldehyde and osmium fixed samples give perfectly acceptable results for scanning electron microscopy. The best volume retention with freeze dried material is matched by the best morphological appearance of the specimen surface, except when ice crystal damage occurs due to a failure to freeze the tissue rapidly enough. For CPD tissues, perforation of the plasmalemma may occur in glutaraldehyde-only fixed tissue, this being prevented by post-osmication if the glutaraldehyde fixation is not unduly prolonged. This perforation may be due to the extraction of some plasmalemma component during dehydration or further solvent substitution on critical point drying. Solvent evaporation drying usually causes recognizable distortion due to shrinkage: this is minimal in the case of solvent evaporation drying in a nearly saturated atmosphere of the same solvent if this is a very volatile solvent. The examples of Freon 113 and diethyl ether are given here. Swelling during early stages of ethanol dehydration can be prevented by using 70% or 100% ethanol as the first step, with marginal reduction in the post CPD volume and no apparent differences in the SEM. The severe swelling causing sample disruption which can occur with GA + OsO4 fixed tissue can also be prevented by treating the sample with divalent cations, such as Ca++ or Cu++, at any stage.

Animals↗

PubMed1980

Treatment with lithium salts reduces ethanol dehydration shrinkage of glutaraldehyde fixed tissue.

Changes in the area of glutaraldehyde fixed 15 day p.c. mouse embryo limbs were recorded using a Quantimet 720 image analysing computer attached to a light microscope: during a period of treatment with an isotonic salt solution (mostly halides of the alkali or alkaline earth metals); a subsequent wash with distilled water; and dehydration through a 30, 50, 70, 80, 90, and 100% ethanol series. Pretreatment with NaCl, KCl, RbCl had no significant effect. Treatment with LiCl, LiNO3, LiF (0.03 M), CsF and CsCl caused an increase (relative to Na, K or Rb treated samples) in the specimen volume during dehydration, which persisted in 100% ethanol. Li treated samples showed the largest post-critical-point-drying (CPD) volumes, followed by Cs treated tissue. Pretreatment with Be, Mg, Ca, Sr and Ba chlorides caused shrinkage and the 100% ethanol and post-CPD volumes of these samples were all lower than those treated with the monovalent cation containing salts.

Aldehydes↗

PubMed1979

A study of digit fusion in the mouse embryo.

During the embryonic development of the mouse limb separation of the digit is followed by their union. This is a true, though temporary, epithelial fusion, a fused layer of epidermal cells remaining intact until separation takes place after birth. The periderm cells in the line of fusion are displaced to the dorsal or ventral surface of the foot. On the dorsal surface these displaced cells form a prominent interdigital ridge of elongated, intertwined cells which remains until the periderm is shed. During the fusion of the eyelids, and also of the pinnae to the scalp, a similar ridge of periderm cells is formed.

Animals↗