Search PubMed⌕ Search

Biomedical subjects

A Boyde

Publications and source records attributed to A Boyde.

At least 145 records · Page 8Linked to original sources

The interface of cells and their matrices in mineralized tissues: a review.

The interface between cells and matrices in mineralized tissues formed in vivo has been studied mainly by looking at the matrix surface, which is easily prepared, and not at the cell surface, which presents problems. Vertebrate calcified tissues range from being acellular to highly cellular, but for all the tissues the formative cells lay down and organise a cell-specific matrix, although this may be deposited initially on a different tissue-type. The formation of hard tissues is a group activity of many cells; resorption is the province of one cell, though it may be controlled by others in the vicinity. Cell-matrix interfaces that develop in vitro have also mainly been studied at the matrix side. The main difficulty with in vitro studies of hard tissue interfaces is that the cells do not have the same activity or even cellular functions as they had in vivo under the complex control of physiological regulation. The question of osteoblastic osteoclasis falls into this category. It is possible to provide new substrata for both formative and resorptive hard tissue cells to test for the interaction between the cells and the 'matrix' on to which they are seeded. The changing cell-matrix interface may also be modelled using computer simulation of osteoclastic movement across a substrate based on known patterns exhibited by other cell types in vitro. Comparison with the shapes of complex resorption pits shows a surprising match. This suggests that the track of the osteoclast due to cell motility and the bone resorptive mechanism resulting in pits along that track are likely to be separately controlled phenomena.

Animals↗

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↗

Stereoscopic images in confocal (tandem scanning) microscopy.

Stereoscopic pair images with parallel projection geometry are obtained by through-focusing along two inclined axes while recording two (summed and stacked) images with a microscope with a very shallow depth of field. The two stack images sample the same depth slice of translucent or reflective specimens. The method will work most conveniently with a tandem scanning microscope (a direct-view, confocal scanning optical microscope). This is a direct method for recording stereo images that can be used to the limit of resolution in optical microscopy. It demonstrates a previously unrealized advantage of confocal optical microscopy.

Animals↗

Bone modelling in the implantation bed.

We consider results from tissue culture studies and the comparative histology of mineralized tissues and other natural tissue interfaces which may have some relevance in understanding the abnormal biology of the immediate environment of an implant in bone. We discuss factors influencing setting, colonization, and migration on natural and artificial substrates by various cell types which may make or remove matrix near the implant. A knowledge of mechanisms of mineral and organic matrix destruction by osteoclasts and other cells must be important in addition to an understanding of the interaction of local and systemic hormones with bone cells. More studies of the role of the immune system in implant failure are urgently required.

Animals↗

The pyroantimonate reaction and transcellular transport of calcium in rat molar enamel organs.

The distribution of calcium in the cells of the enamel organ of developing rat molar tooth germs was studied by the pyroantimonate method. It was found that there was a specific localization to the inner leaflet of the plasma membrane of both secretory and maturation phase ameloblasts. This information can be used to support the model for transcellular transport of calcium involving membrane fluidity, with phosphatidylserine as a carrier (Reith 1983). It can also support an alternative model involving movement of calcium ions over a surface of acidic phospholipids on the inner leaflet of the plasma membrane, without involving the necessity for membrane fluidity.

Animals↗

Unbiased estimation of particle density in the tandem scanning reflected light microscope.

The tandem scanning reflected light microscope has the property of being able to obtain information from 'inside' solid objects by taking a thin optical section at the focal plane of the objective lens. This plane can be focused up and down through the specimen. We describe an unbiased 3-D counting rule for the TSRLM, which is applied to the estimation of osteocyte lacunar density in whole bone. This is shown to be an extremely efficient way of making such an estimate. Further possibilities for the application of the microscope in the field of stereology are discussed.

Animals↗

Optical and scanning electron microscopy in the single osteoclast resorption assay.

The present studies relate to the single or isolated osteoclastic resorption function assay which we introduced in 1983 to overcome objections to assays based upon measurements of calcium release from bones, in which it was never strictly controlled whether the mechanism involved the destruction of bone with the formation of classical Howship's lacunae. The method may prove to be quite popular in the near future and has already been adopted by other research groups. In previous work, we had utilised stereophotogrammetry of scanning electron micrographs to measure the depth, volume and other parameters of the individual lacunae. However, increasing experience with the method has suggested that we can await a wide range of biological variability in single cell function in any one experiment. We have therefore tested other methods from which data could be obtained more rapidly to permit a better statistical analysis, albeit with reduced accuracy, of each resorption complex. The main aim of the studies reported here was to evaluate various methods of optical and scanning electron microscopy that can be used for the visualization of osteoclasts and their associated resorption lacunae generated in vitro in slabs of dentine and bone. Optical microscopy was found to be complementary to SEM, enabling vital microscopy of unstained and stained cells. In particular, oblique illumination LM and tandem scanning reflected LM (TSRLM) proved to be of paramount value for this purpose. Fixed coated specimens could be most rapidly scanned for resorption lacunae using darkfield reflected LM or TSRLM.

Animals↗

Microscopic criteria for the determination of directionality of cutmarks on bone.

Shipman and Rose (1983) have recently attracted attention to the fact that the determination of directionality of cutmarks could possibly provide additional evidence for interpretation of early hominid butchering practices and handedness. They found no criteria of directionality, however. We have previously recognized directionality in cut dental tissues and more recently in bone, and so we undertook a study of over 200 experimentally produced cutmarks on bovine bone by scanning electron microscopy. Three criteria of directionality were observed: bone smears, oblique faulting, and oblique chipping. In our analysis we considered properties of bone as a relevant variable in the microscopic appearance of cutmarks, which has also led to the new finding that not all marks made by a single tool under similar conditions are the same. We observed that bone smears occurred in most specimens, relatively low density forming bone surfaces facilitated oblique fault production, whereas oblique chipping frequently occurred in plexiform bone tissue common to growing artiodactyls. In this study, handedness of the operator could be determined from cutmarks, but much further experimental work will be required in order to detail the criteria.

Animals↗

The resorption of biological and non-biological substrates by cultured avian and mammalian osteoclasts.

Mammalian and avian osteoclasts were isolated mechanically from long bones, seeded on to either untreated, unmineralized, anorganic or surface-demineralized mammalian dental tissues, and cultured for 1-6 h or up to 9 days in medium with added serum (10% heat-inactivated FCS). All substrates showed Howship's resorption lacunae which varied in detail with the composition and structural organization of the tissue. There was no species or substrate specificity. Osteoclasts also adhered, spread, migrated and resorbed in the absence of serum. In addition, osteoclasts resorbed avian egg shell and mollusc shell containing calcite and aragonite. When given the opportunity, osteoclasts are thus biochemically competent to resorb a much wider range of substrates than they usually do in vivo. Access to the substrate and attraction or deliverance of osteoclast precursors to it must be curtailing factors in in vivo resorption.

Absorption↗

Monocyte-enriched cells on calcified tissues.

Monocyte-enriched human blood cells seeded on to sperm whale dentine and cultured for up to 20 days failed to produce any morphological signs of resorbtive activity, although multinucleate giant cells were formed. In contrast, preparations containing known osteoclasts derived from bone resorbed the same substrate within hours.

Animals↗

Motility and resorption: osteoclastic activity in vitro.

Rabbit osteoclasts (OCs), separated mechanically from long bones, were seeded on to glass or plastic substrates or slabs of sperm whale dentine (SWD). Cells were cultured in MEM with 10% FCS with or without added salmon calcitonin (SCT) at dosages of 0.001, 0.1 and 1 IU/ml. Although most rabbit SCT-treated OCs on the non-biological substrates showed inhibition of peripheral ruffling activity and motility at dosages that stop rat OC movement, resorption still occurred on the dentine. Thus such inhibition is unreliable as a general indicator for resorptive capability. Resorption lacunae were observed at all times from 6 h onwards. Using stereophotogrammetric techniques, the following minimum values were obtained from 24 h cultures: highest hourly rate of resorption of dentine for single OC, 570 micron3/h; average rate 165 micron3/h; mean total volume dentine removed per Howship's lacuna complex, 3,885 micron3; average value for plan area of surface attacked per OC, 1,450 micron2.

Animals↗

Dependence of rate of physical erosion on orientation and density in mineralised tissues.

Bone, dentine and enamel samples were treated with a gas-propelled jet of an abrasive, NaHCO3, which is physically much softer than any of these tissues in their fully mineralised condition. It was nevertheless found that they are all eroded by this treatment, which can therefore be used as a new kind of qualitative test of physical properties relating to wear resistance. General correlations were found between both degree of mineralisation and between structure orientation and erosion rate, surface-parallel-feature zones being worn more rapidly. Bone domains with surface-parallel collagen were eroded faster than those with perpendicular lamellae even if they were more densely mineralised. Rates of dentine wear depended on both density and tubule orientation, with peritubular zones and better mineralised incremental layers being more resistant. Enamel tufts wear more rapidly than the surrounding well mineralised regions. Enamel diazones wear less than parazones (areas with surface parallel prisms). At the prism scale, enamel is removed more rapidly near prism boundary discontinuities and in tubular enamel, at tubule walls. As regards the common orientation dependent effects seen in these three tissues, a cohesive explanation would be that structure discontinuities can be better exploited in a wear process if they allow cleavage from the surface; which tendency will increase with parallelism to the surface.

Animals↗

Cyclical phenomena occurring during the maturation of the enamel of rat incisor teeth. Their manifestation during drying.

A pattern of obliquely oriented bands has been demonstrated at the surface of the maturation zone enamel of freshly dissected rat incisor teeth as they dry. This pattern, of which there is no evidence in the fresh, wet, or completely dry teeth, consists of up to 4 or 5 pale grey, translucent lines separated by wider, whiter, more opaque bands and has been shown to correlate directly with a similar pattern seen on the same teeth after staining with toluidine blue and previously described as the maturation cycle banding pattern (Boyde and Reith 1982). A second pattern comprised of much more closely spaced bands is also described. In this pattern, which again correlates with a similar pattern seen after toluidine blue staining, translucent and opaque bands cross the maturation zone more transversely and have a width of about 200 microns, approximating to 8 h tooth growth. It is postulated that these banding patterns reflect alternately different drying rates of the maturation zone enamel and that they may correspond to cyclical changes in the hydrophobicity and hydrophilicity of enamel matrix both on a daily basis and on a larger time scale.

Animals↗

Rapid uptake of calcium in maturing enamel of the rat incisor.

Six rats were given intravascular injections containing 45Ca, killed by perfusion with fixative and the incisor teeth removed within 2 min. Direct autoradiography of the maturing enamel surface showed bands of 45Ca uptake at this short interval.

Animals↗

Collagen orientation in compact bone: I. A new method for the determination of the proportion of collagen parallel to the plane of compact bone sections.

Previous work has shown that different types of osteons that have different mechanical properties with regard to tension and compression can be identified and studied by polarized light microscopy. This study reports the development of an automated method for the analysis of that proportion of collagen in transverse sections of human femurs that lies parallel with the plane of section. The method is based upon the use of circularly polarized light to detect collagen with a major component lying in the plane of the section, correcting the measurement for bright collagen under these circumstances with a corresponding darkfield measurement to determine the area of bone within each field of the section analyzed. The method has been validated in several different ways. The measured value of bright collagen under circularly polarized light depends upon the section thickness, since a linear relationship was found between section thickness and bright areas for a large number of microscopic fields. Plane parallel sections are therefore recommended for this study. The pattern of distribution of bright areas was the same for different observers and different set detection levels in any one section. The pattern found in adjacent sections from the same region of the shaft was identical.

Adult↗

Collagen orientation in compact bone: II. Distribution of lamellae in the whole of the human femoral shaft with reference to its mechanical properties.

Starting from a previous personal investigation (Portigliatti Barbos et al., 1983) indicating that the distribution of osteons and interstitial bone in the middle of the femoral shaft is related to their structure, a new procedure has been devised to allow information of the same kind to be gathered from the whole of the femoral diaphysis. Twenty-three exactly plane parallel cross-sections, 100 micron thick, each located 1 cm from the next, were prepared using an annular blade microtome. The distribution of longitudinal lamellae (whose fiber bundles and crystallites have longitudinal course and withstand loading by tension) and transverse lamellae (whose fiber bundles and crystallites have a transverse course and withstand loading by compression) was determined for both osteonic and interstitial bone, using circularly polarized light as the illuminating source and a Quantimet 720 image analyzing computer. The results show that along the femoral shaft the transverse and longitudinal lamellae from osteonic and interstitial bone have a characteristic rotational distribution consistent with the distribution of the bending forces normally operative in bone.

Adult↗