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Biomedical subjects

C W Archer

Publications and source records attributed to C W Archer.

At least 55 records · Page 3Linked to original sources
PubMed1993

Cell and matrix changes associated with pathological calcification of the human rotator cuff tendons.

The causative mechanism of tendon calcification ('calcifying tendinitis') is unknown. In this report, pathological human tendon samples were examined to give morphological and ultrastructural detail of the calcified regions and these findings were compared with those from normal tendon. Selected specimens were cryosectioned to enable histochemical and immunohistochemical comparison of the occurrence and distribution of specific matrix molecules in diseased and normal tendon tissues. The lack of collagen type II and alkaline phosphatase in the pathological regions suggests that the calcification process is not mediated through an endochondral transition. In contrast, the pathological areas were characterised by widespread labelling for chondroitin-4-sulphate/dermatan sulphate and intense pericellular localisation of chondroitin-6-sulphate.

Adult↗
PubMed1992

Myogenic potential of chick limb bud mesenchyme in micromass culture.

The myogenic potential of chick limb mesenchyme from stages 18-25 was assessed by micromass culture under conditions conductive to myogenesis, and was measured as the proportion of differentiated (muscle myosin-positive) mononucleated cells detected. It was found that similar myogenic potentials existed in mesenchyme from whole limbs between stages 18 and 19, but this potential was halved by stage 20. At stage 21, proximal mesenchyme showed significantly more myogenesis than distal mesenchyme, but this difference was abolished by stage 22. Thereafter, myogenesis was increasingly restricted from the distal mesenchyme, whilst the potential in more proximal regions did not significantly increase after stage 23. When the ratio between total limb myoblasts which differentiated on days 1 and 4 of culture was analysed, it was found that two distinct peaks existed at stages 20 and 23. The significance of these ratio peaks is unclear, but may be related to different proliferative potentials of the pre-myoblasts at these stages.

Animals↗
PubMed1992

Human articular surface chondrocytes initiate alkaline phosphatase and type X collagen synthesis in suspension culture.

The type X collagen is a short chain collagen associated with calcific cartilage and/or the expression of the hypertrophic chondrocyte phenotype. In articular cartilage, type X collagen is restricted to the basal zone of calcified cartilage adjacent to the subchondral bone. However, during pathological change such as in osteoarthritis, the synthesis of type X collagen becomes more widespread but never extends to the articular surface. Using immunocytochemistry and fluorography of newly synthesised collagens, we report that surface articular chondrocytes (which occupy the uppermost 10-15% of the tissue depth) from normal human cartilage initiate de novo synthesis of both type X collagen and alkaline phosphatase when maintained in suspension culture.

Alkaline Phosphatase↗
PubMed1992

The development of the perichondrium in the avian ulna.

The development of the perichondrium surrounding the ulna of chick embryos was examined between stages 26 and 34 (Hamburger & Hamilton, 1952), i.e. 5.5-8 d of incubation. The first appearance of a barrier between cartilage and adjacent tissue was observed at stage 27 when cells were arranged circumferentially around the central flattened core, but no true perichondrium was seen until chondrocyte hypertrophy began at stage 30. At this stage, cells in the barrier became elongated parallel to the long axis of the rudiment and overlapped. With further cell hypertrophy, the perichondrium consolidated and by stage 31 a distinct, compact structure, several cells thick and exhibiting cell-cell contacts had formed around the hypertrophic cell region. By stage 32, overlapping perichondrial cells could be seen surrounding the flattened cell region and this structure consolidated as cell hypertrophy spread from the centre towards the ends of the rudiment. The rounded cell zone at the epiphyses never formed a distinct perichondrium during the time course of this study. This differential structure of a compact perichondrium surrounding the hypertrophic cell region, a loose perichondrium surrounding the rounded cell region and an intermediate perichondrium surrounding the flattened cell region may facilitate the morphogenesis of the rudiment by acting as a constraining sheath restricting radial expansion but favouring longitudinal expansion.

Animals↗
PubMed1991

Isolation and characterisation of a chicken gelatinase (type IV collagenase).

The proform of chick gelatinase (type IV collagenase) was isolated and purified to a high specific activity of 12,071 U/mg from cultured embryonic skin fibroblasts stimulated with cytochalasin-B. The enzyme was activated in the presence of 4-aminophenylmercuric acetate with a fall in molecular weight from 66,000-58,000 on non-reducing polyacrylamide gel electrophoresis and was active over the pH range of 6.0-8.9 against a number of substrates. Further biochemical characterisation showed that the organomercurial activated form of the enzyme behaved like a typical mammalian gelatinase, actively degrading gelatin, soluble type I collagen, collagenase generated type I fragments, type IV collagen (producing 3/4 and 1/4 fragments) and type V collagen, whilst having little effect on laminin. The enzyme was inhibited by metal chelators such as EDTA and 1,10-phenanthroline, but not by inhibitors is suggested that this may be TIMP-2. An antiserum was raised to the proenzyme and was found to localise intra- and extra-cellularly in both tissue sections and cell cultures.

Animals↗
PubMed1991

Three-dimensional collagen architecture in bovine articular cartilage.

The three-dimensional architecture of bovine articular cartilage collagen and its relationship to split lines has been studied with scanning electron microscopy. In the middle and superficial zones, collagen was organised in a layered or leaf-like manner. The orientation was vertical in the intermediate zone, curving to become horizontal and parallel to the articular surface in the superficial zone. Each leaf consisted of a fine network of collagen fibrils. Adjacent leaves merged or were closely linked by bridging fibrils and were arranged according to the split-line pattern. The surface layer (lamina splendens) was morphologically distinct. Although ordered, the overall collagen structure was different in each plane (anisotropic) a property described in previous morphological and biophysical studies. As all components of the articular cartilage matrix interact closely, the three-dimensional organisation of collagen is important when considering cartilage function and the processes of cartilage growth, injury and repair.

Animals↗
PubMed1990

Effects of ascorbate on myogenesis in micromass culture.

Micromass cultures from stage 23 and 24 chick wing mesenchyme were grown in serum-containing medium with or without additional ascorbic acid. It was found that ascorbic acid administered as a single pulse or present continuously throughout culture, in concentrations as low as 25 micrograms/ml, was sufficient to abolish 80% of myogenesis as assessed by immunolocalization using muscle-specific antibodies. This effect was not significantly altered when cultures were maintained in a serum-free medium that promotes myogenesis. In contrast to the above findings, spectrophotometric analysis of accumulated sulphated glycosaminoglycans, an indicator of chondrogenesis, was elevated by ascorbate treatment. Furthermore, a similar level of glycosaminoglycan stimulation was found in ascorbate treated stage 23 distal-tip limb cultures that were essentially free of myogenic cells. We conclude, therefore, that the presence of myoblasts in whole-limb cultures has no appreciable inhibitory effects on chondrogenesis.

Animals↗
PubMed1990

Phenotypic modulation in sub-populations of human articular chondrocytes in vitro.

Human articular cartilage has been separated into surface (approx. 15% of tissue depth) and deep zones (remaining tissue) and the constituent chondrocytes released by enzymic digestion. Subsequent culture either as a low density monolayer or as a suspension over agarose revealed distinct morphological and synthetic behaviour in the two populations. Whilst in monolayer these morphological differences disappeared with time in culture, over agarose they remained. Surface zone cells formed two types of cell cluster; one that was highly cellular with little extracellular matrix, and the other less frequent, which formed copious amounts of fibrillar matrix. Both types of cluster were surrounded by a layer of flattened chondrocytes. In contrast, deep cells formed a single cluster type that lacked a surrounding cell layer, but formed large amounts of sparse cartilage-like matrix and comprised morphologically typical chondrocytes. In monolayer, both populations gradually ceased to synthesise cartilage matrix components with the exception of link protein. In suspension, whilst the chondrogenic phenotype per se was preserved, there was, nevertheless, a loss in qualitative synthetic heterogeneity, which exists between surface and deep cells, that was not accompanied by changes in the differential rate of 35S incorporation into proteoglycan. Under these conditions, surface cells that normally do not synthesise keratan sulphate initiated de novo synthesis of this glycosaminoglycan. Consequently, it appears that the observed modulation in synthetic ability of the cell sub-populations is independent of the cluster morphology, which, once established, remains constant throughout the culture period.

Cartilage, Articular↗
PubMed1990

A role for hyaluronan in joint development.

Hyaluronan, a common connective tissue component, influences cell adhesion, migration and cytodifferentiation in vitro, and because of these properties has been postulated to have a role in morphogenesis. Its role in the development of cartilage-associated structures, such as joints, has yet to be defined. Using a biotinylated hyaluronan-binding region-link protein complex, free hyaluronan binding sites have been localised in the joint region concomitant with the first signs of cavitation (Stage 37), whereafter it is localised in the joint space and is maintained here as this enlarges. The application of our results is discussed in the context of a primary role for hyaluronan in joint cavity formation.

Animals↗
PubMed1987

MZ15, a monoclonal antibody recognizing keratan sulphate, stains chick tendon.

Biochemical, morphometric and electron histochemical methods have failed to demonstrate the presence in tendon of keratan sulphate, a common component of connective tissue proteoglycans. Using a monoclonal antibody specific for keratan sulphate, a positive localization of this molecule was made in the gastrocnemius tendon of stage 44 chicken embryos both at the light and electron microscopical levels.

Animals↗
PubMed1987

The differentiation of normal and muscle-free distal chick limb bud mesenchyme in micromass culture.

Distal chick wing bud mesenchyme from stages 19 to 27 embryos has been grown in micromass culture. The behavior of cultures comprising mesenchyme located within 350 microns of the apical ectodermal ridge (distal zone mesenchyme) was compared to that of cultures of the immediately proximal mesenchyme (subdistal zone cultures). In cultures of the distal mesenchyme from stages 21-24 limbs, all of the cells stained immunocytochemically for type II collagen within 3 days, indicating ubiquitous chondrogenic differentiation. At stage 19 and 20, this behavior was only observed in cultures of the distal most 50-100 microns of the limb bud mesenchyme. Between stages 25 and 27, distal zone cultures failed to become entirely chondrogenic. At all stages, subdistal zone cultures always contained substantial areas of nonchondrogenic cells. The different behavior observed between distal zone and corresponding subdistal zone cultures appears to be a consequence of the presence of somite-derived presumptive muscle cells in the latter, since no such difference was observed in analagous cultures prepared from muscle-free wing buds. The high capacity of the distal zone for cartilage differentiation supports a view of pattern formation in which inhibition of cartilage is an important component. However, its consistent behavior in vitro indicates that micromass cultures do not reflect the in vivo differences between the distal zones at different stages. The subdistal region retains a high capacity of cartilage differentiation and the observed behavior in micromass reflects interactions with a different cell population.

Animals↗
PubMed1987

Cell sorting and chondrogenic aggregate formation in micromass culture.

A fundamental feature of cartilage differentiation in the developing limb is the formation of a prechondrogenic cell condensation. An apparently similar process of prechondrogenic cell aggregation occurs in micromass cultures of limb bud mesenchyme with the formation of cellular aggregates which often differentiate into cartilage nodules. We have investigated the process of aggregate formation in micromass culture using chimaeric mixtures of potentially chondrogenic and nonchondrogenic cell types. Two systems were studied: mixtures of distal and proximal limb mesenchyme cells and mixtures of distal limb cells with avian tendon fibroblasts. In both cases cultures of varying proportions of each cell type have been prepared. The results demonstrate that aggregate formation in vitro is the consequence of a cell sorting process which can involve prechondrogenic cells of widely different spatial origins within the developing limb. This contrasts with in vivo prechondrogenic condensation in which there is no evidence of cell sorting (Searls, R.L. (1967), J. Exp. Zool. 166, 39-50). However, our findings do indicate that cell surface differences occur in apparently undifferentiated limb mesenchyme. The results also suggest that mesenchymal cell aggregates must achieve a threshold size before chondrogenesis can proceed. In addition, the results show that under some culture conditions nonchondrogenic cells will form aggregates.

Animals↗
PubMed1987

The spatial and temporal pattern of collagens I and II and keratan sulphate in the developing chick metatarsophalangeal joint.

Both intrinsic and extrinsic factors are known to be involved in the morphogenesis of diarthrodial joints. The use of specific antibodies to collagens I and II and keratan-sulphate-containing proteoglycans (KSPG) has enabled the distributions of these macromolecules to be followed during the development of the third metatarsophalangeal joint in the chicken embryo. Our study shows that cartilage differentiation occurs as a continuous rod, which is then subsequently divided into separate elements. Further development also reveals that, unlike the matrix of the cartilaginous elements, there is a differential distribution of collagen (type II) and KSPG in the presumptive joint region. It is proposed that a decrease in KSPG in the presumptive joint region at stages 28/30 may be involved in the mechanism for the flattening of cells in formation of the interzone. Whereas, a decrease in collagen across the joint interzone region may provide an area of weakness, which might allow forces produced by the developing musculature to cause cavitation.

Animals↗
PubMed1985

Cartilage morphogenesis in vitro.

The morphogenetic capacity of prechondrogenic mesenchyme from two developmentally distinct sources was investigated in high density micromass cultures. We confirmed an earlier report (Weiss & Moscona, 1958) that scleral mesenchyme formed cartilage sheets whilst limb bud mesenchyme formed distinct cartilage nodules. It was thus suggested by these authors that this morphogenesis was tissue type specific. However, by varying cell density at inoculation (which controls cell configuration) and by varying the relative amount of prechondrogenic mesenchyme present in cultures we found that dramatic changes in morphogenesis could be brought about. Viewed in these terms we suggest that cartilage morphogenesis in vitro is dependent on cell configuration and the presence of non-chondrogenic cell types and hence is not necessarily a function of an intrinsic morphogenetic potential of the constituent cells.

Animals↗
PubMed1983

The effects of pulsed magnetic fields on chick embryo cartilaginous skeletal rudiments in vitro.

The biological response of cultured 7-day embryonic chick tibiae to small alternating currents induced by pulsed magnetic fields (PMFs) was investigated. It was found that continuous exposure to PMFs over 7 days did not alter the overall DNA content of rudiments nor the incorporation of 3H-thymidine when compared with control tibiae. The overall collagen content of explants was slightly reduced by PMF exposure whilst the incorporation of 3H-proline was significantly suppressed. The synthesis of sulphated glycosaminoglycans was also measured in terms of 35SO4--incorporation, but PMF treatment failed to alter the levels of isotope incorporation. These results suggest that, whereas noncollagenous, and possibly collagenous, protein synthesis is affected by PMF exposure, glycosaminoglycan synthesis is not. Histological and electron microscopical observations substantiated this interpretation and revealed selective inhibition of matrix secretion in the periphery of the proliferative epiphyseal zones in experimental explants. High-power electron microscope examination of these zones showed that PMF-exposed matrix was sparsely invested with fibrous protein while elements of the stellate reticulum had formed large aggregates which were often clumped about the cell membrane. The results are discussed in terms of the possible role of naturally occurring potentials in the development and maintenance of connective tissue systems such as cartilage and bone.

Animals↗
PubMed1983

Growth and morphogenesis of the fibula in the chick embryo.

The development of the avian fibula was studied both histologically and experimentally. It was found that from the onset of chondrogenesis, the fibula possessed a smaller diameter than the neighbouring tibia. The truncated growth of the fibula was a result of the loss of its distal epiphysis between stages 27-31. This epiphysis subsequently became fused to the tibia and formed the fibulare of the tibiotarsus. The experiment of Hampé (1960) was repeated by inserting tantalum barriers into the limb between stages 18 and 23: this sometimes prevented the separation of the fibula distal epiphysis, thus giving rise to an elongated element. A similar result was obtained from grafts of polarizing region into the leg bud at stages 18-20. It was concluded that there was no evidence for competitive interaction between the blastemata of the tibia and fibula. In addition, the differential growth in diameters between the tibia and fibula was largely a result of differential osteogenesis rather than chondrogenesis as previously thought.

Animals↗