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Biomedical subjects

S Downes

Publications and source records attributed to S Downes.

53 records · Page 3Linked to original sources

Comparison of the release of growth hormone from hydroxyapatite, heat-treated hydroxyapatite, and fluoroapatite coatings on titanium.

Hydroxyapatite (HA), heat-treated hydroxyapatite (HAH), and fluorapatite (FA) coatings on titanium were loaded with human growth hormone (GH), and the subsequent release was monitored in vitro. The amount of GH released was significantly increased from the HA coating that had received a post-plasma-spraying heat treatment prior to incorporation of the growth hormone. Scanning electron microscopy was used to assess the surfaces of the ceramic coatings prior and postincubation with GH. Surface changes were observed on the HA and HAH coatings but not on the FA coatings after incubation with GH. Osteoblast-like cells were grown on the coatings and maintained in culture. Scanning electron microscopy was used to study the morphology of the cells and the interaction of the cells with the ceramic coatings. We used thymidine uptake and DNA content to determine the relative rates of cell division on the different coatings; the optimum rate of cell proliferation was observed on the HAH coating loaded with 0.1 IU/mL GH.

Apatites↗

Blends of synthetic and natural polymers as drug delivery systems for growth hormone.

In order to overcome the biological deficiencies of synthetic polymers and to enhance the mechanical characteristics of natural polymers, two synthetic polymers, poly(vinyl alcohol) (PVA) and poly(acrylic acid) (PAA) were blended, in different ratios, with two biological polymers, collagen (C) and hyaluronic acid (HA). These blends were used to prepare films, sponges and hydrogels which were loaded with growth hormone (GH) to investigate their potential use as drug delivery systems. The GH release was monitored in vitro using a specific enzyme-linked immunosorbent assay. The results show that GH can be released from HA/PAA sponges and from HA/PVA and C/PVA hydrogels. The initial GH concentration used for sample loading affected the total quantity of GH released but not the pattern of release. The rate and quantity of GH released was significantly dependent on the HA or C content of the polymers.

Acrylic Resins↗

Modifications of the hydrophilicity of heterocyclic methacrylate copolymers for protein release.

A series of copolymers comprising ethyl methacrylate (EM) and tetrahydrofurfuryl methacrylate (THFMA) gelled with either THFMA monomer or hydroxyethyl methacrylate (HEMA) monomer have been developed. In this paper, we examine the water uptake characteristics of the polymer systems and address the possibility of increasing the hydrophilicity of the systems by changing the ratios of the copolymers. We have investigated whether protein release from the polymers is related to the composition of the polymer systems. More protein was released from the polymers gelled with the more hydrophilic monomer (HEMA) than with THFMA. This was consistent with the calculated diffusion coefficients, which were 10 times greater for the polymers gelled with HEMA than those gelled with THFMA. Interestingly, the water uptake and protein release profiles were not dependent on the ratio of EM and THFMA in the copolymers. This is probably due to the conflicting roles of THFMA in the copolymer; it is both the more hydrophilic component as well as a cross-linking agent. In addition, it would appear that the structural and surface topography of these polymers had more significant effects on protein release than copolymer composition.

Absorption↗

Comparison of the early production of extracellular matrix on dense hydroxyapatite and hydroxyapatite-coated titanium in cell and organ culture.

In order to investigate the earliest changes at the bone-hydroxyapatite (HA) interface, two materials, dense HA and HA-coated titanium alloy, were investigated for their ability to stimulate production of extracellular matrix both in cell and organ culture. Osteoblast-like cells were obtained from rabbit calvariae after preliminary culture for 3 d and seeded out onto the materials. When these cells were grown in culture on the dense HA no matrix production was seen with the scanning electron microscope after 4 d, but there did appear to be matrix produced on the rougher surface of the HA coating. In contrast, extracellular matrix production was greatly increased in organ culture where the material was placed on a section of bone, particularly on the HA coating, where it formed an organized layer of collagen within 4 d. These results showed that extracellular matrix production by both osteoblastic cells and in calvarial culture was greater on the HA coatings than dense HA. In addition, organ culture allows the investigation of the earliest changes in the bone-HA interface without having to undertake in vivo implantation studies of such short duration.

Alloys↗

Biodegradable microspheres: a new delivery system for growth hormone.

A drug delivery system for biologically active agents targeted to specific cells could be used to improve tissue repair in orthopaedics. The system should be controllable and capable of drug release over an extended period of time. Biodegradable, membrane-moderated, monolithic microspheres for the controlled release of growth hormone (GH) were developed and the release of GH was monitored in vitro. Cross-linked gelatin microspheres were used as the vehicle, with the drug dispersed within the gelatin. The amount of GH released from the microspheres was increased following ultrasonication. The release of growth hormone was monitored in phosphate buffered saline and horse serum. Interestingly, a higher level of GH was detected in the phosphate buffered saline than in serum. In addition, both pH and enzyme-induced degradation had an effect on the swelling kinetics of the gelatin microspheres. The release of GH from the microspheres was diffusion controlled, during the time period studied.

Biocompatible Materials↗

Validation and optimization of a polymer system for potential use as a controlled drug-delivery system.

A room temperature polymerizing system consisting of polyethylmethacrylate (PEM) and tetrahydrofurfuryl methacrylate (THFMA) monomer was validated for use as a drug-delivery system. The effect of gelling PEM/THFMA copolymers with a more hydrophilic monomer, hydroxyethyl methacrylate (HEMA), was also investigated. The release of growth hormone (GH) and bovine serum albumin (BSA) protein from this polymer has been studied. The polymer has the advantage of high water absorption and low shrinkage properties. Changes in release profiles have been studied by introducing structural differences in the polymer by changing the mixing technique. The stability and bioactivity of the GH incorporated have been examined at various temperatures. To optimize release profiles further, the possibility of attaining a more sustained and controllable release by varying the ratios of polymer used has also been investigated.

Drug Delivery Systems↗

Methods for improving drug release from poly(methyl)methacrylate bone cement.

Poly(methylmethacrylate) (PMMA) is a widely used material with both dental and orthopaedic applications. The acrylic cement is produced by the combination of polymethylacrylate beads with methylmethacrylate monomer. After polymerisation, a heterogeneous and porous matrix is formed which can be used to deliver therapeutic agents. In this work, the release of antibiotic, growth hormone and serum albumin is demonstrated. The mechanism is similar for all agents; a rapid release followed by a slow continuous release. The quantity of drug released depends upon the formulation of both the PMMA and the drug. The polymer-to-monomer ratio can greatly affect the ratio of drug release; increased polymer-to-monomer ratio leads to increased release of antibiotic. Optimum release is achieved if a crystalline formulation of the drug is used rather than a fine powder. Experimental methods to improve the drug release performance of bone cements are presented.

Bone Cements↗

Results of case-control study of leukaemia and lymphoma among young people near Sellafield nuclear plant in West Cumbria.

OBJECTIVE: To examine whether the observed excess of childhood leukaemia and lymphoma near the Sellafield nuclear plant is associated with established risk factors or with factors related to the plant. DESIGN: A case-control study. SETTING: West Cumbria health district. SUBJECTS: 52 Cases of leukaemia, 22 of non-Hodgkin's lymphoma, and 23 of Hodgkin's disease occurring in people born in the area and diagnosed there in 1950-85 under the age of 25 and 1001 controls matched for sex and date of birth taken from the same birth registers as the cases. MAIN OUTCOME MEASURES: Antenatal abdominal x ray examinations, viral infections, habit factors, proximity to and employment characteristics of parents at Sellafield. RESULTS: Expected associations with prenatal exposure to x rays were found, but little information was available on viral illnesses. Relative risks for leukaemia and non-Hodgkin's lymphoma were higher in children born near Sellafield and in children of fathers employed at the plant, particularly those with high radiation dose recordings before their child's conception. For example, the relative risks compared with area controls were 0.17 (95% confidence interval 0.05 to 0.53) for being born further than 5 km from Sellafield 2.44 (1.04 to 5.71) for children of fathers employed at Sellafield at their conception, and 6.42 (1.57 to 26.3) for children of fathers receiving a total preconceptual ionising radiation dose of 100 mSv or more. Other factors, including exposure to x rays, maternal age, employment elsewhere, eating seafood, and playing on the beach did not explain these relationships. Focusing on Seascale, where the excess incidence has predominantly been reported, showed for the four out of five cases of leukaemia and one case of non-Hodgkin's lymphoma whose fathers were employed at Sellafield and for whom dose information was obtained that the fathers of each case had higher radiation doses before their child's conception than all their matched control fathers; the father of the other Seascale case (non-Hodgkin's lymphoma) was not employed at the plant. These results seem to explain statistically the geographical association. For Hodgkin's disease neither geographical nor employment associations with Sellafield were found. CONCLUSIONS: The raised incidence of leukaemia, particularly, and non-Hodgkin's lymphoma among children near Sellafield was associated with paternal employment and recorded external dose of whole body penetrating radiation during work at the plant before conception. The association can explain statistically the observed geographical excess. This result suggests an effect of ionising radiation on fathers that may be leukaemogenic in their offspring, though other, less likely, explanations are possible. There are important potential implications for radiobiology and for protection of radiation workers and their children.

Case-Control Studies↗

Methods and basic data of case-control study of leukaemia and lymphoma among young people near Sellafield nuclear plant in West Cumbria.

OBJECTIVE: To examine whether the observed excess of childhood leukaemia and lymphoma near the Sellafield nuclear plant is associated with established risk factors or with factors related to the plant. DESIGN: A case-control study. SETTING: West Cumbria health district. SUBJECTS: 52 Cases of leukaemia, 22 of non-Hodgkin's lymphoma, and 23 of Hodgkin's disease occurring in people born in the area and diagnosed there in 1950-85 under the age of 25 and 1001 controls matched for sex and date of birth taken from the same birth registers as the cases. MAIN OUTCOME MEASURES: Antenatal abdominal x ray examinations, viral infections, habit factors, proximity to and employment characteristics of parents at Sellafield. RESULTS: Ascertainment of cases through multiple sources was as complete as possible, and the diagnosis was established for nearly all cases from hospital records and by independent pathological review when suitable material (60% (58) of cases) was available. Identification and tracing of the parents of cases and controls enabled questionnaires to be forwarded to 730 (66%), and 467 (64%) of the questionnaires were returned completed. Obstetric records were located for 481 (44%) of the relevant births, more frequently in recent years. Linkage of study subjects to the Sellafield workforce file enabled dates of employment and records on external doses of whole body ionising radiation to be obtained. Concordance of information from duplicate sources (when available) was reasonably high with no indications of bias. CONCLUSION: Overall the collected data were sufficiently reliable for detailed analysis and careful interpretation.

Case-Control Studies↗

Dopamine1-receptor blockade inhibits ANP-induced phosphaturia and calciuria in rats.

Atrial natriuretic peptide (ANP) is known to enhance the excretion of Pi and Ca, solutes reabsorbed primarily by the proximal tubule. Previous studies have shown that proximal tubule Na transport is inhibited by dopamine (DA), and that the natriuretic action of ANP is blunted by DA-receptor blockade. However, alterations in Na reabsorption cannot localize ANP or DA action to a specific nephron site. Therefore, the possibility that DA mediates the apparent proximal tubule effects of ANP was investigated with the use of Pi and Ca as proximal tubule markers. ANP was infused into normal rats in the presence and absence of specific DA-receptor antagonists, and Na, Pi, and Ca excretion rates were determined. ANP enhanced Na, Pi, and Ca excretion at doses that failed to alter glomerular filtration rate and mean arterial pressure (MAP). DA1-receptor blockade significantly blunted the influence of ANP on urinary Na, Pi, and Ca excretion, whereas DA2-receptor blockade was without effect. MAP and inulin and p-aminohippurate (PAH) clearances remained stable during DA-receptor blockade. Because endogenous ANP levels are elevated in rats with remnant kidneys, and because blockade of endogenous ANP reduces Pi and Ca as well as Na excretion in this model, the effect of DA1-receptor blockade on solute excretion was also examined in rats with 5/6 nephrectomy. DA1-receptor blockade significantly reduced absolute and fractional Na, Pi, and Ca excretion in rats with 5/6 nephrectomy, in the absence of measurable changes in MAP, inulin, or PAH clearance.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Growth hormone in polymethylmethacrylate cement.

The in vitro release of human growth hormone (hGH) from polymethylmethacrylate bone cement was measured for up to 40 days using a specific enzyme-linked immunosorbent assay. In rabbits, hGH-loaded bone cement was inserted into one distal femur and plain cement was inserted into the contralateral distal femur as a control. The rabbits were killed at time intervals of one, two, and four months after surgery and the distal femurs embedded whole for histology. Quantitative histomorphometry indicated there was a greater percentage of osteoid present at the hGH-loaded cement surface than at the plain cement surface one month after implantation. The hGH released from bone cement may stimulate osteogenesis and new bone formation, thus improving the strength of the bone-cement interface.

Animals↗

Follow up study of children born elsewhere but attending schools in Seascale, West Cumbria (schools cohort).

Records on 1546 children who were identified as having attended schools in Seascale up to November 1984 and were born since 1950 but not in the civil parish were studied. These children lived in or near Seascale for a period of time while they were attending one or more of three local schools and are an additional group to the 1068 children who were identified as born to mothers resident in Seascale in an accompanying study. Even though some of the schoolchildren apparently remained in the village for a short period only all but 7% were followed up through the National Health Service Central Register. Mortality among these children to 30 June 1986 is comparable to that expected at national rates. From all causes there were 10 observed deaths compared with 12.69 expected--a ratio of 0.79 (95% confidence interval 0.38 to 1.45)--and from cancer one observed death compared with 2.04 expected--a ratio of 0.49 (95% CI 0.01 to 2.73). No deaths from leukaemia or lymphoma were reported, but only 0.83 was expected. Since 1971 (the year when cases of cancer were first notified to the NHS Central Register) three non-fatal cases of cancer were reported, including two lymphomas, compared with 2.04 expected and two cases of carcinoma in situ of the cervix compared with 1.79 expected. In addition, there was a case of leukaemia among the schoolchildren which was known previously and had been diagnosed in 1968. There is an interesting difference between the results of this study and the results of the study of children born to mothers who were resident in Seascale. In the latter study there was an excess of leukaemia and of other cancers, but a similar finding is not apparent among children who spent some time at schools in Seascale but were born elsewhere. This raises the question of whether one or more aetiological factors in childhood cancer were acting on a locality specific basis before birth or early in life. This cannot be answered from these cohort studies, but it is hoped that the case-control study that is under way in West Cumbria will provide relevant information.

Adolescent↗

Follow up study of children born to mothers resident in Seascale, West Cumbria (birth cohort).

Records on 1068 children who were born to mothers resident in Seascale Civil Parish during 1950-83 were studied. There was a large degree of mobility among the families, and nearly half of the children did not subsequently attend the main local school. Use of the National Health Service Central Register, however, enabled us to follow up the children's records regardless of place of residence. The excess of leukaemia among Seascale children first supported from the analysis of geographical areas is confirmed. There were five deaths from leukaemia identified to 30 June 1986 compared with 0.53 expected at national rates--a ratio of 9.36 (95% confidence interval 3.04 to 21.84). One of these deaths occurred after the child had left Seascale. There were four deaths from other cancers compared with 1.06 expected--a ratio of 3.76 (95% CI 1.02 to 9.63). In addition, three further cases of cancer, apart from the deaths, were reported compared with 1.19 expected since 1971--a ratio of 2.53 (95% CI 0.52 to 7.40). For other causes of death, including stillbirths and infant mortality, there was a reported deficit compared with national rates, some of which at least was to be expected on the basis of the social class composition of the population of Seascale. In view of the importance of this cohort of births continued follow up is planned, with the possibility of extending it to include births since 1983, and the methods available for this type of study will be examined further.

Adolescent↗

The clearance of gentamicin from antibiotic-loaded bone cement.

The concentrations of Gentamicin in the serum, drainage fluid and urine were determined by radioimmunoassay in 40 patients after total hip replacement and 10 after knee replacement. Palacos R bone cement with added Gentamicin had been used to fix the prostheses in all cases. Two types of response were observed; in Group I the Gentamicin levels rose very rapidly, whereas in Group II a more gradual increase was seen. In all cases the level of Gentamicin had fallen to a negligible amount by the fifth day after operation.

Bone Cements↗

Osteoblast responses to collagen-PVA bioartificial polymers in vitro: the effects of cross-linking method and collagen content.

A range of 'bioartificial' collagen/poly(vinyl alcohol) blends have been produced, cast as films and cross-linked using either glutaraldehyde or a dehydrothermal treatment (DHT). Films were used as substrates for the culture of osteoblast-like cells. The attachment, adhesion and proliferative responses of these cells to the range of films were examined using proliferation assays, light, electron and confocal microscopy. There was an inverse relationship between collagen content of gluataraldehyde cross-linked films and the extent of cell proliferation on them. A cytotoxicity assay demonstrated no toxic effect related to increasing collagen content. The greatest differences in cell responses observed were associated with the choice of cross-linking method. Films cross-linked with glutaraldehyde showed variation related to collagen content in cell adhesion, proliferation and morphology. Such differences were not apparent with the DHT cross-linked films. Collagen/PVA 'bioartificial' films can be dehydrothermally cross-linked to increase biological stability and reduce water solubility. The method of cross-linking employed is the greater influence in determining osteoblast compatibility with these materials. The DHT cross-linking method is a preferable alternative to the use of glutaraldehyde. Collagen/PVA bioartificial films cross-linked by the DHT method have shown potential for biocompatibility with osteoblasts.

Biocompatible Materials↗