Search PubMed⌕ Search

Biomedical subjects

C M Holt

Publications and source records attributed to C M Holt.

30 records · Page 2Linked to original sources
PubMed1994

Release of platelet-derived growth factor activity from pig venous arterial grafts.

Intimal smooth muscle cell proliferation and superimposed atheroma are the main causes of late failure of saphenous vein bypass grafts. It has been suggested that these reactions are caused by the production of growth factors from the cells of the vessel wall. To test this hypothesis, we cultured segments of pig venous arterial grafts, removed 1 and 4 weeks after implantation, in serum-free medium for 24 hours. Tissue viability as assessed by adenosine triphosphate concentration was maintained throughout the 24-hour culture period (239 +/- 21 nmol/gm wet weight [standard error of the mean], n = 26, 0 hours; 240 +/- 24 nmol/gm wet weight, n = 17, 24 hours). Cell proliferation occurred and autoradiography showed proliferating cells to be located in the neointimal and medial layers. These cells were identified as smooth muscle cells by means of a monoclonal antibody to alpha-actin. Graft-conditioned media were tested for mitogenic activity by means of a fibroblast proliferation assay. Media conditioned for 24 hours produced significant stimulation of cell growth (284% +/- 30%, n = 17) above that obtained in culture medium alone (100%). This mitogenic activity was inhibited by 61% +/- 9%, n = 8, with a polyclonal-neutralizing antibody to platelet-derived growth factor. Reverse-transcription polymerase chain reaction analysis and Northern blots demonstrated platelet-derived growth factor B messenger ribonucleic acid (mRNA) in vein grafts but not in ungrafted vein. Analysis of graft tissue sections by in situ hybridization demonstrated an abundance of platelet-derived growth factor B mRNA positive cells in the endothelial and neointimal layers, as well as in the endothelial cells of the adventitial vessels. These data constitute direct evidence for active growth factor production within the cells of the vein graft. They also suggest that endogenously produced platelet-derived growth factor may play a role in regulating smooth muscle cell proliferation in this model.

Animals↗
PubMed1993

Comparison of response to injury in organ culture of human saphenous vein and internal mammary artery.

Autologous saphenous vein grafts, unlike internal mammary artery grafts, suffer many late occlusions as a result of excessive proliferation of vascular smooth muscle cells and the superimposition of atheroma on the resulting thickened intima. We investigated the possible basis of this difference using organ cultures. Internal mammary artery segments and freshly isolated and surgically prepared saphenous vein segments were obtained from patients undergoing coronary artery bypass grafting. Internal mammary artery and freshly isolated vein segments showed a high degree of endothelial coverage and medial cell viability that were maintained during culture. Surgically prepared veins showed partial endothelial denudation and medial cell injury, both of which tended to be reversed during culture. Neointimal thickening was greater in surgically prepared vein (72 +/- 13 microns; n = 11) than in freshly isolated vein (44 +/- 8 microns; n = 10) or internal mammary artery (34 +/- 4 microns; n = 13) segments. The occurrence of proliferating cells in the medial layer was also significantly greater in surgically prepared vein (2.8 +/- 1.0/mm; n = 11) than in freshly isolated vein (0.8 +/- 0.3/mm; n = 9) or internal mammary artery (0.6 +/- 0.3/mm; n = 10) segments. The data show that although the smooth muscle proliferation was similar in undamaged saphenous vein and internal mammary artery, it was significantly greater in damaged vein. This implies that the greater intimal proliferation seen in saphenous vein grafts may arise not from intrinsic differences in arterial and venous smooth muscle cells but from a greater susceptibility to injury.

Cell Division↗
PubMed1992

Intimal proliferation in an organ culture of human internal mammary artery.

OBJECTIVE: Intimal smooth muscle cell proliferation is an early feature of atherosclerosis. Its progression is difficult to monitor in humans and previous studies have mostly relied on necropsy material. The aim of this study was therefore to establish whether intimal proliferation occurred in an organ culture of human internal mammary artery. METHODS: Segments of freshly isolated internal mammary artery were maintained din standard tissue culture medium containing 30% calf serum for 14 d. Tissue viability (measured by ATP concentration) was maintained during processing and throughout the culture period [211(SEM 28) nmol ATP.g-1 wet weight on d 1 v 208(27) on d 14]. RESULTS: Histological transverse sections of cultured internal mammary artery showed the development of a neointima containing smooth muscle cells identified by immunocytochemistry for alpha actin. Pulse labelling of cultures with [3H]-thymidine showed proliferating cells predominantly in a neointimal layer with few dividing cells in the media. Cultured de-endothelialized vessels showed less neointimal thickening than cultured freshly isolated vessels [16(3) v 36(5) microns, p < 0.0025] as well as a reduced number of dividing cells per mm of neointimal length [3.1(0.6) v 5.5(1.1), p < 0.05]. CONCLUSIONS: Intimal proliferation occurred in organ culture of internal mammary artery. There is evidence for a factor derived from the endothelium, which may be important in the development of intimal proliferation.

Aged↗
PubMed1989

Prostacyclin production by human umbilical vein endothelium in response to serum from patients with systemic sclerosis.

Sera from 29 patients with systemic sclerosis and 30 normal controls were examined for their effect on prostacyclin release by human umbilical vein endothelial cells during periods of response of 15 min and 72 h and for their effect on endothelial growth and 3H-thymidine uptake during a 72-h culture period. In contrast to previous reports, no significant differences were detected between patient and control sera in their effect on endothelial cell prostacyclin release, growth or 3H-thymidine uptake.

Adult↗
PubMed1989

Antibody-dependent cellular cytotoxicity of vascular endothelium: characterization and pathogenic associations in systemic sclerosis.

Ten sera from 48 patients with systemic sclerosis were found to be capable of producing cytotoxicity of human umbilical venous and arterial endothelium when co-cultured with peripheral blood mononuclear cells. Fractionation of sera on Ultrogel and the preparation of monomeric IgG by ion exchange and affinity chromatography suggested that the cytotoxicity was mediated by anti-endothelial antibodies capable of pre-sensitizing target cells in a mechanism that resembled antibody-dependent cellular cytotoxicity. These anti-endothelial antibodies together with C1q-binding immune complexes and anti-cardiolipin antibodies were found in 18 of 28 patients so investigated, suggesting that multiple immunological mechanisms may be involved in the pathogenesis of the vascular lesion of systemic sclerosis.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1988

Elevated von Willebrand factor antigen in systemic sclerosis: relationship to visceral disease.

Plasma levels of the factor VIII complex (von Willebrand factor antigen, factor VIII coagulant and ristocetin co-factor) were measured in 28 patients with systemic sclerosis. Elevated von Willebrand factor antigen was found in 12 patients overall and in 10 of 16 patients characterized by severe extensive visceral disease, with a resulting positive correlation between the extent of visceral involvement and the plasma level of von Willebrand factor antigen (r = 0.60, p less than 0.001). Factor VIII coagulant and ristocetin co-factor levels, however, frequently failed to parallel the increases of von Willebrand factor antigen, supporting the view that these increases were due to in vivo endothelial damage. The findings suggest that vascular damage is an important aspect of the visceral lesions of systemic sclerosis.

Adult↗
PubMed1988

Anticardiolipin antibodies in systemic sclerosis: immunological and clinical associations.

Anticardiolipin antibodies of IgG/IgM class were detected in seven of 28 patients with systemic sclerosis including five of 16 patients severely affected by extensive visceral disease. This severely affected sub-group also showed significant elevations of plasma levels of von Willebrand factor antigen in 10 cases and serum C1q binding activity in seven cases respectively. This triple association raises the possibility that multiple immunological mechanisms are involved in the pathogenesis of systemic sclerosis and its vascular lesions.

Adult↗
PubMedDate not supplied

Development of an ex vivo model to investigate the effects of altered haemodynamics on human bypass grafts.

The insertion of vein grafts into the arterial circulation may contribute to vessel wall thickening and accelerated atherosclerosis, a common feature of late vein graft failure. We aimed to develop a model suitable for investigation of the effects of altered haemodynamics on human saphenous vein following its implantation into the arterial circulation. Segments of human saphenous vein obtained from patients undergoing coronary artery bypass surgery were sutured at each end to PTFE and placed into a flow system. Pressure and flow rates to stimulate the arterial and venous systems were achieved. A theoretical model of the flow chamber was created and computational fluid dynamics software (FLOTRAN, Swanson Analysis Systems) was used to determine the flow profile within the model. In summary, a flow model has been developed to investigate the effect of altered haemodynamics on the molecular and pathological changes that occur in vein grafts incorporated into the arterial circulation.

Blood Flow Velocity↗
PubMedDate not supplied

Biomimicry 1: PC.

The surface properties of stents can be modified by coating them, for example with a polymer. Phosphorylcoline (PC) is the major component of the outer layer of the cell membrane. The haemo- and biocompatibility of a PC-containing polymer is thus based on biomimicry, and has been confirmed by several experiments showing much reduced thrombogenicity of PC-coated surfaces, and porcine coronary artery implants showing no sign of adverse effect. Clinical experience with the PC-coated BiodivYsio appears favourable. The PC coating can be tailored for take up and controlled elution of various drugs for stent-based local delivery, a property which is being actively explored.

Animals↗