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Biomedical subjects

L M Graham

Publications and source records attributed to L M Graham.

82 records · Page 5Linked to original sources
PubMed1980

Endothelial cell seeding of prosthetic vascular grafts: early experimental studies with cultured autologous canine endothelium.

Thirteen adult dogs underwent thoracoabdominal bypass operations with 6-mm, double-velour Dacron grafts 25 to 30 cm long. Experimental grafts were seeded with cultured autologous endothelial cells (n = 7). Unseeded grafts served as controls (n = 6). Endothelial cells were harvested from external jugular vein segments using 0.1% trypsin and 0.5% collagenase solutions. Grafts were studied at weeks 2 and 4. Endothelial cell coverage of experimental graft surfaces after two weeks was 60% to 70%, and approximately 80% after four weeks. Immunofluorescence using factor VIII-related antigen confirmed the graft's inner surface to be endothelium. Endothelial cell coverage in control grafts occurred as pannus ingrowth, and never exceeded more than 10% of the conduit surface. Generation of an early endothelial surface in prosthetic grafts is possible in a canine model using cultured autologous cells.

Animals↗
PubMed1980

Immediate seeding of enzymatically derived endothelium in Dacron vascular grafts. Early experimental studies with autologous canine cells.

Twenty-eight adult dogs underwent thoracoabdominal bypass with 6-mm double-velour Dacron grafts. The experimental grafts were seeded immediately prior to implantation with enzymatically harvested endothelial cells. Endothelial cells were procured from autologous external jugular vein segments using 0.1% trypsin and 0.5% collagenase solutions. Unseeded grafts served as controls. The grafts were studied from one to 28 days after implantation. The seeded grafts exhibited greater than 80% uniform luminal coverage with endothelial cells at day 28. Immunofluorescent microscopy was used to confirm the presence of endothelium. The experimental grafts studied at the two time periods of one to seven days and 14 to 28 days had 94.6% and 88.5% clot-free surfaces, respectively. The control grafts studied during similar periods had 81.0% and 40.1% clot-free surfaces. These differences were significant at the 14- to 28-day period.

Animals↗
PubMed1979

Abdominal aortic coarctation and segmental hypoplasia.

Functionally significant, nonatherosclerotic, noninflammatory, concentric and tubular stenoses of the abdominal aorta, 4 to 16 cm in length, were encountered in five male and five female patients 11 to 49 years old. Seven patients were younger than 19 years of age. Aortic branch stenoses were common, affecting splanchnic vessels in seven patients and renal arteries in eight patients. The pathogenesis of the aortic constrictive lesions remains unknown, but it may be related to developmental error or aortic growth arrest. Existence of multiple renal arteries in 70% of these patients lends support to the developmental hypothesis. Intimal fibroplasia characterized stenotic aortic tissue. Severe hypertension was common, with the mean preoperative arterial pressure being 200/119 mm Hg. Thoracoabdominal bypass was undertaken in eight patients, being combined with renovascular reconstruction on five occasions. Two patients underwent patch graft aortoplasty with bilateral renal revascularization. Therapeutic results were classified as excellent six times and as good four times. Single-stage arterial reconstructions are the preferred method of treatment for abdominal aortic coarctation or segmental hypoplasia with associated aortic branch disease.

Adolescent↗
PubMedDate not supplied

Effects of duration of acetylsalicylic acid administration on patency and anastomotic hyperplasia of ePTFE grafts.

Effects of duration of acetylsalicylic acid (ASA) administration on the patency and development of distal anastomotic intimal hyperplasia (DAIH) of endothelial cell (EC) seeded and unseeded prosthetic aortoiliac grafts were studied in a canine model. ASA, 5 gr po qd, was administered to dogs 1 day prior to placement of bilateral, 12 to 17 cm long, 5 mm inside diameter expanded polytetrafluoroethylene (ePTFE) aortoiliac grafts and continued for 2 wk (Group 1, n = 12 dogs) or 16 wk (Group 2, n = 12 dogs). Six dogs in each group received autologous EC seeded grafts, while the others received unseeded grafts. Prosthesis patency was assessed weekly. At the conclusion of the study, DAIH was measured on serial sections using a computer-linked digitizer. The 16 wk patency for Group 1 grafts was 67%, while that for Group 2 grafts was 88% (p less than 0.09). Luminal narrowing due to DAIH was not significantly different between Groups 1 and 2 (7.7 +/- 8.3% [means +/- SD] and 9.0 +/- 7.8% respectively). EC seeding improved the 16 wk combined patency from 62 to 92% (p less than 0.02). A more complete luminal endothelial cell lining was correlated with reduced DAIH (r = -0.4, p less than 0.05). Chronic ASA administration prevented graft thrombosis between 2 and 4 wk postimplantation in this study but was not associated with decreased DAIH.

Anastomosis, Surgical↗
PubMedDate not supplied

Differential monocytic cell adherence to specific anatomic regions of the canine aorta.

Circulating monocytes and vascular endothelial cells (EC) interact in a complex and dynamic manner that varies between vascular beds. The objective of this study was twofold: to ascertain if monocytic cell adhesion to vascular endothelium differed between specific anatomic regions of the canine aorta, and to investigate the effect of known EC stimulators on monocytic cell adhesion to cells from these regions. Initial in vitro studies measuring adherence of U937 cells, a human monocytic cell line, to canine jugular vein and aortic EC monolayers revealed a dose-dependent increase in adhesion to EC stimulated with interleukin-1 (IL-1), lipopolysaccharide (LPS), phorbol 12-myristate 13-acetate (PMA), or thrombin. While there was no regional difference in monocytic cell adherence to unstimulated EC in tissue culture, studies demonstrated greater monocytic cell adhesion to stimulated EC cultured from the distal versus proximal aorta. In organ culture, unstimulated adhesion of U937 cells or autologous monocytes was significantly greater to the distal aorta than the proximal aorta. Although monocytic cell adhesion to both the proximal and distal aorta increased with stimulation, the percentage increase in the proximal aorta, 1,086% with IL-1, 237% with PMA, 209% with LPS, and 174% with thrombin, was greater than in the distal aorta, demonstrating a significant functional difference in the endothelium from separate anatomic regions of a single vessel. This may have a direct relevance to the regional specificity of vascular disease.

Animals↗
PubMedDate not supplied

Regional differences in platelet-derived growth factor production by the canine aorta.

Platelet-derived growth factor (PDGF) is a potent mitogen and chemotactic agent which may be involved in the formation of proliferative lesions of the arterial system, such as intimal hyperplasia and atherosclerosis. To examine the regional variation in vessel wall production of this mitogen, PDGF production and PDGF A chain mRNA expression by normal arterial wall was studied as a function of vessel location. PDGF production by canine aortic segments was measured after 72 h in organ culture, revealing significantly more PDGF produced by the distal compared to proximal aorta at 77 +/- 10 versus 14 +/- 6 pg/cm2/72 h (p<0.05). Endothelial cells (EC) and smooth muscle cells (SMC), isolated from analogous aortic sites, were grown in tissue culture and the conditioned medium was assayed for PDGF. EC in vitro demonstrated a similar geographic trend in PDGF production (distal=1,501 +/- 389 pg/microgram DNA/72 h, proximal=759 +/- 230 pg/microgram DNA/72 h; p=0.17). PDGF production by SMC in cell culture had a similar pattern with cells from the distal aorta producing 58 +/- 28 pg PDGF/microgram DNA/72 h, compared to cells from the proximal aorta producing 37 +/- 15 pg PDGF/microgram DNA/72 h (p=0.13). Freshly harvested EC and SMC, isolated from the same aortic sites, were subjected to quantitation of PDGF mRNA levels using a coupled reverse transcriptase and polymerase chain reaction amplification method, with glyceraldehyde-phosphate dehydrogenase (GAPDH) as a control. The ratio of PDGF A chain:GAPDH mRNA was significantly greater in distal aortic SMC, 2.30 +/- 0.99, compared to proximal aortic SMC, 1.27 +/- 0.46 (p=0.05), but was not significantly different between proximal and distal aortic EC (p=0.86). These findings demonstrate significant regional differences in PDGF production in the normal canine aorta. Additionally, SMC are implicated as a significant contributor to the regional variation in PDGF production.

Animals↗
PubMedDate not supplied

Effect of complement and arachidonic acid pathway inhibition on white blood cell count and deposition on vascular grafts.

To determine the role of complement and arachidonic acid metabolites in the decrease in peripheral white blood cell count (pWBC) observed with graft implantation, Dacron aortic grafts were implanted in control rabbits (Group I, n = 13), or rabbits pretreated with cobra venom factor (80 U/kg) to deplete complement (Group II, n = 13), indomethacin (2.5 mg/kg) to inhibit cyclooxygenase (Group III, n = 7), or diethylcarbamazine (DEC, 90 mg/kg) to inhibit leukotriene synthesis (Group IV, n = 7). pWBC was measured 15 min and 1 hr after graft implantation. After graft removal, the WBC count on grafts (gWBC) was determined by light microscopy (LM) and scanning electron microscopy (SEM). One hr after graft implantation, pWBC decreased significantly in Groups I-IV to 46%, 52%, 40%, and 45% of preoperative pWBC, respectively. There was no significant difference among the groups. LM revealed gWBC per 63x field of 8.0, 12.3, 5.8, and 6.8 in Groups I-IV, respectively. Similarly, SEM showed gWBC per 2000x field of 2.5, 5.6, 0.7, and 1.5 in Groups I-IV, respectively. SEM gWBC was significantly greater in Group II than I (p less than 0.01), and significantly less in Group III than I (p less than 0.05). Results suggested that complement and arachidonic acid pathways alone do not affect the fall in pWBC, but may influence gWBC.

Animals↗
PubMedDate not supplied

Karyotype analysis of cell sex to determine the source of vascular graft luminal linings following autologous and nonautologous endothelial cell seeding.

Endothelial cell-seeded bilateral carotid ePTFE grafts were studied in 11 dogs to determine the source of their eventual luminal linings. One graft was prepared with autologous cells and the other with nonautologous cells. Immunosuppression was accomplished with cyclosporin and prednisolone. At 2 and 3 weeks post implantation, grafts were removed, revealing 9 of 11 autologous and 8 of 11 nonautologous seeded conduits to be patent. Linings were harvested enzymatically and grown in tissue culture, with karyotype analysis limited to cultures that exhibited greater than or equal to 50% endothelium (n5 autologous, n7 nonautologous). Sex type was classified in 20 cells from each graft. All five autologous cell-seeded grafts revealed host cells. Among the seven nonautologous cell-seeded grafts, one harbored only non-autologous cells, three revealed only autologous cells, and three exhibited a predominance of autologous cells with lesser numbers of nonautologous endothelium. All 100 cells from the autologous-seeded grafts were the same sex as seeded cells, while only 28 of the 140 cells from the nonautologous seeded grafts were the same sex as seeded cells (P less than 0.001). Thus, nonautologous endothelial cell seeding with immunosuppression did not uniformly produce nonautologous cell linings, but was more often associated with autologous endothelial cell linings that may have been promoted by the nonautologous cell seeding.

Animals↗