Sally Baughman

Endothelium develops on seeded human arterial prosthesis: A brief clinical note

Despite a considerable and growing body of evidence that endothelial cell seeding accelerates the healing of arterial prostheses in laboratory animals, there has been no histologic evidence thus far to indicate that a similar process occurs in human beings. A case is reported of histologically confirmed, extensive endothelial healing on a polytetrafluoroethylene femoropopliteal bypass graft 90 days after it was seeded and implanted.

Patency in canine inferior vena cava grafting: Effects of graft material, size, and endothelial seeding

We studied 117 inferior vena cava (IVC) replacements in dogs to determine the effects of graft material, graft size, endothelial seeding, and cultured endothelial linings on graft patency. As a control, the IVC was removed and reimplanted in 11 dogs. Dacron (n = 7) and expanded polytetrafluoroethylene (e-PTFE) grafts (n = 12) were seeded immediately with the use of enzymatically derived autogenous jugular vein endothelium. Cultured linings were prepared for e-PTFE grafts (n = 9) by inoculating the graft with jugular endothelium and nurturing the lining in tissue culture for 14 to 30 days before implantation. Unseeded grafts (n = 27) were prepared according to the manufacturers recommendations. These six methods of preparation were tested in grafts measuring 6 mm I.D. and 60 mm in length. Other sizes were tested with a Latin square study design. After 30 to 60 days the grafts were perfusion fixed and studied with light and transmission electron microscopy. Patency was determined by contrast cavography after 7 and 30 days. Patency in the IVC reimplantation was 100% compared with 28.0% of the e-PTFE (p = 0.001) and none of the Dacron grafts that measured 6 mm I.D. and 60 mm long. e-PTFE and Dacron graft patency also differed significantly (p = 0.035). Seeded and culture-lined e-PTFE grafts in that same size were patent in 31.6% compared with 16.7% of unseeded e-PTFE. With grafts measuring 80 mm long, three of the five e-PTFE grafts were patent between 3 and 7 days. All progressed to occlusion by 30 days and compared poorly with all other graft sizes tested (2.6% progression to occlusion [p = 3 X 10(-8)]). Recanalization was not seen in 10 occluded grafts that were followed for 60 days. The histologic features of seeded grafts differed remarkably from grafts previously studied in the arterial circulation and from culture-lined and unseeded venous prostheses in that 60% had prominent large, random, endothelium-lined channels within the inner capsule. Larger graft diameters (p = 0.009) and the omission of an endothelial surface treatment (p = 0.004) were associated with anastomotic subendothelial fibrous hyperplasia. We conclude that graft material is the major determinant of patency in IVC replacements, that an extensive endothelial surface promotes patency, but that simply seeding e-PTFE or Dacron grafts with 10(5) endothelial cells does not provide sufficient endothelium to alter early patency.(ABSTRACT TRUNCATED AT 400 WORDS)

Leukocyte depletion enhances cultured endothelial retention on vascular prostheses

Approximately 90% of endothelial cells that are seeded or cultured onto vascular prostheses are lost from the flow surface within 24 hours of implantation. To determine the contribution of leukocytes to endothelial cell loss, 111In-labeled, cultured canine jugular venous endothelial cells were grown to confluence on fibronectin-coated polyester elastomer tubes measuring 4 mm inner diameter and 30 mm in length. Autogenous cell-lined tubes were implanted as bilateral carotid replacement grafts in six dogs made leukopenic by cyclophosphamide. Similar unilateral grafts were placed in 12 control dogs. Grafts were removed and perfusion-fixed from six control animals after 2 hours of in vivo arterial perfusion and from the other six animals after 6 hours of perfusion. One graft was removed and perfusion-fixed from each leukopenic animal after 2 hours of implantation and the other after 6 hours. Attachment of endothelial cells to the grafts was measured by indium-labeling technique. Retention of endothelium on grafts removed after 2 hours was measured by planimetric counting with scanning electron microscopy and on those removed after 6 hours by radioisotope quantification. Endothelial cell retention after 2 hours was 37.6% +/- 27.0% in control dogs and 97.0% +/- 3.4% in leukopenic animals (p less than 0.0007). After 6 hours retention was 35.9% +/- 23.2% in control animals and 86.5% +/- 6.0% in leukopenic animals (p less than 0.0009). Leukocyte surface activity was present in less than 1% of the leukopenic dogs compared with 8.5% of the other in vivo midgrafts after 2 hours. These results suggest that leukocytes play a significant role in the loss of seeded endothelium from vascular prostheses.

Incorporation of fucose and leucine into PNS myelin proteins in nerves undergoing early Wallerian degeneration

The simultaneous incorporation of [3H]fucose and [1-14C]leucine into normal rat sciatic nerve was examined using an in vitro incubation model. A linear rate of protein precursor uptake was found in purified myelin protein over 1/2–6 hr of incubation utilizing a supplemented medium containing amino acids. This model was then used to examine myelin protein synthesis in nerves undergoing degeneration at 1–4 days following a crush injury. Data showed a statistically significant decrease in the ratio of fucose to leucine at 2, 3, and 4 days of degeneration, which was the consequence of a significant increase in leucine uptake. These results, plus substantial protein recovery in axotomized nerves, are indicative of active synthesis of proteins that purify with myelin during early Wallerian degeneration.

Short-term in vivo stability of endothelial-lined polyester elastomer and polytetrafluoroethylene grafts

A fibronectin substrate will significantly enhance the strength of endothelial cell attachment on grafts constructed of polyester elastomer (PE) and polytetrafluoroethylene (e-PTFE). This experiment was undertaken to determine the short-term in vivo stability of endothelium on these fibronectin coated surfaces. Eight mongrel dogs underwent bilateral carotid artery replacement with both graft materials. All grafts were inoculated with 2,000 cells/mm2 using cultured autogenous venous endothelium labelled with Indium-111-oxine. The Indium-111 label in the grafts was measured immediately prior to implantation, after 1 hour of in vivo perfusion, and at explantation after 24 hours. The percentage of inoculated cells attached to the grafts before perfusion was similar for both materials, 93.3 +/- 3.0% versus 92.2 +/- 7.2%, for PE and e-PTFE respectively. All grafts were patent at one hour after implantation. PE grafts were found to have 93.8 +/- 3.9% of the attached cells present at one hour while e-PTFE grafts had only 54.5 +/- 10.8% remaining, p less than .001. After 24 hours, 5/8 (62.5%) e-PTFE grafts and 2/8 (25.0%) PE grafts remained patent, p = .13. Of the patent grafts however, endothelial cell retention was still superior on the PE grafts with 78.0 +/- 0.6% of the attached cells remaining compared to only 24.5 +/- 6.1% on e-PTFE, p less than .001. Occluded PE grafts had fewer cells remaining at 24 hours than patent ones, 78.0 +/- 0.6% versus 31.1 +/- 32.8%, respectively, p = .13. Histologically, patent PE grafts demonstrated nearly confluent endothelial monolayers while e-PTFE had patches of endothelial cells surrounded by a platelet-fibrin carpet.(ABSTRACT TRUNCATED AT 250 WORDS)

Short-termin vivo stability of endothelial-lined polyester elastomer and polytetrafluoroethylene grafts@@@Estabilidad «in vivo» a corto plazo de los injertos de elastomero de poliester endotelizado

A fibronectin substrate will significantly enhance the strength of endothelial cell attachment on grafts constructed of polyester elastomer (PE) and polytetrafluoroethylene (e-PTFE). This experiment was undertaken to determine the short-termin vivo stability of endothellum on these fibronectin coated surfaces. Eight mongrel dogs underwent bilateral carotid artery replacement with both graft materlals. All grafts were inoculated with 2,000 cells/mm2 using cultured autogenous venous endothelium labelled with Indium-111-oxine. The Indium-111 label in the grafts was measured immediately prior to implantation, after 1 hour ofin vivo perfusion, and at explantation after 24 hours. The percentage of inoculated cells attached to the grafts before perfusion was simillar for both materials, 93.3±3.0% versus 92.2±7.2%, for PE and e-PTFE respectively. All grafts were patent at one hour after implantation. PE grafts were found to have 93.8±3.9 % of the attached cells present at one hour while e-PTFE grafts had only 54.5 ± 10.8 % remaining, p<.001. After 24 hours, 5/8 (62.5%) e-PTFE grafts and 2/8 (25.0 %) PE grafts remained patent, p=.13. Of the patent grafts however, endothelial cell retention was still superior on the PE grafts with 78.0±0.6% of the attached cells remaining compared to only 24.5±6.1% on e-PTFE, p<.001. Occluded PE grafts had fewer cells remaining at 24 hours than patent ones, 78.0±0.6% versus 31.1±32.8%, respectively, p=.13. Histologically, patent PE grafts demonstrated nearly confluent endothelial monolayers while e-PTFE had patches of endothelial cells surrounded by, a platelet-fibrin carpet. We conclude that short-term patency appears to be determined by the extent of endothelial retention on PE but not e-PTFE.ResumenLa estabilidad de las células endoteliales sembradas, después de una implantación del injerto parecen ser de importancia para la mejora de las propriedades hemodinámicas de las prótesis. En la actualidad se sabe que la fibronectina, que es una glicoproteína de 450.000 daltones de peso molecular, proporciona propiedades de adherencia de células endoteliales en injertos de elastómero de poliester y PTFE expandido. Las células endoteliales fueron extraídos de forma enzimática de la venas yugulares de 8 perros mongrel y cultivadas en placas de Petri. El grueso del grupo celular fue marcado con Indio-111 oxina según la técnica de Sharefkin. A cada perro se le sustituyeron las 2 carótidas con elastómero de poliester (EP) cubierto con fibronectina y PTFE expandido sembrados con células autógenas marcadas con Indio-111. Se permitió el paso del flujo por estas prótesis y a las 24 h fueron sacrificados los animales. Los resultados muestran que las pérdidas más importantes de células en ambos injertos tuvo lugar en los primeros minutos después del desclampaje, siendo estas pérdidas sólo iniciales en el caso del elastómero de poliester, mientras que en el PTFE hubo una segunda fase de pérdida, aunque más lenta que la inicial. A las 24 h el 62.5% de los PTFE expandidos estaban permeables, mientras que sólo el 25% de los EP. Sin embargo, la capacidad de retención celular de estos últimos fue mayor cuando se les comparaba con los PTFE, con una p<0.001. Es curioso destacar que en los injertos permeables existía una zona monolineal de endotelio en las zonas centrales, mientras que las zonas perianastomóticas carecían apenas de endotelio. Si bien es verdad, que la permeabilidad protésica es baja, la tasa de retención celular sobre los injertos de elastomero de poliester fue muy alta, creyéndose pues que desempeñaría un papel profiláctico en la trombosis. El hallazgo de leucocitos debajo de las células endoteliales nos hace sospechar que la lesión endotelial puede ser el resultado de un ataque inmunológico a la fibronectina o del poliester del injerto, siendo la lisis de este substrato de fibronectina el último evento en la secuencia de este proceso inmunologico. Esta hipótesis explicaría la primera fase de pérdida rápida. Por el contrario, las fuerzas de rozamiento serían las responsables de la fase lenta de pérdidas celulares en el PTFE impregnado de fibronectina. Como es conocido las prótesis arteriales de pequeño calibre tienden a la trombosis. En teoría si la prótesis ya posee un lecho endotelial debe poseer cualidades antitrombóticas.