Method of determining endothelial cell coverage of a prosthetic surface

• Main Authors: WILLIAMS; STUART K, JARRELL; BRUCE E
• Format: Patent
• Language: English
• Published: 16.03.1993
• Subjects: BANDAGES, DRESSINGS OR ABSORBENT PADS; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS, ORSURGICAL ARTICLES; DEVICES PROVIDING PATENCY TO, OR PREVENTING COLLAPSING OF,TUBULAR STRUCTURES OF THE BODY, E.G. STENTS; DISINFECTION, STERILISATION, OR DEODORISATION OF AIR; FILTERS IMPLANTABLE INTO BLOOD VESSELS; FIRST-AID KITS; FOMENTATION; HUMAN NECESSITIES; HYGIENE; INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS, OR SURGICALARTICLES; MEASURING; MEDICAL OR VETERINARY SCIENCE; METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS INGENERAL; ORTHOPAEDIC, NURSING OR CONTRACEPTIVE DEVICES; PHYSICS; PROSTHESES; TESTING; TREATMENT OR PROTECTION OF EYES OR EARS

Determination of effectiveness of microvascular endothelial cell seeding upon a vascular graft surface within the operating room environment would be desirable to maintain quality control in any clinical trial. A number of fluorescent dyes including mithramycin, Hoechst 33342, sulfofluorescein diacetate, Nile Red, rhodamine 123, and PKH26-GL were evaluated for their ability to fluorescently label uncultured microvascular endothilial cells on graft material and subsequently allow determination of seeded cell number and cell spreading. Rhodamine 123 and PKH26-GL produced the most desirable characteristics. The selected non-toxic fluorescent dyes allowed excellent cell visualization after a 30 minute incubation. Unlike the other fluorescent dyes evaluated, the selected non-toxic fluorescent dyes caused the cellular cytoplasm to fluoresce bright orange at a 510 nm excitation wavelength while the underlying polyethyleneterephthalate polyester or expanded polytetrafluorethylene demonstrated minimal autofluorescence. No inhibitory effect on cell attachment to plastic or subsequent cell growth in culture was observed. This technique is useful in the operating room to visualize part or all of an microvascular endothelial cell-seeded graft and to permit a quantitative as well as qualitative evaluation of the seeding process to enhance graft patency.