The cytocompatibility of compound polyester-protein surfaces using an in vitro technique

Summary To avoid the need to preclot porous polyester (Dacron) vascular prostheses, we have proposed the use of a protein coating that will promote the growth and adhesion of endothelial cells. This study assessed the relative advantages of coating woven, knitted, and velour polyester fabrics with albumin, collagen, and a albumin-collagen mixture after preservation in saline or drying by a commercial dehydration process. Preclotted fabrics were used as controls. The cytocompatibility of these biopolymers was measured by an organotypic culture technique which relies on the migration of chick embryonic endothelial cells. After 7 d of culture the cytocompatibility was quantified by counting the cells in the area of migration and the morphology of the endothelial cells was observed by scanning electron microscopy. In general, the knitted and velour fabrics showed superior compatibility than the woven one. The results confirmed that collagen, either alone or combined with albumin, provides in most cases a more cytocompatible surface than albumin alone. A cell morphology most closely resembling that of natural arterial endothelial cells was observed on the albumin-collagen substrate. This study was supported by INSERM Grant C. R. L. 82 30 16, the Medical Research Council of Canada, and the Canadian Heart Foundation.     

Self-assembling supramolecular complexes by single-stranded extension from plasmid DNA

Self-assembling supramolecular complexes are of great interest for bottom-up research like nanotechnology. DNA is an inexpensive building block with sequence-specific self-assembling capabilities through Watson-Crick and/or Hoogsteen base pairing and could be used for applications in surface chemistry, material science, nanomechanics, nanoelectronics, nanorobotics, and of course in biology. The starting point is usually single-stranded DNA, which is rather easily accessible for base pairing and duplex formation. When long stretches of double-stranded DNA are desirable, serving either as genetic codes or electrical wires, bacterial expansion of plasmids is an inexpensive approach with scale-up properties. Here, we present a method for using double-stranded DNA of any sequence for generating simple structures, such as junctions and DNA lattices. It is known that supercoiled plasmids are strand-invaded by certain DNA analogs. Here we add to the complexity by using "Self-assembling UNiversal (SUN) anchors" formed by DNA analog oligonucleotides, synthesized with an extension, a "sticky-end" that can be used for further base pairing with single-stranded DNA. We show here how the same set of SUN anchors can be utilized for gene therapy, plasmid purification, junction for lattices, and plasmid dimerization through Watson-Crick base pairing. Using atomic force microscopy, it has been possible to characterize and quantify individual components of such supra-molecular complexes.     

A modular perfused chamber for low- and normal-temperature microscopy of living cells

An inexpensive modular perfused chamber (MPC) designed for low- and normal-temperature live-cell imaging is presented. The device consists of four lathed pieces of stainless steel assembled as a cylindrical open chamber that can hold either round or square glass coverslips. The chamber is connected to a thermal-bath operating with recirculation. For image acquisition at 4°C, cooled air is blown toward the coverslip surface to prevent condensation. Principal advantages of this device are thermal stability in the sample environment, rapid response to changes in temperature set point, and easy sample insertion. The device enables the study of dynamic processes in cells governed by large temperature differences such as those imposed by hypothermic preservation of cells (0-4°C) followed by rewarming to normothermia (37°C). The capabilities of the MPC were demonstrated by monitoring the internalization of fluorescent quantum dots (QDs) in rat hepatocytes after hypothermic storage and during rewarming with an inverted microscope.