Reduction of blood activation in patients receiving aprotinin during cardiopulmonary bypass for coronary artery surgery.

Aprotinin reduces blood loss after cardiac surgery, particularly in patients taking aspirin. This study was performed to evaluate whether the reduction of contact phase activation by aprotinin is related to decreased complement activation during blood activation. Two hundred patients were prospectively operated on for coronary artery bypass. Aprotinin was used in the cardiopulmonary bypass (CPB) prime if aspirin was not discontinued 10 days before surgery and in patients undergoing second operation (n = 102). Blood loss was significantly reduced in patients receiving aprotinin (596 +/- 309 ml vs 754 +/- 329 ml without aprotinin; p = 0.0001), as was the need for transfusion (13% vs 34% without aprotinin; p = 0.0001) after surgery. Blood activation has been studied in 60 patients. Multivariate analysis showed that contact phase activation, as assessed by maximum values of C1 inhibitor/kallikrein complexes, was reduced by aprotinin treatment (p < 0.0001). Fibrinolytic activity decreased with aprotinin treatment, as reflected by lower values of D-dimers at the end of CPB (p < 0.0001). In addition, thrombin generation, as assessed by F1 + 2 scission peptide, was reduced by aprotinin (p = 0.01). However, the stepwise regression model emphasized that activation of the alternative and classic complement pathways, as reflected by C3b/c and C4b/c levels, was not affected by aprotinin; neither was leukocyte activation, as reflected by elastase release. These results suggest that aprotinin does not combine the reduction of complement activation with the reduced activation of the contact phase, fibrinolysis, or coagulation during CPB for coronary artery surgery.

Stromal cells from subcutaneous adipose tissue seeded in a native collagen/elastin dermal substitute reduce wound contraction in full thickness skin defects.

BACKGROUND: Dermal substitutes seeded with cultured fibroblasts have been developed to improve dermal regeneration in full thickness wounds. Because of cell cultivation, 3 weeks are required before patients can be treated with these autologous adipose tissue. This substitute is easily fabricated within hours, which allows immediate treatment of full thickness defects. EXPERIMENTAL DESIGN: Porcine full thickness wounds were substituted with native collagen/alpha-elastin hydrolysate matrices. One group of matrices was left unseeded as negative control. The second was seeded with cultured dermal fibroblasts as positive control. The third was seeded with a stromal-vascular-fraction of adipose tissue, and the fourth was seeded with a stromal fraction with few vascular fragments (SF). All substitutes were covered with split skin mesh grafts and were protected against dehydration and infection with a microporous polyether urethane membrane. For 8 weeks, weekly biopsies were taken, myofibroblasts and fibroblasts were counted, thickness of the granulation tissue band was measured, and wound contraction and histology were evaluated. RESULTS: Negative control and stromal-vascular-fraction substitutes were invaded by high numbers of myofibroblasts and fibroblasts. They did not reduce wound contraction, and scar tissue was formed. SF substitutes reduced the accumulation of myofibroblasts and fibroblasts and prevented the formation of granulation tissue. As a result, dermal regeneration improved, and wound contraction was less than by the other substitutes. CONCLUSIONS: Adipose tissue cell isolates included vascular fragments containing endothelial cells. Seeded in dermal substitutes, these vascular fragments induced hypergranulation tissue formation and caused wound contraction. SF substitutes contained few endothelial cells. As a result, the contraction arresting effect of the seeded stromal cell fraction was effective. Our concept of a cellular dermal substitute seeded with stromal cells from adipose tissue is feasible and allows immediate treatment of full thickness skin defects.

Reduced wound contraction and scar formation in punch biopsy wounds. Native collagen dermal substitutes. A clinical study.

In full-thickness skin wounds dermal regeneration usually fails, resulting in scar formation and wound contraction. We studied dermal regeneration by implantation of collagenous matrices in a human punch biopsy wound model. Matrices were made of native bovine collagen I fibres, and either hyaluronic acid, fibronectin, or elastin was added. Matrices were placed in 6-mm punch biopsy holes in seven patients (biopsies were used for the grafting of leg ulcers), and covered with a protective semi-permeable polyether urethane membrane. Histology, wound contraction and dermal architecture were studied. Dermal architecture was evaluated using a recently developed laser scatter technique. All collagen matrices showed a tendency to reduce wound contraction, compared with control wounds; elastin- and fibronectin-treated matrices showed significantly less contraction than control wounds. Only the addition of elastin had a clear beneficial effect on dermal architecture; collagen bundles were more randomly organized, compared with control wounds, and wounds treated with collagen matrices coated with fibronectin or hyaluronic acid, or without coating. We conclude that the punch biopsy wound model provides important information on dermal regeneration in humans. Native collagen matrices with elastin contributed to dermal regeneration and reduced wound contraction, in contrast with matrices coated with fibronectin or hyaluronic acid, or without coating. Future clinical studies of large-area, full-thickness wounds will be required to establish their clinical relevance for leg ulcer and burn treatment.

Adherence, proliferation and collagen turnover by human fibroblasts seeded into different types of collagen sponges.

We describe an in vitro model that we have used to evaluate dermal substitutes and to obtain data on cell proliferation, the rate of degradation of the dermal equivalent, contractibility and de novo synthesis of collagen. We tested three classes of collagenous materials: (1) reconstituted non-crosslinked collagen, (2) reconstituted collagen that was chemically crosslinked with either glutaraldehyde, aluminium alginate or acetate, and (3) native collagen fibres, with or without other extracellular matrix molecules (elastin hydrolysate, hyaluronic acid or fibronectin). The non-crosslinked reconstituted collagen was degraded rapidly by human fibroblasts. The chemically crosslinked materials proved to be cytotoxic. Native collagen fibres were stable. In the absence of ascorbic acid, the addition of elastin hydrolysate to this type of matrix reduced the rate of collagen degradation. Both elastin hydrolysate and fibronectin partially prevented fibroblast-mediated contraction. Hyaluronic acid was only slightly effective in reducing the collagen degradation rate and more fibroblast-mediated contraction of the material was found than for the native collagen fibres with elastin hydrolysate and fibronectin. In the presence of ascorbate, collagen synthesis was enhanced in the native collagen matrix without additions and in the material containing elastin hydrolysate, but not in the material with hyaluronic acid. These results are indicative of the suitability of tissue substitutes for in vivo application.

Dermal regeneration in native non-cross-linked collagen sponges with different extracellular matrix molecules.

Collagenous dermal templates can prevent scarring and wound contraction in the healing of full-thickness defects. In a porcine wound model, full-thickness wounds were substituted by reconstituted and native collagen sponges in combination with autologous split-skin mesh grafts and covered with a semipermeable wound membrane. Native collagen sponges were also linked with either hyaluronic acid, elastin, or fibronectin. Reconstituted collagen matrixes, composed of cross-linked small collagen fibrils, disintegrated within a week and did not contribute to dermal regeneration, whereas native collagen matrixes, composed of intact collagen fibers, disintegrated within 2 weeks and did contribute to dermal regeneration. Addition of extracellular matrix proteins retarded the disintegration to 4 weeks. However, fibronectin-treated matrixes caused aberrant epithelization. When hyaluronic acid was added, matrixes were invaded by more fibroblasts and myofibroblasts. This process correlated with fibrosis and wound contraction. In contrast, the native collagen/elastin matrix reduced the amount of fibroblasts and myofibroblasts. This latter matrix resulted in optimal dermal regeneration and little wound contraction.

Incorporation of microporous Teflon tracheal prostheses in rabbits: evaluation of surgical aspects.

Because tracheal prostheses made of nonporous silicone rubber develop granulation tissue at the anastomoses, we tested a prosthesis made of a microporous material (polytetrafluoroethylene, Teflon) to see whether this problem could be avoided and the prosthesis could be successfully incorporated (luminal side covered by connective tissue and epithelium). At various times after implantation in the cervical trachea of rabbits, the prostheses were inspected macroscopically for obstruction of the prosthesis lumen (lumen reduced by one-third or more) and microscopically for incorporation and inflammatory reaction (concentration of inflammatory cells) of the prosthesis. The prosthesis was successfully incorporated within 2-4 weeks in most rabbits without granulation tissue at the anastomoses. Two complications were infection of the prosthesis before incorporation was completed and obstruction of the lumen in the center of the prosthesis by granulation tissue or a deformed prosthesis wall. Both problems can be overcome, the first by giving an appropriate antibiotic for a longer period and the second by making a stiffer prosthesis. Thus, the microporous Teflon prosthesis is potentially useful as a tracheal prosthesis in rabbits.

Biodegradability and tissue reaction of random copolymers of L-leucine, L-aspartic acid, and L-aspartic acid esters.

A series of copoly(alpha-amino acids) with varying percentages of hydrophilic (L-aspartic acid) and hydrophobic monomers (L-leucine, beta-methyl-L-aspartate, and beta-benzyl-L-aspartate) were implanted subcutaneously in rats and the macroscopic degradation behavior was studied. Three groups of materials (A,B,C) with different ranges of hydrophilicity were distinguished: A) hydrophobic materials showed no degradation after 12 weeks; B) more hydrophilic materials revealed a gradual reduction in size of the samples, but were still present after 12 weeks; and C) hydrophilic copolymers disappeared within 24 hr. The tissue reactions caused by the materials of group A resembled that of silicone rubber, whereas those of group B showed a more cellular reaction.