Use of a fibrin preparation in the engineering of a vascular graft model.

OBJECTIVE Morphological and functional characterization of cocultured endothelial cells (EC) and myofibroblasts (MFB) seeded on a matrix composed of a fibrin preparation mimicking the microenvironment of a vascular wall. METHODS MFB and EC were isolated from human saphenous veins and expanded separately in vitro. MFB were seeded on a composite matrix consisting of a fibrin preparation (with or without transforming growth factor-beta2) and a polyglactin-mesh to form a 3-dimensional structure, which was consecutively reseeded with EC. Seeded matrices were incubated in a bioreactor. Characterization was done including fluorescence staining, live-/dead-assay and immunohistochemistry. RESULTS High density cocultures in hierarchical structure mimicking the formation of a vascular wall were obtained with nearly complete coverage of the surface with EC. Distribution of preseeded MFB in a 519+/-27 microm thick layer (day 14) was achieved. Cell viability was shown in fluorescence staining for at least 19 days. In deeper layers, no viable cells could be detected within the fibrin preparation. EC covered the surface, had uniform morphology, and their preserved viability was shown for at least 5 days. No EC-ingrowth was found into the fibrin preparation. Neoformation of the matrix proteins laminin and collagen IV was observed. CONCLUSION A structured coculture of MFB and EC was obtained mimicking the formation of a vascular wall with preserved viability utilizing a fibrin preparation. Nutrition problems seem to limit the maximal extent of MFB in the matrix.

[1]  H. Greisler,et al.  Angiogenic effect of fibroblast growth factor-1 and vascular endothelial growth factor and their synergism in a novel in vitro quantitative fibrin-based 3-dimensional angiogenesis system. , 2002, Surgery.

[2]  K. Fujikawa,et al.  α1-Proteinase Inhibitor, α1-Antichymotrypsin, or α2-Macroglobulin Is Required for Vascular Smooth Muscle Cell Spreading in Three-dimensional Fibrin Gel* , 2000, The Journal of Biological Chemistry.

[3]  C. Perka,et al.  Matrix engineering for osteogenic differentiation of rabbit periosteal cells using alpha-tricalcium phosphate particles in a three-dimensional fibrin culture. , 2002, Journal of biomedical materials research.

[4]  S Jockenhoevel,et al.  Fibrin gel as a three dimensional matrix in cardiovascular tissue engineering. , 2000, European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery.

[5]  B J Messmer,et al.  Tissue engineering: complete autologous valve conduit--a new moulding technique. , 2001, The Thoracic and cardiovascular surgeon.

[6]  B. S. Gow,et al.  The Elasticity of Canine and Human Coronary Arteries with Reference to Postmortem Changes , 1979, Circulation research.

[7]  T. Matsuda,et al.  An integrated approach to the design and engineering of hybrid arterial prostheses. , 1994, Journal of vascular surgery.

[8]  A. Seifalian,et al.  A hybrid compliant vascular graft seeded with microvascular endothelial cells extracted from human omentum. , 2001, Artificial organs.

[9]  J. Vacanti,et al.  Tissue engineering : Frontiers in biotechnology , 1993 .

[10]  B J Messmer,et al.  Fibrin gel -- advantages of a new scaffold in cardiovascular tissue engineering. , 2001, European journal of cardio-thoracic surgery : official journal of the European Association for Cardio-thoracic Surgery.

[11]  R T Tranquillo,et al.  Enhanced fibrin remodeling in vitro with TGF-beta1, insulin and plasmin for improved tissue-equivalents. , 2002, Biomaterials.

[12]  J. Glover,et al.  A single-staged technique for seeding vascular grafts with autogenous endothelium. , 1978, Surgery.

[13]  M. Lynn,et al.  Viability of human corneal endothelium following Optisol-GS storage. , 1995, Archives of ophthalmology.

[14]  S. Schwartz,et al.  Role of α5β1 and αvβ3 Integrins on Smooth Muscle Cell Spreading and Migration in Fibrin Gels , 2000, Thrombosis and Haemostasis.

[15]  M. Sittinger,et al.  Preparation of a pure autologous biodegradable fibrin matrix for tissue engineering , 2000, Medical and Biological Engineering and Computing.

[16]  N. Samani,et al.  Expression of the plasminogen activator system in the human vascular wall. , 2000, Atherosclerosis.

[17]  Axel Haverich,et al.  Tissue engineering of small diameter vascular grafts. , 2002, European journal of vascular and endovascular surgery : the official journal of the European Society for Vascular Surgery.

[18]  V. Nehls,et al.  The effect of fibroblasts, vascular smooth muscle cells, and pericytes on sprout formation of endothelial cells in a fibrin gel angiogenesis system. , 1994, Microvascular research.

[19]  H. Greisler,et al.  Selective stimulation of endothelial cell proliferation with inhibition of smooth muscle cell proliferation by fibroblast growth factor-1 plus heparin delivered from fibrin glue suspensions. , 1995, Surgery.