Influence of different concentrations of fibrinogen on the properties of a fibrin matrix for vascular tissue engineering

Abstract

Aim. To evaluate the potential utility of fibrin matrices containing 10, 20, and 25 mg/ml of fibrinogen (fibrin-10, fibrin-20, and fibrin-30, respectively) in vascular tissue engineering (VTE).
 Materials and Methods. Fibrinogen was isolated using the method of ethanol cryoprecipitation and polymerized using a solution of thrombin and CaCl2. The fibrin structure was studied in a scanning electron microscope, and the physical and mechanical properties of the material were tested on a Zwick/Roell test machine. The metabolic activity of endothelial cells (EC) on the fibrin surface was evaluated by the MTT assay, and the viability of fibroblasts in the thickness of fibrin and possibility for migration by in fluorescent and light microscopy. Percent of fibrin shrinkage was determined from the difference in the sample volumes before and after removal of moisture.
 Results. The fiber diameter did not differ among all fibrin samples, but the pore diameter in fibrin-30 was smaller than those in fibrin-10 and fibrin-20. A possibility for migration of fibroblasts into the depth of the fibrin matrix and preservation of 97-100% viability of cells at a depth 5 mm was confirmed. The metabolic activity of EC on the surface of fibrin-20 and fibrin-30 exceeded that on collagen, fibronectin, and fibrin-10. All fibrin samples shrank in volume to 95.5-99.5%, and the highest shrinkage was seen in fibrin-10. The physical and mechanical properties of fibrin were inferior to those of human A. mammaria by a factor of 10.
 Conclusion. Fibrin with fibrinogen concentrations of 20 and 30 mg/ml maintains a high metabolic and proliferative activity of EC on the surface and also a high viability of fibroblasts in the matrix. Its availability, ease of preparation, and a number of other favorable properties make fibrin a promising material for VTE. However, the problem of insufficient strength requires further investigations.

Advantages of Fibrin Polymerization Method without the Use of Exogenous Thrombin for Vascular Tissue Engineering Applications

“…The ethanol precipitation provides a maximum yield of fibrinogen (up to 80% of its total plasma content) for a minimum period of time, and it does not adversely affect the mechanical and biological properties of fibrin synthesized from this fibrinogen [38,39]. Fibrinogen concentrations were chosen according to the literature data and the data obtained in our previous study that indicate that high fibrinogen content (10-30 mg/mL) is most suitable for the fabrication of cardiovascular tissue-engineered implants [18,40,41].…”

Peroxynitrite Modification as a Way to Improve the Cytocompatibility of Sodium Alginate-Based Hydrogels

“…В качестве контроля использовали 0,05% раствор метиленового синего, в который не добавляли гранулы. Насыщенность модифицированных образцов альгината натрия карбоксильными группами рассчитывали по формуле (2).…”

“…В последнее время данная группа патологий привлекает всё большее внимание специалистов в области регенеративной медицины [1]. Данная наука основывается на применении биоматериалов и технологий клеточного культивирования для создания биоинженерных конструкций, которые могли бы заменить в организме человека органы с нарушенным функционированием или стимулировать процессы регенерации в повреждённых тканях и органах [2,3]. В качестве биоматериалов в регенеративной медицине применяются, в том числе, гидрогели на основе модифицированных альгинатов натрия [4], получение которых осложняется дороговизной реактивов и трудоёмкостью процесса очистки от избытков цитотоксических реагентов.…”