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

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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.

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About the authors

Vera G. Matveeva

Scientific Research Institute of Complex Problems of Cardiovascular Diseases

Author for correspondence.
Email: matveeva_vg@mail.ru
ORCID iD: 0000-0002-4146-3373
SPIN-code: 9914-3705
ResearcherId: I-9475-2017

MD, PhD, Senior Researcher of the Cell Technology Laboratory of the Experimental Medicine Department, Scientific Research Institute of Complex Problems of Cardiovascular Diseases

Russian Federation, Kemerovo

Mariam Yu. Khanova

Scientific Research Institute of Complex Problems of Cardiovascular Diseases

Email: khanovam@gmail.com
ORCID iD: 0000-0002-8826-9244
SPIN-code: 5923-0432
ResearcherId: AAR-7341-2020

Junior Researcher of the Cell Technology Laboratory of the Experimental Medicine Department, Scientific Research Institute of Complex Problems of Cardiovascular Diseases

Russian Federation, Kemerovo

Tatyana V. Glushkova

Scientific Research Institute of Complex Problems of Cardiovascular Diseases

Email: bio.tvg@mail.ru
ORCID iD: 0000-0003-4890-0393
SPIN-code: 3151-6002
ResearcherId: H-7659-2017

PhD in Biological sciences, Researcher of the New Biomaterials Laboratory of the Experimental Medicine Department, Scientific Research Institute of Complex Problems of Cardiovascular Diseases

Russian Federation, Kemerovo

Larisa V. Antonova

Scientific Research Institute of Complex Problems of Cardiovascular Diseases

Email: antonova.la@mail.ru
ORCID iD: 0000-0002-8874-0788
SPIN-code: 8634-3286
ResearcherId: I-8624-2017

MD, PhD, Head of the Cell Technology Laboratory of the Experimental Medicine Department, Scientific Research Institute of Complex Problems of Cardiovascular Diseases

Russian Federation, Kemerovo

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Supplementary files

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1. Fig. 1. Photos of fibrin matrices from the side of the surface and in the thickness

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2. Fig. 2. Structural types of the sphenoidal sinus

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3. Fig. 3. Photographs of fibroblasts on the surface and in the thickness of fibrin on the 14th day of cultivation (light microscopy, magnification × 100)

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4. Fig. 4. Results of MTT test on various surfaces

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5. Fig. 5. Shrinkage of fibrin depending on different concentrations of fibrinogen

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Copyright (c) 2021 Matveeva V., Khanova M., Glushkova T., Antonova L.



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