Cryogenically structured mimetic of extracellular matrix based on a concentrated collagen-containing solution
Authors:   V.I. Shumakov Federal Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation, Basok Yu. B. , Grigoriev A. M. ,   V.I. Shumakov Federal Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation, Lozinsky V. I. ,   A.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Kirsanova L. A. ,   V.I. Shumakov Federal Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation, Kulakova V. K. ,   A.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Podorozhko E. A. ,   A.N. Nesmeyanov Institute of Organoelement Compounds, Russian Academy of Sciences, Novikov I. A. ,   Scientific Research Institute of Eye Diseases, Sevastianov V. I. ,   The Institute of Biomedical Research and Technology,   V.I. Shumakov Federal Research Center of Transplantology and Artificial Organs of the Ministry of Healthcare of the Russian Federation
Published: April 2024

Viscoelastic hydrogels based on animal tissue extracts are considered promising biomimetics of the extracellular matrix (ECM) due to their proven effectiveness for stimulating the regeneration of the liver, pancreas and articular cartilage. Cryostructuring is an approach that makes it possible to give polymer scaffolds macroporosity and provide mechanical strength. preparation of a new macroporous cryogenically structured biomimetic of ECM based on a commercially available concentrated collagen-containing solution and evaluation of the possibilities of its application in tissue engineering. The target spongy collagen-containing material was obtained by sequential freezing of a concentrated collagen-containing solution, its subsequent lyophilization and chemical tanning by treatment with an alcohol solution of carbodiimide. The morphology of the cryostructured multicomponent collagen-containing material was studied using optical and scanning electron microscopy (SEM) using lanthanide contrast. The cytotoxicity of the scaffold was studied on the culture of human adipose-derived stem cells (hADSCs). Adhesion and proliferation of hADSCs on the scaffold surface were studied on the 7th day of cultivation. The compression modulus of elasticity of the obtained collagen–containing material in the swollen state in water was 35,3 ± 2,2 kPa, the total water-holding capacity of the material was 45.80 ± 0.46 ml/g of polymer, and the degree of swelling of the walls of macropores was 3.99 ± 0.31 ml/g. During SEM examination and histological staining with hematoxylin and eosin, a broad-pored structure was observed on the surface and cross-section of the disc. The pores in the upper part are larger (the average diameter is not less than ~ 30 µm) than the pores in the lower part of the sponge (the average diameter is not more than ~ 30 µm) due to the occurrence of a vertical temperature gradient. The matrix did not have cytotoxicity relative to the hADSCs. In the sample, active proliferation of hADSCs was observed on the surface of the scaffold. It was shown that the developed cryostructurates based on a concentrated collagen-containing solution had supermacroporosity and a compression modulus of elasticity of 35.3 ± 2.2 kPa. The absence of cytotoxicity and the ability to maintain adhesion and proliferation of hADSCs indicate the possibility of using cryogenically structured biomimetic of the extracellular matrix in tissue engineering and regenerative medicine.