COMPARATIVE ASSESSMENT OF THE POSSIBILITY OF CELLULAR MATERIAL COLLECTION TO MATRIXES OBTAINED BY METHODS OF ELECTROSPINNING AND AERODYNAMIC FORMATION IN A TURBULENT GAS FLOW

Introduction. Methods of electrospinning and aerodynamic formation in the gas stream allow the fabrication of synthetic structures similar to intercellular matrices. But it is important to assess the suitability of such structures for colonization by cellular material. Materials and Methods. Porous matrices of polylactic acid are used. Matrix No. 1 was obtained by the method of electrospinning, matrix No. 2 — by the method of aerodynamic formation. The structure of the matrices was examined on an electron microscope. The matrices were dynamically populated with stromal cells from the rabbit’s bone marrow, then matrices were cultured for 3 days in a CO2 incubator at 37 °C. Cells were detected by fluorescence microscopy. Data were presented as mean ± standard deviation. Results. The matrices differed in structure. Matrix No. 1 was formed by fibers of regular cylindrical shape (diameter 1.5±0.7 μm) without its own relief. The average porosity was 67±3%. In the structure of matrix No.2 it was possible to single out a macrolevel represented by wisps (diameter 27.5±17.4 μm) formed by directed fibers (diameter 0.44±0.14 μm) with developed surface relief. The average porosity was 55±3% (p<0.05 as compared with matrix No. 1). The area of the free surface of matrix No. 2 exceeded this figure by 8 times for matrix No. 1. The matrices significantly (p><0.05) differed in the average number of cells: 56±9 cells and 120±40 cells for the matrix No. 1 against 81±6 cells and 215±18 cells for the matrix No. 2 in 2D and 3D regimes, respectively. Discussion. The best cell adhesion result obtained for the polylactic acid matrix formed by the aerodynamic formation method does not contradict other studies and probably was caused by its more optimal spatial organization. Keywords: nonwoven materials, electrospinning, aerodynamic formation, cell culture><0.05 as compared with matrix No. 1). The area of the free surface of matrix No. 2 exceeded this figure by 8 times for matrix No. 1. The matrices significantly (p<0.05) differed in the average number of cells: 56±9 cells and 120±40 cells for the matrix No. 1 against 81±6 cells and 215±18 cells for the matrix No. 2 in 2D and 3D regimes, respectively. Discussion. The best cell adhesion result obtained for the polylactic acid matrix formed by the aerodynamic formation method does not contradict other studies and probably was caused by its more optimal spatial organization.

1. Попов С.В., Рябов В.В., Суслова Т.Е. и соавт. Фундаментальные и прикладные аспекты клеточных технологий в кардиологии и кардиохирургии // Бюллетень СО РАМН. — 2008. — Т. 28(4). — С. 5–15.

2. Aranovich A., Popkov A., Barbier D., Popkov D. Femoral lengthening by combined technique in melorheostosis: a case report // Eur. Orthop. Traumatol. — 2014. — Vol. 5. — P. 175–179 [Electronic resource] — doi: 10.1007/s12570-013-0220-4.

3. Popkov D., Journeau P., Popkov A. et al. Ollier’s disease limb lenghtening: Should intramedullary nailing be combined with circular external fixation? // Orthop. Traumatol. Surg. Res. — 2010. — Vol. 96. — P. 348–353 [Electronic resource] — doi: 10.1016/j.otsr.2010.01.002.

4. Petite H., Viateau V., Bensaïd W. et al. Tissue-engineered bone regeneration // Nat. Biotechnol. — 2000. — Vol. 18. — P. 959–963 [Electronic resource] — doi: 10.1038/79449.

5. Boccaccini A.R., Blaker J.J. Bioactive composite materials for tissue engineering scaffolds // Expert Rev. Med. Devices. — 2005. — No. 2. — P. 2303–2317 [Electronic resource] — doi: 10.1586/17434440.2.3.303.

6. Santoro M., Shah S.R., Walker J.L., Mikos A.G. Poly(lactic acid) nanofibrous scaffolds for tissue engineering // Adv. Drug Deliv. Rev. — 2016. — Vol. 107. — P. 206–212 [Electronic resource] — doi: 10.1016/j.addr.2016.04.019.

7. Tamayol A., Akbari M., Annabi N. et al. Fiber-based tissue engineering: Progress, challenges, and opportunities // Biotechnol. Adv. — 2013. — Vol. 31. — P. 669–687 [Electronic resource] — doi: 10.1016/j.biotechadv.2012.11.007.

8. Jiang T., Carbone E.J., Lo K.W.-H., Laurencin C.T. Electrospinning of Polymer Nanofibers for Tissue Regeneration // Prog. Polym. Sci. — 2014. — Vol. 46. — P. 1–24 [Electronic resource] — doi: 10.1016/j.progpolymsci.2014.12.001.

9. Daristotle J.L., Behrens A.M., Sandler A.D., Kofinas P. A Review of the Fundamental Principles and Applications of Solution Blow Spinning // ACS Appl. Mater. Interfaces. — 2016. — No. 8. — P. 34951–34963 [Electronic resource] — doi: 10.1021/ acsami.6b12994.

10. Нащекина Ю.А., Никонов П.О., Михайлов В.М. и др. Зависимость заполнения стромальными клетками костного мозга трехмерной матрицы от способа посева клеток и типа модификации поверхности матрицы // Цитология. — 2014. — Т. 56(4). — С. 283–290.

11. Nashchekina Y.A., Nikonov P.O., Mikhailov V.M. et al. Distribution of bone-marrow stromal cells in a 3D scaffold depending on the seeding method and the scaffold inside a surface modification // Cell tissue biol. — 2014. — Vol. 8(4). — P. 313–320.

12. Filatov Y., Budyka A., Kirichenko V. Electrospinning of Micro- and Nanofibers: Fundamentals in Separation and Filtration Processes. — New York : Begell House Inc., 2007. — 404 p.

13. Reneker D.H., Chun I. Nanometre diameter fibres of polymer, produced by electrospinning // Nanotechnology. — 1996. — Vol. 7(3). — P. 216–223.

14. Daristotle J.L., Behrens A.M., Sandler A.D. et al. A Review of the Fundamental Principles and Applications of Solution Blow Spinning // ACS Appl. Mater. Interfaces. — 2016. — Vol. 8(51). — P. 34951–34963.

15. Tomecka E., Wojasinski M., Jastrzebska E. et al. Poly(l-lactic acid) and polyurethane nanofibers fabricated by solution blow spinning as potential substrates for cardiac cell culture // Mater. Sci. Eng. C. — 2017. — Vol. 75. — P. 305–316.

16. Faia-Torres A.B., Charnley M., Goren T. et al. Osteogenic differentiation of human mesenchymal stem cells in the absence of osteogenic supplements: A surface-roughness gradient study // Acta Biomater. — 2015. — Vol. 28. — P. 64–75.