Year 2020 Vol. 28 No 4

SCIENTIFIC PUBLICATIONS
EXPERIMENTAL SURGERY

I.V. MAIBORODIN 1, A.A. SHEVELA 1, S.V. MARCHUKOV 1, V.V. MOROZOV 1, V.A. MATVEEVA 1, V.I. MAIBORODINA 2, A.M NOVIKOV 3, A.I. SHEVELA 1

REGENERATION OF THE BONE DEFECT AT EXPERIMENTAL APPLICATION OF EXTRACELLULAR MICROVESICLES FROM MULTIPOTENT STROMAL CELLS

Institute of Chemical Biology and Fundamental Medicine, The Russian Academy of Sciences, Siberian Branch 1,
Federal Research Center of Fundamental and Translational Medicine of the Ministry of Science and Higher Education of the Russian Federation, Institute of Molecular Pathology and Pathomorphology 2,
JSC Medical Center “Avicenna” 3, Novosibirsk,
The Russian Federation

Objective. To study the results of application extracellular microvesicles from rat multipotent mesenchymal stromal cells of the bone marrow origin (EMV) for the regeneration of rabbit bone defects, as well as to obtain data about the EMV preservation after their introduction into tissues.
Methods. The bone defect (2 mm diameter and 4 mm depth) was created in the proximal condyles of the outbred rabbit tibia. On the left limb the bone defect was filled with saline, on the right – the 50 μg EMV were introduced into defect.
Results. By the 12th day all control rabbits had retained a defect in the bone tissue with forming bone structures and hypertrophic scar in the border with intact areas.
In the most cases after the EMV introduction, no bone damage was found; the scar was thin with ordered structures of the intercellular matrix. 12 days after the application of Vybrant® CM-Dil-labeled EMV, in the periosteum and in adjacent bone marrow, vessels of bone tissue the single, very small, dust-like objects were found fluorescent in red on the background of rhodamine filter use. Sometimes the clear red tint of inclusions was noted in large cellular elements – macrophages. By the 21st day on the right (experimental), in 4 cases out of 5, only scar structures were found at the site of the bone tissue defect, while on the left (control) – only in 2 cases out of 5.
Conclusion. The preliminary results of the EMV application for the bone regeneration defect revealed the faster healing, the frequency of successful regeneration of the damaged bone and the formation of a less rough bone callus. In rabbits EMV are present in tissues at the site of application for at least 12 days. The number of the introduced EMV was found in macrophages.

Keywords: bone tissue, bone tissue regeneration, extracellular microvesicles, macrophages, luminescent microscopy
p. 359-369 of the original issue
References
  1. Takeda YS, Xu Q. Neuronal differentiation of human mesenchymal stem cells using exosomes derived from differentiating neuronal cells. PLoS One. 2015;10(8):e0135111. doi: 10.1371journal.pone.0135111
  2. Wu Y, Huang Q, Bu S. Cross talk between exosomes and pancreatic β-cells in diabetes. Arch Physiol Biochem. 2020 Jul 8:1-10. doi: 10.1080/13813455.2020.1760303. Online ahead of print.
  3. Kumagai Y, Murakami T, Kuwahara-Arai, Iba T, Reich J, Nagaoka I. Antimicrobial peptide LL-37 ameliorates a murine sepsis model via the induction of microvesicle release from neutrophils. Innate Immun. 2020 Jun 29:1753425920936754. doi: 10.1177/1753425920936754. Online ahead of print.
  4. Silachev DN, Goryunov KV, Shpilyuk MA, Beznoschenko OS, Morozova NY, Kraevaya EE, Popkov VA, Pevzner IB, Zorova LD, Evtushenko EA, Starodubtseva NL, Kononikhin AS, Bugrova AE, Evtushenko EG, Plotnikov EY, Zorov DB, Sukhikh GT. Effect of mscs and msc-derived extracellular vesicles on human blood coagulation. Cells. 2019 Mar 19;8(3). pii: E258. doi: 10.3390/cells8030258
  5. Lener T, Gimona M, Aigner L, Börger V, Buzas E, Camussi G, Chaput N, Chatterjee D, Court FA, Del Portillo HA, O’Driscoll L, Fais S, Falcon-Perez JM, Felderhoff-Mueser U, Fraile L, Gho YS, Görgens A, Gupta RC, Hendrix A, Hermann DM, Hill AF, Hochberg F, Horn PA, de Kleijn D, Kordelas L, Kramer BW, Krämer-Albers EM, Laner-Plamberger S, Laitinen S, Leonardi T, Lorenowicz MJ, Lim SK, Lötvall J, Maguire CA, Marcilla A, Nazarenko I, Ochiya T, Patel T, Pedersen S, Pocsfalvi G, Pluchino S, Quesenberry P, Reischl IG, Rivera FJ, Sanzenbacher R, Schallmoser K, Slaper-Cortenbach I, Strunk D, Tonn T, Vader P, van Balkom BW, Wauben M, Andaloussi SE, Théry C, Rohde E, Giebel B. Applying extracellular vesicles based therapeutics in clinical trials – an ISEV position paper. J Extracell Vesicles. Dec 31;4:30087. doi: 10.3402/jev.v4.30087. eCollection 2015.
  6. Narayanan R, Huang CC, Ravindran S. Hijacking the cellular mail: exosome mediated differentiation of mesenchymal stem cells. Stem Cells Int. 2016;2016:3808674. doi: 10.1155/2016/3808674
  7. Furuta T, Miyaki S, Ishitobi H, Ogura T, Kato Y, Kamei N, Miyado K, Higashi Y, Ochi M. Mesenchymal stem cell-derived exosomes promote fracture healing in a mouse model. Stem Cells Transl Med. 2016 Dec;5(12):1620-30. doi: 10.5966/sctm.2015-0285
  8. Maiborodin IV, Matveeva VA, Kolesnikov IS, Drovosekov MN, Toder MS, Shevela AI. The effect of autologous mesencbymal stem cells of bone marrow origin on regeneration of damaged rat bottom jaw bone. Stomatologiia. 2012;91(1):5-9. https://www.mediasphera.ru/issues/stomatologiya/2012/1/030039-1735201211 ( In Russ.)
  9. Zhang S, Chu WC, Lai RC, Lim SK, Hui JH, Toh WS. Exosomes derived from human embryonic mesenchymal stem cells promote osteochondral regeneration. Osteoarthritis Cartilage. 2016 Dec;24(12):2135-40. doi: 10.1016/j.joca.2016.06.022
  10. Wang KX, Xu LL, Rui YF, Huang S, Lin SE, Xiong JH, Li YH, Lee WY, Li G. The effects of secretion factors from umbilical cord derived mesenchymal stem cells on osteogenic differentiation of mesenchymal stem cells. PLoS One. 2015 Mar 23;10(3):e0120593. doi: 10.1371/journal.pone.0120593. eCollection 2015.
  11. Torreggiani E, Perut F, Roncuzzi L, Zini N, Baglìo SR, Baldini N. Exosomes: novel effectors of human platelet lysate activity. Eur Cell Mater. 2014 Sep 22;28:137-51; discussion 151. doi: 10.22203/ecm.v028a11
  12. Lei L, Tzekov R, Tang S, Kaushal S. Accumulation and autofluorescence of phagocytized rod outer segment material in macrophages and microglial cells. Mol Vis. 2012;18:103-13. http://www.molvis.org/molvis/v18/a13
  13. Tang XD, Shi L, Monsel A, Li XY, Zhu HL, Zhu YG, Qu JM. Mesenchymal Stem Cell Microvesicles Attenuate Acute Lung Injury in Mice Partly Mediated by Ang-1 mRNA. Stem Cells. 2017 Jul;35(7):1849-59. doi: 10.1002/stem.2619
Address for correspondence:
630090, Russian Federation,
Novosibirsk, pr. Acad. Lavrenteva, 8,
Institute of Chemical Biology
and Fundamental Medicine,
Center for New Medical Technologies
tel. +7-913-753-0767
e-mail: imai@mail.ru,
Maiborodin Igor V.
Information about the authors:
Maiborodin Igor V., MD, Professor, Chief Researcher, Laboratory of Health Management Technologies, the Center of New Medical Technologies, Institute of Chemical Biology and Fundamental Medicine, the Russian Academy of Sciences, Siberian Branch, Novosibirsk, Russian Federation.
http://orcid.org/0000-0002-8182-5084
Shevela Aleksandr A., PhD, Doctoral Student of Laboratory of Health Management Technologies, the Center of New Medical Technologies, Institute of Chemical Biology and Fundamental Medicine, the Russian Academy of Sciences, Siberian Branch, Novosibirsk, Russian Federation.
http://orcid.org/0000-0001-9235-9384
Marchukov Sergey V., PhD, Doctoral Student of Laboratory of Health Management Technologies, the Center of New Medical Technologies, Institute of Chemical Biology and Fundamental Medicine, the Russian Academy of Sciences, Siberian Branch, Novosibirsk, Russian Federation.
https://orcid.org/0000-0002-7381-5820
Morozov Vitaly V., MD, Professor, Head of Laboratory of Health Management Technologies, the Center of New Medical Technologies, Institute of Chemical Biology and Fundamental Medicine, the Russian Academy of Sciences, Siberian Branch, Novosibirsk, Russian Federation.
https://orcid.org/0000-0002-9810-5593
Matveeva Vera A., PhD, Senior Researcher of Laboratory of Health Management Technologies, the Center of New Medical Technologies, Institute of Chemical Biology and Fundamental Medicine, the Russian Academy of Sciences, Siberian Branch, Novosibirsk, Russian Federation.
https://orcid.org/0000-0002-8706-4853
Maiborodina Vitalina I., MD, Leading Researcher, Laboratory of Ultrastructural Basis of Pathology, Institute of Molecular Pathology and Pathomorphology, Federal State Budget Scientific Institution “Federal Research Center of Fundamental and Translational Medicine”, Novosibirsk, Russian Federation.
http://orcid.org/0000-0002-5169-6373
Novikov Alexey M., Gynecologist of the Department of Gynecology, JSC Medical Center “Avicenna”, Novosibirsk, Russian Federation.
https://orcid.org/0000-0003-1371-7492
Shevela Andrey I., MD, Professor, Head of Department the Center of New Medical Technologies, Institute of Chemical Biology and Fundamental Medicine, the Russian Academy of Sciences, Siberian Branch, Novosibirsk, Russian Federation.
http://orcid.org/0000-0002-3164-9377
Contacts | ©Vitebsk State Medical University, 2007-2023