Localization of vascular endothelial growth factor and transforming growth factor-β in tissues in perifractural zone after fractures of long bones of limbs in humans

Authors

  • Vitaliy Grigoryev
  • Olexii Popsuishapka
  • Nataliya Ashukina
  • Fedor Galkin

DOI:

https://doi.org/10.15674/0030-59872017262-69

Keywords:

bone fracture, regeneration, fibrin-blood clot, endothelial growth factor of suction, transforming growth factor

Abstract

Objective: to study, using immunohistochemical methods, the content of the vascular endothelial growth factor and trans­forming growth factor (TGF-β) in tissues adjacent to bone frag­ments after fracture in humans, as well as in autofibrin obtained in vitro.

Methods: biopsy samples (interfragmental fibrin-blood clot, periosteum, muscular and fatty tissue), 9 of the victims recovered from the near fracture zone with fractures of limb bones during their open comparison were studied. In addition, fibrin-blood clots from the venous blood of the same patients ob­tained during the operation by the method of J. Choukroun were analyzed.

Results: 1–2 days after the fracture, vascular endothe­lial growth factor concentrates in fibrin, forming a fibrin-blood clot in the near-fracture zone. On the 5th–12th day, the intensity of the reaction to vascular endothelial growth factor in fibrin decreases, or it disappears, and expression of the factor mani­fests itself in the cells of the regenerate — osteocytes, osteo­blasts, fibroblasts, endotheliocytes. 10–12 days after injury, a positive reaction to vascular endothelial growth factor is de­fined in endotheliocytes and osteoblasts. The reaction to TGF-β in fibrin-blood clot and regenerate tissues was observed exclu­sively in cells. In autologous fibrin clots, vascular endothelial growth factor expression was observed in fibrin, and TGF was observed in the blood cells that remained in it. However, the in­tensity of the response to vascular endothelial growth factor in fibrin formed in vitro was uneven.

Conclusions: vascular en­dothelial growth factorconcentration on the 1–2 day after frac­ture in the fibrin-blood clot in the near fracture zone is a strong signal for the restoration of blood supply in the area of inju­ry. Expression of TGF-β at all times was found only in cells. The uneven distribution of vascular endothelial growth factor in fibrin formed in vitro should be considered when it is used to optimize bone repair.

Author Biographies

Vitaliy Grigoryev

CI «Cherkasy City Emergency Hospital № 3». Ukraine

grigoriev.doc@gmail.com

 

Olexii Popsuishapka

Kharkiv Medical Academy for Postgraduate Education of the Ministry of Health of Ukraine

MD, Prof. in Orthopaedics and Traumatology

alexecorn@gmail.com

 

 

 

Nataliya Ashukina

Sytenko Institute of Spine and Joint Pathology, Kharkiv. Ukraine

PhD in Biol. Sci.

nataliya.ashukina@gmail.ru

 

 

Fedor Galkin

Cherkasy Regional Oncology Center. Ukraine

MD

jackdow.f@gmail.com

 

 

References

  1. Popsuishapka O, Uzhigova O, Litvishko V. Rate of nonunion and delayed union of fragments in isolated diaphyseal fractures of long bones of the extremities/ Orthopaedics, Traumatology and Prosthetics. 2013;(1): 39–43. doi: 10.15674/0030-59872013139-43. (in Russian)
  2. Korzh NA, Dedukh NV. Reparative regeneration of bone: modern view of the problem (report 1). Orthopaedics, Traumatology and Prosthetics. 2006;(1):77–84. (in Russian)
  3. Sfeir C, Ho L, Doll BA, Azari K, Hollinger JO. Fracture Repair. In: Bone regeneration and repair. Biology and clinical applications. Eds. JR Lieberman, GE Friedlaender. Humana Press, 2005. p. 21–44.
  4. Marti RK, van Heerwaarden RJ. Osteotomies for posttraumatic deformities. Georg Thieme Verlag, 2008. 704 p.
  5. Marsell R, Einhorn TA. The biology of fracture healing. Injury. 2011; 42(6): 551–5. doi: 10.1016/j.injury.2011.03.031.
  6. Kanczler JM, Oreffo RO. Osteogenesis and angiogenesis: the potential for engineering bone. Eur Cell Mater. 2008;15:100–14. doi: 10.22203/ eCM.v015a08.
  7. Sivaraj KK, Adams RH. Blood vessel formation and function in bone. Development. 2016;143(15):2706–15. doi: 10.1242/dev.136861.
  8. Hankenson KD, Dishowitz M, Gray C, Schenker M. Angiogenesis in bone regeneration. Injury. 2011;42(6):556–61. doi: 10.1016/j.injury.2011.03.035.
  9. Tomlinson RE, Silva MJ. Skeletal blood flow in bone repair and maintenance. Bone Research. 2013;4:311–22. doi: 10.4248/BR201304002.
  10. Maes C, Kobayashi T, Selig MK, Torrekens S, Roth SI, Mackem S, Carmeliet G, Kronenberg HM. Osteoblast precursors, but not mature osteoblasts, move into developing and fractured bones along with invading blood vessels. Dev Cell. 2010;19(2):329–44. doi: 10.1016/ j.devcel.2010.07.010.
  11. Ramasamy SK, Kusumbe AP, Schiller M, Zeuschner D, Bixel MG, Milia C, Gamrekelashvili J, Limbourg A, Medvinsky A, Santoro MM, Limbourg FP, Adams RH. Blood flow controls bone vascular function and osteogenesis. Nat Commun. 2016 Dec 6;7:13601. doi: 10.1038/ncomms13601.
  12. Hu K, Olsen BR. The roles of vascular endothelial growth factor in bone repair and regeneration. Bone. 2016;91:30–8. doi: 10.1016/j.bone.2016.06.013.
  13. Van Meeteren LA, Goumans MJ, ten Dijke P. TGF-β receptor signaling pathways in an¬giogenesis; emerging targets for anti-angiogenesis therapy. Curr Pharm Biotechnol. 2011;12(12):2108–20. doi: 10.2174/138920111798808338.
  14. Moreno-Miralles I, Schisler JC, Patterson C. New insights into bone morphogenetic protein signaling: focus on angiogenesis. Curr Opin Hematol. 2009;16(3):195–201. doi: 10.1097/MOH.0b013e32832a07d6.
  15. Raica M, Cimpean Am. Platelet-derived growth factor (PDGF)/PDGF receptors (PDGFR) axis as target for antitumor and antiangiogenic therapy. Pharmaceuticals. 2010;3(3):572–99. doi: 10.3390/ph3030572.
  16. Bastami F, Khojasteh A. Use of leukocyte-and platelet-rich fibrin for bone regeneration: a systematic review. Regeneration, Reconstruction & Restoration. 2016;1(2):47–68. doi: 10.7508/rrr.2016.02.001.
  17. Aydemir Turka H, Demirer S, Dolgun A, Keceli HG. Evaluation of the adjunctive effect of platelet-rich fibrin to enamel matrix derivative in the treatment of intrabony defects. Six-month results of a randomized, split-mouth, controlled clinical study. J Clin Periodontol. 2016;43(11):955–64. doi: 10.1111/jcpe.12598.
  18. Grigoryev V. The use of autofibrinum for the stimulation of osteoreparation in the treatment of long bones fractures. Orthopaedics, Traumatology and Prosthetics. 2015;(4):5–10. doi: 10.15674/0030-5987201545-10. (in Ukrainian)
  19. Choukroun J, Abba F, Schoeffler C, Vervelle A. An opportunity in perio-implantology: the PRF. Implantodontie. 2000;42:55–62.
  20. Sarkisov DS, Perov JL. Microscopic techniques. Moskow: Medicine, 1996. 542 p. (in Russian).
  21. Dong LQ, Yin H, Wang CX, Hu WF. Effect of the timing of surgery on the fracture healing process and the expression levels of vascular endothelial growth factor and bone morphogenetic protein-2. Exp Ther Med. 2014;8(2):595–9. — doi: 10.3892/etm.2014.1735.
  22. Janmey PA, Winer JP, Weisel JW. Fibrin gels and their clinical and bioengineering applications. J R Soc Interface. 2009;6:1–10. doi: 10.1098/rsif.2008.0327.
  23. Kobayashi M, Kawase T, Horimizu M, Okuda K, Wolff LF, Yoshie H. A proposed protocol for the standardized preparation of PRF membranes for clinical use. Biologicals. 2012;40(5):323–9. doi: 10.1016/j.biologicals.2012.07.004.
  24. Kobayashi M, Kawase T, Okuda K, Wolff LF, Yoshie H. In vitro immunological and biological evaluations of the angiogenic potential of platelet-rich fibrin preparations: a standardized comparison with PRP preparations. Int J Implant Dent. 2015;1(1):31. doi: 10.1186/s40729-015-0032-0.
  25. Poniatowski ŁA, Wojdasiewicz P, Gasik R, Szukiewicz D. Transforming growth factor beta family: insight into the role of growth factors in regulation of fracture healing biology and potential clinical applications. Mediators Inflamm. 2015;2015:137823. doi: 10.1155/2015/137823.
  26. Sarahrudi K, Thomas A, Mousavi M, Kaiser G, Köttstorfer J, Kecht M, Hajdu S, Aharinejad S. Elevated transforming growth factor-beta 1 (TGF-β1) levels in human fracture healing. Injury. 2011 Aug;42(8):833-7. doi: 10.1016/j.injury.2011.03.055.

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How to Cite

Grigoryev, V., Popsuishapka, O., Ashukina, N., & Galkin, F. (2017). Localization of vascular endothelial growth factor and transforming growth factor-β in tissues in perifractural zone after fractures of long bones of limbs in humans. ORTHOPAEDICS TRAUMATOLOGY and PROSTHETICS, (2), 62–69. https://doi.org/10.15674/0030-59872017262-69

Issue

No. 2 (2017)

Section

ORIGINAL ARTICLES

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Copyright (c) 2017 Vitaliy Grigoryev, Olexii Popsuishapka, Nataliya Ashukina, Fedor Galkin

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