Differentiation mechanisms of regeneration blastema cells during bone fracture healing

Olexii Popsuishapka, Valeriy Litvishko, Nataliya Ashukina, Vitaliy Grigoryev, Olga Pidgaiska


For understanding of reasons of bone nonunion after fractures we have to study the mechanisms which are on the base of cells differentiation.

Objective: on the base of clinical and morphologic examination we studied the mechanisms of cells differentiation at regeneration process after shaft fractures.

Methods: for histological study we took fibrin-blood clots from perifracture area and adjacent soft tissues in 25 patients after closed shaft fractures at open reduction. Biopsy samples of 9 patients were studied additionally with imunohistochemistry methods for the analysis of vessels endothelial growth factor and transforming growth factor-β (TGF-β). For assessment of fracture union we used X-rays.

Results: in 1–5 days after trauma we have found fibrin-blood clots, where thickened fibrin partitions were located parallel to each other. Cells were formed by fibrin partitions, they had oval shape, it depicted the presence of fluid pressure inside of them. Expressed reaction on the vessels endothelial growth factorwas found in fibrin. During 7–18 days after fracture the fibrin-blood clot was reorganizes with forma­tion of granular, soft tissue and osteogenic tissues. Expression of vessels endothelial growth factorand TGF-β was registered in cells.

Conclusions: osteogenic differentiation of mesenchimal cells in bone callous after the fracture can appear in case of coincide in time and space key factors — presence of fibrin matrix saturated vessels endothelial growth factor. It initiates vessels formation. Also there is need of close contact with alive mature tissues (bone, periosteum, muscles) which are the sources of slightly differentiated cells; tensions in fibrin-collagen blastema.


bone shaft fracture; fibrin-blood clot; histology; immunohistochemistry; functional therapy

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Antonova, E., Le, T. K., Burge, R., & Mershon, J. (2013). Tibia shaft fractures: costly burden of nonunions. BMC Musculoskeletal Disorders, 14(1). doi:10.1186/1471-2474-14-42

Panteli, M., Pountos, I., Jones, E., & Giannoudis, P. V. (2015). Biological and molecular profile of fracture non-union tissue: current insights. Journal of Cellular and Molecular Medicine, 19(4), 685-713. doi:10.1111/jcmm.12532

Dedukh, N. V., Romanenko, K. K., Horidova, L.D., & Ashukina, N. A. (2003). Morphological examination of biopsy materials from bone disregeneration areas. Ukrainian Medical Almanakh, 5(2), 69–72.

Reed, A. A., Joyner, C. J., Brownlow, H. C., & Simpson, A. H. (2002). Human atrophic fracture non-unions are not avascular. Journal of Orthopaedic Research, 20(3), 593-599. doi:10.1016/s0736-0266(01)00142-5

Iwakura, T., Miwa, M., Sakai, Y., Niikura, T., Lee, S. Y., Oe, K., … & Kurosaka, M. (2009). Human hypertrophic nonunion tissue contains mesenchymal progenitor cells with multilineage capacity in vitro. Journal of Orthopaedic Research, 27(2), 208–215. doi:10.1002/jor.20739

Krompecher, I. (1971). Local tissue metabolism and biological particulars of bone regenerate. Osseous tissue regeneration mechanisms. Moscow: Medicine.

Kanczler, J. M., & Oreffo, R. O. (2008). Osteogenesis and angiogenesis: the potential for engineering bone. European Cells & Materials, 15, 100–114. doi:10.22203/ eCM.v015a08

Hankenson, K. D., Dishowitz, M., Gray, C., & Schenker, M. (2011). Angiogenesis in bone regeneration. Injury, 42(6), 556-561. doi:10.1016/j.injury.2011.03.035

Oksymets, V. M. (2014). Cellular-tissue technologies in treatment of reparative osteogenesis disorders and osseous tissue defects: theoretical justification and potential of clinical use (experimental-diagnostic study) (Thesis for …Doctor of Medical Sciences,Donetsk).

Moreno-Miralles, I., Schisler, J. C., & Patterson, C. (2009). New insights into bone morphogenetic protein signaling: focus on angiogenesis. Current Opinion in Hematology, 16(3), 195-201. doi:10.1097/moh.0b013e32832a07d6

Sarkisov, D. S., & Perov, Yu. L. (1996). Microscopic Technique. Moscow: Medicine.

Litvishko, V., & Popsuishapka, O. (2015). The functional treatment of the diaphyseal tibial fractures using plaster cast or external fixator. Orthopaedics, traumatology and prosthetics, 4, 91-102. doi:

Wang, X., Friis, T. E., Masci, P. P., Crawford, R. W., Liao, W., & Xiao, Y. (2016). Alteration of blood clot structures by interleukin-1 beta in association with bone defects healing. Scientific Reports, 6(1). doi:10.1038/srep35645

Street, J., Winter, D., Wang, J. H., Wakai, A., McGuinness, A., & Redmond, H. P. (2000). Is human fracture hematoma inherently angiogenic? Clinical Orthopaedics and Related Research, 378, 224-237. doi:10.1097/00003086-200009000-00033

Martino, M. M., Briquez, P. S., Ranga, A., Lutolf, M. P., & Hubbell, J. A. (2013). Heparin-binding domain of fibrin(ogen) binds growth factors and promotes tissue repair when incorporated within a synthetic matrix. Proceedings of the National Academy of Sciences, 110(12), 4563-4568. doi:10.1073/pnas.1221602110

Mosesson, M. W. (2005). Fibrinogen and fibrin structure and functions. Journal of Thrombosis and Haemostasis, 3(8), 1894-1904. doi:10.1111/j.1538-7836.2005.01365.x

Hu, K. W., & Olsen, B. R. (2016). The roles of vascular endothelial growth factor in bone repair and regeneration. Bone, 91, 30-38. doi:10.1016/j.bone.2016.06.013

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. doi:

Hu, K., & Olsen, B. R. (2016). Osteoblast-derived VEGF regulates osteoblast differentiation and bone formation during bone repair. Journal of Clinical Investigation, 126 (2), 509-526. doi:10.1172/jci82585

Leek, R. D., Hunt, N. C., Landers, R. J., Lewis, C. E., Royds, J. A., & Harris, A. L. (2000). Macrophage infiltration is associated with VEGF and EGFR expression in breast cancer. The Journal of Pathology, 190 (4), 430-436. doi:10.1002/(sici)1096-9896(200003)190:4<430::aid-path538>;2-y

Serov, V. V., & Shekhter, A. B. (1981). Connective tissue. Moscow: Medicine.

Claes, L. E., Heigele, C. A., Neidlinger-Wilke, C., Kaspar, D., Seidl, W., Margevicius, K. J., & Augat, P. (1998). Effects of Mechanical Factors on the Fracture Healing Process. Clinical Orthopaedics and Related Research, 355S, S132-S147. doi:10.1097/00003086-199810001-00015

Litvishko, V., Popsuishapka, O., & Yaresko, O. (2016). Stress-strain state of fibrin-blood clot and periosteum in the area of diaphyseal fracture under different conditions of fracture fragments fixation and its impact on the structural organization of regenerate. Orthopaedics, traumatology and prosthetics, 1, 62-71. doi:

Copyright (c) 2018 Alexey Popsuishapka, Valeriy Litvishko, Nataliya Ashukina, Vitaliy Grigoryev, Olga Pidgaiska

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