Bone regeneration on the background of the local impact of insulin-like growth factor-1 (experimental study)
Keywords:insulin-like growth factor-1, bone repair, experimental model
The problem of the optimization and stimulation of bone regeneration in orthopedics and traumatology is particularly relevant in the presence of large size and complex bone defects. Among the numerous growth factors insulin-like growth factor-1 (IGF-1) is one of the most important hormonal mediators and regulators of growth hormone. IGF-1 may act on bone tissue both systemically and locally. IGF-1 regulates the growth, development and differentiation of cells and tissues and in particular of bone tissue. However, it should be noted that most existing studies consider of the IGF-1 influence on the bone tissue growth, development and metabolism with study of the mechanism of its action at the molecular level. Action of IGF-1 on bone regeneration, especially in the early stages of the fracture healing was shown insufficient.
Purpose: to study peculiarities of bone osteogenesis in bone defect created in rats’ femur after local impact of IGF-1 with using of morphological methods.
Methods: experimental study was performed on 30 white outbred laboratory rats (2 month old, weight 150 ±15 g). Two groups of experiments were carried out:
1st group – modeling perforated defect in the distal metaphysis of the femur (control, 15 animals);
2nd group – modeling perforated defect in the distal metaphysis of the femur and the treatment of animals with IGF-1 (experimental, 15 animals).
0.08 microgram of IGF-1 was introduced in the defects of experimental animals at 1st and 3rd days after surgery.
For morphological study the animals were sacrificed in 3, 5, 7, 14 and 21 days after surgery.
We used morphological, morphometric and transmission electron microscopic methods for assessing of bone repair process. Morphometry of areas of newly formed bone in the defect was performed.
Results: during electron microscopic analysis of the bone defect area at the distal metaphysis of the femur of rats treated with IGF-1 (3rd day after surgery) it was shown that functionally active macrophages with numerous secondary lysosomes and phagosomes in the cytoplasm were located in hematoma. That reflects more active process of bone wound cleaning from fibrin degradation products than in the control group. During ultrastructural analysis of the bone defect of rats treated with IGF-1 on the 5th day after surgery functional activity of fibroblasts was demonstrated. On the 7th day density of osteoblasts increased in the experimental animals as compared with control group. The improvement of quality of the adjacent bone was shown. Apposition formation of bone in lysis areas, cracks and fissures was founded. Bone area in the defect of experimental animals was increased by 19.86 % on the 14th day and by 24.78 % on the 21st day as compared with control animals.
Conclusions: IGF-1 stimulates bone osteogenesis.
Yakar S. IGF-1 and bone: new discoveries from mouse models review / S. Yakar, H.-W. Courtland, D. Clemmons // J. Bone Mineral Research. — 2010. — Vol. 25, № 12. — Р. 2543–2552.
Giustina A. Growth Hormone, insulin-like growth factors, and the skeleton / A. Giustina, Gh. Mazziotti, E. Canalis // Endocrine Reviews. — 2008. — Vol. 29, № 5. — P. 535–559.
Canalis E. Insulin-like growth-factors and their role in osteoporosis / E. Canalis, D. Agnusdei // Calcified tissue international. — 1996. — Vol. 58, № 3. — P. 133–134.
Jadlowiec J. A. Bone tissue engineering: recent advances and promising therapeutic agents / J. A. Jadlowiec, A. B. Celil, J. O. Hollinger // Expert Opin. Biol. Ther. — 2003. — Vol. 3, № 3. — P. 409–423.
Salgado A. J. Bone tissue engineering: state of the art and future trends / A. J. Salgado, O. P. Coutinho, R. L. Reis // Macromol. Biosci. — 2004. — Vol. 4. — Р. 743–765.
Targeted overexpression of insulin-like growth factor I to osteoblasts of transgenic mice: increased trabecular bone volume without increased osteoblast proliferation / G. Zhao, M. C. Monier-Faugere, M. C. Langub [et al.] // Endocrinology. — 2000. — Vol. 141. — P. 2674–2682.
Growth factors regulate the synthesis of insulin-like growth factor-I in bone cell cultures / E. Canalis, J. Pash, B. Gabbitas [et al.] // Endocrinology. — 1993. — Vol. 133. — P. 33–38.
European Convention for the Protection of Vertebrate Animals used for experimental and other scientific purposes. Strasbourg, 18 March 1986: official translation [electronic resource] / Parliament of Ukraine. - Off. website. - (International Council of Europe documents). - Access to document: http:zakon.rada.gov.ua/cgi-bin/laws/main.cgi?nreg=994_137.
Ukraine. Laws. On the protection of animals from abuse (Article 26) [№ 3447-IV of 21.02.2006].
Sarkisov D. S. Microscopic technique / D. S. Sarkisov, L. Yu. Perova. — M.: Medical, 1996. — 542 p.
Weekly B. Electron microscopy for Beginners / B. Weekley. — New York: Wiley, 1975. — 324 p.
Reynolds E. S. The use of lead citrate at high ph an electrono-paque stain in electron microscopy / E. S. Reynolds // J. Cell Biol. — 1963. — Vol. 17. — P. 208–212.
Avtandilov G. G. Medical morphometry / G. G. Avtandilov. — M.: Medical, 1990. — 356 p.
Macrophages accumulate in the early phase of tendon-bone healing / S. Kawamura, L. Ying, H. Kim [et al.] // J. Orthop. Res. — 2005. — Vol. 23, Iss. 6. — P. 1425–1432.
Impaired fracture healing in macrophage migration inhibitory factor-deficient mice / T. Kobayashi, S. Onodera, E. Kondo [et al.] // Osteoporosis International. — 2011. — Vol. 22, № 6. — P. 1955–1965.
Molecular Biology of the cell / B. Alberts, A. Johnson, J. Lewis [et al.] — 4th ed. — New York: Garland Science; 2002.
Schindeler A. Pre-clinical fracture repair studies [Электронный ресурс] / A. Schindeler: meeting Report from the 31st Annual Meeting of the American Society for Bone and Mineral Research (September 11–15, 2009). — Denver, Colorado, 2009. — Режим доступа: http://www.nature.com/bonekey/knowledgeenvironment/2009/0912/bonekey20090418/full/bonekey20090418.html.
Grundnes O. Effects of macrophage activation on bone healing / O. Grundnes, O. Reikeraas // J. Orthop. Sci. — 2000. — Vol. 5, № 3. — P. 243–247.
Pat. 2090888 C2 Russian Federation, МПК G01N33/48. A method for predicting the course of matching a long bone fracture / Chelnokov A. N., Kutepov S.M., Novitskaya N. V., Osipova A. V. — № 94005486/14; application 15.02.1994, published 20.09.1997.
Bone defect repair in rat tibia by TGF-beta1 and IGF-1 released from hydrogel scaffold / S. Srouji, I. Blumenfeld, A. Rachmiel, E. Livne // Cell Tissue Bank. — 2004. — Vol. 5, № 4. — P. 223–230.
Copyright (c) 2014 Olexander Tykhonenko, Ninel Diedukh, Lyudmila Bengus, Sergey Khmyzov
This work is licensed under a Creative Commons Attribution 4.0 International License.
The authors retain the right of authorship of their manuscript and pass the journal the right of the first publication of this article, which automatically become available after 6 months from the date of publication under the terms of Creative Commons Attribution License, which allows others to freely distribute the published manuscript with mandatory linking to authors of the original research and the first publication of this one in this journal.
Authors have the right to enter into a separate supplemental agreement on the additional non-exclusive distribution of manuscript in the form in which it was published by the journal (i.e. to put work in electronic storage of an institution or publish as a part of the book) while maintaining the reference to the first publication of the manuscript in this journal.
The editorial policy of the journal allows authors and encourages manuscript accommodation online (i.e. in storage of an institution or on the personal websites) as before submission of the manuscript to the editorial office, and during its editorial processing because it contributes to productive scientific discussion and positively affects the efficiency and dynamics of the published manuscript citation (see The Effect of Open Access).