Riabenko T. V., Korenkov O. V., Ponyrko A. A., Teslyk T. P., Hula V. I.

MORPHOLOGY

Scentific article

Abstract. Patients with oncological diseases have a high percentage of bone fractures due to the development of disorders of bone metabolism in the form of metastatic bone lesions, osteoporosis and pathological fractures. Due to the need for long-term courses of anticancer chemotherapy to treat cancer, fracture healing in such patients often occurs with cytotoxic therapy. Therefore, the aim of our study was to study the activity of markers of resorption and bone formation in the area of bone regeneration under the influence of antitumor chemotherapeutics. The study was performed on laboratory rats, which were inflicted a perforated defect with a diameter of 2 mm in the middle third of the femoral shaft. Animals were divided into control and three experimental groups, which after injury were injected intraperitoneally with anticancer chemotherapeutics: I – doxorubicin (60 mg / m²), II-5 – fluorouracil (600 mg/m²), III – methotrexate (40 mg / m²). Every 21 days, the animals of the experimental groups were repeated administration of the appropriate chemotherapy throughout the experiment. On the 15th, 30th, 45th, 60th day after injury, the animals were removed from the experiment. Using immunohistochemistry, the expression of cathepsin K and osteopontin was determined by the cellular elements of the regenerate. It was found that antitumor chemotherapeutics increase the expression of cathepsin K in the area of bone regenerate: doxorubicin – 25.72% (p˂0.005), 5-fluorouracil – 17.64% (p˂0.005), methotrexate – 22.76% (p˂0,005) compared with the control group on the 60th day of the experiment. In addition, antitumor chemotherapeutics caused a decrease in osteopontin activity in bone regenerate: doxorubicin-by 20.62% (p˂0.005), 5-fluorouracil-by 18.87% (p˂0.005) and methotrexate-by 21.89% ( p˂0,005) compared with the control on the 60th day after injury. The most pronounced negative effect on the formation of callus was observed with the use of methotrexate and doxorubicin. The appointment of antitumor chemotherapeutics leads to increased resorption processes in bone regenerate due to the activation of osteoclastogenesis and reduced functional activity of osteoblasts, which slows mineralization in the fracture site.

bone regenerate, cathepsin K, osteopontin, antitumor chemotherapeutics.

1. Bahney CS, Zondervan RL, Allison P, Theologis A, Ashley JW, Ahn J, et al. Cellular biology of fracture healing. Journal of Orthopaedic Research. 2019;37(1):35-50. doi: https://doi.org/10.1002/jor.24170.
2. Giannoudis PV, editor. Fracture Reduction and Fixation Techniques. Switzerland: Springer; 2018. Chapter 1, Fracture Healing: Back to Basics and Latest Advances; p. 3-17. doi: https://doi:10.1007/978-3-319-68628-8_1.
3. Sturgeon KM, Mathis KM, Rogers CJ, Schmitz KH, Waning DL. Cancer and Chemotherapy Induced Musculoskeletal Degradation. JBMR plus. 2019:3(3);e10187. doi: https://doi.org/10.1002/jbm4.10187.
4. Handforth C, D’Oronzo S, Coleman R, Brown J. Cancer Treatment and Bone Health. Calcif Tissue Int. 2018;102:251-264. doi: https://doi. org/10. 1007/s00223-017-0369-x.
5. Riabenko TV, Ponyrko AO. Analiz vplyvu zloiakisnykh novoutvoren’ na stan kistkovoi systemy ta poiava dii protypukhlynnykh khimiopreparativ na reheneratsiiu kistkovoi tkanyny (ohliad literatury). Bukovyns’kyi medychnyi visnyk. 2020;24.1(93):242-249. [in Ukrainian].
6. D’Oronzo S, Stucci S, Tucci M, Silvestris F. Cancer treatment-induced bone loss (CTIBL): Pathogenesis and clinical implications. Cancer Treatment Reviews. 2015;41(9):798-808. doi: https://doi:10.1016/j.ctrv.2015.09.003.
7. Greenblatt MB, Tsai JN, Wein MN. Bone Turnover Markers in the Diagnosis and Monitoring of Metabolic Bone Disease, Clinical Chemistry. 2017;63(2):464-474. doi: https://doi.org/10.1373/clinchem.2016.259085.
8. Ferreira A, Alho I, Casimiro S, Costa L. Bone remodeling markers and bone metastases: From cancer research to clinical implications. BoneKEy Reports. 2015;4:668. doi: https://dx.doi.org/ 10.1038 / bonekey.2015.35.
9. Konukoglu D. Bone markers. Int J Med Biochem. 2019;2(2):65-78. doi: 10.14744/ijmb.2019.60362.
10. Takito J, Inoue S, Nakamura M. The Sealing Zone in Osteoclasts: A Self-Organized Structure on the Bone. International Journal of Molecular Sciences. 2018;19(4):984. doi: https://doi.org/10.3390/ijms19040984.
11. Dai R, Wu Z, Chu HY, Lu J, Lyu A, Liu J, et al. Cathepsin K: The action in and beyond bone. Frontiers in Cell and Developmental Biology. 2020;8:433. doi: https://doi.org/10.3389/fcell.2020.00433.
12. Costa AG, Cusano NE, Silva BC, Cremers S, Bilezikian JP. Cathepsin K: its skeletal actions and role as a therapeutic target in osteoporosis. Nat Rev Rheumato. 2011;17:447-456. doi: https://doi.org/10.1038/nrrheum.2011.77.
13. Christensen J, Vonwil D, Shastri VP. Non-invasive in vivo imaging and quantification of tumor growth and metastasis in rats using cells expressing far-red fluorescence protein. PloS one. 2015;10(7):e0132725. doi: https: //doi. org/10.1371/journal.pone.0132725.
14. Drake MT, Clarke BL, Oursler MJ, Khosla S. Cathepsin K inhibitors for osteoporosis: biology, potential clinical utility, and lessons learned. Endocrine reviews. 2017;38(4):325-350. doi: https://doi.org/10.1210/er. 2015-1114.
15. Okamoto K, Takayanagi H. Osteoimmunology: the conceptual framework unifying the immune and skeletal systems. Physiological reviews. 2017;97(4):1295-1349. doi: https://doi.org/10.1152/physrev.00036.2016.
16. Ono T, Nakashima T. Recent advances in osteoclast biology. Histochemistry and cell biology. 2018;149(4):325-341. doi: https://doi. org/10.1007/s00418-018-1636-2.
17. Depalle B, McGilvery CM, Nobakhti S, Aldegaither N, Shefelbine SJ, Porter AE. Osteopontin regulates type I collagen fibril formation in bonetissue. Acta biomaterialia. 2021;120:194-202. doi: https://doi.org/10.1016/j.actbio. 2020.04.040.
18. Rodriguez DE, Thula-Mata T, Toro EJ, Yeh YW, Holt C, Holliday LS, et al. Multifunctional role of osteopontin in directing intrafibrillar mineralization of collagen and activation of osteoclasts. Acta biomaterialia. 2014;10(1):494-507. doi: https://doi.org/10.1016/j.actbio.2013.10.010.
19. Si J, Wang C, Zhang D, Wang B, Hou W, Zhou Y. Osteopontin in bone metabolism and bone diseases. Medical science monitor: international medical journal of experimental and clinical research. 2020;26:e9191591. doi: https://doi.org/10.12659/MSM.919159.
20. Linder HC, Ek-Rylander B, Krumpel M, Norgard M, Narisawa S, Millan JL, et al. Bone Alkaline Phosphatase and Tartrate-Resistant Acid Phosphatase: Potential Co-regulators of Bone Mineralization. Calcified tissue international. 2017;101(1):92-101. doi: https://doi. org/10.1007/s00223-017-0259-2.
21. Verhovna Rada Ukrayni. Evropeyska konventsiya pro zahist hrebetnih tvarin, scho vikoristovuyutsya dlya doslidnih ta inshih naukovih tsiley. Strasburg, 18 bereznya 1986 roku: ofltsiyniy pereklad [Internet]. Dostupno: http:zakon.rada.gov.ua/cgi-bin/laws/main.cgi?nreg=994_137. [in Ukrainian].
22. Verhovna Rada Ukrayni. Zakon Ukrayini № 3447-IV vіd 21.02.2006 «Pro zahist tvarin vid zhorstokogo povodzhennya» [Internet]. 2006. Dostupno: https://zakon.rada.gov.ua/laws/show/3447-15#Text. [in Ukrainian]
23. Kukharuk VR. Alhorytmy heneruvannia imunohistokhimichnykh zobrazhen. Ternopil: Zakhidnoukrayinsʹkyy natsionalʹnyy universytet; 2020. 74 s. [in Ukrainian].
24. Bonnet N, Douni E, Perréard Lopreno G, Besse M, Biver E, Ferrari S. RANKL induced increase in cathepsin K levels restricts cortical expansion in a periostin dependent fashion: a potential new mechanism of bone fragility. Journal of Bone and Mineral Research. 2021;2021:1. doi: https://doi.org/10.1002/jbmr.4307.
25. Duvall CL, Taylor WR, Weiss D, Wojtowicz AM, Guldberg RE. Impaired Angiogenesis, Early Callus Formation, and Late Stage Remodeling in Fracture Healing of Osteopontin-Deficient Mice. Journal of Bone and Mineral Research. 2006;22(2):286-297. doi: 10.1359/jbmr.061103.
26. Bailey S, Karsenty G, Gundberg C, Vashishth D. Osteocalcin and osteopontin influence bone morphology and mechanical properties. Ann N Y Acad Sci. 2017;1409(1):79-84. doi: 10.1111/nyas.13470.
27. Wei R, Wong JPC, Kwok HF. Osteopontin – a promising biomarker for cancer therapy. J Cancer. 2017;8(12):2173-2183. doi: 10.7150/ jca.20480.

«Bulletin of problems biology and medicine» Issue 2 (160), 2021 year, 236-241 pages, index UDK 616.7-006-092.18-085.277.3

10.29254/2077-4214-2021-2-160-236-241