Yıl: 2021 Cilt: 55 Sayı: 2 Sayfa Aralığı: 94 - 101 Metin Dili: İngilizce DOI: 10.5152/j.aott.2021.18480 İndeks Tarihi: 29-09-2021

Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy

Öz:
Objective: This study aims to determine expression profiles of relevant genes in the early stages of post-traumatic heterotopicossification (HO) in a rat model of Achilles tenotomy.Methods: A total of 80 male Sprague-Dawley rats were randomly assigned to two groups: the HO group and the control group.Tenotomy was performed in the Achilles tendon of the rats in the HO group, and no intervention was conducted in the controlgroup. On the 3rd, 5th, 8th, and 14th days after the operation, 8 rats were taken from each group at each time point, and the Achillestendon and surrounding tissue specimens were collected. Gene expressions of TGF-β, BMP, GDF, IL, and MMP families as wellas TNF-α, HIF-1α chordin, gremlin, noggin, and NODAL were analyzed by qRT-PCR. The relevant genes that were highly expressed at different time points were screened, and immunohistochemical staining was then used to verify their expression. Atthe 10th week, HO formation was explored by radiographic and histological examination in the remaining 8 rats of each group.Results: Both the radiographic and histological analyses indicated that all the rats developed HO in the HO group (100%),whereas no HO occurred in the control group. Surrounding tissues obtained from the HO group showed significantly highergene expressions of TGF-β1, BMP-1, IL1β, HIF-1α, and MMP-2 but lower expressions of BMP-4, GDF-8, and TNF-α comparedwith the control group. In addition, immunohistochemical staining confirmed the higher protein expression levels of relevantgenes in the HO group.Conclusion: TGF-β1, BMP-1, IL-1β, HIF-1α, MMP-2, BMP4, GDF-8, and TNF-α may be associated with the formation of traumatic HO; and BMP4, GDF-8, and TNF-α may play a protective role in the early stage of HO. In this study, we investigated theexpression levels of the related cytokines in the early stages of traumatic HO in the Achilles tendon tenotomy rat model to betterunderstand the pathogenesis of HO.
Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Bibliyografik
  • 1. Downey J, Lauzier D, Kloen P, et al. Prospective heterotopic ossification progenitors in adult human skeletal muscle. Bone 2015; 71: 164-70.
  • 2. Winkler S, Niedermair T, Fuchtmeier B, et al. The impact of hypoxia on mesenchymal progenitor cells of human skeletal tissue in the pathogenesis of heterotopic ossification. Int Orthop 2015; 39: 2495-501.
  • 3. Anthonissen J, Ossendorf C, Hock JL, et al. The role of muscular trauma in the development of heterotopic ossification after hip surgery: An animal-model study in rats. Injury 2016; 47: 613-6.
  • 4. Burd TA, Hughes MS, Anglen JO. Heterotopic ossification prophylaxis with indomethacin increases the risk of long-bone nonunion. J Bone Joint Surg Br 2003; 85: 700-5.
  • 5. Tozun R, Pınar H, Yeşiller E, Hamzaoğlu A. Indomethacin for prevention of heterotopic ossification after total hip arthroplasty. J Arthroplasty 1992; 7: 57- 61.
  • 6. Mazonakis M, Berris T, Lyraraki E, Damilakis J. Cancer risk estimates from radiation therapy for heterotopic ossification prophylaxis after total hip arthroplasty. Med Phys 2013; 40: 101702.
  • 7. Pavey GJ, Polfer EM, Nappo KE, Tintle SM, Forsberg JA, Potter BK. What risk factors predict recurrence of heterotopic ossification after excision in combat-related amputations? Clin Orthop Relat Res 2015; 473: 2814-24.
  • 8. Toom A, Arend A, Gunnarsson D, et al. Bone formation zones in heterotopic ossifications: histologic findings and increased expression of bone morphogenetic protein 2 and transforming growth factors beta2 and beta3. Calcif Tissue Int 2007; 80: 259-67.
  • 9. Sawyer JR, Myers MA, Rosier RN, Puzas JE. Heterotopic ossification: clinical and cellular aspects. Calcif Tissue Int 1991; 49: 208-15.
  • 10. Shen Q, Zhu S, Hu J, Geng N, Zou S. Recombinant human bone morphogenetic protein-4 (BMP-4)-stimulated cell differentiation and bone formation within the expanding calvarial suture in rats. J Craniofac Surg 2009; 20: 1561-5.
  • 11. Sampath TK, Maliakal JC, Hauschka PV, et al. Recombinant human osteogenic protein-1 (hOP-1) induces new bone formation in vivo with a specific activity comparable with natural bovine osteogenic protein and stimulates osteoblast proliferation and differentiation in vitro. J Biol Chem 1992; 267: 20352-62.
  • 12. Evans KN, Forsberg JA, Potter BK, et al. Inflammatory cytokine and chemokine expression is associated with heterotopic ossification in high-energy penetrating war injuries. J Orthop Trauma 2012; 26: e204-e13.
  • 13. Rodenberg E, Azhdarinia A, Lazard ZW, et al. Matrix metalloproteinase-9 is a diagnostic marker of heterotopic ossification in a murine model. Tissue Eng Part A 2011; 17: 2487-96.
  • 14. Zimmermann SM, Wurgler-Hauri CC, Wanner GA, Simmen HP, Werner CM. Echinomycin in the prevention of heterotopic ossification - an experimental antibiotic agent shows promising results in a murine model. Injury 2013; 44: 570-5.
  • 15. Lin L, Shen Q, Leng H, Duan X, Fu X, Yu C. Synergistic inhibition of endochondral bone formation by silencing Hif1alpha and Runx2 in trauma-induced heterotopic ossification. Mol Ther 2011; 19: 1426-32.
  • 16. Pfander D, Cramer T, Schipani E, Johnson RS. HIF-1alpha controls extracellular matrix synthesis by epiphyseal chondrocytes. J Cell Sci 2003; 116: 1819-26.
  • 17. Bordeleau F, Mason BN, Lollis EM, et al. Matrix stiffening promotes a tumor vasculature phenotype. Proc Natl Acad Sci U S A 2017; 114: 492-7.
  • 18. Oliveira CO, Ikuta N, Regner A. Outcome biomarkers following severe traumatic brain injury. Rev Bras Ter Intensiva 2008; 20: 411-21.
  • 19. McClure J. The effect of diphosphonates on heterotopic ossification in regenerating Achilles tendon of the mouse. J Pathol 1983; 139: 419-30.
  • 20. Lin L, Shen Q, Xue T, Yu C. Heterotopic ossification induced by Achilles tenotomy via endochondral bone formation: expression of bone and cartilage related genes. Bone 2010; 46: 425-31.
  • 21. Lounev VY, Ramachandran R, Wosczyna MN, et al. Identification of progenitor cells that contribute to heterotopic skeletogenesis. J Bone Joint Surg Am 2009; 91: 652-63.
  • 22. Jackson WM, Aragon AB, Onodera J, et al. Cytokine expression in muscle following traumatic injury. J Orthop Res 2011; 29: 1613-20.
  • 23. Maeda S, Hayashi M, Komiya S, Imamura T, Miyazono K. Endogenous TGF-beta signaling suppresses maturation of osteoblastic mesenchymal cells. EMBO J 2004; 23: 552-63.
  • 24. Thomas M, Langley B, Berry C, et al. Myostatin, a negative regulator of muscle growth, functions by inhibiting myoblast proliferation. J Biol Chem 2000; 275: 40235-43.
  • 25. Oberpichler-Schwenk H. [Increased medical knowledge through and for the MMP]. Med Monatsschr Pharm 2015; 38: 287.
  • 26. Fujiki M, Shineha J, Yamanokuchi K, Misumi K, Sakamoto H. Effects of treatment with polysulfated glycosaminoglycan on serum cartilage oligomeric matrix protein and C-reactive protein concentrations, serum matrix metalloproteinase-2 and -9 activities, and lameness in dogs with osteoarthritis. Am J Vet Res 2007; 68: 827-33.
  • 27. Majmundar AJ, Wong WJ, Simon MC. Hypoxia-inducible factors and the response to hypoxic stress. Mol Cell 2010; 40: 294-309.
  • 28. Kutscher C, Lampert FM, Kunze M, Markfeld-Erol F, Stark GB, Finkenzeller G. Overexpression of hypoxia-inducible factor-1 alpha improves vasculogenesis-related functions of endothelial progenitor cells. Microvasc Res 2016; 105: 85-92.
  • 29. Wang H, Lindborg C, Lounev V, et al. Cellular Hypoxia Promotes Heterotopic Ossification by Amplifying BMP Signaling. J Bone Miner Res 2016; 31: 1652- 65.
  • 30. Agarwal S, Loder S, Brownley C, et al. Inhibition of Hif1alpha prevents both trauma-induced and genetic heterotopic ossification. Proc Natl Acad Sci U S A 2016; 113: E338-E47.
  • 31. Makowski GS, Ramsby ML. Autoactivation profiles of calcium-dependent matrix metalloproteinase-2 and -9 in inflammatory synovial fluid: effect of pyrophosphate and bisphosphonates. Clin Chim Acta 2005; 358: 182-91.
  • 32. Wozney JM, Rosen V, Celeste AJ, et al. Novel regulators of bone formation: molecular clones and activities. Science 1988; 242: 1528-34.
APA yu d, 鞠 鞠, Xue F, zhao y, Shi W, Xiao H (2021). Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy. , 94 - 101. 10.5152/j.aott.2021.18480
Chicago yu du,鞠 鞠金勇,Xue Feng,zhao yong,Shi Weizhe,Xiao Haijun Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy. (2021): 94 - 101. 10.5152/j.aott.2021.18480
MLA yu du,鞠 鞠金勇,Xue Feng,zhao yong,Shi Weizhe,Xiao Haijun Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy. , 2021, ss.94 - 101. 10.5152/j.aott.2021.18480
AMA yu d,鞠 鞠,Xue F,zhao y,Shi W,Xiao H Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy. . 2021; 94 - 101. 10.5152/j.aott.2021.18480
Vancouver yu d,鞠 鞠,Xue F,zhao y,Shi W,Xiao H Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy. . 2021; 94 - 101. 10.5152/j.aott.2021.18480
IEEE yu d,鞠 鞠,Xue F,zhao y,Shi W,Xiao H "Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy." , ss.94 - 101, 2021. 10.5152/j.aott.2021.18480
ISNAD yu, du vd. "Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy". (2021), 94-101. https://doi.org/10.5152/j.aott.2021.18480
APA yu d, 鞠 鞠, Xue F, zhao y, Shi W, Xiao H (2021). Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy. Acta Orthopaedica et Traumatologica Turcica, 55(2), 94 - 101. 10.5152/j.aott.2021.18480
Chicago yu du,鞠 鞠金勇,Xue Feng,zhao yong,Shi Weizhe,Xiao Haijun Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy. Acta Orthopaedica et Traumatologica Turcica 55, no.2 (2021): 94 - 101. 10.5152/j.aott.2021.18480
MLA yu du,鞠 鞠金勇,Xue Feng,zhao yong,Shi Weizhe,Xiao Haijun Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy. Acta Orthopaedica et Traumatologica Turcica, vol.55, no.2, 2021, ss.94 - 101. 10.5152/j.aott.2021.18480
AMA yu d,鞠 鞠,Xue F,zhao y,Shi W,Xiao H Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy. Acta Orthopaedica et Traumatologica Turcica. 2021; 55(2): 94 - 101. 10.5152/j.aott.2021.18480
Vancouver yu d,鞠 鞠,Xue F,zhao y,Shi W,Xiao H Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy. Acta Orthopaedica et Traumatologica Turcica. 2021; 55(2): 94 - 101. 10.5152/j.aott.2021.18480
IEEE yu d,鞠 鞠,Xue F,zhao y,Shi W,Xiao H "Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy." Acta Orthopaedica et Traumatologica Turcica, 55, ss.94 - 101, 2021. 10.5152/j.aott.2021.18480
ISNAD yu, du vd. "Expression and significance of related genes in the early stage of post-traumatic heterotopic ossification in a rat model of Achilles tenotomy". Acta Orthopaedica et Traumatologica Turcica 55/2 (2021), 94-101. https://doi.org/10.5152/j.aott.2021.18480