Open Access

Molecular profiling of a simple rat model of open tibial fractures with hematoma and periosteum disruption

  • Authors:
    • Jose Rafael Villafan‑Bernal
    • Lorenzo Franco‑de la Torre
    • Ana Soledad Sandoval‑Rodriguez
    • Juan Armendariz‑Borunda
    • Juan Luis Alcala‑Zermeno
    • Jose Alfonso Cruz‑Ramos
    • Gabriela Lopez‑Armas
    • Blanca Estela Bastidas-Ramirez
    • Gracia Viviana González‑Enríquez
    • Emerson Armando Collazo‑Guzman
    • Raigam Jafet Martinez‑Portilla
    • Sergio Sánchez‑Enríquez
  • View Affiliations

  • Published online on: September 29, 2016     https://doi.org/10.3892/etm.2016.3758
  • Pages: 3261-3267
  • Copyright: © Villafan‑Bernal et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Bone fractures are a worldwide public health concern. Therefore, improving understanding of the bone healing process at a molecular level, which could lead to the discovery of potential therapeutic targets, is important. In the present study, a model of open tibial fractures with hematoma disruption, periosteal rupture and internal fixation in 6-month-old male Wistar rats was established, in order to identify expression patterns of key genes and their protein products throughout the bone healing process. A tibial shaft fracture was produced using the three‑point bending technique, the hematoma was drained through a 4‑mm incision on the medial aspect of the tibia and the fracture stabilized by inserting a needle into the medullary canal. Radiographs confirmed that the induced fractures were diaphyseal and this model was highly reproducible (kappa inter‑rater reliability, 0.82). Rats were sacrificed 5, 14, 21, 28 and 35 days post‑fracture to obtain samples for histological, immunohistochemical and molecular analysis. Expression of interleukin‑1β (Il‑1β), transforming growth factor‑β2 (Tgf‑β2), bone morphogenetic protein‑6 (Bmp‑6), bone morphogenetic protein‑7 (Bmp‑7) and bone γ‑carboxyglutamic acid‑containing protein (Bglap) genes was determined by reverse transcription quantitative polymerase chain reaction and protein expression was evaluated by immunohistochemistry, while histological examination allowed characterization of the bone repair process. Il‑1β showed a biphasic expression, peaking 5 and 28 days post‑fracture. Expression of Tgf‑β2, Bmp‑6 and Bmp‑7 was restricted to the period 21 days post‑fracture. Bglap expression increased gradually, peaking 21 days post‑fracture, although it was expressed in all evaluated stages. Protein expression corresponded with the increased expression of their corresponding genes. In conclusion, a clear and well‑defined expression pattern of the evaluated genes and proteins was observed, where their maximal expression correlated with their known participation in each stage of the bone healing process.
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November-2016
Volume 12 Issue 5

Print ISSN: 1792-0981
Online ISSN:1792-1015

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Spandidos Publications style
Villafan‑Bernal JR, Franco‑de la Torre L, Sandoval‑Rodriguez AS, Armendariz‑Borunda J, Alcala‑Zermeno JL, Cruz‑Ramos JA, Lopez‑Armas G, Bastidas-Ramirez BE, González‑Enríquez GV, Collazo‑Guzman EA, Collazo‑Guzman EA, et al: Molecular profiling of a simple rat model of open tibial fractures with hematoma and periosteum disruption. Exp Ther Med 12: 3261-3267, 2016.
APA
Villafan‑Bernal, J.R., Franco‑de la Torre, L., Sandoval‑Rodriguez, A.S., Armendariz‑Borunda, J., Alcala‑Zermeno, J.L., Cruz‑Ramos, J.A. ... Sánchez‑Enríquez, S. (2016). Molecular profiling of a simple rat model of open tibial fractures with hematoma and periosteum disruption. Experimental and Therapeutic Medicine, 12, 3261-3267. https://doi.org/10.3892/etm.2016.3758
MLA
Villafan‑Bernal, J. R., Franco‑de la Torre, L., Sandoval‑Rodriguez, A. S., Armendariz‑Borunda, J., Alcala‑Zermeno, J. L., Cruz‑Ramos, J. A., Lopez‑Armas, G., Bastidas-Ramirez, B. E., González‑Enríquez, G. V., Collazo‑Guzman, E. A., Martinez‑Portilla, R. J., Sánchez‑Enríquez, S."Molecular profiling of a simple rat model of open tibial fractures with hematoma and periosteum disruption". Experimental and Therapeutic Medicine 12.5 (2016): 3261-3267.
Chicago
Villafan‑Bernal, J. R., Franco‑de la Torre, L., Sandoval‑Rodriguez, A. S., Armendariz‑Borunda, J., Alcala‑Zermeno, J. L., Cruz‑Ramos, J. A., Lopez‑Armas, G., Bastidas-Ramirez, B. E., González‑Enríquez, G. V., Collazo‑Guzman, E. A., Martinez‑Portilla, R. J., Sánchez‑Enríquez, S."Molecular profiling of a simple rat model of open tibial fractures with hematoma and periosteum disruption". Experimental and Therapeutic Medicine 12, no. 5 (2016): 3261-3267. https://doi.org/10.3892/etm.2016.3758