Experimental Induction of Physeal Injuries by Fracture, Drill, and Ablation Techniques: Analyses of Immunohistochemical Findings

J Pediatr Orthop. 2019 Oct;39(9):479-486. doi: 10.1097/BPO.0000000000001093.

Abstract

Background: Although physeal fractures and physeal bars can result in significant clinical consequences to growth and development of the injured physis, little orthopaedic research has focused upon this topic. Our objective was to extend a previously developed rat model to examine the immunohistochemical features following surgical application of techniques disrupting the physis.

Methods: Physes were surgically disrupted using fracture (control), epiphyseal scrape (ES), or epiphyseal drill (ED). After 1, 3, 6, 10, or 21 days, animals were euthanized, sites processed for histology and immunohistochemical localization of vascular endothelial growth factor (VEGF), Factor VIII, Sox-9, PTHrP (parathyroid hormone-related protein) and PTHrP-R (parathyroid hormone-related protein receptor) in resting, proliferative, and hypertrophic physeal zones. Incidence of physeal bars, vertical septa and islands within the metaphysis was quantified. Semiquantitative analysis of immunohistochemistry was performed.

Results: Physeal bars, vertical septa, and displaced cartilage islands were present each of the surgical treatments. Fisher's exact test showed a statistically significant increase in the presence of physeal bars (P=0.002) and vertical septa (P=0.012) in the ED group at 10 and 21 days. Analysis of VEGF showed significant differences among the surgical treatments involving the resting zone, and the proliferative zone for days 1, 6, and 21 (P≤0.02) with greater mean scores present in the fracture (control) group, followed by the ED group; the lowest scores were present in the ES group. PTHrP-R immunolocalization showed significant differences among treatments in the hypertrophic zone at days 6 and 21 (P=0.022 and 0.044, respectively).

Conclusions: On the basis of the type of surgical treatment, results show significant differences in the presence of VEGF (reflecting the vascular bed) in the resting and proliferating zones at days 1, 6, and 21. VEGF localization was less abundant in the ED group (which had more physeal bars), suggesting that lack of vascular ingrowth plays a role in physeal bar formation.

Clinical relevance: Basic science data presented here provide insight into the importance of the various regions of the physis and its repair and continued growth after physeal fracture. We suggest that a better understanding of the cellular basis of physeal arrest following physeal fracture may have future relevance for the development of treatments to prevent or correct arrest.

MeSH terms

  • Ablation Techniques
  • Animals
  • Epiphyses / injuries
  • Epiphyses / metabolism
  • Factor VIII / metabolism
  • Growth Plate / metabolism*
  • Growth Plate / surgery
  • Immunohistochemistry
  • Parathyroid Hormone-Related Protein / metabolism
  • Rats
  • Receptor, Parathyroid Hormone, Type 1 / metabolism
  • SOX9 Transcription Factor / metabolism
  • Salter-Harris Fractures / metabolism*
  • Salter-Harris Fractures / surgery
  • Vascular Endothelial Growth Factor A / metabolism

Substances

  • Parathyroid Hormone-Related Protein
  • Pth1r protein, rat
  • Pthlh protein, rat
  • Receptor, Parathyroid Hormone, Type 1
  • SOX9 Transcription Factor
  • Sox9 protein, rat
  • Vascular Endothelial Growth Factor A
  • vascular endothelial growth factor A, rat
  • Factor VIII