Elsevier

World Neurosurgery

Volume 105, September 2017, Pages 702-708
World Neurosurgery

Original Article
The Histological Effects of Ozone Therapy on Sciatic Nerve Crush Injury in Rats

https://doi.org/10.1016/j.wneu.2017.05.161Get rights and content

Objective

Peripheral nerve injury is a common, important problem that lacks a definitive, effective treatment. It can cause neurologic deficits ranging from paresthesia to paralysis. This study evaluated the effect of ozone therapy on sciatic nerve crush injury in rats.

Materials and Methods

Twenty-four male rats were divided into control sham surgery, sciatic nerve injury, and sciatic nerve injury with ozone groups (each n = 8). The sciatic nerve injury was inflicted via De Koning's crush-force method. The sciatic nerve injury group received medical air and the sciatic nerve injury ozone group received 0.7 mg/kg ozone. Sciatic nerve samples were obtained 4 weeks after injury. Vascular congestion, vacuolization, edema formation, S100 expression, and the thicknesses of the perineurium and endoneurium and diameter of the injured sciatic nerves were evaluated.

Results

The diameter of the sciatic nerve and thicknesses of the perineurium and epineurium were significantly greater in the sciatic nerve injury group (P < 0.05) and significantly less in the sciatic nerve injury with ozone group (P < 0.001). High S100 immunoreactivity was seen in the sciatic nerve injury group compared with the other 2 groups (P < 0.05). The distributions of vascular congestion and vacuolization were significantly less in the sciatic nerve injury with ozone group (P < 0.05).

Conclusions

Ozone therapy improved sciatic nerve injury recovery without causing an increase in fibrotic tissue. Ozone reduced fibrosis, vascular congestion, vacuolization, and edema in rodents. Ozone treatment might be used to assist in sciatic nerve injury.

Introduction

Peripheral nerve injury is a common problem that often has a poor outcome that results in disability. The incidence of peripheral nerve injury in all trauma cases is 2.8%.1 Issues related to crush injury include increased endoneurial edema, free oxygen radicals, inflammatory reactions, de- and remyelination, axonolysis, regeneration, and degeneration.2, 3, 4 However, there is no definitive treatment for peripheral nerve damage; at present, surgical exploration is the only treatment that can be offered to such patients, with varying results reported. Many drugs have been used in the treatment of experimentally induced peripheral nerve injury, including nonsteroidal anti-inflammatory agents, steroids, nerve growth factors, erythropoietin, thyroid hormone, growth hormone, adrenocorticotropic hormone, and insulin-like peptides.2, 3, 5

Ozone has antioxidant, antiapoptotic, and anti-inflammatory effects in the body by activating heme oxygenase-1.6, 7, 8 Neuroprotective effects of ozone were reported in an in vitro model of brain ischemia via up-regulation of intracellular antioxidant enzymes.9 In an experimental study, Ozbay et al.10 showed the beneficial effect of ozone on the regeneration of crushed facial nerves in rats; however, no study has evaluated ozone therapy for sciatic nerve injury, to our knowledge.

Schwann cell proliferation is an early nerve response to injury.11 The newly proliferated Schwann cells replace dead or dying cells. The S100 proteins are a group of low-molecular-weight acidic proteins that are found in glia and Schwann cells; they are thought to have a role in nervous system development in rats.12 The amount of S100 immunoreactivity is correlated with the thickness of the myelin sheath in Schwann cells.

We postulated that the anti-inflammatory and antioxidant effects of ozone might improve sciatic nerve recovery after crush injury in rats without causing an increase in fibrotic tissue in the traumatized sciatic nerve. Therefore, to evaluate the effects of ozone in sciatic nerve injury in rats, this study measured vascular congestion, vacuolization, edema formation, S100 expression, and the thicknesses of the perineurium and endoneurium and diameter of the injured sciatic nerves.

Section snippets

Materials and Methods

Twenty-four male rats were divided into sham surgery (group S), sciatic nerve injury (group SNI), and sciatic nerve injury with ozone (group SNO) groups with 8 rats per group. Sciatic nerve injury was inflicted via De Koning's crush-force method.13 In the sciatic nerve injury groups, the animals were anesthetized intraperitoneally and placed on an operating board in the prone position. The surgical field was cleaned and draped with sterile towels. The skin was incised at the proximal half of

Histologic Analysis

The sciatic nerve was examined for edema in the epineurium and hemorrhage around the epineurium in groups SNI and SNO (Figure 1). Mast cells and macrophages between the nerve fascicles were counted. Comparing fibrosis in the nerve fibers, group SNI had more fibrosis than group SNO (Figure 2). The vascular congestion and vacuolization were significantly lower in group SNO (P < 0.05) (Tables 1 and 2).

Histomorphometric Analysis

The sciatic nerve diameters ranged from 137.1 to 550.8 μm (mean 338.91 ± 97.98 μm), the thickness

Discussion

Peripheral nerve injury is an important common problem that lacks a definitive effective treatment. It can cause neurologic deficits ranging from paresthesia to paralysis. The injured neurons and cells of peripheral nerves fail to maintain an effective growth-promoting response, which commonly results in incomplete functional recovery in humans.15 Despite the advances in microsurgical techniques and rehabilitation facilities, the achievement of desired functional peripheral nerve regeneration

References (29)

  • Z.Z. Rodríguez et al.

    Preconditioning with ozone/oxygen mixture induces reversion of some indicators of oxidative stress and prevents organic damage in rats with fecal peritonitis

    Inflamm Res

    (2009)
  • H. Sahin et al.

    The acute effects of preoperative ozone therapy on surgical wound healing

    Acta Cir Bras

    (2016)
  • M. Frosini et al.

    Selective ozone concentrations may reduce the ischemic damage after a stroke

    Free Radic Res

    (2012)
  • I. Ozbay et al.

    Effects of ozone therapy on facial nerve regeneration

    Braz J Otorhinolaryngol

    (2016)
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    Conflict of interest statement: The authors declare that the article content was composed in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

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