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Pre-required Condition of Schwann Cells Proliferation in the Injured Nerves
Kai J Yang, MD; Ji-Geng Yan, MD, PhD; Hani Matloub, MD; Robert Havlik, MD
Medical College of Wisconsin, Milwaukee, WIIntroduction: Following peripheral nerve injury, there is an increase in cellular calcium concentration and degree of accumulation correlates directly with severity of injury. In addition, calcium resorption is associated with nerve electrophysiological recovery. Furthermore, Schwann cells play a pivotal role in peripheral nerve regeneration and recovery. Thus, in this study, we aim to examine cultured Schwann cell survivals in various concentration of calcium containing growth mediums. We also examined the effect of calcitonin on Schwann cells in such mediums.
Materials & Methods: To establish baseline calcium concentration in a post nerve injury state, crush injury is induced on eight Sprague Dawley rats’ sciatic nerves. Calcium concentrations inside and outside the nerve fibers were measured to establish baseline values of 1.8 mM and 16.8 mM respectively. To study Schwann cell survival in different calcium concentration mediums, uninjured sciatic nerve segment was harvested and cultured in mediums containing 1.8 mM, 5.0 mM, 7.5 mM, 10.0 mM and 16.8 mM calcium. Cells were fixed, blocked and incubated with Anti-S-100, Goat-Anti-Mouse, and Propidium Iodide and Schwann cells were studied and analyzed under fluorescent conditions.
To study the effect of calcitonin, healthy sciatic nerve segments were again harvested and cultured in four different kind of calcium containing mediums: 1) normal control (N) (1x105 mM Ca2+); 2) normal control with calcitonin added (N-CT); 3) high calcium medium (HC) (7.5 mM Ca2+); 4) high calcium medium with calcitonin added (HC-CT). Cells were again incubated with Anti-S-100, Goat-Anti-Mouse, and Propidium Iodide and analyzed under fluorescent conditions.
Results: In the control group, there were a large number of viable Schwann cells in the normal growth medium. With increasing calcium concentration in the medium, there was a significant decrease in the number of Schwann cells present. And the amount decreased correlates directly with the Calcium concentration increased. However, when calcitonin was added to the growth medium, there were similar amount of Schwann cells present in the high calcium-containing medium comparing to the low calcium medium control group. (N-CT: N=97.4±11.8; HC-CT: N=95.8±9.3)
Conclusion: Schwann cells play pivotal role in peripheral nerve recovery following injuries. These cells are very sensitive to the increase in calcium concentration in the post-injury environment. Calcitonin counteracts the detrimental effects of increasing calcium on Schwann cell survival. This can have significant future clinical implications for patients with peripheral nerve injuries.
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