Investigation of therapeutic effect of bevacizumab on rat trigeminal nerve cells as an in vitro model of trigeminal neuralgia

Introduction&Goal:Trigeminal neuralgia (TGN) is a neuropathic disorder characterized by severe pain that reduces the quality of life. It is estimated that the incidence in the community is quite high compared to the reported one, since its diagnosis and differential diagnosis are difficult. Trigeminal neuralgia (TGN) is still a major problem today and continues to be one of the most challenging diseases to treat. Therefore it creates both a psychological and financial burden for the individual as well as the society. For this reason, molecules that are newly released and are in the experimental stage can be hope in the treatment of trigeminal neuralgia. The causes of trigeminal neuralgia, although it is not certain, are primarily damage to the trigeminal nerve cells and surrounding neurons as a result of the compression of any lesion or structure on the nerve. This initiates inflammatory cascades at the cellular level, disrupting the perfusion and the resulting hypoxic state creates structural damage to the neurons. A series of cellular changes that begin after primary injury and continue over time. At the end of these processes, the myelination of the trigeminal nerve is impaired and edema develops. As a result of demyelination, hyperexcitability occurs in neurons and as a result, pain develops. Attempts to regenerate nerve cells begin, albeit weakly. In general, experimental treatments are aimed at preventing secondary damage and contributing to neuroregeneration. Studies have shown that one of the most important conditions in the preventing secondary damage and recovery period is to reduce the pressure and edema, and providing revascularization and nutrition to the cell as well as removing free radicals and harmful substances in the damaged area. The goal is eliminating the microcirculation disorder of the trigeminal nerve and improving perineural perfusion to eliminate the circulatory disorder. However, the revascularization that occurs during this period must be in a certain balance, and insufficient or excessive perfusion is also an issue. VEGF (Vascular Endothelial Growth Factor) type drugs have been previously used experimentally in these damage models, and their effects on reducing secondary damage by increasing vascularization. Additionally neuroprotective effects on damaged neural tissues have been shown. However, it has recently been thought that anti-VEGF drugs may also have a positive effect by preventing excessive vascularization. It is thought that it does these effects simply by reducing the post-damage edema and facilitating the transmission of oxygen and energy resources to the cells. Furthermore it may have neuroprotective effects by providing a limited angiogenesis, and increasing the efficiency in the nutrition of nerve cells; however in the literature, there is not enough scientific data. Material&Methods: Ten rats were used in the experiment. Cell culture protocols were performed after trigeminal ganglia (TG) were removed after decapitation. After the cells were separated, sterile normoglycemic (NG), hyperosmolar (HPO) and hyperglycemic (HPG) environments were created and placed on the plates in the xcelligence system with 100-200 cells. For the adhesion, growth and proliferation of cells, impedance measurements were carried out using biosensor electrodes for up to 24 hours according to RTCA (Real Time Cell Analysis) software. After 24 hours, NGF (Nerve Growth Factor) was given and impedance measurements continued for the next 24 hours. At the end of the 48th hour, treatment doses as bevacizumab dose 1 (0.025 mg/ml), bevacuzimab dose 2 (0.25 mg/ml), bevacuzimab dose 3 (1 mg/ml), bevacuzimab dose 4 (2.5 mg/ml) and impedance measurements were continued every 15 minutes with RTCA software until the end of 160 hours. Results: It was shown that all bevacizumab treatment doses did not have negative efficacy in NG medium, however, there was a positive significant difference in cell index at dose 4 (p<0.05). Although there was a statistically significant difference between only dose 4 and other groups, it was observed that there has been a dose-dependent positive effect in cell index. It has been shown that bevacizumab treatment in HPO medium has not a harmful effect in all treatment modalities, but a significant difference was found in cell indices in a dose-dependent manner and positive efficacy was demonstrated. Although there was no statistically significant difference among all doses, it was observed that there was a dose-dependent positive effect in cell index. It has been shown that bevacizumab treatment in HPG media has not a harmful effect in all treatment modalities, but a significant difference was found in cell indices in a dose-dependent manner and positive efficacy was demonstrated. Although there was no statistically significant difference among all doses, it was observed that there was a dose-dependent positive effect in cell index. Conclusions: It has been thought that anti VEGF treatment may contribute to neuronal regeneration in trigeminal neuralgia, and may have a reducing or limiting effect on secondary damage by protecting functional neurons. Key Words: trigeminal neuralgia, trigeminal neuropathy, bevacizumab, anti vegf, neuroregeneration, xcelligence