The Effect of Two Different Botulinum Neurotoxin A On The Cortical Neuron Cells In Terms of Apoptosis and MMP 2, MMP 7, and MMP9 Localizations
Abstract
This study aimed to associate the possible cytotoxic and apoptotic effects of Botox (Allergan) and Dysport (Ipsen) and immunolocalization of matrix metalloproteinase (MMP) proteins with HCN2 cortical neuron cell line. Accordingly, cytotoxic potentials of Botox and Dysport were determined on different concentrations. Then, the apoptosis rates of these cells were evaluated by TUNEL method. MMP2, MMP7, and MMP9 proteins were also visualized using immunofluorescence method. There was a significant difference in cytotoxicity between those treated with 3.2, 6.4, and 12.8IU compared with the control. The Dysport 12.8 IU group was statistically more toxic than Botox group at the same concentration. Therefore, the number of apoptotic cells increased from 0.1 IU in Botox and 0.01 IU in Dysport groups compared to the control. The number of apoptotic cells was significantly higher in Dysport group at 1.6, 3.2, 6.4, and 12.8 IU concentrations than in Botox group. It was determined that MMPs increased gradually at the concentrations where the number of apoptotic cells was highest compared to the control group. As a result, we consider that it may be necessary to deal with the dose adjustment in Botox and Dysport applications, together with detailed studies to be carried out in the future.
Keywords
Apoptosis, Botulinum neurotoxin A, HCN2, MMP, TUNEL.
Supporting Institution
CÜBAP
Project Number
T-756
References
- [1] Li Z., Lu J., Tan X., Wang R., Xu Q., Yu Y., Yang Z., Functional EL-HN Fragment as a Potent Candidate Vaccine for the Prevention of Botulinum Neurotoxin Serotype E, Toxins, 14 (2022) 135.
- [2] Alberts T., Antipova V., Holzmann C., Hawlitschka A., Schmitt O., Kurth J., Stenzel J., Lindner T., Krause B.J., Wree A., Witt, M., Olfactory Bulb D2/D3 Receptor Availability after Intrastriatal Botulinum Neurotoxin-A Injection in a Unilateral 6-OHDA Rat Model of Parkinson’s Disease, Toxins, 14 (2022) 94.
- [3] Carlı L., Montecucco C., Rossetto O., Assay of diffusion of different Botulinum neurotoxin type A formulations injected in the mouse leg, Muscle Nerve, 40 (2009) 374-380.
- [4] Curra A., Trompetto C., Abbruzzese G., Berardelli A., Central Effects of Botulinum Toxin Type A: Evidence and Supposition, Movement Disorders, 19 (8) (2004) 60-64.
- [5] Aoki K.R., Ranoux D., Wissel J., Using translational medicine to understand clinical differences between botulinum toxin formulations, Eur. J. Neurol., 13 (4) (2006) 10–19.
- [6] Wiegand H., Erdmann G., Wellhoner H.H., 125I-labelled botulinum A neurotoxin: pharmacokinetics in cats after intramuscular injection, Naunyn Schmiedebergs Arch. Pharmacol., 292 (1976) 161-165.
- [7] Hagenah R., Benecke R., Wiegand H., Effects of type A botulinum toxin on the cholinergic transmission at spinal Renshaw cells and on the inhibitory action at Ia inhibitory interneurones, Naunyn Schmiedebergs Arch. Pharmacol., 299 (1977) 267-272.
- [8] Boroff D.A., Chen G.S., On the question of permeability of the blood-brain barrier to BoNT, Int. Arch. Allergy Appl. Immunol., 48 (1975) 495-504.
- [9] Welch M.J., Purkiss J.R., Foster K.A., Sensitivity of embryonic rat dorsal root ganglia neurons to Clostridium botulinum neurotoxins, Toxicon, 38 (2000) 245-258.
- [10] Berliocchi L., Fava E., Leist M., Horvat V., Dinsdale D., Read D., Nico P., Botulinum neurotoxin C initiates two different programs for neurite degeneration and neuronal apoptosis, J. Cell Biol., 168 (2005) 607-618.