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In Vitro Effects of Topical Neuromodulatory Medication on Orofacial Tissue

Lookup NU author(s): Dr Mustafa Al-Musawi, Professor John Whitworth, Dr Simon StoneORCiD, Professor Justin DurhamORCiD, Professor Ruth Valentine


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Objective: Systemic neuromodulatory medication (NM) such as Amitriptyline are used in the management of neuropathic orofacial pain. Topical administration of NM, as well as topical anaesthesia has also been reported, but the effect of exposure of oral mucosa to topical NMs has not been examined fully.Aim:The aim of this study was to investigate the effects of amitriptyline on an oral mucosal model.Methods: Methods: Amitriptyline was applied to a commercially available (MatTek) and in-house constructed 3D oral mucosal model at 226┬ÁM. Cell viability was measured at different time-points, using lactate dehydrogenase (LDH). Cell death related to apoptosis was investigated by measuring Caspase-3 levels. High-Performance Liquid Chromatography (HPLC) as well as immunofluorescence was used to investigate tissue infiltration of the drug.Results: Results: Tissue exposure to amitriptyline for 30 min and 24 h resulted in significant levels of LDH, at 30 minutes and 24 hours compared with untreated control tissue samples in the in-house model (1.30 + 0.0501, 1.32 + 0.033 versus 1.00 + 0.072 at 30 min and 24 h respectively; p<0.05). In the MatTek model, levels of LDH were increased at both time points, but only significant after 24 hours exposure (1.24 + 0.0088 versus 1.0 + 0.0009; p < 0.05). The indirect measurement of apoptosis using Caspase-3 levels identified significantly elevated levels in the in-house model at both time points compared with the untreated control model (36 + 1.3 vs 18; 30 minutes, 30 + 2.6 vs 10 at 24 hours; p<0.05), whereas significant levels were only identified at 24 hours in the MatTek model (19 + 0.99 vs 11; p< 0.05). Amitriptyline passage through both models was confirmed using immunofluorescence and chemical analysis.Conclusions: Conclusions: Long term exposure to topical Amitriptyline in vitro is was associated with increased apoptotic activity in tissue models. Further research will investigate the effect of topical anaesthetic in this model system.

Publication metadata

Author(s): Al-Musawi M, Sayeed Z, Colley H, Murdoch C, Whitworth J, Stone S, Nixdorf D, Durham J, Valentine R

Publication type: Conference Proceedings (inc. Abstract)

Publication status: Published

Conference Name: IADR/PER General Session

Year of Conference: 2018

Pages: 3420

Online publication date: 25/07/2018

Acceptance date: 28/07/2018

Publisher: IADR