Acute cytotoxic effects of marketed ophthalmic formulations on human corneal epithelial cells

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• 作者：Jenni J. Hakkarainen^a ; ^{jenni@experimentica.com" class="auth_mail" title="E-mail the corresponding author} ; Mika Reinisalo^a ; ^b ; Symantas Ragauskas^a ; ^c ; Aila Seppä ; nen^a ; Simon Kaja^a ; ^d ; ^e ; Giedrius Kalesnykas^a ; ^f
• 关键词：BAC ; benzalkonium chloride ; MGHS40 ; macrogolglycerol hydroxystearate ; PS80 ; polysorbate 80 ; HCE-T ; human corneal epithelial cell line ; HBSS ; Hank⿿s balanced salt solution ; DPBS ; Dulbecco⿿s phosphate buffered saline ; DAPI ; 4⿲ ; 6-Diamidino-2-phenylindole dihydrochloride ; RFU ; relative fluorescence units ; IC50 value ; concentration of compound giving 50% cell viability
• 刊名：International Journal of Pharmaceutics
• 出版年：2016
• 出版时间：10 September 2016
• 年：2016
• 卷：511
• 期：1
• 页码：73-78
• 全文大小：1093 K

文摘

The purpose of the study was to devise a fast, reliable and sensitive cell viability assay for assessment of acute cytotoxicity on human corneal epithelial cells by using a clinically relevant exposure time. Acute cytotoxic effects of the pharmaceutical excipients benzalkonium chloride (BAC), macrogolglycerol hydroxystearate (MGHS40), polysorbate 80 (PS80) and marketed ophthalmic formulations (Lumigan^®, Monoprost^®, Taflotan^®, Travatan^®, Xalatan^®) containing these excipients were tested. Human corneal epithelial cell (HCE-T) viability was assessed by measuring the reduction of resazurin to highly fluorescent resorufin. Expression of the tight junction proteins in HCE-T cells were characterized by immunofluorescence staining. Presence of tight junction proteins in HCE-T cells was demonstrated. BAC preserved ophthalmic formulations showed concentration-dependent and time-dependent cytotoxicity to human corneal epithelium. In contrast, no acute cytotoxicity of non-ionic stabilizing/solubilizing excipients (MGSH40 and PS80) or ophthalmic formulation containing these excipients was observed. Marketed ophthalmic formulations used for glaucoma medication show differential toxicity on human corneal epithelial cells. The present study revealed that BAC-preserved ophthalmic formulations were able to induce acute cytotoxic effects even during a clinically relevant exposure time, which was not observed with MGSH40 and PS80 excipients or ophthalmic formulations containing these excipients.