Gallagher Andrew G, McLean Keri, Stewart Rosalind M K, Wellings Don A, Allison Heather E, Williams Rachel L
Department of Eye and Vision Science, Institute of Ageing and Chronic Diseases, University of Liverpool, Liverpool, United Kingdom.
SpheriTech Ltd., Runcorn, Cheshire, United Kingdom.
Invest Ophthalmol Vis Sci. 2017 Sep 1;58(11):4499-4505. doi: 10.1167/iovs.17-22301.
The purpose of this study was to develop a more efficient drug delivery device to overcome the limitations of current drop therapy for the treatment of fungal keratitis.
Amphotericin B (AmpB), 0 to 30 μg/mL, was associated with a poly-ε-lysine (pεK) hydrogel. Fungicidal effect against Candida albicans was assessed at 18 and 42 hours by optical density (OD600) and growth on agar. Tear film dilution effect was mimicked by storage of AmpB pεK gels in 3.4 mL sterile PBS for 24 hours prior to fungal incubation. Drug elution over 96 hours was evaluated by HPLC, and drug stability was tested while associated with the gel by OD600 up to 48 hours. Lack of cytotoxicity toward the HCE-T corneal epithelial cell line was assessed over 7 days.
AmpB pεK gels show fungicidal activity in normal conditions (0.057 OD600, SD 0.003, P < 0.005) and in the presence of horse serum (0.048 OD600, SD 0.028 P < 0.005) at 18 hours. The drug release profile was above therapeutic levels (0.188 μg/mL) for up to 72 hours. Tear dilution had no significant effect at higher concentrations of AmpB (3 to 10 μg/mL). AmpB pεK gels were not cytotoxic to the HCE-T cell line.
We demonstrated that AmpB pεK gels confer sustained therapeutic antifungal activity for at least 48 hours without corneal epithelial cell line cytotoxicity, suggesting their potential for in vivo use as an antifungal bandage contact lens. This could avoid the need for intensive topical medication in the treatment of fungal keratitis.
本研究的目的是开发一种更高效的药物递送装置，以克服当前滴眼疗法在治疗真菌性角膜炎方面的局限性。
将0至30μg/mL的两性霉素B（AmpB）与聚-ε-赖氨酸（pεK）水凝胶结合。在18小时和42小时时，通过光密度（OD600）和琼脂上的生长情况评估对白色念珠菌的杀菌效果。在真菌孵育前，将AmpB pεK凝胶在3.4 mL无菌磷酸盐缓冲盐水中储存24小时，以模拟泪膜稀释效应。通过高效液相色谱法评估96小时内的药物洗脱情况，并通过OD600测试与凝胶结合时长达48小时的药物稳定性。在7天内评估对HCE-T角膜上皮细胞系的细胞毒性。
AmpB pεK凝胶在正常条件下（OD600为0.057，标准差0.003，P < 0.005）和18小时时在存在马血清的情况下（OD600为0.048，标准差0.028，P < 0.005）显示出杀菌活性。药物释放曲线在长达72小时内高于治疗水平（0.188μg/mL）。在较高浓度的AmpB（3至10μg/mL）下，泪液稀释没有显著影响。AmpB pεK凝胶对HCE-T细胞系没有细胞毒性。
我们证明AmpB pεK凝胶可提供至少48小时的持续治疗性抗真菌活性，且对角膜上皮细胞系无细胞毒性，表明其有作为抗真菌绷带隐形眼镜体内使用的潜力。这可以避免在治疗真菌性角膜炎时频繁局部用药的需要。
