Controlled surface morphology and hydrophilicity of polycaprolactone toward human retinal pigment epithelium cells.
Controlled surface morphology and hydrophilicity of polycaprolactone toward human retinal pigment epithelium cells.
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DOI：
10.1016/j.msec.2016.11.076
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发表时间：
2017-04
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通讯作者：
Saleheh Shahmoradi;F. Yazdian;F. Tabandeh;Z. Soheili;Ashraf Sadat Hatamian Zarami;M. Navaei-Nigjeh
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文献类型：
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作者：
Saleheh Shahmoradi;F. Yazdian;F. Tabandeh;Z. Soheili;Ashraf Sadat Hatamian Zarami;M. Navaei-Nigjeh
应用支架作为床促进细胞增殖甚至分化是治疗老年性黄斑变性(AMD)等视网膜疾病的方法之一，AMD会恶化光感受器，最终导致失明。本研究采用电纺法制备了取向聚己内酯(PCL)纳米纤维，并在不同条件下对其进行了扫描电子显微镜(SEM)和接触角测试。采用响应面方法(RSM)优化了纳米纤维的直径。以溶液浓度和电压值两个因素为自变量，采用中心复合设计法考察了它们对纳米纤维直径的影响，得到最佳工艺条件分别为0.12 mg/mL和20 kV。为了降低聚己内酯的疏水性，采用碱性水解法对其表面进行了改性。采用Box Behnken设计优化了支架与碱性溶液的接触时间和碱性溶液的浓度(分别为120min和5min)。接触角测量表明，处理后的支架具有较高的亲水性(接触角为7.48°)。采用等离子体表面处理方法，比较了两种表面改性方法同时对水解性支架材料的改性效果。对培养在支架上的RPE细胞进行免疫细胞化学(ICC)、四甲基偶氮唑盐(四甲基偶氮唑盐)和细胞形态连续检测。有趣的是，人RPE细胞在水解支架上很好地显示了其特征形态。因此，我们引进了一种直径小(185.8 nm)、孔隙率高(82%)、亲水性好(接触角7.48°)的培养基质，有望用于人视网膜色素上皮细胞移植。
Applying scaffolds as a bed to enhance cell proliferation and even differentiation is one of the treatment of retina diseases such as age-related macular degeneration (AMD) which deteriorating photoreceptors and finally happening blindness. In this study, aligned polycaprolactone (PCL) nanofibers were electrospun and at different conditions and their characteristics were measured by scanning electron microscope (SEM) and contact angle. Response surface methodology (RSM) was used to optimize the diameter of fabricated nanofibers. Two factors as solution concentration and voltage value were considered as independent variables and their effects on nanofibers' diameters were evaluated by central composite design and the optimum conditions were obtained as 0.12 g/mL and 20 kV, respectively. In order to decrease the hydrophobicity of PCL, the surface of the fabricated scaffolds was modified by alkaline hydrolysis method. Contact time of the scaffolds and alkaline solution and concentration of alkaline solution were optimized using Box Behnken design and (120 min and 5 M were the optimal, respectively). Contact angle measurement showed the high hydrophilicity of treated scaffolds (with contact angle 7.48°). Plasma surface treatment was applied to compare the effect of using two kinds of surface modification methods simultaneously on hydrolyzed scaffolds. The RPE cells grown on scaffolds were examined by immunocytochemistry (ICC), MTT and continuous inspection of cellular morphology. Interestingly, Human RPE cells revealed their characteristic morphology on hydrolyzed scaffold well. As a result, we introduced a culture substrate with low diameter (185.8 nm), high porosity (82%) and suitable hydrophilicity (with contact angle 7.48 degree) which can be promising for hRPE cell transplantation.