CN115466426B - 用于治疗角膜水肿的后弹力层植入型薄膜的制备方法 - Google Patents

用于治疗角膜水肿的后弹力层植入型薄膜的制备方法 Download PDF

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CN115466426B
CN115466426B CN202210945029.1A CN202210945029A CN115466426B CN 115466426 B CN115466426 B CN 115466426B CN 202210945029 A CN202210945029 A CN 202210945029A CN 115466426 B CN115466426 B CN 115466426B
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杜亚楠
王丽强
黄一飞
李俊阳
方逸凡
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Chinese PLA General Hospital
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Abstract

本发明涉及角膜植入体领域,提供一种用于治疗角膜水肿的后弹力层植入型薄膜的制备方法,能够获得具有良好的生物相容性的植入型薄膜,可以在房水以及PBS等液体中具有良好的可塑性,折叠后经2.4mm左右的小切口植入眼内并展开;另外,植入型薄膜的亲水面与角膜基质以及后弹力层具有良好的亲和力,且疏水效果确切,可以阻挡房水进入角膜基质,可以达到实现减轻角膜水肿的技术效果。

Description

用于治疗角膜水肿的后弹力层植入型薄膜的制备方法
技术领域
本发明涉及角膜植入体技术领域,涉及一种用于治疗角膜水肿的后弹力层植入型薄膜的制备方法。
背景技术
角膜是位于眼球前表面的最外层透明结构,是视觉形成的主要屈光介质,其透明性的维持主要依赖于角膜内皮细胞的离子泵、脱水能力及屏障功能。病理状态下,例如Fuchs角膜内皮营养不良、青光眼等因素导致内皮细胞丢失或死亡(<500/mm2)会产生。大泡性角膜炎由于基质水肿刺激角膜神经,房水大量聚集在角膜后弹力层,由于基质水肿刺激角膜神经,轻者导致的患者疼痛,重则最终导致角膜炎、角膜浑浊甚至失明。通过角膜移植治疗角膜大泡性水肿具有较高成功率,成为角膜大泡性水肿患者的福音。但是,由于角膜大泡性水肿患者群体的体量较大,角膜供源严重不足。
随着生物材料的发展,通过使用生物相容性较好的高分子材料,通过改变亲疏水的性质逆转角膜水肿,减少房水积累,使角膜恢复透明,达到了减轻患者疼痛的效果。然而,现有技术中的高分子材料薄膜,具有的弊端如下:
1)现有的薄膜与角膜基质的贴合性,有待提高;
2)现有的薄膜材料为了提升贴合性,会损失薄膜自身的光学特性和力学性能,最终影响手术整体效果。
因此,亟需一种在保证现有光学特性和力学性能的基础上,与角膜基质贴合性高的高分子材料薄膜。
发明内容
本发明提供一种用于治疗角膜水肿的后弹力层植入型薄膜的制备方法,以获得在保证现有光学特性和力学性能的基础上,可与角膜基质贴合性高的后弹力层植入型薄膜的技术效果。
为实现上述目的,本发明提供的一种用于治疗角膜水肿的后弹力层植入型薄膜的制备方法,方法包括:将单体分子和交联剂按照10~50:1的比例混合,在氮气环境下进行减压至0.005~0.010MPa;其中,所述单体分子为丙烯酸或者HEMA,所述交联剂为PEGDA或者EGDMA;
加入预设规格的交联引发剂混匀后,倒入设定直径的玻璃模具中,在4℃的温度条件下,过夜固化,获得后弹力层植入型薄膜;
在利用后弹力层植入型薄膜进行手术前1h,通过PVP溶液浸泡所述后弹力层植入型薄膜的一侧,实现所述后弹力层植入型薄膜的单侧亲疏水改性,以获得可折叠的后弹力层植入型薄膜。
进一步,优选的,所述加入预设规格的交联引发剂混匀后的方法,包括:
首先加入浓度为0.5%-5%(w/v)的APS混匀后;
然后加入浓度为0.01%-0.8%(v/v)的TEMED混匀。
进一步,优选的,所述PVP溶液为经过0.22μm微孔滤膜过滤的溶液。
进一步,优选的,在获得后弹力层植入型薄膜之后,还包括,
将所述后弹力层植入型薄膜在医用酒精中浸泡2h后,用生理盐水冲洗。
进一步,优选的,所述后弹力层植入型薄膜的厚度为10μm到200μm。
本发明还保护利用上述用于治疗角膜水肿的后弹力层植入型薄膜的制备方法制备的后弹力层植入型薄膜。
本发明的用于治疗角膜水肿的后弹力层植入型薄膜的制备方法,能够获得具有良好的生物相容性的植入型薄膜,可以在房水以及PBS等液体中具有良好的可塑性,折叠后经2.4mm左右的小切口植入眼内并展开;另外,植入型薄膜的亲水面与角膜基质以及后弹力层具有良好的亲和力,且疏水效果确切,可以阻挡房水进入角膜基质,可以达到实现减轻角膜水肿的技术效果。
附图说明
图1为本发明的实施例提供的后弹力层植入型薄膜的成品图;
图2为本发明提供的用于治疗角膜水肿的后弹力层植入型薄膜的制备方法的疾病模型原理示意图;
图3为本发明提供的植入型薄膜通过人工晶体推注器植入前方的状态图;
图4为本发明一实施例提供的植入型薄膜在眼内展开的场景图;
图5为通过本发明提供的植入型薄膜的贴合度效果图;
图6为本发明提供的植入型薄膜的光学透明性状态图;
图7为本发明一实施例提供的植入型薄膜在角膜基质层间植入后生物相容性效果示意图;
图8为本发明提供的植入型薄膜与对照组的阻挡房水进入角膜的效果图;
图9为本发明提供的植入型薄膜与对照组的OCT表征角膜厚度效果对比图;
本发明目的的实现、功能特点及优点将结合实施例,参照附图做进一步说明。
具体实施方式
应当理解,以下的实施例便于更好地理解本发明,但并不限定本发明。下述实施例中的实验方法,如无特殊说明,均为常规方法。下述实施例中所用的试验材料,如无特殊说明,均为自常规生化试剂商店购买得到的。以下实施例中的定量试验,均设置三次重复试验,结果取平均值。
在本发明化合物的制备说明中使用的缩写:
HEMA:甲基丙烯酸2-羟乙酯(Bimax)
PVP:聚(N-乙烯基吡咯烷酮)(ISP Ashland)
EGDMA:二甲基丙烯酸乙二醇酯(Ethylene dimethacrylate)
PEGDA:聚乙二醇(二醇)二丙烯酸酯[Poly(ethylene glycol)diacrylate]
PEGDA可以为PEGDA 250,PEGDA 575,PEGDA 4000;
HEMA:甲基丙烯酸羟乙酯单体(2-Hydroxyethyl methacrylate)
APS:过硫酸铵(Ammonium persulphate)
TEMED:四甲基乙二胺(N,N,N',N'-Tetramethylethylenediamine)
OTS:十八烷基三氯硅烷(Octadecyltrichlorosilane)
实施例1
盖玻片预处理:将直径为25mm的盖玻片使用OTS混合液进行浸泡30分钟后,在60℃下进行烘干;其中,OTS混合液为10%的OTS与环己烷溶液的混合溶液;
后弹力层植入型薄膜制备:将HEMA和EGDMA按照10:1的比例混合,通入氮气30min,在氮气环境下进行减压至0.005MPa;加入浓度为0.5%(w/v)的APS混匀;加入浓度为0.8%(v/v)的TEMED混匀后获得50μL预混液,倒入预处理后的盖玻片中,在4℃下,过夜固化,获得厚度为60μm的后弹力层植入型薄膜;将所述后弹力层植入型薄膜在医用酒精中浸泡2h后,用生理盐水冲洗。在利用后弹力层植入型薄膜进行手术前1h,将PVP溶液经过0.22μm微孔滤膜过滤后,利用1%的PVP溶液浸泡所述后弹力层植入型薄膜的一侧,实现所述后弹力层植入型薄膜的单侧亲疏水改性,以获得可折叠的后弹力层植入型薄膜。
在具体的实施过程中,可以根据实际需求,在盖玻片中加入不同体积的预混液最终得到不同厚度的后弹力层植入型薄膜。以直径为25mm的盖玻片为例,若加入8.3μL预混液,则可获得厚度为10μm的后弹力层植入型薄膜;若加入167μL预混液,则可获得厚度为200μm的后弹力层植入型薄膜。
图1为本发明的实施例1提供的后弹力层植入型薄膜的成品图;根据图1所示,后弹力层植入型薄膜具有较好的可折叠性。
实施例2
模具预处理:将模具使用OTS混合液进行浸泡30分钟后,在60℃下进行烘干;其中,OTS混合液为10%的OTS与环己烷溶液的混合溶液;模具为使用AutoCAD设计3d打印模具,打印模具的材料可以选择树脂,金属,玻璃等。在本实施例中,打印模具的材料为玻璃。
后弹力层植入型薄膜制备:将丙烯酸和EGDMA按照50:1的比例混合,在氮气环境下进行减压至0.010MPa;加入浓度为5%(w/v)的APS混匀;加入浓度为0.01%(v/v)的TEMED混匀后获得预混液100μL,倒入直径为25μm的模具中,在4℃下,过夜固化,获得厚度为120μm的后弹力层植入型薄膜;将所述后弹力层植入型薄膜在医用酒精中浸泡2h后,用生理盐水冲洗。在利用后弹力层植入型薄膜进行手术前1h,所述PVP溶液为经过0.22μm微孔滤膜过滤的溶液,通过PVP溶液浸泡所述后弹力层植入型薄膜的一侧,实现所述后弹力层植入型薄膜的单侧亲疏水改性,以获得可折叠的后弹力层植入型薄膜。
实施例3
盖玻片预处理:将直径为25mm盖玻片使用OTS混合液进行浸泡30分钟后,在60℃下进行烘干;其中,OTS混合液为10%的OTS与环己烷溶液的混合溶液;
后弹力层植入型薄膜制备:将HEMA和PEGDA按照30:1的比例混合,通入氮气30min在氮气环境下进行减压至0.008MPa;加入浓度为0.2%(w/v)的APS混匀;加入浓度为0.3%(v/v)的TEMED混匀后获得预混液25μL,倒入设定直径的玻璃模具中,在4℃下,过夜固化,获得厚度为30μm后弹力层植入型薄膜;将所述后弹力层植入型薄膜在医用酒精中浸泡2h后,用生理盐水冲洗。在利用后弹力层植入型薄膜进行手术前1h,所述PVP溶液为经过0.22μm微孔滤膜过滤的溶液,通过PVP溶液浸泡所述后弹力层植入型薄膜的一侧,实现所述后弹力层植入型薄膜的单侧亲疏水改性,以获得可折叠的后弹力层植入型薄膜。
实验例
图2为本发明提供的用于治疗角膜水肿的后弹力层植入型薄膜的制备方法的疾病模型原理示意图;如图2所示,要对实验小兔进行麻醉后,对兔角膜切口,使用角膜内皮刮除器刮去角膜内皮细胞;然后,将折叠后的植入型薄膜推注进入前房,进行缝合。
具体步骤如下:
将实验动物小兔全身麻醉(赛拉嗪+咪达唑仑1.2mL);麻醉效果满意后,常规碘伏消毒眼部,使用开睑器开眼睑,再使用奥布卡因滴眼液表面麻醉,妥布霉素生理盐水冲洗结膜囊;2.4mm角膜穿刺刀经角膜缘做穿刺口,肾上腺素使用5iu/mL肝素+BSS水(平衡盐溶液)冲洗前房以减少渗出;注入黏弹剂撑起前房,使用自制角膜内皮刮除器械(siliconneedle)刮除中央8mm内皮;手动注吸黏弹剂,0.04%台盼蓝染色确定刮除范围;取一可折叠的后弹力层植入型薄膜,折叠后放入推注器中,经2.4mm角膜切口推入前房,虹膜恢复器辅助展开植片后缝合主切口;卡巴胆碱收缩瞳孔至正常大小并形成前房。
图3为本发明提供的植入型薄膜通过人工晶体推注器植入前方的状态图;如图3所示,植入型薄膜具有良好的可折叠性,可以通过人工晶体推注器植入前房。
图4为本发明提供的植入型薄膜在眼内展开的场景图;如图4所示,植入型薄膜具有可塑性强,在眼内易于展开的特点。
随后注入过滤20%SF6顶压植片,泪道冲洗针调整植片位置至角膜中央而后保持动物术眼朝上体位90-120分钟,保证植片贴附。
术后每天以妥布霉素地塞米松滴眼液点眼3次,持续14d。第1,4,7d,观察术眼,第1d时前房由于气体存在中央未发生明显水肿,周边角膜轻度水肿;4d时周边角膜水肿,中央角膜植片区域角膜透明;7d时,周边角膜内皮移行,周边角膜水肿逐渐减轻;14d时,角膜完全透明,植片完全贴合与于角膜后弹力层。
图5~图9对本发明的植入型薄膜的实验效果进行了整体说明;其中,图5为通过本发明提供的植入型薄膜的贴合度效果图;图6为本发明提供的植入型薄膜的光学透明性状态图;图7为本发明一实施例提供的植入型薄膜在角膜基质层间植入后生物相容性效果示意图;图8为本发明提供的植入型薄膜与对照组的OCT表征角膜厚度效果对比图;图9为本发明提供的植入型薄膜与对照组的阻挡房水进入角膜的效果图。
通过图5和图6所示,本发明的植入型薄膜具有与角膜具有较好的相容性和透光性。通过图7所示,本发明的植入型薄膜与后弹力层贴合良好。通过图8所示,本发明的植入型薄膜与对照组相比,本发明的后弹力层植入型薄膜阻挡房水进入角膜基质的效果确切。通过图9所示,本发明的植入型薄膜移植后的OCT表征角膜厚度与对照组的OCT表征角膜厚度相比,具有明显优势。
最后应说明的是,以上实施例仅用以说明本发明的技术方案而非限制,尽管参照较佳实施例对本发明进行了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或等同替换,而不脱离本发明技术方案的精神和范围。

Claims (5)

1.一种用于治疗角膜水肿的后弹力层植入型薄膜的制备方法,其特征在于,方法包括:
将单体分子和交联剂按照10~50:1的比例混合后,在氮气环境下进行减压至0.005~0.010MPa;其中,所述单体分子为丙烯酸或者HEMA,所述交联剂为PEGDA或者EGDMA;当所述单体分子为HEMA时,所述交联剂为PEGDA或者EGDMA;当所述单体分子为丙烯酸时,所述交联剂为EGDMA;
加入预设规格的交联引发剂混匀后,倒入设定直径的玻璃模具中,在4℃的温度条件下,过夜固化,获得后弹力层植入型薄膜;其中,所述加入预设规格的交联引发剂混匀后的方法,包括:首先加入浓度为0.5%-5%(w/v)的APS混匀后;然后加入浓度为0.01%-0.8%(v/v)的TEMED混匀;
在利用后弹力层植入型薄膜进行手术前1h,通过PVP溶液浸泡所述后弹力层植入型薄膜的一侧,实现所述后弹力层植入型薄膜的单侧亲疏水改性,以获得可折叠的后弹力层植入型薄膜。
2.根据权利要求1所述的用于治疗角膜水肿的后弹力层植入型薄膜的制备方法,其特征在于,
所述PVP溶液为经过 0.22μm微孔滤膜过滤的溶液。
3.根据权利要求1所述的用于治疗角膜水肿的后弹力层植入型薄膜的制备方法,其特征在于,
在获得后弹力层植入型薄膜之后,还包括,
将所述后弹力层植入型薄膜在医用酒精中浸泡2h后,用生理盐水冲洗。
4.根据权利要求3所述的用于治疗角膜水肿的后弹力层植入型薄膜的制备方法,其特征在于,所述后弹力层植入型薄膜的厚度为10μm到200μm。
5.一种后弹力层植入型薄膜,其特征在于,利用所述权利要求1~4中任一项所述的用于治疗角膜水肿的后弹力层植入型薄膜的制备方法进行制备。
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