[1]李涛,廖灵珊,朱胜兰,等.丝裂霉素C-全氟溴辛烷脂质体纳米药物对人翼状胬肉成纤维细胞治疗安全性及效果评价[J].眼科新进展,2024,44(2):100-105.[doi:10.13389/j.cnki.rao.2024.0020]
　LI Tao,LIAO Lingshan,ZHU Shenglan,et al.Evaluation of the safety and efficacy of mitomycin C-perfluorooctyl bromide liposome nanoparticles in the treatment of human pterygium fibroblasts[J].Recent Advances in Ophthalmology,2024,44(2):100-105.[doi:10.13389/j.cnki.rao.2024.0020]
点击复制

丝裂霉素C-全氟溴辛烷脂质体纳米药物对人翼状胬肉成纤维细胞治疗安全性及效果评价/HTML
分享到：

《眼科新进展》[ISSN:1003-5141/CN:41-1105/R]

卷:
44卷
期数:
2024年2期
页码:
100-105
栏目:
实验研究
出版日期:
2024-01-26

文章信息/Info

Title:
Evaluation of the safety and efficacy of mitomycin C-perfluorooctyl bromide liposome nanoparticles in the treatment of human pterygium fibroblasts
作者:
李涛; 廖灵珊; 朱胜兰; 唐娟; 吴小利; 方其林; 李盈; 李彪; 田琴; 万俊梅; 阳毅; 谭越月; 李佳倩; 杜鹃; 周燕; 张丹; 刘兴德
641300　四川省资阳市，资阳市第一人民医院眼科（李涛，吴小利，方其林，李盈，李彪，田琴，万俊梅，阳毅，谭越月，李佳倩，杜鹃，周燕，张丹，刘兴德）；641300　四川省资阳市，资阳第一人民医院眼科重点实验室（李涛，吴小利，方其林，李盈，李彪，田琴，万俊梅，阳毅，谭越月，李佳倩，杜鹃，周燕，张丹，刘兴德）；400030　重庆市，重庆大学附属肿瘤医院肿瘤放射治疗中心（廖灵珊）；641300　四川省资阳市，资阳口腔职业学院基础教学部（朱胜兰）；641300　四川省资阳市，资阳市第一人民医院内分泌科（唐娟）
Author(s):
LI Tao1; 2; LIAO Lingshan3; ZHU Shenglan4; TANG Juan5; WU Xiaoli1; 2; FANG Qilin1; 2; LI Ying1; 2; LI Biao1; 2; TIAN Qin1; 2; WAN Junmei1; 2; YANG Yi1; 2; TAN Yueyue1; 2; LI Jiaqian1; 2; DU Juan1; 2; ZHOU Yan1; 2; ZHANG Dan1; 2; LIU Xingde1; 2
1.Department of Ophthalmology,the First People’s Hospital of Ziyang
2.Key Laboratory of Ophthalmology,the First People’s Hospital of Ziyang
3.Department of Tumor Radiotherapy Center,Chongqing University Cancer Hospital
4.Department of Basic Teaching Department,Ziyang College of Dental Technology
5.Department of Endocrinology,the First People’s Hospital of Ziyang
关键词:
翼状胬肉; 丝裂霉素C; 脂质体; 纳米医学; 抗炎
Keywords:
pterygium;  mitomycin C;  liposome;  nanomedicine;  anti-inflammation
分类号:
R777.33
DOI:
10.13389/j.cnki.rao.2024.0020
文献标志码:
A
摘要:
目的　制备一种以脂质体为载体，以液态的全氟溴辛烷（PFOB）为内核，丝裂霉素C（MMC）搭载于脂质体壳的纳米药物PFOB@Lip-MMC，研究其对人翼状胬肉成纤维细胞（HPF）增殖的抑制作用。
方法　采用薄膜分散-水化超声法制备PFOB@Lip-MMC，并检测其物理、化学性质。通过CCK-8、Cam-PI细胞活/死染色、流式细胞术等方法检测不同浓度PFOB@Lip-MMC对HPF活性的影响。在激光共聚焦显微镜下采用DiI荧光标记的PFOB@Lip-MMC观察纳米药物对于HPF的药物渗透性。建立HPF炎症细胞模型后，按照实验要求分为对照组（加入无菌PBS溶液）、PFOB@Lip组（加入PFOB@Lip）、MMC组（加入MMC）、PFOB@Lip-MMC组（加入PFOB@Lip-MMC）、正常组（加入新鲜培养基），共孵育24 h后，流式细胞仪检测炎症细胞的凋亡率以及从PCR角度分析细胞中白细胞介素（IL）-1β、前裂腺素E2（PGE2）、肿瘤坏死因子（TNF）-α及血管内皮生长因子（VEGF）基因表达水平。
结果　PFOB@Lip-MMC平均粒径和Zeta电位分别为（103.45±2.17）nm和（27.34±1.03）mV，其包封率和载药率分别为（72.85±3.28）%和（34.27±2.04）%。体外24 h眼表环境下载药纳米微球缓释MMC可达（78.34±2.92）%。PFOB@Lip-MMC的生物安全性较MMC明显提高。DiI荧光标记的PFOB@Lip-MMC与炎症化HPF共同孵育2 h后，DiI荧光标记弥漫性分布于炎症化HPF中。PFOB@Lip-MMC组炎症化HPF细胞凋亡率［（77.23±4.93）%］明显高于MMC组［（51.62±3.28）%］；PCR检查结果显示，其余各组IL-1β、PGE2、TNF-α及VEGF基因基因转录水平与对照组和PFOB@Lip组相比均明显下降，其中PFOB@Lip-MMC组降低最明显，差异均有统计学意义（均为P<0.05）。
结论　本研究成功合成了一种新型的抗HPF增殖的纳米药物PFOB@Lip-MMC，且细胞毒性较原药明显降低，具有良好的生物相容性和抗炎效果，为降低翼状胬肉术后复发率提供了一种新的治疗思路。
Abstract:
Objective　To prepare a nano drug (PFOB@ Lip-MMC) with liposome as the carrier, liquid perfluorooctyl bromide (PFOB) as core and mitomycin C (MMC) loading on the liposome shell and study its inhibitory effect on the proliferation of human pterygium fibroblasts (HPFs).
Methods　The thin film dispersion-hydration ultrasonic method was used to prepare PFOB@ Lip-MMC and detect its physical and chemical properties. Cell Counting Kit-8, Cam-PI cell viability staining and flow cytometry were employed to detect the impact of different concentrations of PFOB@ Lip-MMC on the viability of HPFs. DiI fluorescence labeled PFOB@ Lip-MMC was used to observe the permeability of the nano drug to HPFs under a laser confocal microscope. After establishing HPF inflammatory cell models, they were divided into the control group (with sterile phosphate-buffered saline solution added), PFOB@ Lip group (with PFOB@ Lip added), MMC group (with MMC added), PFOB@ Lip-MMC group (with PFOB@ Lip-MMC added) and normal group (with fresh culture medium added) according to the experimental requirements. After co-incubation for 24 h, flow cytometer was used to detect the apoptosis rate of inflammatory cells, and the gene expression levels of interleukin (IL)-1β, prostaglandin E2 (PGE2), tumor necrosis factor (TNF)-α and vascular endothelial growth factor (VEGF) in cells were analyzed by PCR.
Results　The average particle size and Zeta potential of PFOB@ Lip-MMC were (103.45±2.17)nm and (27.34±1.03)mV, respectively, and its entrapped efficiency and drug loading rate were (72.85±3.28)% and (34.27±2.04)%, respectively. The sustained-release MMC of drug-loaded nanospheres reached (78.34±2.92)% in vitro in a 24-hour ocular surface environment. The biological safety of PFOB@ Lip-MMC significantly improved compared to MMC. In terms of the DiI fluorescence labeled PFOB@ Lip-MMC, after co-incubation with inflammatory HPFs for 2 h, DiI fluorescence labeling was diffusely distributed in the cytoplasm of inflammatory HPFs. The apoptosis rate of inflammatory HPFs in the PFOB@ Lip-MMC group ［(77.23±4.93)%］ was significantly higher than that in the MMC group ［(51.62±3.28)%］. The PCR examination results showed that the gene transcription levels of IL-1β, PGE2, TNF-α and VEGF in other groups were significantly reduced compared to the control group and PFOB@ Lip group, with the most significant decrease in the PFOB@ Lip-MMC group (all P<0.05).
Conclusion　In this study, a novel nano drug (PFOB@ LIP-MMC) that inhibited the proliferation of HPFs was successfully synthesized, and its cytotoxicity was significantly reduced compared to the original drugs. It has good biocompatibility and anti-inflammatory effects, providing a new treatment approach for reducing the recurrence rate after pterygium surgery.

参考文献/References:

［1］　CHALKIA A K,SPANDIDOS D A,DETORAKIS E T.Viral involvement in the pathogenesis and clinical features of ophthalmic pterygium［J］.Int J Mol Med,2013,32(3):539-543.
［2］　CHUI J,CORONEO M T,TAT L T,CROUCH R,WAKEFIELD D,DI GIROLAMO N.Ophthalmic pterygium:a stem cell disorder with premalignant features［J］.Am J Pathol,2011,178(2):817-827.
［3］　YUE X L,GAO Z Q.Identification of pathogenic genes of pterygium based on the Gene Expression Omnibus database［J］.Int J Ophthalmol,2019,12(4):529-535.
［4］　WANG Y C,ZHAO F K,LIU Q,YU Z Y,WANG J,ZHANG J S.Bibliometric analysis and mapping knowledge domain of pterygium:2000-2019［J］.Int J Ophthalmol,2021,14(6):903-914.
［5］　ZHENG B,LIU Y,HE K,WU M,JIN L,JIANG Q,et al.Research on an intelligent lightweight-assisted pterygium diagnosis model based on anterior segment images［J］.Dis Markers,2021,2021:7651462.
［6］　VAN ACKER S I,VAN DEN BOGERD B,HAAGDORENS M,SIOZOPOULOU V,Ní DHUBHGHAILL S,PINTELON I,et al.Pterygium-the good,the bad,and the ugly［J］.Cells,2021,10(7):1567.
［7］　DADEYA S,FATIMA S.Corneoscleral perforation after pterygium excision and intraoperative mitomycin C［J］.Ophthalmic Surg Lasers Imaging,2003,34(2):146-148.
［8］　MANNING C A,KLOESS P M,DIAZ M D,YEE R W.Intraoperative mitomycin in primary pterygium excision.A prospective,randomized trial［J］.Ophthalmology,1997,104(5):844-848.
［9］　BAHAR I,KAISERMAN I,LANGE A P,SLOMOVIC A,LEVINGER E,SANSANAYUDH W,et al.The effect of mitomycin C on corneal endothelium in pterygium surgery［J］.Am J Ophthalmol,2009,147(3):447-452.
［10］　GAO Y,WU Y.Recent advances of chitosan-based nanoparticles for biomedical and biotechnological applications［J］.Int J Biol Macromol,2022,203:379-388.
［11］　KULKARNI A D,PATEL H M,SURANA S J,VANJARI Y H,BELGAMWAR V S,PARDESHI C V.N,N,N-Trimethyl chitosan:An advanced polymer with myriad of opportunities in nanomedicine［J］.Carbohydr Polym,2017,157:875-902.
［12］　RATHOD S,DESHPANDE S G.Design and evaluation of liposomal formulation of pilocarpine nitrate［J］.Indian J Pharm Sci,2010,72(2):155-160.
［13］　STAFFEY K S,DENDI R,BROOKS L A,PRETORIUS A M,ACKERMANN L W,ZAMBA K D,et al.Liquid ventilation with perfluorocarbons facilitates resumption of spontaneous circulation in a swine cardiac arrest model［J］.Resuscitation,2008,78(1):77-84.
［14］　安美霞,吴开力,林少春.初发和复发翼状胬肉成纤维细胞分泌基质金属蛋白酶的比较及汉防己甲素对其的影响［J］.眼科新进展,2011,31(8):708-710．
AN M X,WU K L,LIN S C.Comparison of matrix metalloproteinase secreted by primary and recurrent pterygium fibroblasts and inhibitory effect of tetrandrine［J］.Rec Adv Ophthalmol,2011,31(8):708-710．
［15］　HAN S,CHEN Y,GAO Y,SUN B,KONG Y.MicroRNA-218-5p inhibit the migration and proliferation of pterygium epithelial cells by targeting EGFR via PI3K/Akt/mTOR signaling pathway［J］.Exp Eye Res,2019,178:37-45.
［16］　HUANG Y,HE H,SHEHA H,TSENG S C.Ocular demodicosis as a risk factor of pterygium recurrence.Ophthalmology［J］.2013,120(7):1341-1347.
［17］　YUE X L,GAO Z Q.Identification of pathogenic genes of pterygium based on the Gene Expression Omnibus database［J］.Int J Ophthalmol,2019,12(4):529-535.
［18］　WADHWA S,PALIWAL R,PALIWAL S R,VYAS S P.Hyaluronic acid modified chitosan nanoparticles for effective management of glaucoma:development,characterization,and evaluation［J］.J Drug Target,2010,18(4):292-302.
［19］　NAM H Y,KWON S M,CHUNG H,LEE S Y,KWON S H,JEON H,et al.Cellular uptake mechanism and intracellular fate of hydrophobically modified glycol chitosan nanoparticles［J］.J Control Release,2009,135(3):259-267.
［20］　KUMAR S,DUTTA J,DUTTA P K,KOH J.A systematic study on chitosan-liposome based systems for biomedical applications［J］.Int J Biol Macromol,2020,160:470-481.
［21］　GHOLIZADEH S,WANG Z,CHEN X,DANA R,ANNABI N.Advanced nanodelivery platforms for topical ophthalmic drug delivery［J］.Drug Discov Today,2021,26(6):1437-1449.

相似文献/References:

[1]吴恺 刘二华 谭钢 邵毅.雷公藤多甙对体外培养的人翼状胬肉上皮鳞状化生的影响[J].眼科新进展,2012,32(6):000.
[2]叶芬 田农 薛春燕 施宇华 曹茜 黄振平.翼状胬肉面积与术后残余像差之间的关系[J].眼科新进展,2013,33(1):000.
[3]贾烨 赖兆光.翼状胬肉切除术式对术后眼表修复的影响[J].眼科新进展,2013,33(1):000.
[4]李新宇 王娟 徐玲娟 胡军 胡维琨 李贵刚.绷带型角膜接触镜在翼状胬肉手术中的应用[J].眼科新进展,2013,33(8):000.
[5]李景翠 张雪菲.组织因子在翼状胬肉中的表达[J].眼科新进展,2012,32(2):000.
[6]王淑华 王春艳.Ahmed青光眼阀植入术治疗难治性青光眼的疗效观察[J].眼科新进展,2012,32(10):000.
[7]彭立 谢青 蔡枫 李爱国.同步辐射微束X射线检测翼状胬肉实验条件的探讨[J].眼科新进展,2013,33(11):000.
[8]胡翠霞 刘伟 张在芬.改良复合式小梁切除术治疗原发性青光眼的临床观察[J].眼科新进展,2013,33(11):000.
[9]石一宁,张纯涛.TGF-β1、TGF-βRⅡ及Smad4蛋白在原发性翼状胬肉中的表达[J].眼科新进展,2010,30(5):000.
[10]黄 焱 杨 静 张 娣 马惠杰 廖 凯沈 平 袁 安.自体角膜缘干细胞移植与常规翼状胬肉切除术疗效比较[J].眼科新进展,2009,29(1):000.

备注/Memo

备注/Memo:
四川省资阳市医学科学课题计划项目（编号：KY2023001，KY2023022）；四川省资阳市科学技术局计划项目（编号：zykjjsc20-yyjc-2023-04）；河北省医学科学研究课题计划（编号：20240294）
更新日期/Last Update: 2024-02-05