组织工程与重建外科杂志
組織工程與重建外科雜誌
조직공정여중건외과잡지
JOURNAL OF TISSUE ENGINEERING AND RECONSTRUCTIVE SURGERY
2014年
2期
77-81
,共5页
张铮%沃雁%张振%毛小慧%苏薇洁%濮哲铭%张艳%章一新
張錚%沃雁%張振%毛小慧%囌薇潔%濮哲銘%張豔%章一新
장쟁%옥안%장진%모소혜%소미길%복철명%장염%장일신
醇脂体5-氟尿嘧啶%增生性瘢痕%成纤维细胞%细胞内递送
醇脂體5-氟尿嘧啶%增生性瘢痕%成纖維細胞%細胞內遞送
순지체5-불뇨밀정%증생성반흔%성섬유세포%세포내체송
Ethosomes%5-fluorouracil%Hypertrophic scar%Fibroblast%Intracellular delivery
目的:构建纳米级载5-氟尿嘧啶醇脂体(5-FU ES),观察其对瘢痕成纤维细胞的抑制作用。方法采用Touitou法制备5-FU ES悬液并进行质量评价。体外培养增生性瘢痕成纤维细胞,荧光素RhoB标记醇脂体,进行体外透细胞实验,以水醇溶液为对照。激光共聚焦显微镜(CLSM)观察不同作用时间后细胞内荧光分布和强度。CCK-8检测不同浓度5-FU ES对成纤维细胞活性的影响,计算载5-FU ES对成纤维细胞活性的半数抑制浓度(IC50)。CCK-8检测醇脂体对瘢痕成纤维细胞生长的影响。结果制备所得5-FU ES粒径为(87.5±5.4) nm,分散系数为0.151±0.27,包封率为(10.03±2.12)%。 CLSM图像显示,醇脂体促进RhoB进入细胞内,可见荧光分布和强度均较水醇溶液组增多增强;经Image-Pro Plus 6.0图像分析软件计算单位面积光密度值,RhoB标记醇脂体组(Rho ES)细胞内荧光光密度显著高于水醇溶液组(Rho HA)(P<0.01)。随着5-FU浓度增加,5-FU HA 和5-FU ES对成纤维细胞的生长抑制率逐渐增强;5-FU ES和5-FU HA对成纤维细胞的半数抑制浓度(IC50)分别为(9.2582±1.2329)μg/mL和(18.0352±2.3145)μg/mL,差异具有统计学意义(P<0.05);同时,醇脂体本身对细胞活性无明显抑制作用。结论醇脂体可有效携带5-FU向瘢痕成纤维细胞内递送,促进5-FU对成纤维细胞的活性抑制作用。醇脂体作为经皮给药载体,是一种安全有效的透细胞药物载体。
目的:構建納米級載5-氟尿嘧啶醇脂體(5-FU ES),觀察其對瘢痕成纖維細胞的抑製作用。方法採用Touitou法製備5-FU ES懸液併進行質量評價。體外培養增生性瘢痕成纖維細胞,熒光素RhoB標記醇脂體,進行體外透細胞實驗,以水醇溶液為對照。激光共聚焦顯微鏡(CLSM)觀察不同作用時間後細胞內熒光分佈和彊度。CCK-8檢測不同濃度5-FU ES對成纖維細胞活性的影響,計算載5-FU ES對成纖維細胞活性的半數抑製濃度(IC50)。CCK-8檢測醇脂體對瘢痕成纖維細胞生長的影響。結果製備所得5-FU ES粒徑為(87.5±5.4) nm,分散繫數為0.151±0.27,包封率為(10.03±2.12)%。 CLSM圖像顯示,醇脂體促進RhoB進入細胞內,可見熒光分佈和彊度均較水醇溶液組增多增彊;經Image-Pro Plus 6.0圖像分析軟件計算單位麵積光密度值,RhoB標記醇脂體組(Rho ES)細胞內熒光光密度顯著高于水醇溶液組(Rho HA)(P<0.01)。隨著5-FU濃度增加,5-FU HA 和5-FU ES對成纖維細胞的生長抑製率逐漸增彊;5-FU ES和5-FU HA對成纖維細胞的半數抑製濃度(IC50)分彆為(9.2582±1.2329)μg/mL和(18.0352±2.3145)μg/mL,差異具有統計學意義(P<0.05);同時,醇脂體本身對細胞活性無明顯抑製作用。結論醇脂體可有效攜帶5-FU嚮瘢痕成纖維細胞內遞送,促進5-FU對成纖維細胞的活性抑製作用。醇脂體作為經皮給藥載體,是一種安全有效的透細胞藥物載體。
목적:구건납미급재5-불뇨밀정순지체(5-FU ES),관찰기대반흔성섬유세포적억제작용。방법채용Touitou법제비5-FU ES현액병진행질량평개。체외배양증생성반흔성섬유세포,형광소RhoB표기순지체,진행체외투세포실험,이수순용액위대조。격광공취초현미경(CLSM)관찰불동작용시간후세포내형광분포화강도。CCK-8검측불동농도5-FU ES대성섬유세포활성적영향,계산재5-FU ES대성섬유세포활성적반수억제농도(IC50)。CCK-8검측순지체대반흔성섬유세포생장적영향。결과제비소득5-FU ES립경위(87.5±5.4) nm,분산계수위0.151±0.27,포봉솔위(10.03±2.12)%。 CLSM도상현시,순지체촉진RhoB진입세포내,가견형광분포화강도균교수순용액조증다증강;경Image-Pro Plus 6.0도상분석연건계산단위면적광밀도치,RhoB표기순지체조(Rho ES)세포내형광광밀도현저고우수순용액조(Rho HA)(P<0.01)。수착5-FU농도증가,5-FU HA 화5-FU ES대성섬유세포적생장억제솔축점증강;5-FU ES화5-FU HA대성섬유세포적반수억제농도(IC50)분별위(9.2582±1.2329)μg/mL화(18.0352±2.3145)μg/mL,차이구유통계학의의(P<0.05);동시,순지체본신대세포활성무명현억제작용。결론순지체가유효휴대5-FU향반흔성섬유세포내체송,촉진5-FU대성섬유세포적활성억제작용。순지체작위경피급약재체,시일충안전유효적투세포약물재체。
Objective To establish ethosomes encapsulated with 5-Fluorouracil and to explore the inhibitory effects of 5-FU on human hypertrophic scar fibroblasts. Methods Ethosomes encapsulated with 5-Fluorouracil was prepared by Touitou method and was evaluated. Human hypertrophic scar fibroblasts were cultured in vitro, the penetration of the fluorescent probes into fibroblasts was examined by CLSM, and hydroalcoholic solution was set as control group. The effect of ethosomes encapsulating different concentration of 5-FU on fibroblast viability was tested by cell counting kit-8 (CCK-8), and the median inhibitory concentration (IC50) was calculated. The effect of ethosomes on fibroblast viability was tested by CCK-8. Results The particle size of 5-FU encapsulated ethosomes was (87.5 ±5.4) nm, the polydispersion index was 0.151 ±0.27, and the encapsulation efficiency of 5-FU was (10.03 ±2.12)%. CLSM micrographs showed that ethosomes facilitated the penetration of RhoB into the cell, as evident from the high intensity fluorescence. In comparison, when incorporated in hydroalcoholic solution, very weak fluorescence was detected. The optical density (OD) of unit area was calculated through the Image-Pro Plus 6.0 image analysis software, and the OD value of ethosomal group was distinguished higher than hydroalcoholic group (P<0.01). With the increase of the concentration of 5-FU, growth inhibition rate of fibroblast affected by 5-FU HA and 5-FU ES was gradually enhanced. The IC50 of 5-FU encapsulated ethosomes and 5-FU hydroalcoholic solution to fibroblasts were (9.2582±1.2329) μg/mL and (18.0352±2.3145) μg/mL, respectively, the difference between the two groups was distinguished (P<0.05). Meanwhile, ethosomal carrier was not toxic to the cultured cells. Conclusion Ethosomes can effectively enhance intracellular delivery of 5-FU, as a result can enhance the inhibitory effect of 5-FU on fibroblast viability. Meanwhile, ethosomal carrier was not toxic to the cultured cells. Ethosomes as percutaneous drug delivery carriers, are also a promising candidate for the delivery of biological and chemical compounds to cultured cells.