人表皮细胞生长因子脂质体制备及促大鼠烫伤修复研究

Preparation of Recombinant Human Epidermal Growth Factor Liposomes and the Effects in Burned Rats

目的:制备重组人表皮细胞生长因子(rhEGF)脂质体,并考察其促大鼠烫伤创面愈合的作用。方法:采用pH梯度法制备rhEGF脂质体;超滤-离心法分离rhEGF脂质体混悬液中的游离rhEGF,ELISA法测定rhEGF含量,计算脂质体包封率;采用透射电镜观察脂质体的外观形态;采用纳米粒度及Zeta电位分析仪分别测定脂质体的粒径和Zeta电位;以大鼠烫伤模型观察给药后各试验组创面愈合过程中的形态、愈合时间和愈合率的变化。结果:制备的rhEGF脂质体包封率为57.7±1.1%;脂质体形状较为规则,呈完整圆球形或椭圆形的单室囊泡;脂质体粒度分布均匀,呈正态分布,平均粒径为63.7 nm;脂质体的Zeta电位为+9.2mV,带正电荷;rhEGF脂质体高、中剂量组能显著性促进大鼠烫伤创面愈合,使创面愈合时间明显提前,低剂量组促烫伤修复效应不明显。结论:pH梯度法制备的rhEGF脂质体包封率较高,rhEGF脂质体对大鼠烫伤创面的愈合有明显促进作用。

Objective： To prepare recombinant human epidermal growth factor （rhEGF） liposome, and research the effects in burned rats. Methods： The rhEGF liposomes were prepared by pH-gradient, rhEGF was separated from the liposome suspention with ultrafiltration. The encapsulation efficiency was determined by ELISA. The morphological examination of rhEGF liposomes was performed using transmission electron microscopy. The particle size and Zeta potential of the liposomes were measured. The changes of wound appearance, wound healing time and wound healing rates were observed. Results： The encapsulation efficiency of optimal formulation was 57.72± 1.1%. The mean value of particle size and Zeta potential of the rhEGF liposomes were 63.7 nm, ＋9.2 mV. The wound appearance results indicated that the high dosage group, middle dosage group of rhEGF liposomes could significantly promote the wound healing, and low dosage group was not obvious. Conclusion： The rhEGF liposomes were prepared by pH-gradient had high encapsulation efficiency, and the rhEGF lioposomes visibly promoting the rats＇bum wound healing.