【摘要】 为提高重组人源Ⅲ型胶原蛋白（RHC）经皮递送性能，采用大豆卵磷脂（简称磷脂）、甘油、吐温80构建了甘油体，然后对RHC进行包封，制备了包载RHC的甘油体（RHC甘油体），并建立了Folch结合体积排阻色谱法（Folch-HPLC法）用于检测RHC包封率。通过多角度粒度及高灵敏Zeta电位分析仪，考察了甘油和吐温80质量浓度对RHC甘油体的粒径和多分散性指数（PDI）的影响。通过荧光探针探究了甘油、吐温80质量浓度对RHC甘油体微黏度的影响及其稳定机理。结合激光共聚焦显微镜（CLSM）测试，评价了RHC甘油体的透皮性能。采用鸡胚绒毛尿囊膜实验评估了RHC甘油体的刺激性。结果表明，Folch-HPLC法能够完全去除磷脂的干扰，有效保留RHC，且RHC的提取回收率为100.44%±0.84%。当磷脂质量浓度150 g/L、RHC质量浓度2 g/L、甘油质量浓度250 g/L、吐温80质量浓度25 g/L时，制备的RHC甘油体为黄色澄清透明液体，稳定性最好，平均粒径为(92.34±1.22) nm，多分散性指数为0.233±0.01，包封率为12.78%±0.13%。与RHC的磷酸盐缓冲液相比，RHC甘油体具有更强的皮肤渗透性，能够有效促进RHC的经皮递送，并表现出对鸡胚绒毛尿囊膜的无刺激性。更多还原

【Abstract】 To enhance the transdermal delivery performance of recombinant human collagen Ⅲ(RHC), glycerosomes were constructed from soybean lecithin(for short lecithin), glycerol and Tween 80, and subsequently used for encapsulation of RHC to form RHC-loaded glycerosomes(RHC glycerosomes). A Folch-based high-performance liquid chromatography method(Folch-HPLC method) combining the Folch extraction protocol with size exclusion chromatography was established to measure the RHC encapsulation efficiency. The effects of glycerol and Tween 80 mass concentrations on the particle size and polydispersity index(PDI) of RHC glycerosomes were investigated by multi-angle particle size and high sensitivity Zeta potential analyzer, while the influence of glycerol and Tween 80 mass concentration on the micro-viscosity and stabilization mechanism of RHC glycerosomes was explored via fluorescence probe analysis. The transdermal delivery capability of RHC glycerosomes was evaluated by confocal laser scanning microscopy(CLSM), with its irritancy assessed by chorioallantoic membrane assay in chicken embryos. The results demonstrated that the Folch-HPLC method effectively removed interference from lecithin while retained the RHC, achieving an RHC recovery rate of 100.44%±0.84%. Under the conditions of lecithin mass concentration 150 g/L, RHC mass concentration 2 g/L, glycerol mass concentration 250 g/L, and Tween 80 mass concentration 25 g/L, the prepared RHC glycerosomes appeared as a yellow, clear, and transparent solution, showed the best stability, displayed a particle size of(92.34±1.22) nm, a PDI of 0.233±0.01 and an encapsulation efficiency of 12.78%±0.13%. Compared with RHC solution(phosphate buffer solution as solvent), RHC glycerosomes exhibited significantly enhanced skin penetration, effectively promoted the transdermal delivery of RHC, and showed no irritation in chorioallantoic membrane.更多还原