重组人源Ⅲ型胶原蛋白乳膏剂的制备及其促创伤愈合药效学评价

Preparation of Rhc-Ⅲ Cream and its Pharmacodynamic Evaluation of Promoting Wound Healing

以O/W型软膏基质作为载体,制备Rhc-Ⅲ(Recombinant human collagenⅢ,简称Rhc-Ⅲ)乳膏剂并对其质量和促创伤愈合作用进行评价。采用乳化法制备Rhc-Ⅲ乳膏剂,BCA法测定其总胶原蛋白的含量,并对其外观性状、pH、黏度、重金属和微生物等进行质量检查;采用切除全层皮肤法建立大鼠伤口模型,以伤口的愈合率和伤口组织切片的HE染色、Masson染色结果为评价指标,对Rhc-Ⅲ乳膏剂促进伤口愈合作用进行药效学评价。乳化法制备的Rhc-Ⅲ乳膏剂外观为乳白色、黏稠、无异味且易于涂布的半固体,其pH为(6.66±0.03),黏度为(48 136±595)mPa·s,总胶原蛋白的含量为(1.38±0.15) mg/g,重金属和微生物限度符合要求;耐寒耐热试验后稳定性良好;模型大鼠给药后在3,7和14 d, Rhc-Ⅲ乳膏剂组的伤口愈合率均大于模型组和空白乳膏组;HE染色和Masson染色结果显示,Rhc-Ⅲ乳膏剂组肉芽组织出现早,成熟快,胶原纤维沉积量多,伤口愈合效果好。Rhc-Ⅲ乳膏剂制备方法简便、质量可控且稳定,具有促进伤口愈合的作用。

With O/W ointment as a carrier, Rhc-Ⅲ(recombinant human collagen-Ⅲ) cream was prepared and its quality and effect on pharmacodynamics for promoting wound healing were evaluated.First, Rhc-Ⅲ cream was prepared by emulsification method, and the content of total collagen was determined by BCA method, its appearance was observed, its pH and viscosity was measured, and the limits of heavy metals and microorganisms was checked;then preliminary evaluation of its stability was carried out by cold and heat resistance test;finally, the full-thickness skin was removed to establish a rat wound model.The wound healing rate and the results of HE staining and Masson staining of wound tissue sections were used as evaluation indicators to evaluate the effect of Rhc-Ⅲ cream on promoting wound healing.The appearance of Rhc-Ⅲ cream prepared by emulsification method was milky white, viscous, odorless and easy to spread.Its pH was(6.66±0.03) and viscosity was(48136±595) mPa·s.The content of collagen is(1.38±0.15) mg/g, and the limits of heavy metals and microorganisms meet the requirements;the results of cold tolerance and heat resistance showed that the cream was stable;after being treated with Rhc-Ⅲ cream for 3,7 and 14 days,the wound healing rate of the ointment group was higher than that of the model group and the blank cream group.The HE staining results showed that the capillaries and granulation tissues of the Rhc-Ⅲ cream group appeared early and matured faster.The Masson staining results showed that the amount of collagen fibers of the Rhc-Ⅲ cream deposited was larger than that of the model group and the blank cream group.The preparation method of Rhc-Ⅲ cream is simple and convenient, the quality of the Rhc-Ⅲ cream is controllable and stable, and it has the effect of promoting wound healing.