牛脐带提取物的特定成分分析及细胞功效评价

Specific Component Analysis and Cell Efficacy Evaluation of Bovine Umbilical Extract

为开发动物源功能性化妆品原料,采用检验合格的牛脐带组织,利用一系列生物学分离提取技术手段,摸索并建立稳定可重复的牛脐带提取物的提取制备工艺,并开展细胞功效评价。研究结果如下:(1)建立了一种稳定的牛脐带提取物的提取制备工艺,通过切向流浓缩,牛脐带提取物蛋白质量浓度可达(9.21±1.95)mg/mL;(2)小细胞外囊泡成分检测显示产物中富集小细胞外囊泡成分;(3)基于细胞模型的功效评价结果显示,牛脐带提取物促进人皮肤成纤维细胞(HSF)模型的细胞增殖活性,并提高细胞划痕修复率,显著下调小鼠巨噬细胞炎性细胞模型的IL-6、TNF-α表达水平,减少人永生化表皮(HaCaT)细胞凋亡模型的凋亡细胞百分率,减少HSF衰老模型的β-半乳糖苷酶染色阳性细胞数量。建立了一种富含小细胞外囊泡成分且具有多种细胞功效的牛脐带提取物的标准化制备工艺。

To develop animal derived functional cosmetic raw materials,qualified bovine umbilical cords were used as subject,a series of biological separation and extraction techniques were used to explore and establish a standardized and repeatable extraction and preparation process of bovine umbilical cord extract.The cell efficacy of bovine umbilical cord extract was evaluated.The experimental results were as follows.(1)A stable extraction and preparation process of bovine umbilical cord extract was established.The protein concentration of bovine umbilical cord extract was up to(9.21±1.95)mg/mL by tangential flow concentration.(2)The detection of small extracellular vesicle components showed that the products were enriched in small extracellular vesicles.(3)The efficacy evaluation based on cell models showed that bovine umbilical cord extract promoted the cell proliferation activity of human skin fibroblasts(HSF)model and increased the cell scratch repair rate,significantly down-regulated the expression levels of IL-6 and TNF-α in mouse macrophage inflammatory cell model,reduced the percentage of apoptotic cells in human immortalized epidernal cells(HaCaT)apoptotic cell model and reduced β-galactosidase staining positive cells of HSF cell senescence model.A standardized preparation process of bovine umbilical cord extract rich in small extracellular vesicles with multiple cell efficacy was established.