Page 74 - 《精细化工》2023年第11期

第 40 卷第 11 期 精 细 化 工 Vol.40, No.11
2 023 年 11 月 FINE CHEMICALS Nov. 2023

功能材料
冷冻研磨对胶原蛋白分子结构及性能的调控

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刘佳林，戴 蕾，徐玉玲 ，汪海波，许承志，未本美
（武汉轻工大学 化学与环境工程学院，湖北 武汉 430023）

摘要：采用冷冻研磨技术将商品化天然牛跟腱胶原蛋白（海绵状）处理成粉末，采用凝胶电泳、FTIR、圆二色
谱、AFM、SEM、DSC 考察了冷冻时间对胶原蛋白分子结构和理化性能的影响，并评价了其对小鼠成纤维细胞
的促细胞增殖性能。结果显示，冷冻时间会影响胶原蛋白的三螺旋结构及分子间相互作用力。短时间的冷冻研
磨（12 h 以内）会对胶原蛋白分子的肽链结构产生一定影响，冷冻 24 h 后（样品 COL24h）该影响可降至最低。
COL24h 的三螺旋结构最完整，且其体外组装成纤维的能力未受影响。研磨样品的水溶性（4 ℃）随预冻时间
的增加而增加，与对照组（COL）相比，COL24h 在水中的溶解度提高了近 3 倍。冷冻研磨对胶原蛋白分子的
热变性温度影响不大，对其热变焓值影响较大，但随着冷冻时间的延长，产生的影响也逐渐降低。研磨后胶原
蛋白溶液整体黏度降低，流动性更好。COL24h 具有更强的促细胞增殖能力。
关键词：胶原蛋白；冷冻研磨；溶解度；性能；黏度；三螺旋结构；功能材料
中图分类号：O636 文献标识码：A 文章编号：1003-5214 (2023) 11-2386-08

Effects of cryo-grinding pretreatment on molecular
structure and properties of collagen

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LIU Jialin, DAI Lei, XU Yuling , WANG Haibo, XU Chengzhi, WEI Benmei
（School of Chemical and Environmental Engineering, Wuhan Polytechnic University, Wuhan 430023, Hubei, China）

Abstract: Commercial natural bovine tendon collagen (sponge-like) was processed into powder via
cryo-grinding technique. The effects of freezing time on the molecular structure and physicochemical
properties of collagen were evaluated by gel electrophoresis, FTIR, circular dichroism, AFM, SEM and
DSC, followed by assessment on the influence of collagen on proliferative properties of mouse
fibroblasts. The results revealed that that freezing time exhibited impact on the triple-helix structure
and molecular interaction force of collagen, while short frozen grinding time (within 12 h) showed a
certain effect on the peptide chain structure of collagen protein molecules, which could be minimized
after 24 h of freezing (sample COL24h). Sample COL24h displayed the most intact triple-helix
structure and its ability to assemble fibers in vitro was not affected. The water solubility (4 ℃) of the
ground samples increased with prolonged pre-freezing time, with that of COL24h nearly three times
larger than that of the control group (COL). In addition, cryo-grinding showed little effect on the
thermal denaturation temperature of collagen molecules, but great influence on the thermal change
enthalpy value. However, the influence was gradually reduced with the extension of freezing time. The
collagen after grinding exhibited reduced viscosity and a better fluidity. The COL24h showed more
substantial capacity in promoting cell proliferation.
Key words: collagen; cryo-grinding; solubility; properties; viscosity; triple-helical structure; functional materials

Ⅰ型胶原蛋白由 3 条肽链右手螺旋而成，是细 织中，具有良好的生物活性 [1-3] 。以胶原蛋白为基质
胞外基质的主要成分，大量存在于动物体的结缔组 或主要构件的海绵、凝胶、人造角膜、皮肤替代物、

收稿日期：2022-12-08; 定用日期：2023-05-29; DOI: 10.13550/j.jxhg.20221133
基金项目：国家自然科学基金项目（22178277）。
作者简介：刘佳林（1996—），女，硕士生，E-mail：916032823@qq.com。联系人：徐玉玲（1978—），女，副教授，E-mail：xyling619@126.com