Fluorescent labeling with 5-DTAF reduces collagen fiber uncrimping in loaded tendons
Fluorescent labeling with 5-DTAF reduces collagen fiber uncrimping in loaded tendons
复制标题
DOI：
10.1016/j.jmbbm.2022.105646
复制
发表时间：
2022-12-27
影响因子：
3.9
通讯作者：
Michalek，Arthur J.
中科院分区：
文献类型：
--
作者：
Voce，Gianna T.;Michalek，Arthur J.
一种通常用于组织成像的荧光染料（5-DTAF）先前已被证明可以使肌腱变硬。本研究假设5-DTAF染色通过减少纤维滑动使肌腱变硬，从而改变卷曲的胶原纤维在负载下伸直的速度。采用反射交叉偏振光显微镜测量颈椎指长伸肌腱标本在轴向载荷下的纤维卷曲周期。标本分别用磷酸盐缓冲盐水（阴性对照）、吉尼平（阳性对照）或5-DTAF处理。在盐水处理的样品中，卷曲时间（相对于未拉伸）增加了大约2.5倍的轴向应变，表明纤维有很大的滑动。在5-DTAF和genipin处理的标本中，这一比例降至1:1，表明纤维没有滑动。这些结果进一步证明，在使用5-DTAF染色组织以研究组织中的微尺度变形时应谨慎。
A fluorescent dye commonly used to image tissues under load (5-DTAF) has previously been shown to stiffen tendons. This study hypothesized that 5-DTAF staining stiffens tendons through reduced fiber sliding, altering the rate at which crimped collagen fibers straighten under load. This was tested by using reflected cross-polarized light microscopy to measure fiber crimp period of cervine extensor digitorum longus tendon specimens under axial load. Specimens were treated with either phosphate buffered saline (negative control), genipin (positive control), or 5-DTAF. In saline treated specimens, crimp period (relative to unstretched) increased at approximately 2.5 times the applied axial strain, indicating substantial fiber sliding. In both 5-DTAF and genipin treated specimens, this ratio was reduced to 1:1, indicating no fiber sliding. These results add further evidence that care should be taken when using 5-DTAF to stain tissue for studying microscale deformations in tissues.