[基于激光显微手术细胞球体修复过程的细胞模型研究]。

Ontogenez Pub Date : 2017-01-01
N V Kosheleva, I V Ilina, K V Kozhina, I M Zurina, A E Roskova, A A Gorkun, A V Ovchinnikov, M B Agranat, S G Morozov, I N Saburina
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引用次数: 0

摘要

本研究利用现代激光显微手术技术,建立了一种新的简单、可重复的细胞球体修复和体外再生机制研究模型。应用纳秒激光脉冲对真皮成纤维细胞的球状体的内外区进行显微解剖。为了实现对球体的有效解剖和保存,选择了最佳的实验参数:波长355 nm,频率100 Hz,脉冲时间2 ns,激光脉冲范围7 ~ 9 μ j。显微解剖后,观察到球体的损伤:创面边缘开放,角度开放达到180°以上。早在激光对球体进行显微解剖后的第一个小时内,存活细胞的形状就发生了变化:球体表面和直接受损区域的细胞变成了圆形。显微解剖1天后,球体结构开始部分恢复,表层细胞开始呈现原来的扁平形态;死亡受损细胞的碎片及其碎片逐渐从球体成分中清除。在该模型中,首次获得了用aP199合成多肽刺激真皮成纤维细胞损伤球体结构恢复的数据,aP199合成多肽在美容中用于启动皮肤的抗衰老和再生作用。显微解剖后,在P199肽存在的情况下,含有少量平面层叠叠排列细胞和内带多边形细胞的球状结构恢复速度比对照组快,在7天内完成,可能与存活细胞的重塑有关。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Cellular Model Based on Laser Microsurgery of Cell Spheroids to Study the Repair Process].

In this study, modern techniques of laser microsurgery of cell spheroids have been used to develop a new simple, reproducible model for studying the mechanisms of repair and regeneration in vitro. Nanosecond laser pulses were applied to perform a microdissection of the outer and the inner zones of the spheroids from dermal fibroblasts. To achieve effective dissection and preservation of spheroid viability, the optimal parameters were chosen: 355 nm wavelength, 100 Hz frequency, 2 ns pulse duration, laser pulses in the range of 7–9 μ J. After microdissection, we observed injury of the spheroids : the edges of the wound surface opened and the angular opening reached a value of more than 180°. As early as during the first hour after spheroid microdissection with laser radiation, the surviving cells changed their shape: cells on the spheroid surface and directly in the damaged area became rounded. One day after microdissection, the structure of the spheroids began to partially recover, the cells in the surface layers began to take the original flattened shape; debris of dead damaged cells and their fragments was gradually cleared from the spheroid composition. In the proposed model, the first data on stimulation of structure recovery of injured spheroids from dermal fibroblasts with a P199 synthetic polypeptide, which is used in cosmetology for the initiation of antiaging and regenerative effects in the skin, were received. After microdissection, recovery of the spheroids structure with a few surface layers of flattened imbricated arranged cells and polygonal cells of the inner zone in the presence of P199 peptide was faster than in the control group, and was completed within 7 days, presumably due to the remodeling of the survived cells.

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