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钬激光照射对椎间盘细胞的影响。

The influence of Ho:YAG laser irradiation on intervertebral disc cells.

作者信息

Sato Masato, Ishihara Miya, Kikuchi Makoto, Mochida Joji

机构信息

Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Kanagawa, Japan.

出版信息

Lasers Surg Med. 2011 Nov;43(9):921-6. doi: 10.1002/lsm.21120.

Abstract

BACKGROUND AND OBJECTIVE

Various types of laser have been reported for percutaneous laser disc decompression (PLDD). The aim of this study was to understand the effects on intervertebral disc cells following Ho:YAG laser irradiation, using a three-dimensional culture model, and consider appropriate irradiation conditions.

STUDY DESIGN/MATERIALS AND METHODS: Intervertebral discs from the lumbar spine were obtained from 36 female Japanese white rabbits and processed to obtain isolated cells in three-dimensional cultures. Photoacoustic and photothermal effects were investigated by irradiating three-dimensional cultures with Ho:YAG laser at 27 or 54 J. Residual cell counts after irradiation were estimated based on DNA content according to fluorometric assay. Lactate dehydrogenase levels were also investigated as a marker of damage to cell plasma membranes. Finally, proteoglycan synthesis was measured by rapid filtration assay of (35) S incorporation, as an index of matrix synthesis.

RESULTS

Residual cell count tended to be higher in the 27-J group. Plasma membrane damage was higher and remained high longer after irradiation in the 54-J group. Proteoglycan synthesis was higher in the 27-J group than in the 54-J group, with some conditions (e.g., 90 mJ/pulse condition) showing marked activation of proteoglycan synthesis maintained for a long time after irradiation.

CONCLUSIONS

Three-dimensional culture models of intervertebral disc cells are useful for clarifying relationships between cell reactions and photoacoustic and photothermal effects after laser irradiation. Total energy is closely related to optimization of irradiation conditions, which may allow optimization of cytoprotection and promotion of matrix synthesis in clinical practice.

摘要

背景与目的

已有多种类型的激光用于经皮激光椎间盘减压术(PLDD)。本研究的目的是利用三维培养模型了解钬激光照射后对椎间盘细胞的影响,并探讨合适的照射条件。

研究设计/材料与方法:从36只日本雌性白兔的腰椎获取椎间盘,处理后在三维培养中获得分离细胞。用27或54焦耳的钬激光照射三维培养物,研究光声和光热效应。根据荧光分析法,基于DNA含量估算照射后的残留细胞计数。还研究了乳酸脱氢酶水平,作为细胞膜损伤的标志物。最后,通过快速过滤法检测(35)S掺入量来测量蛋白聚糖合成,作为基质合成的指标。

结果

27焦耳组的残留细胞计数往往更高。54焦耳组照射后细胞膜损伤更高且持续时间更长。27焦耳组的蛋白聚糖合成高于54焦耳组,在某些条件下(如90毫焦/脉冲条件),照射后蛋白聚糖合成有明显激活且持续时间长。

结论

椎间盘细胞的三维培养模型有助于阐明激光照射后细胞反应与光声和光热效应之间的关系。总能量与照射条件的优化密切相关,这可能有助于在临床实践中优化细胞保护和促进基质合成。

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