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利多卡因对牛关节软骨细胞的体外细胞毒性:细胞活力和蛋白聚糖代谢的变化。

Lidocaine cytotoxicity to the bovine articular chondrocytes in vitro: changes in cell viability and proteoglycan metabolism.

机构信息

Department of Orthopaedics and Rehabilitation Medicine, Faculty of Medical Sciences, The University of Fukui, Shimoaizuki 23, Matsuoka, Fukui 910-1193, Japan.

出版信息

Knee Surg Sports Traumatol Arthrosc. 2011 Jul;19(7):1198-205. doi: 10.1007/s00167-010-1369-9. Epub 2011 Jan 13.

Abstract

PURPOSE

A lot of studies on the effect of intra-articular injections are clinical, but many questions on the effect of lidocaine to articular chondrocytes remain unanswered. This study was performed to determine the effects of varying concentrations and exposure times of lidocaine on the viability and proteoglycan metabolism of chondrocytes in vitro.

METHOD

Cartilage was obtained from metatarsal joints of adult bovines. Chondrocytes in alginate beads were cultured in medium containing 6% fetal calf serum at 370 mOsmol at cell densities of 4 million cells/ml. They were then cultured for 24 h under 21% oxygen with 0.125, 0.25, 0.5, and 1% lidocaine and without lidocaine as control. The cell viability profile across intact beads was determined by manual counting using fluorescent probes and transmission electron microscopy.

RESULT

Lactate production was measured enzymatically as a marker of energy metabolism. Glycosaminoglycan (GAG) accumulation was measured using a modified dimethylmethylene blue assay. Cell viability decreased in a time- and dose-dependent manner in the concentration range of 0.125-1.0% lidocaine under the confocal microscope. Under the electron microscope, apoptosis increased as the concentration of lidocaine increased. GAG accumulation/tissue volume decreases as the concentration of lidocaine increased. However, GAG produced per million cells and the rate of lactate production per live cell were significantly higher for cells cultured at 0.5 and 1% lidocaine than the control group. Bovine chondrocytes cultured in alginate beads under high oxygen pressure are negatively influenced by increasing concentrations of lidocaine.

CONCLUSION

Cell viability and proteoglycan production (GAG accumulation/tissue volume) decreased as the concentration of lidocaine increased. These data suggest caution in prolonged exposure of cartilage to high concentration lidocaine. Repeated joint injection of lidocaine potentially worsens osteoarthrosis by accelerating cartilage degradation.

摘要

目的

许多关于关节内注射效果的研究都是临床研究,但关于利多卡因对关节软骨细胞的影响仍有许多问题尚未得到解答。本研究旨在确定不同浓度和暴露时间的利多卡因对体外软骨细胞活力和蛋白聚糖代谢的影响。

方法

从成年牛的跖骨关节中获取软骨。将软骨细胞在含有 6%胎牛血清的海藻酸盐珠中培养,在 370 毫渗摩尔渗透压下以 400 万细胞/ml 的细胞密度进行培养。然后在 21%氧气下培养 24 小时,加入 0.125%、0.25%、0.5%和 1%的利多卡因,不加利多卡因作为对照。通过手动计数使用荧光探针和透射电子显微镜来确定完整珠粒中的细胞活力分布。

结果

通过酶法测定乳酸盐产生作为能量代谢的标志物。使用改良的二甲亚甲基蓝测定法测量糖胺聚糖(GAG)积累。在共聚焦显微镜下,在 0.125-1.0%利多卡因浓度范围内,细胞活力随时间和剂量的增加呈时间和剂量依赖性下降。在电子显微镜下,随着利多卡因浓度的增加,细胞凋亡增加。随着利多卡因浓度的增加,GAG 积累/组织体积减少。然而,在 0.5%和 1%利多卡因培养的细胞中,每百万细胞产生的 GAG 和每活细胞产生的乳酸盐的速率均明显高于对照组。在高氧压下培养的牛软骨细胞,随着利多卡因浓度的增加,受到负面影响。

结论

随着利多卡因浓度的增加,细胞活力和蛋白聚糖产生(GAG 积累/组织体积)下降。这些数据表明,在长时间暴露于高浓度利多卡因时需要谨慎。重复关节内注射利多卡因可能通过加速软骨降解而使骨关节炎恶化。

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