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通过荧光激活细胞分选分析评估洗必泰和印楝提取物对培养的人牙龈成纤维细胞的细胞毒性:一项研究。

Cytotoxicity of chlorhexidine and neem extract on cultured human gingival fibroblasts through fluorescence-activated cell sorting analysis : An study.

作者信息

Verma Umesh Pratap, Gupta Abhaya, Yadav Rakesh Kumar, Tiwari Rini, Sharma Ramesh, Balapure Anil Kumar

机构信息

Department of Periodontology, Faculty of Dental Sciences, King George's Medical University, Lucknow, Uttar Pradesh, India.

Department of Conservative and Endodontics, Faculty of Dental Sciences, King George's Medical University, Lucknow, Uttar Pradesh, India.

出版信息

Eur J Dent. 2018 Jul-Sep;12(3):344-349. doi: 10.4103/ejd.ejd_149_17.

Abstract

OBJECTIVE

To assess the influence of chlorhexidine (CHX), neem vehicle control (NVC), and neem extract (NE) on cultured human gingival fibroblasts (hGFs) using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and fluorescence-activated cell sorting (FACS) analysis.

MATERIALS AND METHODS

Fibroblasts were derived from healthy gingival biopsy specimens harvested aseptically. The effects of CHX, NVC, and NE were evaluated on cultured hGFs through FACS and MTT assay.

RESULTS

MTT assay with hGFs indicated altered morphology with maximum cell death at 10% CHX, while NVC and NE showed similar results at a concentration of 75% and above. On FACS analysis, beyond 1%, CHX adversely affected the cell cycle phase distribution whereas NE exerted a detrimental effect only at 100%. Moreover, both with NVC and NE cells were well differentiated in all the three phases of the cell cycle, with distinction getting lost at 50% to finally causing cell death at 100%.

CONCLUSIONS

CHX beyond 1% concentration exhibited a toxic effect on hGFs at 1, 5, and 15 min time exposure. However, NE did not adversely affect the fibroblasts even up to 50% concentration showing a less toxic effect in comparison with CHX on these cells. The cytoprotective and oral friendly quality of NE emphaisze the superiority of NE over CHX.

摘要

目的

使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐(MTT)法和荧光激活细胞分选(FACS)分析,评估洗必泰(CHX)、印楝赋形剂对照(NVC)和印楝提取物(NE)对培养的人牙龈成纤维细胞(hGFs)的影响。

材料与方法

成纤维细胞来源于无菌采集的健康牙龈活检标本。通过FACS和MTT法评估CHX、NVC和NE对培养的hGFs的影响。

结果

hGFs的MTT法显示,10%的CHX导致细胞形态改变且细胞死亡最多,而NVC和NE在75%及以上浓度时显示出相似结果。FACS分析表明,超过1%时,CHX会对细胞周期阶段分布产生不利影响,而NE仅在100%时产生有害作用。此外,NVC和NE处理的细胞在细胞周期的所有三个阶段均分化良好,在50%时差异消失,最终在100%时导致细胞死亡。

结论

浓度超过1%的CHX在暴露1、5和15分钟时对hGFs表现出毒性作用。然而,NE即使在浓度高达50%时也不会对成纤维细胞产生不利影响,与CHX相比,对这些细胞的毒性作用较小。NE的细胞保护和口腔友好特性强调了其优于CHX的优势。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e355/6089059/bc77e2bec9f5/EJD-12-344-g008.jpg

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