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大蒜提取物对人牙髓成纤维细胞的细胞毒性评估。

Evaluation of Cytotoxicity of (Garlic Extract) against Human Dental Pulp Fibroblasts.

作者信息

Devaraju Raghu, Reddy Divya, Paul Santhosh T, Azher Umme, Umashankar Keerthy, Srinivas Likhith

机构信息

Department of Paedodontics and Preventive Dentistry, Sri Rajiv Gandhi College of Dental Sciences and Hospital, Bengaluru, Karnataka, India.

Department of Periodontology and Implantology, Priya Multispeciality Dental Clinic, Sainagar, Bengaluru, Karnataka, India.

出版信息

Int J Clin Pediatr Dent. 2024 Feb;17(2):143-148. doi: 10.5005/jp-journals-10005-2781.

Abstract

BACKGROUND

Vital pulp therapy procedures in primary dentition focuses on preservation and maintenance of pulp tissue that has been compromised due to caries, trauma, etc. Several pulp dressing materials have been used in primary teeth and some natural materials from the field of traditional medicine have also been introduced as medicaments in vital pulp therapy. The understanding of biologic and cytotoxic properties of newer materials is important for safe clinical usage. The biologic compatibility of these newer materials is imperative to limit or avoid tissue irritation or degeneration.

AIM

To evaluate the cytotoxic effects of on cultured human primary dental pulp fibroblasts.

MATERIALS AND METHODS

Primary pulp fibroblasts were cultured from the pulp tissue obtained from extracted deciduous primary canines and central incisor teeth. The freshly prepared concentrations of 1000, 500, 250, 125, and 62.5 µg/mL extract were added to the 96-well plate in triplicates to which culture medium containing fourth passage cell suspension was added previously. Cells without treatment served as control, while cells treated with 5% dimethyl sulfoxide (DMSO) served as toxic control. After the addition of experimental and control agents, cells were incubated for 24 and 48 hours at 37°C in 5% CO atmosphere. After the incubation period, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide (MTT) assay was performed to determine the number of viable cells. Absorbance was read with a microplate reader at 570 nm wavelength and the relative viability of dental pulp fibroblasts at various concentrations was expressed as color intensity of the experimental wells relative to that of control. The percentage of cell viability was also calculated accordingly.

RESULTS

The MTT assay results revealed that extract, in all the concentrations tested at both the time intervals maintained a cell viability of greater than 90%. At 24 hours, the mean absorbance value of untreated control wells was recorded as 0.84400 ± 0.00916 with 100% cell viability. Among all the concentrations of garlic extract tested, highest mean absorbance value of 0.83933 ± 0.00550 with 99.44% cell viability was recorded for 62.5 µg/mL concentration. At 48 hours, the mean absorbance value of untreated control wells was recorded as 1.22767 ± 0.01106 with 100% cell viability, and the highest mean absorbance value of 1.22567 ± 0.01006 with 99.83% cell viability was recorded for 62.5 µg/mL concentration. The cell viability did not seem to be affected by the concentration of extract at 24 hours. However, at 48 hours, the sensitivity of the cells was observed to be dependent on the concentration of with a decrease in the viability of cells noted with the increase in concentration.

CONCLUSION

extract is noncytotoxic in nature and preserves the vitality of cultured human primary dental pulp fibroblasts making it a suitable material for use in vital pulp therapy procedures of primary teeth.

HOW TO CITE THIS ARTICLE

Devaraju R, Reddy D, Paul ST, Evaluation of Cytotoxicity of (Garlic Extract) against Human Dental Pulp Fibroblasts. Int J Clin Pediatr Dent 2024;17(2):143-148.

摘要

背景

乳牙的活髓治疗程序着重于保存和维持因龋齿、外伤等而受损的牙髓组织。几种牙髓敷料材料已用于乳牙,一些传统医学领域的天然材料也已被引入作为活髓治疗的药物。了解新型材料的生物学和细胞毒性特性对于安全临床应用很重要。这些新型材料的生物相容性对于限制或避免组织刺激或退变至关重要。

目的

评估[具体材料名称]对培养的人原代牙髓成纤维细胞的细胞毒性作用。

材料与方法

从拔除的乳牙尖牙和中切牙的牙髓组织中培养原代牙髓成纤维细胞。将新配制的浓度为1000、500、250、125和62.5μg/mL的[具体材料名称]提取物一式三份加入96孔板中,之前已向其中加入含有第四代细胞悬液的培养基。未处理的细胞作为对照,用5%二甲基亚砜(DMSO)处理的细胞作为毒性对照。加入实验试剂和对照试剂后,将细胞在37°C、5%二氧化碳气氛中孵育24小时和48小时。孵育期结束后,进行3-(4,5-二甲基-2-噻唑基)-2,5-二苯基溴化四氮唑(MTT)试验以确定活细胞数量。用酶标仪在570nm波长下读取吸光度,不同浓度下牙髓成纤维细胞的相对活力以实验孔相对于对照孔的颜色强度表示。相应地还计算了细胞活力百分比。

结果

MTT试验结果显示,在两个时间间隔测试的所有浓度下,[具体材料名称]提取物均保持细胞活力大于90%。在24小时时,未处理对照孔的平均吸光度值记录为0.84400±0.00916,细胞活力为100%。在测试的所有大蒜提取物浓度中,62.5μg/mL浓度的平均吸光度值最高,为0.83933±0.00550,细胞活力为99.44%。在48小时时,未处理对照孔的平均吸光度值记录为1.22767±0.01106,细胞活力为100%,62.5μg/mL浓度的平均吸光度值最高,为1.22567±0.01006,细胞活力为99.83%。在24小时时,细胞活力似乎不受[具体材料名称]提取物浓度的影响。然而,在48小时时,观察到细胞的敏感性取决于[具体材料名称]的浓度,随着浓度增加细胞活力降低。

结论

[具体材料名称]提取物本质上无细胞毒性,并能保持培养的人原代牙髓成纤维细胞的活力,使其成为乳牙活髓治疗程序中适用的材料。

如何引用本文

Devaraju R, Reddy D, Paul ST, [具体材料名称](大蒜提取物)对人牙髓成纤维细胞细胞毒性的评估。《国际临床儿科牙科学杂志》2024;17(2):143 - 148。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4aa2/11339490/652a7ea9e794/ijcpd-17-143-g001.jpg

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