Cytotoxicity of Calcium Hydroxide on Human Umbilical Cord Mesenchymal Stem Cells

Eric Priyo Prasetyo,Ira Widjiastuti,Febriastuti Cahyani,Mefina Kuntjoro,Alexander Patera Nugraha,Ervina Restiwulan Winoto,Helen Susilowati,Fedik Abdul Rantam,Eryk Hendrianto,Sianiwati Goenharto,Ninuk Hariyani,Nike Hendrijantini

doi:10.1590/pboci.2020.141

Abstract

Objective: To examine the cytotoxicity of calcium hydroxide on human umbilical cord mesenchymal stem cells (HUCMSC) to understand the characteristics for use in regenerative dentistry procedures especially regenerative endodontics. Material and Methods: HUCMSC was isolated, cultured, and confirmed by flow cytometry. The biological characteristics, such as cell morphology, proliferation, and protein expression, were screened. To check the cytotoxicity, HUCMSC was cultured and divided into two groups, the control group (cultured in minimum essential medium (MEM) alpha) and calcium hydroxide group (cultured in MEM alpha and calcium hydroxide). Methyl-thiazole-tetrazolium (MTT) assay was done on different concentrations of calcium hydroxide (0.39 to 25 µg/mL) and the cells were observed and counted. One-way ANOVA test was used with a significance level set at 5% . Results: Flow cytometric analysis confirmed positive of CD73, CD90, CD105, negative of CD45 and CD34. A significant difference was found between the concentration of 6.25 and 3.125 µg/mL (p=0.004). There was no significant difference among 6.25, 12.5 and 25 µg/mL concentrations. There was also no significant difference among 0.39, 0.78, 1.56, and 3.125 µg/mL concentrations . Conclusion: Even though calcium hydroxide is a medicament of choice in clinical endodontics, it decreases the viability of HUCMSC. The lower the concentration of calcium hydroxide, the higher the viability of HUCMSC.

Highlights

There are several mesenchymal stem cells (MSC) sources for clinical applications
Human umbilical cord mesenchymal stem cells (HUCMSCs) are MSC found in the umbilical cords and may be attractive candidates for tissue regeneration [1]
The presence of mesenchymal stem cells taken from human umbilical cord was established and identified through antibodies for CD105, CD90, CD73, CD45, and CD34

Summary

Introduction

There are several mesenchymal stem cells (MSC) sources for clinical applications. Human umbilical cord mesenchymal stem cells (HUCMSCs) are MSC found in the umbilical cords and may be attractive candidates for tissue regeneration [1]. Calcium hydroxide (CH), an odorless white powder, is a strong alkali or base [3]. It has long been used in various treatment modalities in conservative dentistry, including intracanal dressings, root canal sealers, and pulp-capping material [4]. Calcium hydroxide is an essential alkaline material used in protocols of a successful regenerative endodontic treatment regimen [5] and the material of choice of all pulp therapy [4]. Its high pH level determines the mineralizing action of calcium hydroxide, and the hydroxyl group is reckoned as the most important part as it gives an alkaline milieu, which promotes healing and mineralization [3]

Methods

Results

Discussion

Conclusion