Prasetyo Eric Priyo, Kuntjoro Mefina, Goenharto Setyabudi, Juniarti Devi Eka, Cahyani Febriastuti, Hendrijantini Nike, Nugraha Alexander Patera, Hariyani Ninuk, Rantam Fedik Abdul
Doctoral Program, Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia.
Department of Conservative Dentistry, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia.
Clin Cosmet Investig Dent. 2021 Mar 8;13:59-65. doi: 10.2147/CCIDE.S284240. eCollection 2021.
Calcium hydroxide is a gold standard dental material generally used for pulpal and periapical therapy including regenerative endodontic procedures because of its positive properties. However, evaluation about this material on stem cells is limited. Human umbilical cord mesenchymal stem cells (HUCMSCs) are potential to be used in regenerative therapy. Regenerative therapy needs a sustainable cell supply to maintain its regenerative capacity. The aim of this study was to ascertain the apoptosis result of calcium hydroxide on HUCMSCs through the expression of apoptotic protease-activating factor-1 (APAF-1), caspase-3, and caspase-9.
This study used a thawed frozen stock of passage 5 HUCMSCs, grown in minimum essential medium (MEM) alpha containing calcium hydroxide at concentration of 0.1 microgram/mL for 1, 3 and 7 days. Polyclonal antibody with fluorescence isothiocyanate (FITC) label was used to evaluate the expressions. APAF-1, caspase-3, and caspase-9 expressions were recorded and compared on every observation day using fluorescence microscope. Analysis of variance was performed to analyze the significance among the results of treatment groups. The results were concluded significant if p<0.05.
The addition of calcium hydroxide in MEM alpha medium increases HUCMSCs expression of APAF-1, caspase-3 and caspase-9 significantly, compared to the control group without calcium hydroxide (p<0.05) in all the times. Day 1 showed the lowest increase followed by higher expressions on day 3 and day 7.
HUCMSCs express increased APAF-1, caspase-3 and caspase-9 after in-vitro calcium hydroxide exposure. This should be considered when using calcium hydroxide on HUCMSCs for regenerative procedures with regard to other positive properties.
氢氧化钙是一种黄金标准的牙科材料，由于其良好的性能，通常用于牙髓和根尖周治疗，包括再生性牙髓治疗程序。然而，关于这种材料对干细胞影响的评估有限。人脐带间充质干细胞（HUCMSCs）有潜力用于再生治疗。再生治疗需要可持续的细胞供应来维持其再生能力。本研究的目的是通过凋亡蛋白酶激活因子-1（APAF-1）、半胱天冬酶-3和半胱天冬酶-9的表达来确定氢氧化钙对HUCMSCs的凋亡结果。
本研究使用第5代HUCMSCs的解冻冻存细胞，在含有浓度为0.1微克/毫升氢氧化钙的α-最低必需培养基（MEM）中培养1、3和7天。使用异硫氰酸荧光素（FITC）标记的多克隆抗体来评估表达情况。使用荧光显微镜记录并比较每个观察日的APAF-1、半胱天冬酶-3和半胱天冬酶-9的表达。进行方差分析以分析各治疗组结果之间的显著性。如果p<0.05，则结果具有显著性。
与不含氢氧化钙的对照组相比，在MEMα培养基中添加氢氧化钙显著增加了HUCMSCs中APAF-1、半胱天冬酶-3和半胱天冬酶-9的表达（p<0.05），在所有时间点均如此。第1天显示增加最少，随后在第3天和第7天表达更高。
体外暴露于氢氧化钙后，HUCMSCs中APAF-1、半胱天冬酶-3和半胱天冬酶-9的表达增加。在将氢氧化钙用于HUCMSCs进行再生程序时，考虑到其其他积极特性，应考虑到这一点。
