A Potential Intracanal Medicament, 2-Hydroxyisocaproic Acid (HICA): Cytotoxicity, Genotoxicity, and Its Effect on SCAP Differentiation

Intracanal medicaments with maximal antimicrobial efficacy and minimal damage to resident stem cells are essential for successful regenerative endodontic procedures. 2-Hydroxyisocaproic acid (HICA) could have the attributes of a potential intracanal medicament. This study evaluates its cytotoxicity,...

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Main Authors: Cristine Smoczer, Yun K. Park, James B. Herrington, Mazin A. Askar, Sarah Plecha, Eric Krukonis, Susan B. Paurazas
Format: Article
Language:English
Published: MDPI AG 2023-11-01
Series:Dentistry Journal
Subjects:
Online Access:https://www.mdpi.com/2304-6767/11/12/270
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author Cristine Smoczer
Yun K. Park
James B. Herrington
Mazin A. Askar
Sarah Plecha
Eric Krukonis
Susan B. Paurazas
author_facet Cristine Smoczer
Yun K. Park
James B. Herrington
Mazin A. Askar
Sarah Plecha
Eric Krukonis
Susan B. Paurazas
author_sort Cristine Smoczer
collection DOAJ
description Intracanal medicaments with maximal antimicrobial efficacy and minimal damage to resident stem cells are essential for successful regenerative endodontic procedures. 2-Hydroxyisocaproic acid (HICA) could have the attributes of a potential intracanal medicament. This study evaluates its cytotoxicity, genotoxicity, and effects on the odontogenic and osteogenic differentiation of the stem cells of the apical papilla (SCAP). Cytotoxicity and cell viability assays were performed on cells treated for 24, 48, and 72 h with varying concentrations of HICA and compared to the standard intracanal medicament, calcium hydroxide. The genotoxicity was assessed via immunofluorescence for two markers of DNA double-strand breaks: phosphorylated γH<sub>2</sub>AX and 53BP1. The SCAP differentiation was evaluated based on the alkaline phosphatase activity, Alizarin Red staining, and expression of odontogenic and osteogenic genes (<i>DSPP1</i>, <i>BSP1</i>, <i>OCN</i>, <i>RUNX2</i>) in the presence of selected HICA concentrations. HICA was not cytotoxic at concentrations up to 10 mg/mL, regardless of the exposure time, although it was cytostatic at all tested concentrations. HICA was not genotoxic at concentrations below 5 mg/mL. No difference in cytotoxicity or genotoxicity was found between HICA and calcium hydroxide at 1 mg/mL. HICA retained about 70% of the osteogenic differentiation potential at 1 mg/mL. Within the limitations of this in vitro study, we show that HICA at 1 mg/mL could be a potential intracanal medicament for REPs.
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spelling doaj.art-e7123f90703a452581ecb9cec3467e702023-12-22T14:02:42ZengMDPI AGDentistry Journal2304-67672023-11-01111227010.3390/dj11120270A Potential Intracanal Medicament, 2-Hydroxyisocaproic Acid (HICA): Cytotoxicity, Genotoxicity, and Its Effect on SCAP DifferentiationCristine Smoczer0Yun K. Park1James B. Herrington2Mazin A. Askar3Sarah Plecha4Eric Krukonis5Susan B. Paurazas6Division of Integrated Biomedical Sciences, University of Detroit Mercy School of Dentistry, Detroit, MI 48208, USAGraduate Endodontics, University of Detroit Mercy School of Dentistry, Detroit, MI 48208, USAGraduate Endodontics, University of Detroit Mercy School of Dentistry, Detroit, MI 48208, USAGraduate Endodontics, University of Detroit Mercy School of Dentistry, Detroit, MI 48208, USADivision of Integrated Biomedical Sciences, University of Detroit Mercy School of Dentistry, Detroit, MI 48208, USADivision of Integrated Biomedical Sciences, University of Detroit Mercy School of Dentistry, Detroit, MI 48208, USAGraduate Endodontics, University of Detroit Mercy School of Dentistry, Detroit, MI 48208, USAIntracanal medicaments with maximal antimicrobial efficacy and minimal damage to resident stem cells are essential for successful regenerative endodontic procedures. 2-Hydroxyisocaproic acid (HICA) could have the attributes of a potential intracanal medicament. This study evaluates its cytotoxicity, genotoxicity, and effects on the odontogenic and osteogenic differentiation of the stem cells of the apical papilla (SCAP). Cytotoxicity and cell viability assays were performed on cells treated for 24, 48, and 72 h with varying concentrations of HICA and compared to the standard intracanal medicament, calcium hydroxide. The genotoxicity was assessed via immunofluorescence for two markers of DNA double-strand breaks: phosphorylated γH<sub>2</sub>AX and 53BP1. The SCAP differentiation was evaluated based on the alkaline phosphatase activity, Alizarin Red staining, and expression of odontogenic and osteogenic genes (<i>DSPP1</i>, <i>BSP1</i>, <i>OCN</i>, <i>RUNX2</i>) in the presence of selected HICA concentrations. HICA was not cytotoxic at concentrations up to 10 mg/mL, regardless of the exposure time, although it was cytostatic at all tested concentrations. HICA was not genotoxic at concentrations below 5 mg/mL. No difference in cytotoxicity or genotoxicity was found between HICA and calcium hydroxide at 1 mg/mL. HICA retained about 70% of the osteogenic differentiation potential at 1 mg/mL. Within the limitations of this in vitro study, we show that HICA at 1 mg/mL could be a potential intracanal medicament for REPs.https://www.mdpi.com/2304-6767/11/12/2702-hydroxyisocaproic acidSCAPcytotoxicitygenotoxicityodonto/osteogenic differentiationregenerative endodontic procedures
spellingShingle Cristine Smoczer
Yun K. Park
James B. Herrington
Mazin A. Askar
Sarah Plecha
Eric Krukonis
Susan B. Paurazas
A Potential Intracanal Medicament, 2-Hydroxyisocaproic Acid (HICA): Cytotoxicity, Genotoxicity, and Its Effect on SCAP Differentiation
Dentistry Journal
2-hydroxyisocaproic acid
SCAP
cytotoxicity
genotoxicity
odonto/osteogenic differentiation
regenerative endodontic procedures
title A Potential Intracanal Medicament, 2-Hydroxyisocaproic Acid (HICA): Cytotoxicity, Genotoxicity, and Its Effect on SCAP Differentiation
title_full A Potential Intracanal Medicament, 2-Hydroxyisocaproic Acid (HICA): Cytotoxicity, Genotoxicity, and Its Effect on SCAP Differentiation
title_fullStr A Potential Intracanal Medicament, 2-Hydroxyisocaproic Acid (HICA): Cytotoxicity, Genotoxicity, and Its Effect on SCAP Differentiation
title_full_unstemmed A Potential Intracanal Medicament, 2-Hydroxyisocaproic Acid (HICA): Cytotoxicity, Genotoxicity, and Its Effect on SCAP Differentiation
title_short A Potential Intracanal Medicament, 2-Hydroxyisocaproic Acid (HICA): Cytotoxicity, Genotoxicity, and Its Effect on SCAP Differentiation
title_sort potential intracanal medicament 2 hydroxyisocaproic acid hica cytotoxicity genotoxicity and its effect on scap differentiation
topic 2-hydroxyisocaproic acid
SCAP
cytotoxicity
genotoxicity
odonto/osteogenic differentiation
regenerative endodontic procedures
url https://www.mdpi.com/2304-6767/11/12/270
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