Effect of calcium hydroxide and adipose-derived stem cell conditioned medium (ADSCs-CM) combination on reparative dentin formation in Wistar rats through RUNX-2 and OPN expression

Background: Vital pulp exposure requires effective pulp capping to preserve the vitality and function of the dental pulp. Calcium hydroxide (Ca(OH)2) is widely used as a pulp capping agent; however, it has significant limitations, including the formation of reparative dentin with tunnel defects. The...

Full description

Saved in:
Bibliographic Details
Main Authors: Radixtio Auzan Fepiosandi, Dian Agustin Wahjuningrum, Galih Sampoerno, Anuj Bhardwaj, Ajinkya M. Pawar
Format: Article
Language:English
Published: Elsevier 2025-09-01
Series:Journal of Oral Biology and Craniofacial Research
Subjects:
Online Access:http://www.sciencedirect.com/science/article/pii/S2212426825001459
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849243732176535552
author Radixtio Auzan Fepiosandi
Dian Agustin Wahjuningrum
Galih Sampoerno
Anuj Bhardwaj
Ajinkya M. Pawar
author_facet Radixtio Auzan Fepiosandi
Dian Agustin Wahjuningrum
Galih Sampoerno
Anuj Bhardwaj
Ajinkya M. Pawar
author_sort Radixtio Auzan Fepiosandi
collection DOAJ
description Background: Vital pulp exposure requires effective pulp capping to preserve the vitality and function of the dental pulp. Calcium hydroxide (Ca(OH)2) is widely used as a pulp capping agent; however, it has significant limitations, including the formation of reparative dentin with tunnel defects. These defects may compromise long-term success. Adipose-derived stem cell conditioned medium (ADSCs-CM) has shown regenerative potential through paracrine mechanisms. Objectives: To evaluate the effect of combining Ca(OH)2 with ADSCs-CM on the expression of odontogenic markers RUNX-2 and osteopontin (OPN), and secondarily to assess the histological characteristics of reparative dentin in healthy Wistar rats. Materials and methods: An in vivo experimental study was conducted using twelve healthy female Wistar rats (Rattus norvegicus), randomly divided into control (Ca(OH)2 only) and treatment (Ca(OH)2 + ADSCs-CM in a 2:1 ratio) groups (n = 6/group). Class V cavities were prepared with standardized pulp exposure. Following hemostasis, the respective materials were applied and sealed with glass ionomer cement. Histological evaluation using hematoxylin and eosin (HE) staining and immunohistochemical assessment of RUNX-2 and OPN expression were performed on days 1, 3, and 6. Statistical analysis was conducted using the Shapiro-Wilk test and independent t-tests (p < 0.05). Results: RUNX-2 and OPN expressions were significantly higher in the treatment group at all time points (p < 0.05), with peak levels observed on day 6. Histologically, the treatment group exhibited more homogeneous and better organized reparative dentin compared to the control group. Conclusion: The addition of ADSCs-CM to Ca(OH)2 enhances reparative dentinogenesis by upregulating RUNX-2 and OPN expression. This combination therapy may serve as a biologically favorable alternative for direct pulp capping procedures.
format Article
id doaj-art-29e9fcb1b6b540e68fb03e1058723e67
institution Kabale University
issn 2212-4268
language English
publishDate 2025-09-01
publisher Elsevier
record_format Article
series Journal of Oral Biology and Craniofacial Research
spelling doaj-art-29e9fcb1b6b540e68fb03e1058723e672025-08-20T03:59:22ZengElsevierJournal of Oral Biology and Craniofacial Research2212-42682025-09-011551162116810.1016/j.jobcr.2025.07.001Effect of calcium hydroxide and adipose-derived stem cell conditioned medium (ADSCs-CM) combination on reparative dentin formation in Wistar rats through RUNX-2 and OPN expressionRadixtio Auzan Fepiosandi0Dian Agustin Wahjuningrum1Galih Sampoerno2Anuj Bhardwaj3Ajinkya M. Pawar4Doctoral Study Program in Dental Science, Faculty of Dental Medicine, Universitas Airlangga ,Jawa Timur, Indonesia; Department of Conservative Dentistry, Faculty of Dental Medicine, Universitas Airlangga, Jawa Timur, IndonesiaDepartment of Conservative Dentistry, Faculty of Dental Medicine, Universitas Airlangga, Jawa Timur, Indonesia; Corresponding author. Department of Conservative Dentistry, Faculty of Dental Medicine, Universitas Airlangga St. Mayjend. Prof. Dr. Moestopo No. 47. Surabaya, Indonesia.Department of Conservative Dentistry, Faculty of Dental Medicine, Universitas Airlangga, Jawa Timur, IndonesiaDepartment of Conservative Dentistry and Endodontics, College of Dental Science and Hospital, Rau, Indore, IndiaDepartment of Conservative Dentistry and Endodontics, Nair Hospital Dental College, Mumbai, Maharashtra, IndiaBackground: Vital pulp exposure requires effective pulp capping to preserve the vitality and function of the dental pulp. Calcium hydroxide (Ca(OH)2) is widely used as a pulp capping agent; however, it has significant limitations, including the formation of reparative dentin with tunnel defects. These defects may compromise long-term success. Adipose-derived stem cell conditioned medium (ADSCs-CM) has shown regenerative potential through paracrine mechanisms. Objectives: To evaluate the effect of combining Ca(OH)2 with ADSCs-CM on the expression of odontogenic markers RUNX-2 and osteopontin (OPN), and secondarily to assess the histological characteristics of reparative dentin in healthy Wistar rats. Materials and methods: An in vivo experimental study was conducted using twelve healthy female Wistar rats (Rattus norvegicus), randomly divided into control (Ca(OH)2 only) and treatment (Ca(OH)2 + ADSCs-CM in a 2:1 ratio) groups (n = 6/group). Class V cavities were prepared with standardized pulp exposure. Following hemostasis, the respective materials were applied and sealed with glass ionomer cement. Histological evaluation using hematoxylin and eosin (HE) staining and immunohistochemical assessment of RUNX-2 and OPN expression were performed on days 1, 3, and 6. Statistical analysis was conducted using the Shapiro-Wilk test and independent t-tests (p < 0.05). Results: RUNX-2 and OPN expressions were significantly higher in the treatment group at all time points (p < 0.05), with peak levels observed on day 6. Histologically, the treatment group exhibited more homogeneous and better organized reparative dentin compared to the control group. Conclusion: The addition of ADSCs-CM to Ca(OH)2 enhances reparative dentinogenesis by upregulating RUNX-2 and OPN expression. This combination therapy may serve as a biologically favorable alternative for direct pulp capping procedures.http://www.sciencedirect.com/science/article/pii/S2212426825001459Calcium hydroxideAdipose-derived stem cells conditioned mediumReparative dentinRUNX-2OsteopontinMedicine
spellingShingle Radixtio Auzan Fepiosandi
Dian Agustin Wahjuningrum
Galih Sampoerno
Anuj Bhardwaj
Ajinkya M. Pawar
Effect of calcium hydroxide and adipose-derived stem cell conditioned medium (ADSCs-CM) combination on reparative dentin formation in Wistar rats through RUNX-2 and OPN expression
Journal of Oral Biology and Craniofacial Research
Calcium hydroxide
Adipose-derived stem cells conditioned medium
Reparative dentin
RUNX-2
Osteopontin
Medicine
title Effect of calcium hydroxide and adipose-derived stem cell conditioned medium (ADSCs-CM) combination on reparative dentin formation in Wistar rats through RUNX-2 and OPN expression
title_full Effect of calcium hydroxide and adipose-derived stem cell conditioned medium (ADSCs-CM) combination on reparative dentin formation in Wistar rats through RUNX-2 and OPN expression
title_fullStr Effect of calcium hydroxide and adipose-derived stem cell conditioned medium (ADSCs-CM) combination on reparative dentin formation in Wistar rats through RUNX-2 and OPN expression
title_full_unstemmed Effect of calcium hydroxide and adipose-derived stem cell conditioned medium (ADSCs-CM) combination on reparative dentin formation in Wistar rats through RUNX-2 and OPN expression
title_short Effect of calcium hydroxide and adipose-derived stem cell conditioned medium (ADSCs-CM) combination on reparative dentin formation in Wistar rats through RUNX-2 and OPN expression
title_sort effect of calcium hydroxide and adipose derived stem cell conditioned medium adscs cm combination on reparative dentin formation in wistar rats through runx 2 and opn expression
topic Calcium hydroxide
Adipose-derived stem cells conditioned medium
Reparative dentin
RUNX-2
Osteopontin
Medicine
url http://www.sciencedirect.com/science/article/pii/S2212426825001459
work_keys_str_mv AT radixtioauzanfepiosandi effectofcalciumhydroxideandadiposederivedstemcellconditionedmediumadscscmcombinationonreparativedentinformationinwistarratsthroughrunx2andopnexpression
AT dianagustinwahjuningrum effectofcalciumhydroxideandadiposederivedstemcellconditionedmediumadscscmcombinationonreparativedentinformationinwistarratsthroughrunx2andopnexpression
AT galihsampoerno effectofcalciumhydroxideandadiposederivedstemcellconditionedmediumadscscmcombinationonreparativedentinformationinwistarratsthroughrunx2andopnexpression
AT anujbhardwaj effectofcalciumhydroxideandadiposederivedstemcellconditionedmediumadscscmcombinationonreparativedentinformationinwistarratsthroughrunx2andopnexpression
AT ajinkyampawar effectofcalciumhydroxideandadiposederivedstemcellconditionedmediumadscscmcombinationonreparativedentinformationinwistarratsthroughrunx2andopnexpression