The Influence of Vitamin C–incorporated Polycaprolactone on Osteogenesis in Osteoblast-Osteoclast Co-culture In Vitro

ABSTRACT: Objectives: Implantation of biomaterials generates reactive oxygen species (ROS) in the peri-implant microenvironment. Bone loss occurs when ROS levels exceed the local antioxidant capacity. The aim of this study was to investigate the influence of vitamin C–incorporated polycaprolactone...

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Main Authors: Elaf Akram Abdulhameed, K.G. Aghila Rani, Ensanya A. Abou Neel, Nadia Khalifa, Yanti Johari, Marzuki Omar, Ab Rani Samsudin
Format: Article
Language:English
Published: Elsevier 2025-10-01
Series:International Dental Journal
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Online Access:http://www.sciencedirect.com/science/article/pii/S0020653925002369
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author Elaf Akram Abdulhameed
K.G. Aghila Rani
Ensanya A. Abou Neel
Nadia Khalifa
Yanti Johari
Marzuki Omar
Ab Rani Samsudin
author_facet Elaf Akram Abdulhameed
K.G. Aghila Rani
Ensanya A. Abou Neel
Nadia Khalifa
Yanti Johari
Marzuki Omar
Ab Rani Samsudin
author_sort Elaf Akram Abdulhameed
collection DOAJ
description ABSTRACT: Objectives: Implantation of biomaterials generates reactive oxygen species (ROS) in the peri-implant microenvironment. Bone loss occurs when ROS levels exceed the local antioxidant capacity. The aim of this study was to investigate the influence of vitamin C–incorporated polycaprolactone (PCL-Vit C) membrane in scavenging ROS and enhancing biomineralisation in osteoblast–osteoclast (OB-OC) co-culture system. Methods: OB-OCs were cultured on polycaprolactone (PCL) and PCL-Vit C membranes under osteogenic conditions to mimic the bone microenvironment. ROS generation was measured using flow cytometry. ALP and the RANKL/OPG ratio were determined by colorimetric and ELISA assays, respectively. Gene expression of ALP, Col1, Runx-2 and OCN and protein expression of Runx-2, BMP-7, Col1 and OCN were determined by real-time PCR and western blotting, respectively. Activation of P38, ERK and JNK and beta-catenin expression was also analysed. Results: OB-OC grown on PCL membrane generated a higher amount of ROS compared to those on PCL-Vit C. Colorimetric assays revealed a significantly higher ALP activity in OB-OC co-cultures on PCL-Vit C membranes. Furthermore, OB-OC grown on PCL-Vit C membrane showed significant upregulation in mRNA levels of ALP, Col1 and OCN with lower RANKL/OPG ratio and higher amounts of mineralisation nodules. Runx-2 expression was comparable in both membranes. Western blotting showed a significant increase in phosphorylation of P38 MAPK, ERK, JNK and beta-catenin expression in OB-OC maintained on PCL membrane, plausibly due to increased ROS levels, compared to PCL-Vit C. Significance: OB-OC grown on PCL-Vit C membrane scavenged ROS and supported a higher osteogenic potential environment for OB-OC co-culture in vitro.
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spelling doaj-art-a05fd0d6399446f7beecc2092e4201832025-08-22T04:54:39ZengElsevierInternational Dental Journal0020-65392025-10-0175510094910.1016/j.identj.2025.100949The Influence of Vitamin C–incorporated Polycaprolactone on Osteogenesis in Osteoblast-Osteoclast Co-culture In VitroElaf Akram Abdulhameed0K.G. Aghila Rani1Ensanya A. Abou Neel2Nadia Khalifa3Yanti Johari4Marzuki Omar5Ab Rani Samsudin6Restorative & Preventive Dentistry Department, College of Dental Medicine, University of Sharjah, Sharjah, United Arab Emirates; School of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, MalaysiaResearch Institute for Medical and Health Sciences, University of Sharjah, Sharjah, United Arab EmiratesRestorative & Preventive Dentistry Department, College of Dental Medicine, University of Sharjah, Sharjah, United Arab Emirates; UCL Eastman Dental Institute, Biomaterials & Tissue Engineering Division, Royal Free Hospital, Rowland Hill Street, London, UKRestorative & Preventive Dentistry Department, College of Dental Medicine, University of Sharjah, Sharjah, United Arab EmiratesSchool of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, MalaysiaSchool of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, Malaysia; Corresponding authors. College of Dental Medicine, Oral & Craniofacial Health Sciences; School of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, Malaysia, Tel: +97165057600.Oral & Craniofacial Health Sciences, College of Dental Medicine, University of Sharjah, Sharjah, United Arab Emirates; Corresponding authors. College of Dental Medicine, Oral & Craniofacial Health Sciences; School of Dental Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, Malaysia, Tel: +97165057600.ABSTRACT: Objectives: Implantation of biomaterials generates reactive oxygen species (ROS) in the peri-implant microenvironment. Bone loss occurs when ROS levels exceed the local antioxidant capacity. The aim of this study was to investigate the influence of vitamin C–incorporated polycaprolactone (PCL-Vit C) membrane in scavenging ROS and enhancing biomineralisation in osteoblast–osteoclast (OB-OC) co-culture system. Methods: OB-OCs were cultured on polycaprolactone (PCL) and PCL-Vit C membranes under osteogenic conditions to mimic the bone microenvironment. ROS generation was measured using flow cytometry. ALP and the RANKL/OPG ratio were determined by colorimetric and ELISA assays, respectively. Gene expression of ALP, Col1, Runx-2 and OCN and protein expression of Runx-2, BMP-7, Col1 and OCN were determined by real-time PCR and western blotting, respectively. Activation of P38, ERK and JNK and beta-catenin expression was also analysed. Results: OB-OC grown on PCL membrane generated a higher amount of ROS compared to those on PCL-Vit C. Colorimetric assays revealed a significantly higher ALP activity in OB-OC co-cultures on PCL-Vit C membranes. Furthermore, OB-OC grown on PCL-Vit C membrane showed significant upregulation in mRNA levels of ALP, Col1 and OCN with lower RANKL/OPG ratio and higher amounts of mineralisation nodules. Runx-2 expression was comparable in both membranes. Western blotting showed a significant increase in phosphorylation of P38 MAPK, ERK, JNK and beta-catenin expression in OB-OC maintained on PCL membrane, plausibly due to increased ROS levels, compared to PCL-Vit C. Significance: OB-OC grown on PCL-Vit C membrane scavenged ROS and supported a higher osteogenic potential environment for OB-OC co-culture in vitro.http://www.sciencedirect.com/science/article/pii/S0020653925002369Osteoblast–osteoclast co-culturePolycaprolactoneVitamin CReactive oxygen speciesOxidative stressOsteogenic potential
spellingShingle Elaf Akram Abdulhameed
K.G. Aghila Rani
Ensanya A. Abou Neel
Nadia Khalifa
Yanti Johari
Marzuki Omar
Ab Rani Samsudin
The Influence of Vitamin C–incorporated Polycaprolactone on Osteogenesis in Osteoblast-Osteoclast Co-culture In Vitro
International Dental Journal
Osteoblast–osteoclast co-culture
Polycaprolactone
Vitamin C
Reactive oxygen species
Oxidative stress
Osteogenic potential
title The Influence of Vitamin C–incorporated Polycaprolactone on Osteogenesis in Osteoblast-Osteoclast Co-culture In Vitro
title_full The Influence of Vitamin C–incorporated Polycaprolactone on Osteogenesis in Osteoblast-Osteoclast Co-culture In Vitro
title_fullStr The Influence of Vitamin C–incorporated Polycaprolactone on Osteogenesis in Osteoblast-Osteoclast Co-culture In Vitro
title_full_unstemmed The Influence of Vitamin C–incorporated Polycaprolactone on Osteogenesis in Osteoblast-Osteoclast Co-culture In Vitro
title_short The Influence of Vitamin C–incorporated Polycaprolactone on Osteogenesis in Osteoblast-Osteoclast Co-culture In Vitro
title_sort influence of vitamin c incorporated polycaprolactone on osteogenesis in osteoblast osteoclast co culture in vitro
topic Osteoblast–osteoclast co-culture
Polycaprolactone
Vitamin C
Reactive oxygen species
Oxidative stress
Osteogenic potential
url http://www.sciencedirect.com/science/article/pii/S0020653925002369
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