Effects of Connective Tissue Growth Factor on the Cell Viability, Proliferation, Osteogenic Capacity and mRNA Expression of Stem Cell Spheroids

Background: Connective tissue growth factor (CTGF) is a cellular communication network factor family protein involved in many cellular functions. The purpose of this study was to determine the effects of CTGF on the proliferation, osteogenic capacity, and mRNA expression of spheroids composed of gin...

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Main Authors: Abdullah Zaki Alnahash, Young-Min Song, Sae-Kyung Min, Hyun-Jin Lee, Min-Ji Kim, Yoon-Hee Park, Je-Uk Park, Jun-Beom Park
Format: Article
Language:English
Published: MDPI AG 2021-07-01
Series:Applied Sciences
Subjects:
Online Access:https://www.mdpi.com/2076-3417/11/14/6572
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spelling doaj-737601b30c7345bcba0ef0e5aca132a32021-07-23T13:30:04ZengMDPI AGApplied Sciences2076-34172021-07-01116572657210.3390/app11146572Effects of Connective Tissue Growth Factor on the Cell Viability, Proliferation, Osteogenic Capacity and mRNA Expression of Stem Cell SpheroidsAbdullah Zaki Alnahash0Young-Min Song1Sae-Kyung Min2Hyun-Jin Lee3Min-Ji Kim4Yoon-Hee Park5Je-Uk Park6Jun-Beom Park7Department of Oral and Maxillofacial Surgery, College of Medicine, The Catholic University of Korea, Seoul 06591, KoreaDepartment of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, KoreaDepartment of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, KoreaDepartment of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, KoreaCollege of Dentistry, Chosun University, Gwangju 61452, KoreaEbiogen, #405, Sungsu A1 Center 48 Ttukseom-ro 17-ga-gil, Seoul 04785, KoreaDepartment of Oral and Maxillofacial Surgery, College of Medicine, The Catholic University of Korea, Seoul 06591, KoreaDepartment of Periodontics, College of Medicine, The Catholic University of Korea, Seoul 06591, KoreaBackground: Connective tissue growth factor (CTGF) is a cellular communication network factor family protein involved in many cellular functions. The purpose of this study was to determine the effects of CTGF on the proliferation, osteogenic capacity, and mRNA expression of spheroids composed of gingiva-derived mesenchymal stem cells (GMSCs). Methods: CTGF was applied at final concentrations of 0, 25, 50, 100, and 200 ng/mL. Qualitative cell viability was determined using Live/Dead kit assay. Metabolic viability was determined with a colorimetric assay kit. Osteogenic activity was analyzed with alkaline phosphatase activity and Alizarin Red S staining. Quantitative polymerase chain reaction (qPCR) was used to assess the expression levels of RUNX2, BSP, OCN, and COL1A1. Results: In general, there was no significant difference in cell viability between the groups on Days 1, 4, and 7. Addition of CTGF produced an increase in Alizarin Red S staining. qPCR results demonstrated that the mRNA expression levels of RUNX2, BSP, OCN, and COL1A1 were significantly increased with the addition of CTGF. Conclusions: Based on these findings, we conclude that CTGF can be applied for increased osteogenic differentiation of stem cell spheroids.https://www.mdpi.com/2076-3417/11/14/6572cellular spheroidscell survivalconnective tissue growth factorgingivaosteogenesisstem cells
collection DOAJ
language English
format Article
sources DOAJ
author Abdullah Zaki Alnahash
Young-Min Song
Sae-Kyung Min
Hyun-Jin Lee
Min-Ji Kim
Yoon-Hee Park
Je-Uk Park
Jun-Beom Park
spellingShingle Abdullah Zaki Alnahash
Young-Min Song
Sae-Kyung Min
Hyun-Jin Lee
Min-Ji Kim
Yoon-Hee Park
Je-Uk Park
Jun-Beom Park
Effects of Connective Tissue Growth Factor on the Cell Viability, Proliferation, Osteogenic Capacity and mRNA Expression of Stem Cell Spheroids
Applied Sciences
cellular spheroids
cell survival
connective tissue growth factor
gingiva
osteogenesis
stem cells
author_facet Abdullah Zaki Alnahash
Young-Min Song
Sae-Kyung Min
Hyun-Jin Lee
Min-Ji Kim
Yoon-Hee Park
Je-Uk Park
Jun-Beom Park
author_sort Abdullah Zaki Alnahash
title Effects of Connective Tissue Growth Factor on the Cell Viability, Proliferation, Osteogenic Capacity and mRNA Expression of Stem Cell Spheroids
title_short Effects of Connective Tissue Growth Factor on the Cell Viability, Proliferation, Osteogenic Capacity and mRNA Expression of Stem Cell Spheroids
title_full Effects of Connective Tissue Growth Factor on the Cell Viability, Proliferation, Osteogenic Capacity and mRNA Expression of Stem Cell Spheroids
title_fullStr Effects of Connective Tissue Growth Factor on the Cell Viability, Proliferation, Osteogenic Capacity and mRNA Expression of Stem Cell Spheroids
title_full_unstemmed Effects of Connective Tissue Growth Factor on the Cell Viability, Proliferation, Osteogenic Capacity and mRNA Expression of Stem Cell Spheroids
title_sort effects of connective tissue growth factor on the cell viability, proliferation, osteogenic capacity and mrna expression of stem cell spheroids
publisher MDPI AG
series Applied Sciences
issn 2076-3417
publishDate 2021-07-01
description Background: Connective tissue growth factor (CTGF) is a cellular communication network factor family protein involved in many cellular functions. The purpose of this study was to determine the effects of CTGF on the proliferation, osteogenic capacity, and mRNA expression of spheroids composed of gingiva-derived mesenchymal stem cells (GMSCs). Methods: CTGF was applied at final concentrations of 0, 25, 50, 100, and 200 ng/mL. Qualitative cell viability was determined using Live/Dead kit assay. Metabolic viability was determined with a colorimetric assay kit. Osteogenic activity was analyzed with alkaline phosphatase activity and Alizarin Red S staining. Quantitative polymerase chain reaction (qPCR) was used to assess the expression levels of RUNX2, BSP, OCN, and COL1A1. Results: In general, there was no significant difference in cell viability between the groups on Days 1, 4, and 7. Addition of CTGF produced an increase in Alizarin Red S staining. qPCR results demonstrated that the mRNA expression levels of RUNX2, BSP, OCN, and COL1A1 were significantly increased with the addition of CTGF. Conclusions: Based on these findings, we conclude that CTGF can be applied for increased osteogenic differentiation of stem cell spheroids.
topic cellular spheroids
cell survival
connective tissue growth factor
gingiva
osteogenesis
stem cells
url https://www.mdpi.com/2076-3417/11/14/6572
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