Katalin Perczel-Kovách1, Orsolya Hegedűs2, Anna Földes3, Thanyaporn Sangngoen4, Karola Kálló5, Martin C Steward6, Gábor Varga7, Krisztina S Nagy8. 1. Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary. Electronic address: perczel.katalin@dent.semmelweis-univ.hu. 2. Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary. Electronic address: hegedus.orsolya@dent.semmelweis-univ.hu. 3. Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary. Electronic address: foldes.anna@dent.semmelweis-univ.hu. 4. Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary. Electronic address: thanyaporn.sang-ngoen@dent.semmelweis-univ.hu. 5. Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary. 6. Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary; School of Medical Sciences, University of Manchester, Manchester, United Kingdom. Electronic address: martin.steward@me.com. 7. Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary. Electronic address: varga.gabor@dent.semmelweis-univ.hu. 8. Department of Oral Biology, Semmelweis University, Nagyvárad Square 4. H-1089 Budapest, Hungary. Electronic address: nagy.krisztina@dent.semmelweis-univ.hu.
Abstract
OBJECTIVE: Although the osteogenic differentiation potential of mesenchymal stem cells of dental origin is well established, the roles of different marker proteins in this process remain to be clarified. Our aim was to compare the cellular and molecular changes, focusing in particular on mesenchymal stem cell markers, during in vitro osteogenesis in three dental stem cell types: dental follicle stem cells (DFSCs), periodontal ligament stem cells (PDLSCs) and dental pulp stem cells (DPSCs). DESIGN: Human DFSCs, PDLSCs and DPSCs were isolated, cultured and their osteogenic differentiation was induced for 3 weeks. Mineralization was assessed by von Kossa staining and calcium concentration measurements. The expression of mesenchymal and osteogenic markers was studied by immunocytochemistry and qPCR techniques. Alkaline phosphatase (ALP) activity and the frequency of STRO-1 positive cells were also quantified. RESULTS: The three cultures all showed abundant mineralization, with high calcium content by day 21. The expression of vimentin and nestin was sustained after osteogenic induction. The osteogenic medium induced a considerable elevation of STRO-1 positive cells. By day 7, the ALP mRNA level had increased more than 100-fold in DFSCs, PDLSCs, and DPSCs. Quantitative PCR results indicated dissimilarities of osteoblastic marker levels in the three dental stem cell cultures. CONCLUSIONS: DFSCs, PDLSCs and DPSCs have similar functional osteogenic differentiation capacities although their expressional profiles of key osteogenic markers show considerable variations. The STRO-1 positive cell fraction expands during osteogenic differentiation while vimentin and nestin expression remain high. For identification of stemness, functional studies rather than marker expressions are needed.
OBJECTIVE: Although the osteogenic differentiation potential of mesenchymal stem cells of dental origin is well established, the roles of different marker proteins in this process remain to be clarified. Our aim was to compare the cellular and molecular changes, focusing in particular on mesenchymal stem cell markers, during in vitro osteogenesis in three dental stem cell types: dental follicle stem cells (DFSCs), periodontal ligament stem cells (PDLSCs) and dental pulp stem cells (DPSCs). DESIGN:Human DFSCs, PDLSCs and DPSCs were isolated, cultured and their osteogenic differentiation was induced for 3 weeks. Mineralization was assessed by von Kossa staining and calcium concentration measurements. The expression of mesenchymal and osteogenic markers was studied by immunocytochemistry and qPCR techniques. Alkaline phosphatase (ALP) activity and the frequency of STRO-1 positive cells were also quantified. RESULTS: The three cultures all showed abundant mineralization, with high calcium content by day 21. The expression of vimentin and nestin was sustained after osteogenic induction. The osteogenic medium induced a considerable elevation of STRO-1 positive cells. By day 7, the ALP mRNA level had increased more than 100-fold in DFSCs, PDLSCs, and DPSCs. Quantitative PCR results indicated dissimilarities of osteoblastic marker levels in the three dental stem cell cultures. CONCLUSIONS: DFSCs, PDLSCs and DPSCs have similar functional osteogenic differentiation capacities although their expressional profiles of key osteogenic markers show considerable variations. The STRO-1 positive cell fraction expands during osteogenic differentiation while vimentin and nestin expression remain high. For identification of stemness, functional studies rather than marker expressions are needed.
Authors: Anna Földes; Hajnalka Reider; Anita Varga; Krisztina S Nagy; Katalin Perczel-Kovach; Katalin Kis-Petik; Pamela DenBesten; András Ballagi; Gábor Varga Journal: Polymers (Basel) Date: 2021-11-15 Impact factor: 4.329
Authors: Wijin Kim; Eungyeong Park; Hyuk Sang Yoo; Jongmin Park; Young Mee Jung; Ju Hyun Park Journal: Nanomaterials (Basel) Date: 2022-08-25 Impact factor: 5.719