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      A novel hydrogel scaffold for periodontal ligament stem cells

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          Abstract

          Periodontal ligament stem cells (PDLSCs) possess extensive regeneration potential. However, their therapeutic application demands a scaffold with appropriate properties. HydroMatrix (HydM) is a novel injectable peptide nanofiber hydrogel developed recently for cell culture. Our aim was to test whether HydM would be a suitable scaffold for proliferation and osteogenic differentiation of PDLSCs. PDLSCs were seeded on non-coated or HydM-coated surfaces. Both real-time impedance analysis and cell viability assay documented cell growth on HydM. PDLSCs showed healthy, fibroblast-like morphology on the hydrogel. After a 3-week-long culture in osteogenic medium, mineralization was much more intense in HydM cultures compared to control. Alkaline phosphatase activity of the cells grown on the gels reached the non-coated control levels. Our data provided evidence that PDLSCs can adhere, survive, migrate, and proliferate on HydM and this gel also supports their osteogenic differentiation. We first applied impedimetry for dental stem cells cultured on a scaffold. HydM is ideal for in vitro studies of PDLSCs. It may also serve not only as a reference material but also in the future as a promising biocompatible scaffold for preclinical studies.

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          Most cited references29

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          Hydrogels in pharmaceutical formulations.

          N. Peppas (2000)
          The availability of large molecular weight protein- and peptide-based drugs due to the recent advances in the field of molecular biology has given us new ways to treat a number of diseases. Synthetic hydrogels offer a possibly effective and convenient way to administer these compounds. Hydrogels are hydrophilic, three-dimensional networks, which are able to imbibe large amounts of water or biological fluids, and thus resemble, to a large extent, a biological tissue. They are insoluble due to the presence of chemical (tie-points, junctions) and/or physical crosslinks such as entanglements and crystallites. These materials can be synthesized to respond to a number of physiological stimuli present in the body, such as pH, ionic strength and temperature. The aim of this article is to present a concise review on the applications of hydrogels in the pharmaceutical field, hydrogel characterization and analysis of drug release from such devices.
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            In situ gelling hydrogels for pharmaceutical and biomedical applications.

            Since Wichterle et al. introduced hydrogels as novel materials possibly suitable for a variety of biomedical applications, hydrogel research has become a fast-developing and exciting research field. The soft and hydrophilic nature of hydrogels makes them particularly suitable as protein delivery system or as cell-entrapping scaffold in tissue engineering. Traditional hydrogels were formed by chemical crosslinking of water-soluble polymers or by polymerization (of mixtures) of water-soluble monomers. Because of incompatibility of these crosslinking methods with fragile molecules like pharmaceutical proteins and living cells, in recent years research interest has been focused on hydrogels that gel spontaneously under physiological conditions. In these systems, hydrogel formation occurs in situ, at the site of injection, without the aid of potentially toxic or denaturizing crosslinking agents. This review provides an overview of in situ gelling systems and their potential in biomedical applications. Both photopolymerizable as well as self-assembling hydrogels, based on either chemical crosslinks or physical interactions will be addressed.
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              Isolation and characterization of multipotent human periodontal ligament stem cells.

              Periodontal ligament (PDL) repair is thought to involve mesenchymal progenitor cells capable of forming fibroblasts, osteoblasts and cementoblasts. However, full characterization of PDL stem cell (SC) populations has not been achieved. To isolate and characterize PDLSC and assess their capability to differentiate into bone, cartilage and adipose tissue. Human PDL cells were stained for STRO-1, FACS sorted and expanded in culture. Human bone marrow SC (BMSC) served as a positive control. PDLSC and BMSC were cultured using standard conditions conducive for osteogenic, chondrogenic and adipogenic differentiation. Osteogenic induction was assayed using alizarine red S staining and expression of alkaline phosphatase (ALP) and bone sialoprotein (BSP). Adipogenic induction was assayed using Oil Red O staining and the expression of PPAR gamma 2 (early) and LPL (late) adipogenic markers. Chondrogenic induction was assayed by collagen type II expression and toluidine blue staining. Human PDL tissue contains about 27% STRO-1 positive cells with 3% strongly positive. In osteogenic cultures ALP was observed by day-7 in BMSC and day-14 in PDLSC. BSP expression was detectable by day-7; with more intense staining in PDLSC cultures. In adipogenic cultures both cell populations showed positive Oil Red O staining by day-25 with PPAR gamma 2 and LPL expression. By day-21, both BMSC and PDLSC chondrogenic induced cultures expressed collagen type II and glycosaminoglycans. The PDL contains SC that have the potential to differentiate into osteoblasts, chondrocytes and adipocytes, comparable with previously characterized BMSC. This adult PDLSC population can be utilized for potential therapeutic procedures related to PDL regeneration.
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                Author and article information

                Journal
                imas
                IMAS
                Interventional Medicine and Applied Science
                IMAS
                Akadémiai Kiadó (Budapest )
                2061-1617
                2061-5094
                25 July 2018
                September 2018
                : 10
                : 3
                : 162-170
                Affiliations
                [ 1 ]Department of Oral Biology, Semmelweis University , Budapest, Hungary
                [ 2 ]Department of Genetics, Cell- and Immunobiology, Semmelweis University , Budapest, Hungary
                [ 3 ]Department of Community Dentistry, Semmelweis University , Budapest, Hungary
                [ 4 ]Department of Oral Diagnostics, Semmelweis University , Budapest, Hungary
                Author notes
                [* ]Corresponding author: Gábor Varga, PhD, DSc; Department of Oral Biology, Semmelweis University, Nagyvárad tér 4, H-1089 Budapest, Hungary; Phone: +36 1 210 4415; Fax: +36 1 210 4421; E-mail: varga.gabor@ 123456dent.semmelweis-univ.hu
                Article
                10.1556/1646.10.2018.21
                6343580
                30713756
                9ea48e9a-e191-458a-bfb3-f3e4abee6edb
                © 2018 The Author(s)

                This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 International License, which permits unrestricted use, distribution, and reproduction in any medium for non-commercial purposes, provided the original author and source are credited, a link to the CC License is provided, and changes – if any – are indicated.

                History
                : 21 December 2017
                : 05 March 2018
                : 04 April 2018
                Page count
                Figures: 4, Tables: 0, Equations: 2, References: 32, Pages: 9
                Funding
                Funding sources: This work was supported by the Hungarian National Scientific Research Fund (OTKA-NKTH CK-80928) and Hungarian Human Resources Development Operational Programme (EFOP-3.6.2-16-2017-00006).
                Categories
                ORIGINAL PAPER

                Medicine,Immunology,Health & Social care,Microbiology & Virology,Infectious disease & Microbiology
                stem cell,HydroMatrix,impedimetry,osteogenic differentiation,periodontal ligament,cell proliferation

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