A Biomimetic Polynucleotides–Hyaluronic Acid Hydrogel Promotes the Growth of 3D Spheroid Cultures of Gingival Fibroblasts

(1) Background: Three-dimensional cultures are useful tools to evaluate regenerative approaches in vitro, as they may mimic the spatial arrangement of cells more closely to natural tissues than routine 2D culture methods. (2) Methods: We investigated the effects of a polynucleotide, hyaluronic acid...

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Main Authors: Maria Teresa Colangelo, Maria Luisa Vicedomini, Silvana Belletti, Paolo Govoni, Stefano Guizzardi, Carlo Galli
Format: Article
Language:English
Published: MDPI AG 2023-01-01
Series:Applied Sciences
Subjects:
Online Access:https://www.mdpi.com/2076-3417/13/2/743
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author Maria Teresa Colangelo
Maria Luisa Vicedomini
Silvana Belletti
Paolo Govoni
Stefano Guizzardi
Carlo Galli
author_facet Maria Teresa Colangelo
Maria Luisa Vicedomini
Silvana Belletti
Paolo Govoni
Stefano Guizzardi
Carlo Galli
author_sort Maria Teresa Colangelo
collection DOAJ
description (1) Background: Three-dimensional cultures are useful tools to evaluate regenerative approaches in vitro, as they may mimic the spatial arrangement of cells more closely to natural tissues than routine 2D culture methods. (2) Methods: We investigated the effects of a polynucleotide, hyaluronic acid (PN, HA) compound on 3D spheroid cultures of primary gingival fibroblasts, by measuring their morphology over time, cell viability with Calcein-AM, a fluorescent marker, and cell growth potential by re-plating spheroids in attachment-permissive regular culture plates under routine conditions and following them up for 15 days. (3) Results: PN + HA induced an increase in spheroid size and perimeter and a decrease in spheroid circularity, as cells tended to grow and form small peripheral stacks around the spheroid. Levels of cell viability were also higher in this group. After re-plating, only the spheroids previously stimulated with PN + HA dissolved completely during the second week of culture and colonized the plate, thus indicating the retention of a higher level of viability by the cells forming the whole spheroid with this stimulus. (4) Conclusions: Taken together, our data support the idea that the combination of PN and HA has synergic effects on primary fibroblasts and promotes their viability, the growth of 3D cellular constructs, and the retention of a remarkable proliferative potential over the course of the experimental period, making it a promising compound for further investigations.
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spelling doaj.art-fb28ae8af2614639b7cb1fc5b2bdd3372023-11-30T21:00:49ZengMDPI AGApplied Sciences2076-34172023-01-0113274310.3390/app13020743A Biomimetic Polynucleotides–Hyaluronic Acid Hydrogel Promotes the Growth of 3D Spheroid Cultures of Gingival FibroblastsMaria Teresa Colangelo0Maria Luisa Vicedomini1Silvana Belletti2Paolo Govoni3Stefano Guizzardi4Carlo Galli5Histology and Embryology Lab, Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43126 Parma, ItalyHistology and Embryology Lab, Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43126 Parma, ItalyHistology and Embryology Lab, Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43126 Parma, ItalyHistology and Embryology Lab, Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43126 Parma, ItalyHistology and Embryology Lab, Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43126 Parma, ItalyHistology and Embryology Lab, Department of Medicine and Surgery, University of Parma, Via Volturno 39, 43126 Parma, Italy(1) Background: Three-dimensional cultures are useful tools to evaluate regenerative approaches in vitro, as they may mimic the spatial arrangement of cells more closely to natural tissues than routine 2D culture methods. (2) Methods: We investigated the effects of a polynucleotide, hyaluronic acid (PN, HA) compound on 3D spheroid cultures of primary gingival fibroblasts, by measuring their morphology over time, cell viability with Calcein-AM, a fluorescent marker, and cell growth potential by re-plating spheroids in attachment-permissive regular culture plates under routine conditions and following them up for 15 days. (3) Results: PN + HA induced an increase in spheroid size and perimeter and a decrease in spheroid circularity, as cells tended to grow and form small peripheral stacks around the spheroid. Levels of cell viability were also higher in this group. After re-plating, only the spheroids previously stimulated with PN + HA dissolved completely during the second week of culture and colonized the plate, thus indicating the retention of a higher level of viability by the cells forming the whole spheroid with this stimulus. (4) Conclusions: Taken together, our data support the idea that the combination of PN and HA has synergic effects on primary fibroblasts and promotes their viability, the growth of 3D cellular constructs, and the retention of a remarkable proliferative potential over the course of the experimental period, making it a promising compound for further investigations.https://www.mdpi.com/2076-3417/13/2/743polynucleotidesfibroblastshyaluronic acidspheroids3D cultures
spellingShingle Maria Teresa Colangelo
Maria Luisa Vicedomini
Silvana Belletti
Paolo Govoni
Stefano Guizzardi
Carlo Galli
A Biomimetic Polynucleotides–Hyaluronic Acid Hydrogel Promotes the Growth of 3D Spheroid Cultures of Gingival Fibroblasts
Applied Sciences
polynucleotides
fibroblasts
hyaluronic acid
spheroids
3D cultures
title A Biomimetic Polynucleotides–Hyaluronic Acid Hydrogel Promotes the Growth of 3D Spheroid Cultures of Gingival Fibroblasts
title_full A Biomimetic Polynucleotides–Hyaluronic Acid Hydrogel Promotes the Growth of 3D Spheroid Cultures of Gingival Fibroblasts
title_fullStr A Biomimetic Polynucleotides–Hyaluronic Acid Hydrogel Promotes the Growth of 3D Spheroid Cultures of Gingival Fibroblasts
title_full_unstemmed A Biomimetic Polynucleotides–Hyaluronic Acid Hydrogel Promotes the Growth of 3D Spheroid Cultures of Gingival Fibroblasts
title_short A Biomimetic Polynucleotides–Hyaluronic Acid Hydrogel Promotes the Growth of 3D Spheroid Cultures of Gingival Fibroblasts
title_sort biomimetic polynucleotides hyaluronic acid hydrogel promotes the growth of 3d spheroid cultures of gingival fibroblasts
topic polynucleotides
fibroblasts
hyaluronic acid
spheroids
3D cultures
url https://www.mdpi.com/2076-3417/13/2/743
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