Morphological and Biological Evaluations of Human Periodontal Ligament Fibroblasts in Contact with Different Bovine Bone Grafts Treated with Low-Temperature Deproteinisation Protocol

Several types of deproteinised bovine bone mineral (DBBM) are available on the market, and each one is obtained with a thermic and chemical process that can differ, achieving different results. Currently, several protocols using low temperature are suggested to reduce the possible particle crystalli...

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Main Authors: Serena Bianchi, Sara Bernardi, Antonella Mattei, Loredana Cristiano, Leonardo Mancini, Diana Torge, Giuseppe Varvara, Guido Macchiarelli, Enrico Marchetti
Format: Article
Language:English
Published: MDPI AG 2022-05-01
Series:International Journal of Molecular Sciences
Subjects:
Online Access:https://www.mdpi.com/1422-0067/23/9/5273
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author Serena Bianchi
Sara Bernardi
Antonella Mattei
Loredana Cristiano
Leonardo Mancini
Diana Torge
Giuseppe Varvara
Guido Macchiarelli
Enrico Marchetti
author_facet Serena Bianchi
Sara Bernardi
Antonella Mattei
Loredana Cristiano
Leonardo Mancini
Diana Torge
Giuseppe Varvara
Guido Macchiarelli
Enrico Marchetti
author_sort Serena Bianchi
collection DOAJ
description Several types of deproteinised bovine bone mineral (DBBM) are available on the market, and each one is obtained with a thermic and chemical process that can differ, achieving different results. Currently, several protocols using low temperature are suggested to reduce the possible particle crystallisation during the production process. This study aimed to evaluate the biomorphological reaction of periodontal fibroblast cultures in contact with different DBBM particles treated with a low-temperature protocol (Thermagen<sup>®</sup>) and without exposure to sodium hydroxide (NaOH). Morphological evaluation was performed using light, confocal laser, and scanning electron microscopy, and the biological reaction in terms of proliferation was performed using an XTT proliferation assay at 24 h (T1), 72 h (T2), and 7 days (T3). The morphological analysis highlighted how the presence of the materials stimulated a change in the morphology of the cells into a polygonal shape, surface reactions with the thickening of the membrane, and expression of actin. In particular, the morphological changes were appreciable from T1, with a progressive increase in the considered morphological characteristics at T2 and T3 follow-ups. The proliferation assay showed a statistical significance between the different experimental materials and the negative control in T2 and T3 follow-ups. The post hoc analysis did not reveal any differences between the materials. In conclusion, the grafts obtained with the low-temperature extractions protocol and not exposed to NaOH solution showed positive morphological reactions with no differences in the sizes of particles.
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spelling doaj.art-c3320daa616d45669afb6e0547cb108a2023-11-23T08:29:38ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-05-01239527310.3390/ijms23095273Morphological and Biological Evaluations of Human Periodontal Ligament Fibroblasts in Contact with Different Bovine Bone Grafts Treated with Low-Temperature Deproteinisation ProtocolSerena Bianchi0Sara Bernardi1Antonella Mattei2Loredana Cristiano3Leonardo Mancini4Diana Torge5Giuseppe Varvara6Guido Macchiarelli7Enrico Marchetti8Department of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, ItalyDepartment of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, ItalyDepartment of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, ItalyDepartment of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, ItalyDepartment of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, ItalyDepartment of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, ItalyDepartment of Innovative Technologies in Medicine and Dentistry, University of Chieti-Pescara ‘Gabriele d’Annunzio’, Via Dei Vestini 11, 66100 Chieti, ItalyDepartment of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, ItalyDepartment of Life, Health and Environmental Sciences, University of L’Aquila, 67100 L’Aquila, ItalySeveral types of deproteinised bovine bone mineral (DBBM) are available on the market, and each one is obtained with a thermic and chemical process that can differ, achieving different results. Currently, several protocols using low temperature are suggested to reduce the possible particle crystallisation during the production process. This study aimed to evaluate the biomorphological reaction of periodontal fibroblast cultures in contact with different DBBM particles treated with a low-temperature protocol (Thermagen<sup>®</sup>) and without exposure to sodium hydroxide (NaOH). Morphological evaluation was performed using light, confocal laser, and scanning electron microscopy, and the biological reaction in terms of proliferation was performed using an XTT proliferation assay at 24 h (T1), 72 h (T2), and 7 days (T3). The morphological analysis highlighted how the presence of the materials stimulated a change in the morphology of the cells into a polygonal shape, surface reactions with the thickening of the membrane, and expression of actin. In particular, the morphological changes were appreciable from T1, with a progressive increase in the considered morphological characteristics at T2 and T3 follow-ups. The proliferation assay showed a statistical significance between the different experimental materials and the negative control in T2 and T3 follow-ups. The post hoc analysis did not reveal any differences between the materials. In conclusion, the grafts obtained with the low-temperature extractions protocol and not exposed to NaOH solution showed positive morphological reactions with no differences in the sizes of particles.https://www.mdpi.com/1422-0067/23/9/5273biomaterialcell culturedeproteinised bovine bonefibroblastsperiodontal regeneration
spellingShingle Serena Bianchi
Sara Bernardi
Antonella Mattei
Loredana Cristiano
Leonardo Mancini
Diana Torge
Giuseppe Varvara
Guido Macchiarelli
Enrico Marchetti
Morphological and Biological Evaluations of Human Periodontal Ligament Fibroblasts in Contact with Different Bovine Bone Grafts Treated with Low-Temperature Deproteinisation Protocol
International Journal of Molecular Sciences
biomaterial
cell culture
deproteinised bovine bone
fibroblasts
periodontal regeneration
title Morphological and Biological Evaluations of Human Periodontal Ligament Fibroblasts in Contact with Different Bovine Bone Grafts Treated with Low-Temperature Deproteinisation Protocol
title_full Morphological and Biological Evaluations of Human Periodontal Ligament Fibroblasts in Contact with Different Bovine Bone Grafts Treated with Low-Temperature Deproteinisation Protocol
title_fullStr Morphological and Biological Evaluations of Human Periodontal Ligament Fibroblasts in Contact with Different Bovine Bone Grafts Treated with Low-Temperature Deproteinisation Protocol
title_full_unstemmed Morphological and Biological Evaluations of Human Periodontal Ligament Fibroblasts in Contact with Different Bovine Bone Grafts Treated with Low-Temperature Deproteinisation Protocol
title_short Morphological and Biological Evaluations of Human Periodontal Ligament Fibroblasts in Contact with Different Bovine Bone Grafts Treated with Low-Temperature Deproteinisation Protocol
title_sort morphological and biological evaluations of human periodontal ligament fibroblasts in contact with different bovine bone grafts treated with low temperature deproteinisation protocol
topic biomaterial
cell culture
deproteinised bovine bone
fibroblasts
periodontal regeneration
url https://www.mdpi.com/1422-0067/23/9/5273
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