Bio-Morphological Reaction of Human Periodontal Ligament Fibroblasts to Different Types of Dentinal Derivates: In Vitro Study

Abstract

Understanding the biological and morphological reactions of human cells towards different dentinal derivate grafting materials is fundamental for choosing the type of dentin for specific clinical situations. This study aimed to evaluate human periodontal ligament fibroblasts (hPLF) cells exposed to different dentinal derivates particles. The study design included the in vitro evaluation of mineralized dentine (SG), deproteinized and demineralized dentine (DDP), and demineralized dentine (TT) as test materials and of deproteinized bovine bone (BIOS) as the positive control material. The materials were kept with the hPLF cell line, and the evaluations were made after 24 h, 72 h, and 7 days of in vitro culture. The evaluated outcomes were proliferation by using XTT assays, the morphological characteristics by light microscopy (LM) and by the use of scanning electron microscopy (SEM), and adhesion by using confocal microscopy (CLSM). Overall, the experimental materials induced a positive response of the hPLFs in terms of proliferation and adhesion. The XTT assay showed the TT, and the SG induced significant growth compared to the negative control at 7 days follow-up. The morphological data supported the XTT assay: the LM observations showed the presence of densely packed cells with a modified shape; the SEM observations allowed the assessment of how fibroblasts exposed to DDP and TT presented cytoplasmatic extensions; and SG and BIOS also presented the thickening of the cellular membrane. The CLMS observations showed the expression of the proliferative marker, as well as and the expression of cytoskeletal elements involved in the adhesion process. In particular, the vinculin and integrin signals were stronger at 72 h, while the actin signal remained constantly expressed in all the follow-up of the sample exposed to SG material. The integrin signal was stronger at 72 h, and the vinculin and actin signals were stronger at 7 days follow-up in the sample exposed to DDP material. The vinculin and integrin signals were stronger at 72 h follow-up in the sample exposed to TT material; vinculin and integrin signals appear stronger at 24 h follow-up in the sample exposed to BIOS material. These data confirmed how dentinal derivates present satisfying biocompatibility and high conductivity and inductivity properties fundamental in the regenerative processes. Furthermore, the knowledge of the effects of the dentin’s degree of mineralization on cellular behavior will help clinicians choose the type of dentine derivates material according to the required clinical situation.

Autogenous Dentin Particulate Graft for Alveolar Ridge Augmentation with and without Use of Collagen Membrane: Preliminary Histological Analysis on Humans

“…An explanation of this phenomenon could be found by comparing the result with other studies on the subject. A review showed that all preliminary data regarding the Tooth Transformer method demonstrated histomorphometric data, like the results of this study, when a membrane is used [35][36][37].…”

Regeneration of periodontal bone defects with mesenchymal stem cells in animal models. Systematic review and meta-analysis

“…Bianchi et al studied the bio-morphological reaction of human periodontal ligament fibroblasts to different types of dentinal derivates (mineralized dentine, deproteinized and demineralized dentine, and demineralized dentine), and a positive response was observed in terms of proliferation and adhesion, with stronger vinculin and integrin signal. This therefore confirms that dentinal derivates present high conductivity and inductivity properties in the regenerative processes [18].…”

A prospective clinical study with one year follow up of deep caries management using a novel biomaterial

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