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Fulltext Cytotoxicity of gypsum-based biomaterial for direct pulp capping using stem cells from human exfoliated deciduous teeth

Subhi H, Reza F, Husein A, Nurul AA

J Conserv Dent, 2018 4 10;21(1):21-25.
PMID: 29628642 DOI: 10.4103/JCD.JCD_86_17

Aim: The aim of this study was to evaluate the cytotoxicity effects of experimental gypsum-based biomaterial prepared with various concentrations of chitosan (Gyp-CHT).
Materials and Methods: The study was performed using cell viability assay for mitochondrial dehydrogenase activity in stem cells from human exfoliated deciduous teeth (SHED), after 1, 2, and 3 days of exposure to the biomaterial extracts of varying concentrations. Differences in mean cell viability values were assessed by one-way analysis of variance, followed by Dunnett T3 post hoc test for multiple comparisons (P < 0.05).
Results: The cell viability to Gyp-CHT in low extract concentrations was statistically similar to that of the control and different from that of high extract concentrations. Gyp-5% CHT showed the highest percentage of cell viability with 110.92%, 108.56%, and 109.11%. The cell viability showed a tendency toward increment with low extract concentration and no constant effect of CHT on cell viability toward higher or lower.
Conclusions: Gyp-CHT biomaterial has no cytotoxic effects on the cultured SHED.
Fulltext Gypsum-Based Material for Dental Pulp Capping: Effect of Chitosan and BMP-2 on Physical, Mechanical, and Cellular Properties

Subhi H, Reza F, Husein A, Al Shehadat SA, Nurul AA

Int J Biomater, 2018;2018:3804293.
PMID: 30147725 DOI: 10.1155/2018/3804293

Effective pulp capping material must be biocompatible and have the ability to induce dentin bridge formation as well as having suitable physical and mechanical properties; however, many current materials do not satisfy the clinical requirements. This study aimed to assess the physical and mechanical properties of gypsum-based chitosan material (Gp-CT) and to evaluate its effects on cellular properties of stem cells from human exfoliated deciduous teeth (SHED). The experimental material was prepared with different concentrations of chitosan (CT) with or without BMP-2. Then, setting time, compressive strength, and pH were determined. In addition, cell viability, alkaline phosphatase (ALP) activity, and cell attachment were assessed. The setting time, compressive strength, and pH obtained were 4.1-6.6 min, 2.63-5.83 MPa, and 6.5-5.7, respectively. The cell viability to gypsum (Gp) with different CT concentrations was similar to that of the control on day 1 but statistically different from that of Gp alone on day 3. The ALP activity of SHED was significantly higher (p < 0.05) in CT- and BMP-2-containing materials than those in the control and Dycal at days 3 and 14. The scanning electron microscopy (SEM) image revealed that flattened cells were distributed across and adhered to the material surface. In conclusion, Gp-CT material shows promise as a potential material for direct pulp capping.
Chitosan-Based Accelerated Portland Cement Promotes Dentinogenic/Osteogenic Differentiation and Mineralization Activity of SHED

Subhi H, Husein A, Mohamad D, Nik Abdul Ghani NR, Nurul AA

Polymers (Basel), 2021 Sep 30;13(19).
PMID: 34641172 DOI: 10.3390/polym13193358

Calcium silicate-based cements (CSCs) are widely used in various endodontic treatments to promote wound healing and hard tissue formation. Chitosan-based accelerated Portland cement (APC-CT) is a promising and affordable material for endodontic use. This study investigated the effect of APC-CT on apoptosis, cell attachment, dentinogenic/osteogenic differentiation and mineralization activity of stem cells from human exfoliated deciduous teeth (SHED). APC-CT was prepared with various concentrations of chitosan (CT) solution (0%, 0.625%, 1.25% and 2.5% (w/v)). Cell attachment was determined by direct contact analysis using field emission scanning electron microscopy (FESEM); while the material extracts were used for the analyses of apoptosis by flow cytometry, dentinogenic/osteogenic marker expression by real-time PCR and mineralization activity by Alizarin Red and Von Kossa staining. The cells effectively attached to the surfaces of APC and APC-CT, acquiring flattened elongated and rounded-shape morphology. Treatment of SHED with APC and APC-CT extracts showed no apoptotic effect. APC-CT induced upregulation of DSPP, MEPE, DMP-1, OPN, OCN, OPG and RANKL expression levels in SHED after 14 days, whereas RUNX2, ALP and COL1A1 expression levels were downregulated. Mineralization assays showed a progressive increase in the formation of calcium deposits in cells with material containing higher CT concentration and with incubation time. In conclusion, APC-CT is nontoxic and promotes dentinogenic/osteogenic differentiation and mineralization activity of SHED, indicating its regenerative potential as a promising substitute for the commercially available CSCs to induce dentin/bone regeneration.

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