Asma Abdullah Nurul

The Modulation of PPARγ1 and PPARγ2 mRNA Expression by Ciglitazone in CD3/CD28-Activated Naïve and Memory CD4+ T Cells

Given their roles in immune regulation, the expression of the nuclear receptor peroxisome proliferator-activated receptor γ (PPARγ) 1 and 2 isoforms was investigated in human naïve (CD45RA+) and memory (CD45RO+) CD4+ T cells. Stimulation of both types of cells via the CD3/CD28 pathway resulted in high expression of both PPARγ receptors as measured by real-time PCR. Treatment with the PPARγ agonist, ciglitazone, increased PPARγ1 expression but decreased PPARγ2 expression in stimulated naïve and memory cells. Furthermore, when present, the magnitude of both PPARγ receptors expression was lower in naïve cells, perhaps suggesting a lower regulatory control of these cells. Similar profiles of selected proinflammatory cytokines were expressed by the two cell types following stimulation. The induction of PPARγ1 and suppression of PPARγ2 expressions in naïve and memory CD4+ T cells in the presence of ciglitazone suggest that the PPARγ subtypes may have different roles in the regulation of T-cell function.

Gypsum-based biomaterials: Evaluation of physical and mechanical properties, cellular effects and its potential as a pulp liner.

This in vitro study aimed to evaluate setting time and compressive strength of gypsum-based chitosan biomaterials and its effect on proliferation of stem cells from human exfoliated deciduous teeth (SHED) and alkaline phosphatase (ALP) activity. Pure-GYP was mixed with water (2.5 g: 1.9 mL); Gyp-CHT was prepared with gypsum, chitosan, and water (2.5 g: 0.285 g: 1.9 mL). Cell viability and ALP activity were assessed at different periods. Data were analyzed using SPSS (p<0.05). The setting times were 2.7 min and 2.8 min for pure-GYP and Gyp-CHT, respectively. Significantly higher compressive strength was observed with Gyp-CHT. SHED treatments with both materials were not cytotoxic. ALP was consistently higher in the treated groups compared with the control. Cellular attachments were evident with SEM. Excellent cellular viability with pure-GYP and Gyp-CHT, as well as increased ALP activities, suggested the possibility of tertiary dentin formation. Further studies are necessary to evaluate the biomaterials for its pulp protective potentialities.

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

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.

Interleukin-17A promotes osteogenic differentiation by increasing OPG/RANKL ratio in stem cells from human exfoliated deciduous teeth (SHED)

Stem cells derived from human exfoliated deciduous teeth (SHED) represent a promising cell source for bone tissue regeneration. This study evaluated the effects of interleukin‐17A (IL‐17A) on the osteogenic differentiation of SHED. SHED were cultured in complete alpha minimum essential medium supplemented with osteoinducing reagents and treated with recombinant IL‐17A. The cells were quantitatively analysed for proliferative activity by MTS assay, cell markers expression, and apoptotic activity by flow cytometry. For osteogenic differentiation, alkaline phosphatase (ALP) activity was quantified; mineralization assays were carried out using von Kossa and Alizarin red, and expression of osteogenic markers were analysed by real‐time polymerase chain reaction and Western blot. The results showed that treatment with IL‐17A increased proliferative activity in a dose‐dependent manner, but reduced the expression of stem cell markers (c‐Myc and Nanog) as the days progressed. IL‐17A induced osteogenic differentiation in SHED as evidenced by high ALP activity, increased matrix mineralization, and upregulation of the mRNA expression of the osteogenic markers ALP, alpha 1 type 1 collagen (Col1A1), runt‐related transcription factor 2 (RUNX2), osteopontin (OPN), osteocalcin (OCN), and osteoprotegerin (OPG) but downregulation of receptor activator of nuclear factor κB ligand (RANKL) as well as altering the OPG/RANKL ratio. Findings from our study indicate that IL‐17A enhances proliferation and osteogenic differentiation of SHED by regulating OPG/RANKL mechanism thus suggests therapeutic potential of IL‐17A in bone regeneration.

Gypsum-Based Material for Dental Pulp Capping: Effect of Chitosan and BMP-2 on Physical, Mechanical, and Cellular Properties

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.

Cementoblastic lineage formation in the cross-talk between stem cells of human exfoliated deciduous teeth and epithelial rests of Malassez cells.

ObjectivesThe purpose of this study was to evaluate the synergistic effect of epithelial rests of Malassez cells (ERM) and transforming growth factor-β1 (TGF-β1) on proliferation, cementogenic and osteogenic differentiation of stem cells derived from human exfoliated deciduous teeth (SHED).Materials and methodsSHED were co-cultured with ERM with/without TGF-β1. Then, SHED proliferation, morphological appearance, alkaline phosphatase (ALP) activity, mineralization behaviour and gene/protein expression of cemento/osteoblastic phenotype were evaluated.ResultsTGF-β1 enhanced SHED proliferation when either cultured alone or co-cultured with ERM. ERM induced the cementoblastic differentiation of SHED which was significantly accelerated when treated with TGF-β1. This activity was demonstrated by high ALP activity, strong mineral deposition and upregulation of cementum/bone-related gene and protein expressions (i.e. ALP, collagen type I, bone sialoprotein, osteocalcin and cementum attachment protein).ConclusionsERM were able to induce SHED differentiation along the cemento/osteoblastic lineage that was triggered in the presence of TGF-β1.Clinical relevanceThe cemento/osteoblastic differentiation capability of SHED possesses a therapeutic potential in endodontic and periodontal tissue engineering.