Mariam Binti Abdullah

Differentiation of Dental Pulp Stem Cells into Islet-like Aggregates

The post-natal dental pulp tissue contains a population of multipotent mesenchymal progenitor cells known as dental pulp stromal/stem cells (DPSCs), with high proliferative potential for self-renewal. In this investigation, we explored the potential of DPSCs to differentiate into pancreatic cell lineage resembling islet-like cell aggregates (ICAs). We isolated, propagated, and characterized DPSCs and demonstrated that these could be differentiated into adipogenic, chondrogenic, and osteogenic lineage upon exposure to an appropriate cocktail of differentiating agents. Using a three-step protocol reported previously by our group, we succeeded in obtaining ICAs from DPSCs. The identity of ICAs was confirmed as islets by dithiozone-positive staining, as well as by expression of C-peptide, Pdx-1, Pax4, Pax6, Ngn3, and Isl-1. There were several-fold up-regulations of these transcription factors proportional to days of differentiation as compared with undifferentiated DPSCs. Day 10 ICAs released insulin and C-peptide in a glucose-dependent manner, exhibiting in vitro functionality. Our results demonstrated for the first time that DPSCs could be differentiated into pancreatic cell lineage and offer an unconventional and non-controversial source of human tissue that could be used for autologous stem cell therapy in diabetes.

Human platelet lysate permits scale-up of dental pulp stromal cells for clinical applications

BACKGROUND AIMS. Dental pulp stromal cells (DPSC) are considered to be a promising source of stem cells in the field of regenerative therapy. However, the usage of DPSC in transplantation requires large-scale expansion to cater for the need for clinical quantity without compromising current good manufacturing practice (cGMP). Existing protocols for cell culturing make use of fetal bovine serum (FBS) as a nutritional supplement. Unfortunately, FBS is an undesirable additive to cells because it carries the risk of transmitting viral and prion diseases. Therefore, the present study was undertaken to examine the efficacy of human platelet lysate (HPL) as a substitute for FBS in a large-scale set-up. METHODS. We expanded the DPSC in Dulbeccos modified Eagles medium-knock-out (DMEM-KO) with either 10% FBS or 10% HPL, and studied the characteristics of DPSC at pre- (T25 culture flask) and post- (5-STACK chamber) large-scale expansion in terms of their identity, quality, functionality, molecular signatures and cytogenetic stability. RESULTS. In both pre- and post-large-scale expansion, DPSC expanded in HPL showed extensive proliferation of cells (c. 2-fold) compared with FBS; the purity, immune phenotype, colony-forming unit potential and differentiation were comparable. Furthermore, to understand the gene expression profiling, the transcriptomes and cytogenetics of DPSC expanded under HPL and FBS were compared, revealing similar expression profiles. CONCLUSIONS. We present a highly economized expansion of DPSC in HPL, yielding double the amount of cells while retaining their basic characteristics during a shorter time period under cGMP conditions, making it suitable for therapeutic applications.

Pairing as an instructional strategy to promote soft skills amongst clinical dental students

Training dentists today is challenging as they are expected to provide a wide range of dental care. In the provision of good dental care, soft skills are equally important as clinical skills. Therefore in dental education the development of soft skills are of prime concern. This study sought to identify the development of soft skills when dental students are paired in their clinical training. In this perception study, four open-ended items were used to elicit students feedback on the appropriateness of using clinical pairing as an instructional strategy to promote soft skills. The most frequently cited soft skills were teamwork (70%) and communication (25%) skills. However, both negative and positive behaviours were reported. As for critical thinking and problem solving skills, more positive behaviours were reported for abilities such as to explain, analyze, find ideas and alternative solutions, and make decisions. Leadership among peers was not evident as leading without legitimate authority could be a hindrance to its development. If clinical pairing is to be used as an effective instructional strategy to promote soft skills amongst students, clear guidelines need to be developed to prepare students to work in a dental team and the use of appropriate assessment tools can facilitate the development of these soft skills.

Aspirin Enhances Osteogenic Potential of Periodontal Ligament Stem Cells (PDLSCs) and Modulates the Expression Profile of Growth Factor–Associated Genes in PDLSCs

BACKGROUNDnThis study investigates the effects of aspirin (ASA) on the proliferative capacity, osteogenic potential, and expression of growth factor-associated genes in periodontal ligament stem cells (PDLSCs).nnnMETHODSnMesenchymal stem cells (MSCs) from PDL tissue were isolated from human premolars (n = 3). The MSCs identity was confirmed by immunophenotyping and trilineage differentiation assays. Cell proliferation activity was assessed through 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Polymerase chain reaction array was used to profile the expression of 84 growth factor-associated genes. Pathway analysis was used to identify the biologic functions and canonic pathways activated by ASA treatment. The osteogenic potential was evaluated through mineralization assay.nnnRESULTSnASA at 1,000 μM enhances osteogenic potential of PDLSCs. Using a fold change (FC) of 2.0 as a threshold value, the gene expression analyses indicated that 19 genes were differentially expressed, which includes 12 upregulated and seven downregulated genes. Fibroblast growth factor 9 (FGF9), vascular endothelial growth factor A (VEGFA), interleukin-2, bone morphogenetic protein-10, VEGFC, and 2 (FGF2) were markedly upregulated (FC range, 6 to 15), whereas pleotropin, FGF5, brain-derived neurotrophic factor, and Dickkopf WNT signaling pathway inhibitor 1 were markedly downregulated (FC 32). Of the 84 growth factor-associated genes screened, 35 showed high cycle threshold values (≥35).nnnCONCLUSIONSnASA modulates the expression of growth factor-associated genes and enhances osteogenic potential in PDLSCs. ASA upregulated the expression of genes that could activate biologic functions and canonic pathways related to cell proliferation, human embryonic stem cell pluripotency, tissue regeneration, and differentiation. These findings suggest that ASA enhances PDLSC function and may be useful in regenerative dentistry applications, particularly in the areas of periodontal health and regeneration.

Diverse Effects of Lead Nitrate on the Proliferation, Differentiation, and Gene Expression of Stem Cells Isolated from a Dental Origin

Lead (Pb2+) exposure continues to be a significant public health problem. Therefore, it is vital to have a continuous epidemiological dataset for a better understanding of Pb2+ toxicity. In the present study, we have exposed stem cells isolated from deciduous and permanent teeth, periodontal ligament, and bone marrow to five different types of Pb2+ concentrations (160, 80, 40, 20, and 10u2009µM) for 24 hours to identify the adverse effects of Pb2+ on the proliferation, differentiation, and gene expression on these cell lines. We found that Pb2+ treatment altered the morphology and adhesion of the cells in a dose-dependent manner. There were no significant changes in terms of cell surface phenotypes. Cells exposed to Pb2+ continued to differentiate into chondrogenesis and adipogenesis, and a severe downregulation was observed in osteogenesis. Gene expression studies revealed a constant expression of key markers associated with stemness (Oct 4, Rex 1) and DNA repair enzyme markers, but downregulation occurred with some ectoderm and endoderm markers, demonstrating an irregular and untimely differentiation trail. Our study revealed for the first time that Pb2+ exposure not only affects the phenotypic characteristics but also induces significant alteration in the differentiation and gene expression in the cells.

Comparison between gutta-percha and resin-coated gutta-percha using different obturation techniques

Background The aim of this study was to evaluate and compare the obturation quality between canals obturated with gutta-percha/AH Plus sealer (GP group) and resin-coated GP/EndoREZ® sealer (ER group). Methods A total sample of 90 mandibular premolar teeth was divided into 2 groups (2 × 45 canals): the GP group and ER group. Each group was further divided into 3 subgroups (n = 15): cold lateral compaction (CLC), warm lateral compaction (WLC) and single cone (SC). The teeth were subsequently embedded in resin and sectioned horizontally at 1, 3, 6 and 9 mm. All sections were then viewed with a stereomicroscope at ×40 magnification. The area occupied by core filling materials was determined using Cell^D software. Results With CLC, the percentage of core filling materials in the ER group was significantly higher than in the GP group at the 1- and 3-mm levels. Similarly, with WLC, the percentage of core filling material in the ER group was significantly higher than in the GP group at the 1-, 3- and 9-mm levels. With SC, the percentage of core filling materials in the ER group was significantly higher than in the GP group at all levels. Conclusions It can be concluded that the resin-coated GP/EndoREZ® sealer is superior to the gutta-percha/AH Plus in the percentage of core filling material.