Background: The effects of lipopolysaccharide (LPS) on cell proliferation and osteogenic potential (OP) of MSCs have been frequently studied. Objective: to compare the effects of LPS on periodontal-ligament-derived mesenchymal stem cells (PDLSCs) in monolayer and 3D culture. Methods: The PDLSCs were colorimetrically assessed for proliferation and osteogenic potential (OP) after LPS treatment. The 3D cells were manually prepared by scratching and allowing them to clump up. The clumps (C-MSCs) were treated with LPS and assessed for Adenosine triphosphate (ATP) and OP. Raman spectroscopy was used to analyze calcium salts, DNA, and proline/hydroxyproline. Multiplexed ELISA was performed to assess LPS induced local inflammation. Results: The proliferation of PDLSCs decreased with LPS. On Day 28, LPS-treated cells showed a reduction in their OP. C-MSCs with LPS did not show a decrease in ATP production. Principal bands identified in Raman analysis were the P–O bond at 960 cm−1 of the mineral component, 785 cm−1, and 855 cm−1 showing qualitative changes in OP, proliferation, and proline/hydroxyproline content, respectively. ELISA confirmed increased levels of IL-6 and IL-8 but with the absence of TNF-α and IL-1β secretion. Conclusions: These observations demonstrate that C-MSCs are more resistant to the effects of LPS than cells in monolayer cell culture. Though LPS stimulation of C-MSCs creates an early pro-inflammatory milieu by secreting IL-6 and IL-8, PDLSCs possess inactivated TNF promoter and an ineffective caspase-1 activating process.
Epiregulin (EPR) is a novel member of the epidermal growth factor (EGF) family. It has been shown to promote wound healing in oral epithelium, enhance proliferation of other epithelial tissues, and is involved in several epithelial-related malignancies such as colorectal, lung, and bladder carcinoma. More recently, EPR transcripts were found to be high in a study on archival oral squamous cell carcinoma (OSCC) specimens. This implies that EPR may be responsible for the progression of OSCC. The aim of this was to elucidate the effects of EPR on (i) cell morphological changes, (ii) cell proliferation and (iii) receptor expression of the H-series OSCC cell lines. The clinicopathological origin and the expression of the epidermal growth factor receptor (EGFR) and ErbB4 receptors of the H-series cell lines were initially characterised. Based on these parameters, two of the H-series cell lines, namely H103 and H357 were selected for downstream experiments. The cell lines were treated with 1 ng/ml, 10 ng/ml, and 20 ng/ml of EPR for 24 and 48 hours in all subsequent experiments. Untreated cells acted as the control which was used for comparison with each treated group. The cell morphological changes, cell proliferation and receptor expression of the OSCC cell lines were evaluated using phase contrast microscopy, 5-bromo-2’-deoxy-uridine (BrdU) assays and flow cytometry respectively. The results were compared and analysed using the student t-test. There were no appreciable morphological changes in the cells regardless of the dose of EPR tested nor between the different timelines. There were no significant changes in cell proliferation after EPR treatment. As for the effect of EPR on receptor expression, 20 ng/ml of EPR significantly reduced the density of EGFR expression (p value = 0.049) in the H103 cell line after the 24-hour treatment. No other statistically significant changes were detected. The results show that EPR had no effect on the morphology and proliferativity of OSCC cells. However, the significant decline in EGFR expression after EPR treatment suggests that EPR might play an important role in the regulation of EGFR expression and hence OSCC progression.
This study measures the impact of the implementation of a dedicated research semester on various perceived competencies related to research. In 2016, surveys were conducted on final undergraduate Pharmacy students in regard to appraisal and critical thinking skills. Students’ perceptions of the impact of research in enhancing their employment potential were investigated. Our evaluation included students’ self-assessment of their writing, presentation, critical thinking and research skills. To assess qualitative parameters, the data obtained were analysed using the Wilcoxon signed-rank test. A total of 113 responses was received. A majority of students indicated that the research semester prepared them in undertaking their research projects. They acknowledged that research helped in building confidence and to acquire the ability to work independently. Most students perceived that the experience gained in research would enhance their employment potential. Overall, students developed critical thinking skills through their respective research project.