High-Glucose Media Reduced the Viability and Induced Differential Pro-Inflammatory Cytokines in Human Periodontal Ligament Fibroblasts.

Hyperglycemic condition in diabetic patients tends to exacerbate periodontitis severity. Thus, the influence of hyperglycemia on the biological and inflammatory response of periodontal ligament fibroblasts (PDLFs) needs to be elucidated. In this study, PDLFs were seeded in media containing glucose concentrations (5.5, 25, or 50 mM) and stimulated with 1 µg/mL of lipopolysaccharide (LPS). PDLFs' viability, cytotoxicity, and the migration ability were determined. The mRNA expression of Interleukin (IL)-6, IL-10, and IL-23 (p19/p40), and Toll-like receptor (TLR)-4 were analyzed; at 6 and 24 h, protein expression of IL-6 and IL-10 was also determined. PDLFs grown in 50 mM glucose medium showed lower viability. The 5.5 mM glucose led to the highest percentage of wound closure compared to 25 mM and 50 mM glucose with/without LPS. Additionally, 50 mM glucose with LPS exhibited the least migration ability among all groups. The expression of IL-6 was amplified significantly in LPS-stimulated cells in 50 mM glucose medium. IL-10 was constitutively expressed in different glucose concentrations, and LPS stimulation decreased it. IL-23 p40 was up-regulated after LPS stimulation in 50 mM glucose concentration. TLR-4 was highly expressed after LPS stimulation in all glucose concentrations. Hyperglycemic conditions limit PDLF proliferation and migration, and enhance the expression of certain pro-inflammatory cytokines to induce periodontitis.

Cytokine expression and anti-microbial effectiveness of different calcium hydroxide dilutions: An In Vitro study.

Aims: To determine the cytokine expression by human gingival fibroblasts in response to different calcium hydroxide (Ca(OH)2) dilutions and test the effectiveness of these dilutions in root canal dentin infected with Enterococcus faecalis (E. faecalis). Methods: UltraCal XS Ca(OH)2 dilutions were prepared (60, 10, and 1 mg\mL) and co-cultured with gingival fibroblasts for 24 and 48 hours. Untreated cells were used as controls. Expressions of interleukin (IL-1ß), tumour necrosis factor alpha (TNF-α), transforming growth factor beta (TGF-ß), and IL-10 were analysed with real-time polymerase chain reaction (PCR). Root canals of extracted human teeth were inoculated with E. faecalis. After 21 days, canals were medicated with Ca(OH)2 dilutions for 7 days. Samples were taken to determine bacterial reduction using quantitative PCR. Analysis of variance, Tukey post-test, and Wilcoxon matched pair test were used for statistics. Results: IL-1ß and TNF-α expressions of all Ca(OH)2 dilutions were higher at 24 and 48 hours compared to the control. Similarly, all Ca(OH)2 dilutions induced TGF-ß expression at 24 hours compared to the control and continued to be higher in 60 mg/mL groups at 48 hours. In contrast, IL-10 was constitutively expressed by untreated cells in the control group and was down-regulated significantly by all Ca(OH)2 dilutions at 24 and 48 hours. All dilutions demonstrated a significant E. faecalis reduction (P < 0.001) with no significant difference between dilution groups (P > 0.05). Conclusions: All Ca(OH)2 dilutions had a differential inflammatory effect on fibroblasts and had a down-regulation effect to IL-10. All dilutions tested were effective against E. faecalis, with 60 mg/mL having the highest bacterial reduction.

Silver Nanoparticles Alone or in Combination with Calcium Hydroxide Modulate the Viability, Attachment, Migration, and Osteogenic Differentiation of Human Mesenchymal Stem Cells.

This study aimed to evaluate the effect of silver nanoparticles (AgNPs) alone or in combination with calcium hydroxide (Ca(OH)2) on the proliferation, viability, attachment, migration, and osteogenic differentiation of human mesenchymal stem cells (hMSCs). Different concentrations of AgNPs alone or mixed with Ca(OH)2 were prepared. Cell proliferation was measured using AlamarBlue, and hMSCs attachment to dentin disks was evaluated using scanning electron microscopy. Live-dead imaging was performed to assess apoptosis. Wound healing ability was determined using the scratch-migration assay. To evaluate osteogenic differentiation, the expression of Runt-related transcription factor (RUNX2), Transforming growth factor beta-1 (TGF-ß1), Alkaline Phosphatase (ALP), and Osteocalcin (OCN) were measured using real-time reverse transcriptase polymerase chain reaction. ALP staining and activity were also performed as indicators of osteogenic differentiation. AgNPs alone seemed to favor cell attachment. Lower concentrations of AgNPs enhanced cell proliferation. AgNP groups showed markedly less apoptosis. None of the medicaments had adverse effects on wound closure. The expression of TGF-ß1 was significantly upregulated in all groups, and OCN was highly expressed in the AgNP groups. AgNPs 0.06% showed the most enhanced ALP gene expression levels, activity, and marked cytochemical staining. In conclusion, AgNPs positively affect hMSCs, making them a potential biomaterial for various clinical applications.

Bisphosphonate-associated osteonecrosis of jaw reoccurrence after methotrexate therapy: a case report.

INTRODUCTION: Bisphosphonate-related osteonecrosis of the jaw (BRONJ) is a well-known complication caused by amino-bisphosphonate therapy. We document one case of BRONJ associated with oral administration of methotrexate, a known immunosuppressive drug used to treat rheumatoid arthritis. METHODS: A 66-year-old woman was referred for evaluation and endodontic surgery of recently re-treated tooth 13. Tooth 14 was extracted 3 months prior, and the extraction site had not completely healed. Her medical history revealed rheumatoid arthritis and osteoporosis. She had been taking Fosamax (alendronate) 70 mg daily. Because of adequate root canal therapy of tooth 13, endodontic surgery was performed. Five months after apicoectomy, her symptoms had not changed. Tooth 13 was extracted, and the socket healed without complications. The socket of extracted tooth 14 was also healing. At the 3-month recall visit, bone exposure and purulent discharge at the site of extracted tooth 14 were noted. The patient had recently received methotrexate. The methotrexate was discontinued, and she was given course of amoxicillin. RESULTS: At the 18-month follow-up, the healing progressed, and the wound was closed. CONCLUSIONS: A medication that suppresses the immune system such as methotrexate may complicate the management of BRONJ. Once a diagnosis of BRONJ is made, a closely monitored conservative approach is recommended.