Melatonin antagonizes lipopolysaccharide-induced pulpal fibroblast responses.

BACKGROUND: Pulpal inflammation is known to be mediated by multiple signaling pathways. However, whether melatonin plays regulatory roles in pulpal inflammation remains unclear. This study aimed at elucidating an in situ expression of melatonin and its receptors in human pulpal tissues, and the contribution of melatonin on the antagonism of lipopolysaccharide (LPS)-infected pulpal fibroblasts. METHODS: Melatonin expression in pulpal tissues harvested from healthy teeth was investigated by immunohistochemical staining. Its receptors, melatonin receptor 1 (MT1) and melatonin receptor 2 (MT2), were also immunostained in pulpal tissues isolated from healthy teeth and inflamed teeth diagnosed with irreversible pulpitis. Morphometric analysis was subsequently performed. After LPS infection of cultured pulpal fibroblasts, cyclo-oxygenase (COX) and interleukin-1 ß (IL-1 ß) transcripts were examined by using reverse transcription-polymerase chain reaction (RT-PCR). Analysis of mRNA expression was performed to investigate an antagonism of LPS stimulation by melatonin via COX and IL-1 ß induction. Mann-Whitney U test and One-way ANOVA were used for statistical analysis to determine a significance level. RESULTS: Melatonin was expressed in healthy pulpal tissue within the odontoblastic zone, cell-rich zone, and in the pulpal connective tissue. Furthermore, in health, strong MT1 and MT2 expression was distributed similarly in all 3 pulpal zones. In contrast, during disease, expression of MT2 was reduced in inflamed pulpal tissues (P-value< 0.001), but not MT1 (P-value = 0.559). Co-culturing of melatonin with LPS resulted in the reduction of COX-2 and IL-1 ß expression in primary pulpal fibroblasts, indicating that melatonin may play an antagonistic role to LPS infection in pulpal fibroblasts. CONCLUSIONS: Human dental pulp abundantly expressed melatonin and its receptors MT1 and MT2 in the odontoblastic layers and pulpal connective tissue layers. Melatonin exerted antagonistic activity against LPS-mediated COX-2 and IL-1 ß induction in pulpal fibroblasts, suggesting its therapeutic potential for pulpal inflammation and a possible role of pulpal melatonin in an immunomodulation via functional melatonin receptors expressed in dental pulp.

Immunohistochemical and gene expression analysis of stem-cell-associated markers in rat dental pulp.

Stem cells in the dental pulp comprise rare populations lacking definitive cytological markers and thus are poorly characterized in vivo, especially in rat species. To gain more insight into the phenotypical characteristics and tissue distribution of these cells, we examined the distribution of stem-cell-associated marker-expressing cells and mRNA expression levels of stem-cell-associated markers in the rat molar. CD146-positive cells co-expressing microtubule-associated protein 1B were counted following double-labeling immunoperoxidase staining and their density in the coronal pulp, root pulp and periodontal ligament was compared. Moreover, mRNA expression levels of CD146, CD105, CD166 and secreted phosphoprotein 1 (SPP1; also known as osteopontin, a negative regulatory element of the stem cell niche) were analyzed in these regions by using real time polymerase chain reaction. The double-positive cells could be clearly distinguished from non-stem cells single-stained by either of the markers and showed a significantly higher density in the coronal pulp compared with the other regions (P<0.05). Moreover, mRNA expression levels of CD146, CD105 and CD166 were significantly higher in the coronal pulp than in the other regions (P<0.05). On the other hand, SPP1 mRNA expression was significantly higher in the periodontal ligament than in the pulp. Thus, the density of stem-cell-associated marker-expressing cells and stem-cell-associated gene expression levels are higher in the coronal pulp than in the root pulp and periodontal ligament, suggesting that the coronal pulp harbors more stem cells than the other regions.

In vitro resorbability and granular characteristics of 3D-printed hydroxyapatite granules versus allograft, xenograft, and alloplast for alveolar cleft surgery applications.

Three-dimensionally printed hydroxyapatite (3DP HA) was investigated in regards to its functional properties supporting bone regeneration and tooth movement in alveolar cleft applications. Commercially available bovine xenograft (BXG), biphasic calcium phosphate alloplast (BCP), and two types of freeze-dried bone allograft granules (FDBA and FDBA-CMC) were employed as control samples. Degradability was studied by submerging the samples in pH 7.4 buffered solution at 37°C for 28 days and determining subsequent weight loss percentage. The wicking property and granular agglomeration were evaluated by putting the granules in contact with deionized water, blood, and phosphate-buffered saline (PBS). Both of FDBA and FDBA-CMC showed the greatest weight loss at 28 days followed by 3DP HA. In contrast, 3DP HA showed significantly greater wicking ability than other samples for all liquid types. FDBA-CMC exhibited the greatest granular agglomeration for all liquid types followed by 3DP HA. 3DP HA was found to be a favorable candidate for bone grafting in alveolar cleft treatment.