Decreased calcitonin gene-related peptide expression in the dorsal root ganglia of TNF-deficient mice in a monoiodoacetate-induced knee osteoarthritis model

Background: The detailed mechanisms of knee osteoarthritis (OA) pain have not been clarified, but involvement of inflammatory cytokines such as tumor necrosis factor-alpha (TNF) has been suggested. The present study aimed to investigate the more detailed neurological involvement of TNF in joint pain using a TNF-knockout mouse OA model. Methods: The right knees of twelve-week-old C57BL/6J wild and TNF-deficient knockout (TNF-ko) mice (n=15, each group) were given a single intra-articular injection of 10 µg monoiodoacetate in 10 mL sterile saline. The left knees were only punctured as the control. Evaluations were performed immediately after the injection (baseline) and at 7, 14, and 28 days after the injection with a subsequent intra-articular injection of neurotracer into both knees. The animals were evaluated for immunofluorescence of the lumbar dorsal root ganglia (DRG) innervating the knee joints. The injected knees were observed macroscopically and mouse pain-related behaviors were scored. Results: Macroscopic observation showed similar knee OA development in both wild and TNF-ko mice. Calcitonin gene-related peptide (CGRP, a neuropeptide identified as a inflammatory pain-related biomarker) was significantly increased in DRG neurons innervating OA-induced knee joints with significantly less CGRP expression in TNF-ko animals. Pain-related behavior scoring showed a significant increase in pain in OA-induced joints, but there was no significant difference in pain observed between the wild and TNF-ko mice. Conclusions: The result of the present study indicates the possible association of TNF-alpha in OA pain but not OA development.     

Association Between Glucan Synthesis by Streptococcus mutans and Caries Incidence in Schoolchildren Receiving a Fluoride Mouth Rinse

Purpose: To determine whether variation in glucan synthesis by Streptococcus mutans isolates is associated with caries development in children receiving a fluoride mouth rinse (FMR). Materials and Methods: Of 122 children (aged 9 to 10 years), 64 had received FMR (FMR(+) group) and the remaining 58 children had not (FMR(-) group). The number of decayed and filled teeth (DFT) and increases in the number of DFT in 1 year (dDFT) were recorded. Saliva samples were collected to isolate the clinical S. mutans strains. The isolates were incubated in heart infusion broth supplemented with 1% sucrose, then the amount of water-insoluble glucan (WIG) formed on a glass tube surface was evaluated. Results: In the FMR(-) group, children carrying S. mutans had a higher DFT (P = 0.039) and tended to have a higher dDFT (P = 0.080) than the others. In the FMR(+) group, although the differences between children with and without S. mutans were not significant, children carrying S. mutans that produced a high amount of WIG had higher dDFT than the other S. mutans-positive children (P = 0.034). Conclusions: This study revealed that the variation in glucan synthesis by S. mutans is associated with caries development in children receiving a FMR.     

Reprogrammed keratinocytes from elderly type 2 diabetes patients suppress senescence genes to acquire induced pluripotency

Nuclear reprogramming enables patient-specific derivation of induced pluripotent stem (iPS) cells from adult tissue. Yet, iPS generation from patients with type 2 diabetes (T2D) has not been demonstrated. Here, we report reproducible iPS derivation of epidermal keratinocytes (HK) from elderly T2D patients. Transduced with human OCT4, SOX2, KLF4 and c-MYC stemness factors under serum-free and feeder-free conditions, reprogrammed cells underwent dedifferentiation with mitochondrial restructuring, induction of endogenous pluripotency genes - including NANOG, LIN28, and TERT, and down-regulation of cytoskeletal, MHC class I- and apoptosis-related genes. Notably, derived iPS clones acquired a rejuvenated state, characterized by elongated telomeres and suppressed senescence-related p15INK4b/p16INK4a gene expression and oxidative stress signaling. Stepwise guidance with lineage-specifying factors, including Indolactam V and GLP-1, redifferentiated HK-derived iPS clones into insulin-producing islet-like progeny. Thus, in elderly T2D patients, reprogramming of keratinocytes ensures a senescence-privileged status yielding iPS cells proficient for regenerative applications.