Concurrence of replicative senescence and elevated expression of p16(INK4A) with subculture-induced but not calcium-induced differentiation in normal human oral keratinocytes

Primary normal human oral keratinocytes (NHOKs) undergo differentiation in the presence of calcium concentrations higher than 0.15 mM in vitro, which is useful in investigating the mechanisms involved in the differentiation of epithelial cells. Serial subculture of NHOKs to the postmitotic stage also induces terminal differentiation. However, the detailed mechanisms of both differentiation processes remain substantially unknown. To investigate the molecular differences in these processes, NHOKs were induced to differentiate by exposure to 1.2 mM of calcium and by serial subculture to the postmitotic stage. To study whether the cells were induced to differentiate and to undergo replicative senescence, the amount of cellular involucrin and the expression of senescence-associated beta-galactosidase (SA-beta-gal) were measured respectively. The expression of replicative senescence-associated genes and the activity of telomerase from the differentiated cells were also determined. Both calcium treatment and serial subculture to the postmitotic stage notably elevated the cellular involucrin. The percentage of SA-beta-gal-positive cells was significantly elevated by the continued subculture, but such changes were not observed in keratinocytes exposed to calcium. The concentration of cellular p16(INK4A) protein was progressively increased by the continued subculture but was not changed by calcium treatment. On the other hand, the concentrations of cellular p53 were similar in both differentiation processes. However, telomerase activity was lost in NHOKs that had undergone differentiation by both calcium treatment and serial subculture. The results indicate that calcium-induced differentiation of NHOKs has similar characteristics to their serial subculture-induced differentiation, but that the differentiation processes are not identical, because calcium-induced differentiation does not concur with either replicative senescence or the gradually increased concentration of p16(INK4A).     

Eugenol inhibits K+ currents in trigeminal ganglion neurons

Eugenol, a natural capsaicin congener, is widely used in dentistry. Eugenol inhibits voltage-activated Na(+) and Ca(2+) channels in a transient receptor potential vanilloid 1 (TRPV1)-independent manner. We hypothesized that eugenol also inhibits voltage-gated K(+) currents, and investigated this in rat trigeminal ganglion neurons and in a heterologous system using whole-cell patch clamping. Eugenol inhibited voltage-gated K(+) currents, and the inhibitory effects of eugenol were observed in both capsaicin-sensitive and capsaicin-insensitive neurons. Pre-treatment with capsazepine, a well-known antagonist of TRPV1, failed to block the inhibitory effects of eugenol on K(+) currents, suggesting no involvement of TRPV1. Eugenol inhibited human Kv1.5 currents stably expressed in Ltk(-) cells, where TRPV1 is not endogenously expressed. We conclude that eugenol inhibits voltage-gated K(+) currents in a TRPV1-independent manner. The inhibition of voltage-gated K(+) currents is likely to contribute to the irritable action of eugenol. Abbreviations: human Kv1.5 channel, hKv1.5; transient receptor potential vanilloid 1, TRPV1.     

Activation of vanilloid receptor 1 (VR1) by eugenol

The structural similarity of eugenol with capsaicin suggests that these two agents may share molecular mechanisms to produce their effects. We investigated the effects of eugenol in comparison with those of capsaicin using whole-cell patch clamp and Fura-2-based calcium-imaging techniques in a heterologous expression system and with sensory neurons. In vanilloid receptor 1 (VR1)-expressing human embryonic kidney (HEK) 293 cells and trigeminal ganglion (TG) neurons, eugenol activated inward currents, whereas capsazepine, a competitive VR antagonist, and ruthenium red (RR), a functional VR antagonist, completely blocked eugenol-induced inward currents. Moreover, eugenol caused elevation of [Ca(2+)](i), and this was completely abolished by both capsazepine and ruthenium red in VR1-expressing HEK 293 cells and TG neurons. Our results provide strong evidence that eugenol produces its effects, at least in part, via VR1 expressed by the sensory nerve endings in the teeth.     

Malignant tumor involving paranasal sinuses

Carcinoma of the paranasal sinuses is a very serious disease, but its incidence is lower than that of any other oral malignancy. Failure to diagnose it early because of misleading symptoms resembling sinusitis usually leads to its detection at a stage when it has already destroyed the bony walls of the sinus and has spread to surrounding structures (infratemporal fossa, posterior ethmoid cells, orbit, cribriform plate, nasopharynx, sphenoid sinus or base of the skull). It is uncommon for this cancer to spread to the regional lymph nodes as long as it remains confined within the maxillary sinus. CT scanning provides an accurate assessment of the extent of resection required and is usually used as an indicator of operability. A 56-year-old man with a chief complaint of palatal swelling, pain and pus discharge was referred to Seoul National University Dental Hospital. Orthopantomogram and Waters' view revealed an aggressive bone destruction in the right maxillary molar area, ipsilateral sinus opacification and destruction of the lateral wall of the maxillary sinus. The CT fidding showed that the right maxillary sinus was occupied by a soft tissue mass which represented a central low density, with an irregular shape at some levels. The tumor mass also involved the pterygopalatine fossa, right esthmoid sinus, nasal cavity and hard palate. Incisional biopsy was done and the biopsy report was squamous cell carcinoma. He was transferred to Internal Medicine for chemotheraphy because of inoperability.     

Synthesis and preparation of novel 4-arm star-shape poly(carboxylic acid)s for improved light-cured glass-ionomer cements.

OBJECTIVE: The objective of this study was to synthesize and characterize novel 4-arm star-shape poly(acrylic acid)s (poly(AA)s) via atom-transfer radical polymerization (ATRP) technique, tether in situ light-curable methacrylate functionalities onto the poly(AA) backbone, use these star-shape poly(AA)s to formulate the light-cured glass-ionomer cements (LCGICs), and evaluate the mechanical strengths of the formed cements. MATERIALS AND METHODS: The 4-arm poly(AA)s were synthesized using ATRP and tethered with either 2-isocyanatoethyl methacrylate (IEM) or glycidyl methacrylate (GM). The polymers were formulated with 2-hydroxyethyl methacrylte (HEMA) or methacryloyl beta-alanine (MBA), water, initiators, and Fuji II LC filler. Compressive strength (CS) was used as a tool to evaluate the formed cements. The specimens were conditioned in distilled water at 37 degrees C for 24h prior to testing. RESULTS: The 4-arm poly(AA) showed a lower viscosity as compared to its linear counterpart. Both IEM-tethered and GM-tethered 4-arm poly(AA) constructed LCGICs showed significantly high mechanical strengths. Both types of co-monomer and grafting agent dramatically affected the mechanical strengths. The MBA-containing poly(AA) cements exhibited much higher CS than the HEMA-containing cements. The IEM-tethered poly(AA) cements showed much higher CS and DTS than the GM-tethered cements. CONCLUSIONS: This study developed a novel light-curable 4-arm star-shape poly(AA) system. The system was 13% in CS, 178% in DTS and 123% in FS, compared to Fuji II LC.     

Xeroderma pigmentosum: a case report

This paper presents a case study of a child with xeroderma pigmentosum (XP). The disease results in sensitivity to UV radiation as a result of reduced activity in a defective enzyme responsible for DNA repair. Affected individuals have a variety of clinical symptoms, which may include problems of the skin and oral mucosa, ocular manifestations, and neurologic impairment. A number of precautions must be taken when treating these patients, which include proper shielding from damaging light and the selection of suitable dental materials. The necessary measures required when treating patients with XP are reviewed in this report.     

Genome-wide linkage analysis of mandibular prognathism in Korean and Japanese patients

The existence of familial aggregation of mandibular prognathism (MP) suggests that genetic components play an important role in its etiology. In this study, a genome-wide linkage analysis to identify loci susceptible to MP was conducted with 90 affected sibling-pairs in 42 families, comprised of 40 Korean sibling-pairs and 50 Japanese sibling-pairs. Two non-parametric linkage analyses, GENEHUNTER-PLUS and SIBPAL, were applied and detected nominal statistical significance of linkage to MP at chromosomes 1p36, 6q25, and 19p13.2. The best evidence of linkage was detected near D1S234 (maximum Z(lr) = 2.51, P = 0.0012). In addition, evidence of linkage was observed near D6S305 (maximum Z(lr) = 2.23, P = 0.025) and D19S884 (maximum Z(lr) = 1.93, P = 0.0089). Identification of the susceptible genes in the linkage regions will pave the way for insights into the molecular pathways that cause MP, especially overgrowth of the mandible, and may lead to the development of novel therapeutic tools.     

Effect of finish line variants on marginal accuracy and fracture strength of ceramic optimized polymer/fiber-reinforced composite crowns

STATEMENT OF PROBLEM: Ceramic optimized polymer (Ceromer)/fiber-reinforced composite (FRC) crowns have been promoted as alternatives to conventional crowns. However, little is known regarding the ideal tooth preparation for this type of crown. PURPOSE: This in vitro study evaluated the marginal adaptation and fracture strength of ceromer/FRC crowns with respect to the various types of finish lines. MATERIAL AND METHODS: Four metal dies with different finish lines (0.9-mm chamfer, 1.2-mm chamfer, 1.2-mm rounded shoulder, and 1.2-mm shoulder) were prepared. Forty (10 for each finish line) Targis/Vectris crowns were fabricated on duplicated base metal alloy dies. The restorations were stereoscopically evaluated at 56 points along the entire circumferential margin for measuring the margin adaptation before and after cementation with a resin luting agent. The specimens were then compressively loaded to failure using a universal testing machine. The marginal adaptation (microm) was analyzed with the Kruskal-Wallis test and post-hoc Dunnett test (alpha=.05). The fracture load (N) was analyzed with a 1-way analysis of the variance and the Scheffe adjustment (alpha=.05). The fractured surfaces of the crowns were examined with a scanning electron microscope to determine the mode of fracture. RESULTS: The marginal adaptation of crowns with a shoulder finish line was significantly better than crowns with a chamfer finish line before and after cementation (P<.001). The increased marginal gap after cementation was the lowest in the 1.2-mm rounded shoulder group. The fracture strength of the crowns with the 0.9-mm chamfer and crowns with 1.2-mm chamfer was significantly greater than those of the crowns with the 1.2-mm shoulder or rounded shoulder (P=.011, P=.049, respectively). The mean fracture load of all crowns, regardless of the finish line design, was 1646 N. The fractured surface of the crown revealed adhesive failure and 3 types of cohesive failure (fracture of the Targis and Vectris, Targis fracture with a crack in the Vectris layer, and crushing without fracture). CONCLUSION: The marginal gaps were greater for the chamfer finish line specimens than in the shoulder finish line specimens. However, the fracture strength of the chamfer finish line specimens was greater than that of the shoulder finish line specimens.