The curcumin analog DM-1 induces apoptotic cell death in melanoma

The main difficulty in the successful treatment of metastatic melanoma is that this type of cancer is known to be resistant to chemotherapy. Chemotherapy remains the treatment of choice, and dacarbazine (DTIC) is the best standard treatment. The DM-1 compound is a curcumin analog that possesses several curcumin characteristics, such as antiproliferative, antitumor, and antimetastatic properties. The objective of this study was to evaluate the signaling pathways involved in melanoma cell death after treatment with DM-1 compared to the standard agent for melanoma treatment, DTIC. Cell death was evaluated by flow cytometry for annexin V and iodide propide, cleaved caspase 8, and TNF-R1 expression. Hoechst 33342 staining was evaluated by fluorescent microscopy; lipid peroxidation and cell viability (MTT) were evaluated by colorimetric assays. The antiproliferative effects of the drugs were evaluated by flow cytometry for cyclin D1 and Ki67 expression. Mice bearing B16F10 melanoma were treated with DTIC, DM-1, or both therapies. DM-1 induced significant apoptosis as indicated by the presence of cleaved caspase 8 and an increase in TNF-R1 expression in melanoma cells. Furthermore, DM-1 had antiproliferative effects in this the same cell line. DTIC caused cell death primarily by necrosis, and a smaller melanoma cell population underwent apoptosis. DTIC induced oxidative stress and several physiological changes in normal melanocytes, whereas DM-1 did not significantly affect the normal cells. DM-1 antitumor therapy in vivo showed tumor burden decrease with DM-1 monotherapy or in combination with DTIC, besides survival rate increase. Altogether, these data confirm DM-1 as a chemotherapeutic agent with effective tumor control properties and a lower incidence of side effects in normal cells compared to DTIC.     

Bcl-2 family proteins and cytoskeleton changes involved in DM-1 cytotoxic effect on melanoma cells

Melanoma is one of the most aggressive types of skin cancer and its incidence rate is still increasing. All existing treatments are minimally effective. Consequently, new therapeutic agents for melanoma treatment should be developed. The DM-1 compound is a curcumin analog that possesses several curcumin characteristics, such as antiproliferative, antitumor, and anti-metastatic properties. The aim of this study was to evaluate the different signaling pathways involved in the cytotoxic effect of DM-1 on melanoma cells. The apoptotic process and cytoskeletal changes were evaluated by immunoblotting and immunofluorescence, respectively, in melanoma cells. After DM-1 treatment, SK-MEL-5 melanoma cells showed actin filament disorganization with spicule formation throughout the cytoskeleton and significant reduction of focal adhesion as well as they were present only at cell extremities, conferring a poor connection between the cell and the substrate. Besides this, there was significant filopodium retraction and loss of typical cytoskeleton scaffold. These modifications contributed to cell detachment followed by cell death. Furthermore, DM-1-induced apoptosis was triggered by multiple Bcl-2 proteins involved in both the extrinsic and the intrinsic apoptotic pathways. SK-MEL-5 cells showed a death mechanism mainly by Bcl-2/Bax ratio decrease, whereas A375 cells presented apoptosis induction by Mcl-1 and Bcl-xL downregulation. In SK-MEL-5 and A375 melanoma cells, there was a significant increase in the active form of caspase 9, and the inactive form of the effector caspase 3 was decreased in both cell lines. Expression of cleaved poly ADP ribose polymerase was increased after DM-1 treatment in these melanoma cell lines, demonstrating that the apoptotic process occurred. Altogether, these data elucidate the cellular and molecular mechanisms involved in the cytotoxicity induced by the antitumor agent DM-1 in melanoma cells.     

Treatment of gingival fibromatosis associated with Zimmermann-Laband syndrome

BACKGROUND: Gingival fibromatosis is a rare condition characterized by a generalized enlargement of the buccal and lingual aspects of the attached and marginal gingiva. METHODS: This case report describes the periodontal management of a 13-year-old female patient with gingival fibromatosis associated with Zimmermann-Laband syndrome. The patient presented with gingival enlargement involving the maxillary and the mandibular arches, anterior open bite, and non-erupted teeth. Periodontal treatment included gingivectomy in all four quadrants. RESULTS: Histopathologic evaluation of the excised tissue supported the diagnosis of gingival fibromatosis. A significant improvement in esthetic appearance and eruption of the non-erupted teeth were obtained. The patient was referred for appropriate orthodontic treatment and has been closely followed for the earliest signs of recurrence of gingival enlargement. CONCLUSIONS: The successful therapy for gingival fibromatosis depends on correctly identifying the etiological factors and improving the impaired function and esthetic appearance through surgical intervention and adjunctive orthodontics. Maintaining treatment results depends on preservation of periodontal health.     

Microtensile bond strength of self-etching adhesives to enamel and dentin

PURPOSE: To measure the microtensile bond strength to enamel and dentin of three self-etching adhesives in comparison with a total-etch two-step system as a control. MATERIALS AND METHODS: A total of 40 extracted human molars were stored in saline solution until use, then divided into 4 groups of 10 teeth (one group per adhesive system). Half of each of these groups underwent bond strength tests on enamel, and the other half was used for adhesion testing on dentin. The following experimental groups (n = 5) were then formed: E(1) Adper Prompt-L-Pop (AP, 3M ESPE) on enamel; E(2) Xeno CF II (X, Sanking Kogyo) on enamel; E(3) AdheSE (AS, Ivoclar-Vivadent) on enamel; E(4) Excite (EX, Ivoclar-Vivadent) on enamel; D(1) AP on dentin; D(2): X on dentin; D(3) AS on dentin; D(4) EX on dentin. Each tooth yielded 15 to 20 sticks about 0.9 x 0.9 mm in cross-sectional area for microtensile testing. Specimens were loaded in tension at a crosshead speed of 0.5 mm/minute, and bond strength at failure was calculated in MPa. A two-way ANOVA was applied to test for significance of the differences among the groups. RESULTS: The bond strength values of Excite (the control) were significantly higher than those of the test products on enamel (42.92+/-4.8 MPa) and on dentin (45.80+/-5.79 MPa). The self-etching adhesives AdheSE (28.48+/-4.71 MPa) and Xeno CF II (27.22+/-2.74 MPa) revealed significantly stronger adhesion than Adper Prompt-L-Pop (20.16+/-2.07 MPa) on dentin. On enamel, all self-etching test materials performed similarly. The substrate did not appear to have a significant influence on adhesion, as each material reached comparable levels of bond strength on enamel and dentin. CONCLUSION: On both substrates the self-etching adhesives tested performed significantly worse than did the total-etch system.     

The role of parathyroid hormone-related protein in the regulation of osteoclastogenesis by cementoblasts

BACKGROUND: Parathyroid hormone-related protein (PTHrP) promotes osteoclastogenesis by inhibiting expression of osteoprotegerin (OPG), a decoy receptor for the receptor activator of nuclear factor kappa B (RANK), and by enhancing production of RANK ligand (RANKL) by osteoblasts. However, little is known regarding the role of PTHrP in regulating cementoblast-mediated osteoclastogenesis. METHODS: This study determined the impact of PTHrP on osteoclastogenesis using: 1) OCCM-30 (immortalized murine cementoblasts), 2) RAW 264.7 cells (murine myeloid cells), or 3) OCCM-30 plus RAW 264.7 cells. Cells were treated with PTHrP (1-34), RANKL, or PTHrP and RANKL combined. Enzyme-linked immunosorbent assays (ELISAs) for OPG and RANKL were performed on media and cell lysates, and tartrate-resistant acid phosphatase (TRAP) and mRNA detection for the osteoclast associated receptor (OSCAR) were performed. RESULTS: The highest numbers of TRAP-positive cells and cells expressing OSCAR were found in the RAW cell group treated with either RANKL alone or RANKL and PTHrP. TRAP-positive cells were fewer when OCCM cells were co-cultured with RAW, but the greatest numbers were still with both PTHrP and RANKL. OPG levels were highest from OCCM cells and PTHrP decreased these levels. In contrast, RANKL levels were low in OCCM cell lysates and PTHrP increased RANKL. In vivo studies also revealed high osteoclastic activity surrounding developing teeth in mice administered PTH. CONCLUSIONS: These results demonstrate that PTHrP influences the balance of OPG and RANKL production by cementoblasts, and further indicate that this effect, in the context of surrounding cells, might have a significant impact on osteoclastogenesis, root resorption, and tooth eruption.     

Effects of solvents on the early stage stiffening rate of demineralized dentin matrix

OBJECTIVES: To monitor the stiffening rate of demineralized dentin matrix at the early stages after exposure to different neat solvents. METHODS: Dentin beams approximately 0.8x0.7x8.0 mm were obtained from human third molars. After covering their ends with resin composite, the middle exposed length of 4.0mm (gauge-length) was demineralized in 0.5 M EDTA (pH 7.0) for 7 days. The specimens were gripped by a testing machine, pre-loaded to 10 g and cyclically stressed in tension to 5% strain, for 30 repeated cycles (total 20 min) at 0.6 mm/min while immersed in water (control). Then, water was replaced by either 100% acetone, methanol, ethanol, propanol, HEMA or air and the specimens subjected to the same cyclic protocol. The maximum apparent modulus of elasticity (E(Max)) was calculated for every cycle, plotted as a function of time and subjected to regression analysis. Stiffening rate was calculated as changes in E (min). Regression analysis examined the relationship between E and time for each solvent. Data were analyzed by one-way ANOVA and Student-Newman-Keuls test at alpha=0.05. RESULTS: Regression analysis showed that E increased significantly with time in all water-free solvents (R2=0.8-0.99). Stiffening rate was higher for acetone (0.9 MPa/min) and ethanol (0.8 MPa/min), intermediate for air (0.7 MPa/min), methanol (0.6 MPa/min) and propanol (0.5 MPa/min), lower for HEMA (0.2 MPa/min) and practically none for water (0.07 MPa/min) with p<0.05. CONCLUSIONS: The solvent-induced stiffening rate of demineralized dentin matrix is both time and solvent-dependent. The ability of solvents to promptly stiffen the demineralized dentin matrix may be important in maintaining the resin-infiltrated matrix expanded during the solvent evaporation stage of resin bonding.     

Tooth loss and subluxation in the primary dentition: a twelve-year follow-up case report

The case of a five-year-old child is reported, who suffered dento-alveolar injury including subluxation of the right upper lateral incisor and avulsion of the upper central incisors and left upper lateral incisor and laceration in the mucosa. The case was followed for 12 years until complete root formation and alignment of the anterior permanent teeth.     

Microtensile bond strength tests: scanning electron microscopy evaluation of sample integrity before testing

The failure of a certain number of microtensile specimens during their preparation and before loading is a common and undesirable occurrence. This study was aimed at observing, under a scanning electron microscope, enamel and dentin microtensile specimens, in order to find structural faults that might be responsible for their premature failure. In a sample of 80 sticks, none of the specimens was found to be free of defects. These may consist of microcracks in enamel, most often at the periphery of the stick, or in dentin at the level of hybrid layer. Gaps were often seen at the interfaces between the substrates. Voids were sometimes visible within the resin composite thickness. Enamel specimens tended to exhibit more defects than dentin specimens. It is fair to suspect that, because of the brittleness of the tissue, enamel microtensile specimens are intrinsically more prone to failure, thus yielding bond strengths which are not significantly higher than those measured on dentin specimens. This leads one to question the reliability of the microtensile method for testing adhesion on enamel. It seems sensible to develop a method for a quantitative assessment of specimens integrity before loading as a possible predictor for their performance under load.