Dentine tubule occlusion effect of hydrolyzed casein in a bioactive glass-based dental desensitizing gel.

OBJECTIVES: The effectiveness of three different groups of polyethylene glycol (PEG)-based gels containing powders on dentin hypersensitivity (DH) treatment were assessed and compared with Actimins® as commercial reference group. METHODS: Hydroxyapatite nanorods (nHA) and sol-gel-derived 45S5 bioglass (SGD 45S5) powders were synthesized through hydrothermal and sol-gel methods, respectively. First, 25 demineralized dentin disks were divided into five groups. Then, the prepared gels based on 45S5 bioglass with and without hydrolyzed casein (HC) as experimental, nHA gel and Actimins® as positive and commercial reference groups were applied twice a day on disks by a micro applicator. To mimic the oral environment, treated disks were immersed in artificial saliva in a water bath at 37 °C for 7 days. However, in the negative control group, no agent was applied on the samples. FE-SEM, EDS, AFM, and XRD were performed to assess tubule occlusion. One-way ANOVA test was used for statistical analysis and p*<0.05 was set as the significance level. RESULTS: The nHA with an average aspect ratio of 2.77 and the SGD 45S5 powders with a polygonal morphology and the average size of 48.64±11.38 µm were synthesized. After treatment, tubule occlusion in HC-SGD 45S5 and nHA gels were shown to be higher than other groups. The root mean square roughness (Rrms) of the above-mentioned gels showed to be 121.54±9.25 nm, and 312.6 ± 9 nm, respectively. CONCLUSION: The nHA containing group exhibited the highest tubule occlusion efficiency (i.e., tubule diameter of 0.92±0.32 µm) with a superior mineral precipitation. HC as a novel material demonstrates to be potentially beneficial in DH treatment. CLINICAL SIGNIFICANCE: DH as a common issue may be reduced or eliminated by occlusion of patent dentinal tubules. There are various types of desensitizing agents capable of controlling the DH by the occlusion of patent dentinal tubules. The desensitizing gels developed in this study showed to be promising for clinical and home-use applications.

Novel calcified gum Arabic porous nano-composite scaffold for bone tissue regeneration.

The aim of this study was to investigate the biomechanical and biological properties of a nanocomposite scaffold containing both mineral and polysaccharide constituents. Hydroxyapatite nanoparticles (n-HA) was synthesized from dead abra ovata shells using wet chemical methods and was used in different ratios in concert with gum Arabic, a branched plant polysaccharide. N-HA/gum nanocomposite was fabricated with freeze-drying process and characterized by FTIR and SEM for chemical structure and morphology. Porosity was estimated using liquid substitution method. The scaffold mechanical properties were evaluated by compressive test measurement. Osteogenic differentiation was assessed using alkaline phosphatase production and biomineralization was evaluated using Alizarin red assay. Results demonstrated that the hydroxyapatite/gum Arabic nanocomposite had favorable biocompatibility and a similar structure to natural bone matrix. Porous nanocomposite possessed macropore networks with a porosity 87-93% and mean pore size ranging between 164 and 230 µm. The gum/HA with a ratio of 50% w/w HA had the highest compressive modulus of ∼75.3 MPa Pa (MPa) and the ultimate compressive stress of ∼16.6 MPa. C2C12 cells cultured on a scaffold with higher percentage (40 and 50 w/w) of HA demonstrated increased ALP levels and calcium deposition. The data from the present study demonstrated significant changes to the biomechanical properties and osteoconductivity of the nanocomposite scaffold by modulating its mineral content. Nanocomposite scaffolds containing gum and n-HA of 40-50% exhibited highest mechanical properties, as well as supported increased biomineralization.

Oxidative stress level and tyrosinase activity in vitiligo patients.

BACKGROUND: Vitiligo is an acquired pigmentary disorder of the skin. Genetic factors, oxidative stress, autoimmunity, and neurochemical agents might be contributing factors for the development of the disease. AIMS: To evaluate the oxidative stress level and tyrosinase activity in vitiligo patients and to compare them with healthy volunteers. MATERIALS AND METHODS: We used Comet assay to evaluate DNA strand breaks in peripheral blood cells of active vitiligo patients. We then extracted total protein from lesional and nonlesional skin of ten selected patients. Tyrosinase activity was found to play a crucial role in melanogenesis. RESULTS: The basal level of systemic oxidative DNA strand breaks in leukocytes increased in vitiligo patients compared to healthy participants. We observed that tyrosinase activity in lesional skin was lower than in nonlesional skin. CONCLUSION: Our finding suggests that increased levels of oxidative stress might impact tyrosinase activity and eumelanin synthesis via anabolism pathway of melanin synthesis. In sum, we observed a negative correlation between levels of systemic oxidative stress and of tyrosinase activity.

ASSESSMENT OF MC1R AND α-MSH GENE SEQUENCES IN IRANIAN VITILIGO PATIENTS.

BACKGROUND: Vitiligo is an acquired pigmentary disorder of the skin that is caused by unknown factors and is characterized by white and depigmented patches that enlarge and become more numerous with time. Genetic factors, oxidative stress, autoimmunity, and neurochemical agents, such as catecholamines might also contribute to vitiligo. Cutaneous pigmentation is determined by the amounts of eumelanin and pheomelanin synthesized by the epidermal melanocytes and interference of melanocortin-1 receptor (MC1R), a G-protein coupled receptor, its normal agonist, alpha-melanocyte stimulating hormone (α-MSH), and key enzymes, such as tyrosinase, to protect against sun-induced DNA damage. The MC1R, a 7 pass trans-membrane G-protein coupled receptor, is a key control point in melanogenesis. Loss-of-function mutations at the MC1R are associated with a switch from eumelanin to pheomelanin production, resulting in a red or yellow coat color. AIM: In this research, we aim to examine the genetic variety of MC1R and α-MSH gene in 20 Iranian vitiligo patients and 20 healthy controls. MATERIALS AND METHODS: Analysis of the MC1R coding gene was performed with direct sequencing. RESULTS: We found the following 9 MC1R coding region variants: Arg163Gl (G488A), Arg227Leu (G680A), Val 97Phe (G289T), Asp184Asn (G550A), Arg227Lys (G680A), Arg142His (G425A), Val60Leu (G178T), Val247Met (C739A), and Val174Ile (G520A). We also found 2 frameshift changes: one of them was the Insertion of C (frameshift in Pro136, stop at Trp148) and the other, Insertion of G (frameshift in Pro256, stop at Trp 333). Of all the changes, the most common was Val60Leu at 5% in patients vs 20% in controls, Val247Met at 15% in patients vs 0% in controls and Val174Ile at 15% in controls and 0% in patients. The other variants showed a frequency <5% in both patients and controls. Also in this study, we have examined the frequency of single nucleotide polymorphisms within the α-MSH genes with direct sequencing in 20 patients and 20 healthy subjects but found no changes along this gene. CONCLUSION: We could not find any relationship between MC1R and α-MSH genes and their effect on the disease in Iranian vitiligo patients.

Elevated expression of human alpha-1 antitrypsin mediated by yeast intron in Pichia pastoris.

Intron-mediated enhancement has been documented in many cases to involve large positive effect on gene expression. To address this, human Alpha-1 antitrypsin (hAAT) gene was integrated into Pichia pastoris with and without a yeast intron generated from the final plasmid pBlu-exII-int-exIII and ligated into the EcoRI/BamHI multiple cloning site of the yeast shuttle vector pHIL-S1. The chimeric exon-intron complex in the middle of the naturally occurring hAAT exons II and III caused a 23-fold enhancement of hAAT expression in P. pastoris, measured through SDS-PAGE and immunoblot analyses.