Region-Specific Decellularization of Porcine Uterine Tube Extracellular Matrix: A New Approach for Reproductive Tissue-Engineering Applications.

The uterine tube extracellular matrix is a key component that regulates tubal tissue physiology, and it has a region-specific structural distribution, which is directly associated to its functions. Considering this, the application of biological matrices in culture systems is an interesting strategy to develop biomimetic tubal microenvironments and enhance their complexity. However, there are no established protocols to produce tubal biological matrices that consider the organ morphophysiology for such applications. Therefore, this study aimed to establish region-specific protocols to obtain decellularized scaffolds derived from porcine infundibulum, ampulla, and isthmus to provide suitable sources of biomaterials for tissue-engineering approaches. Porcine uterine tubes were decellularized in solutions of 0.1% SDS and 0.5% Triton X-100. The decellularization efficiency was evaluated by DAPI staining and DNA quantification. We analyzed the ECM composition and structure by optical and scanning electronic microscopy, FTIR, and Raman spectroscopy. DNA and DAPI assays validated the decellularization, presenting a significative reduction in cellular content. Structural and spectroscopy analyses revealed that the produced scaffolds remained well structured and with the ECM composition preserved. YS and HEK293 cells were used to attest cytocompatibility, allowing high cell viability rates and successful interaction with the scaffolds. These results suggest that such matrices are applicable for future biotechnological approaches in the reproductive field.

Pressurized liquid extraction followed by high-performance liquid chromatography for determination of beta-ecdysone extracted from Pfaffia glomerata (Spreng.) Pedersen.

Pfaffia glomerata (Spreng.) Pedersen has among its main bioactive compounds saponins, with the phytoestroid ß-ecdysone as its chemical marker. In this study, pressurized liquid extraction (PLE), a green extraction technique used to obtain bioactive compounds from plants, was employed to extract beta-ecdysone from P. glomerata leaves, stems, and roots. The 22 factorial design was used with the variables temperature (333 K and 353 K) and flow rate (1.5 and 2 mL min-1), pressure (300 Bar), time (60 min), and solvent [ethanol and distilled water (70:30 (v/v)] were kept constant for all parts of the plant. The results of experimental responses demonstrated that the factors temperature and flow rate significantly interfere with the yields of leaf (0.499%), root (0.65%) and stem (0.764%) extracts. The latter presented presents the highest yield compared to the other parts of the plant. HPLC results showed the presence of beta-ecdysone in all parts of the plant with concentrations of ß-ecdysone 86.82, 76.53 and 195.86 mg L-1 to leaf, root and stem, respectively. FT Raman results exhibited typical peaks of beta-ecdysone, such as 3310 cm-1, 1654 cm-1, and 1073 cm-1 for all plant parts. Another interesting result was the presence of the peak at 1460 cm-1 in the PLE root extract can be associated with selenium. This foundational knowledge confirms that the PLE extraction process was efficient in obtaining the chemical marker of Pfaffia glomerata in all plant parts.

Optimization and Characterization of Protein Extraction from Asparagus Leafy By-Products.

Asparagus production generates significant amounts of by-products during the summer and post-harvest growth period. By-products can be good sources of nutrients and phytochemicals. The interest in increasing the availability of proteins for human consumption has led to the use of new plant sources rich in proteins. The objective of this study was to use response surface methodology (RSM) to optimize the aqueous extraction process of proteins from asparagus leafy by-products, for the production of new protein ingredients. The optimum extraction condition was at pH 9, with 40 min of extraction at 50 °C, and the concentration was fixed at 5 g·L-1. The isolate obtained presented 90.48% protein with 43.47% protein yield. Amino acids such as alanine, proline, valine, leucine/isoleucine, asparagine, and phenylalanine were identified, and the antioxidant activity for 2,2 AZINO BIS (3-ethylbenzo thiazoline 6 sulfonic acid diammonium salt) was 145.76 equivalent to Trolox µmol.100g-1 and for DPPH 65.21 equivalent to Trolox µmol.100g-1. The product presented favorable technological properties (water absorption capacity 4.49 g·g-1 and oil absorption capacity 3.47 g·g-1) and the color tended towards dark green (L* 31.91, a* -1.01, b* -2.11). The protein isolate obtained through the extraction optimization process showed high potential to be used as a protein ingredient.

Performance of self-etching adhesives on caries-affected primary dentin treated with glutaraldehyde or silver diamine fluoride.

OBJECTIVES: to evaluate the quality and stability of adhesive interfaces established by self-etching adhesives on caries-affected primary dentin (CAD) treated with glutaraldehyde (GA) or silver diamine fluoride (SDF). METHODS: 42 primary molars were exposed to a microbiological caries-inducing protocol and divided into 6 groups according to the adhesive system (Clearfil SE - CL or FL Bond II - FL) and pretreatment (water, GA or SDF) applied on CAD. One tooth from each group was analyzed for surface modification using infrared spectroscopy. Crowns were restored with resin composite (n = 36) and cut into beams and slices. The beams were subjected to microtensile testing, Raman spectroscopy and SEM after 24 h and 6 months of storage. The slices were analyzed using Micro-Raman spectroscopy to determine the diffusion zone thickness (DZ) in each period. Data were analyzed by ANOVA and Tukey or Kruskal-Wallis and Dunn tests (α = 0.05%). RESULTS: SDF reduced the immediate bond strength for both adhesives. The control groups showed a decrease in BS after 6 months in artificial saliva. GA increased immediate DZ for FL, while SDF had the opposite effect on CL. GA decreased the DZ for FL at 6 months. There was a predominance of adhesive failures with areas of cohesive dentin fractures within control groups. SIGNIFICANCE: Modifications caused by dentin surface treatments may directly affect the performance of adhesive systems and the quality and stability of adhesive restorations.

Study of the chemical interaction between a high-viscosity glass ionomer cement and dentin

Abstract Objective To investigate the chemical interactions between a high-viscosity glass ionomer cement (GIC) (KetacTM Molar Easymix, 3M ESPE, Seefeld, Bavaria, Germany) and human dentin. It was also analyzed the dynamics of GIC setting mechanism based on the time intervals required for the GIC and the GIC mixed with dentin to achieve stability. Material and Methods Each constituent of GIC - powder (P) and liquid (L) - and powdered dentin (D), as well as the associations P+L, D+L, and P+L+D in the concentrations of 29%, 50%, 65%, 78%, 82%, and 92% of GIC were analyzed with Fourier transform infrared (FTIR) and Raman spectroscopy. Results New optical absorption bands and/or Raman bands, which were not present in P, L, or D, were observed in the associations. The concentrations of 29% and 50% of GIC showed higher interaction, revealing that the amount of dentin influences the formation of new optical absorption or scattering bands. FTIR bands showed that the setting time to achieve bond stability was longer for the high-viscosity GIC (38±7 min) than for the sample with 29% of GIC (28±4 min). Conclusions The analysis revealed the formation of new compounds or molecular rearrangements resulting from the chemical interactions between GIC and dentin. Moreover, this study provides an effective method to evaluate the dynamics of the setting mechanism of GICs.