Introduction: Biocompatible and biodegradable scaffolds based on natural polymers such as gelatin and chitosan (CS) provide suitable microenvironments in dental tissue engineering. In the present study, we report on the synthesis of injectable thermosensitive hydrogel (PNIPAAm-g-CS copolymer/gelatin hybrid hydrogel) for osteogenic differentiation of human dental pulp stem cells (hDPSCs). Methods: The CS-g-PNIPAAm was synthesized using the reaction of carboxyl terminated PNIPAAm with CS, which was then mixed with various amounts of gelatin solution in the presence of genipin as a chemical crosslinker to gain a homogenous solution. The chemical composition and microstructures of the fabricated hydrogels were confirmed by FT-IR and SEM analysis, respectively. To evaluate the mechanical properties (e.g., storage and loss modulus of the gels), the rheological analysis was considered. Calcium deposition and ALP activity of DPSCs were carried out using alizarin red staining and ALP test. While the live/dead assay was performed to study its toxicity, the real-time PCR was conducted to investigate the osteogenic differentiation of hDPSCs cultured on prepared hydrogels. Results: The hydrogels with higher gelatin incorporation showed a slightly looser network compared to the other ones. The hydrogel with less gelatin indicates a rather higher value of G', indicating a higher elasticity due to more crosslinking reaction of amine groups of CS via a covalent bond with genipin. All the hydrogels contained viable cells with negligible dead cells, indicating the high biocompatibility of the prepared hydrogels for hDPSCs. The quantitative results of alizarin red staining displayed a significant rise in calcium deposition in hDPSCs cultured on prepared hydrogels after 21 days. Further, hDPSCs cultured on hydrogel with more gelatin displayed the most ALP activity. The expression of late osteogenic genes such as OCN and BMP-2 were respectively 6 and 4 times higher on the hydrogel with more gelatin than the control group after 21 days. Conclusion: The prepared PNIPAAm-g-CS copolymer/gelatin hybrid hydrogel presented great features (e.g., porous structure, suitable rheological behavior, and improved cell viability), and resulted in osteogenic differentiation necessary for dental tissue engineering.
Introduction: In this work, a flexible, and wearable point-of-care (POC) device integrated on a pain relief patch as wearable colorimetric sensors have been developed for sweat analysis, such as lactic acid, sodium ions, and pH simultaneously. Herein, the patch has still functioned as pain relief, while it allows for sweat monitoring during exercise, and in daily activities.
Methods: It was constructed on cotton cloth using wax printing technology (batik stamp) as cloth-based microfluidic devices (CMDs). Here, it uses micro volumes of samples to perform the reaction in the sensing zones, where the sensitive reagents are immobilized so that it can collect and analyze the sweat (lactic acid, sodium ions, and pH) as the model for sweat analytes. The colorimetric analysis was conducted via a smartphone camera by using a free app (Color Grab) for a color image analysis that uses for quantitative analysis or naked eye for semi-qualitative analysis.
Results: The ∆RGB value of the CMDS shows the excellent linear correlation vs analytes concentration, where the coefficient of correlations was found for lactic acid (R2 = 0.994), sodium ion (R2 = 0.998), and pH (R2 = 0.994). The ∆RGB value shows the appropriate color value for the linear correlation of the analyte target concentrations in the sweat samples. Here, the limit of detection (LOD) was found at 45.73 µg/mL for lactic acid and 56.46 µg/mL for sodium ions. The reproducibility was found at 0.79% and 0.89%, for lactic acid and sodium ions respectively.
Conclusion: It was applied for sweat analysis during exercise, and the results show in agreement with the standard methods used in a clinical laboratory.