METHOD: DPSC from murine incisors were isolated through either the outgrowth (DPSC-OG) or the enzymatic digestion (DPSC-ED) method. Cells at passage 4 were used in this study. The cells were characterized through morphology and expression of cell surface markers. The cells' doubling time when cultured using different seeding densities was calculated and analyzed using one-way ANOVA and Tukey's multiple comparison post-test. The ability of cells to differentiate to chondrocyte and osteoblast was evaluated through staining and analysis on the matrices secreted.
RESULTS: Gene expression analysis showed that DPSC-OG and DPSC-ED expressed dental pulp mesenchymal stem cell markers, but not hematopoietic stem cell markers. The least number of cells that could have been used to culture DPSC-OG and DPSC-ED with the shortest doubling time was 5 × 10(2) cells/cm(2) (11.49 ± 2.16 h) and 1 × 10(2) cells/cm(2) (10.55 h ± 0.50), respectively. Chondrocytes differentiated from DPSC-ED produced 2 times more proteoglycan and at a faster rate than DPSC-OG. FTIR revealed that DPSC-ED differentiated into osteoblast also secreted matrix, which more resembled a calvaria.
DISCUSSION: Isolation approaches might have influenced the cell populations obtained. This, in turn, resulted in cells with different proliferation and differentiation capability. While both DPSC-OG and DPSC-ED expressed mesenchymal stem cell markers, the percentage of cells carrying each marker might have differed between the two methods. Regardless, enzymatic digestion clearly yielded cells with better characteristics than outgrowth.
METHODS: 1.5% (w/v) chitosan films with Chrysanthemum morifolium essential oil (0% to 6% (v/v)) were produced through homogenization, the casting of a film solution in a petri dish and convection drying. The edible film was evaluated in terms of its physical (color, thickness, water vapor permeability), mechanical (puncture strength, tensile strength, elongation at break) and chemical properties (antioxidant assay, Fourier Transform Infrared Spectroscopy (FTIR)).
RESULTS: With an increasing concentration of Chrysanthemum morifolium in the chitosan film, the test values of physical properties such as tensile strength, puncture force, and elongation at break declined significantly. However, the thickness, water permeability, and color profile (L*, a*, b*) values of the chitosan film increased. Similarly, the scavenging effect of antioxidant assay increased (from 4.97% to 18.63%) with a rise in Chrysanthemum morifolium concentration. 2%, 3%, and 4% of Chrysanthemum morifolium in the chitosan film showed a significant inhibition zone ranging from 2.67 mm to 3.82 mm against Staphylococcus aureus, a spoilage bacterium that is commonly found in chicken and beef products. The storage and pH tests showed that 4% of Chrysanthemum morifolium in the film maintained pH level (safe to consume), and the shelf life was extended from 3 days to 5 days of meat storage.
CONCLUSIONS: This study demonstrated that the incorporation of 4% (v/v) Chrysanthemum morifolium extract into 1.5% (w/v) chitosan film extends the storage duration of raw meat products noticeably by reducing Staphylococcus aureus activity. Therefore, it increases the quality of the edible film as an environmentally friendly food packaging material so that it can act as a substitute for the use of plastic bags. Future studies will be conducted on improving the tensile strength of the edible film to increase the feasibility of using it in the food industry. In addition, the microstructure and surface morphology of the edible film can be further determined.