MATERIALS AND METHODS: Three categories of materials, namely, test group 1 (cGIC or type IX GIC), test group 2 (HA-GIC or hydroxyapatite-added GIC), and positive control (glass cover slips) were incubated with human periodontal ligament fibroblasts. The samples were viewed under scanning electron microscope to study the morphological characteristics of fibroblasts. Additionally, elemental analysis was performed to differentiate between the two test groups based on surface chemical composition.
RESULTS: Test group 1 (cGIC) exhibited cells with curled up morphology, indicative of poor attachment to the substrate. Test group 2 (Ha-GIC) exhibited cells with flattened morphology and numerous cellular extensions such as lamellipodia and blebs, indicative of good attachment to the substrate. The test group 2 (Ha-GIC) demonstrated higher surface elemental percentages of calcium and phosphorus.
CONCLUSION: Within the limitations of this study, it may be concluded that hydroxyapatite-added GIC is more biocompatible than conventional GIC (type IX), probably attributed to high elemental percentages of calcium and phosphorus.
CLINICAL SIGNIFICANCE: The search for an ideal cervical restorative dental material has been ever elusive. Hydroxyapatite-added GIC is a simple and economical dental material to fabricate from basic conventional GIC. The results from this study strengthen its candidature for cervical and root surface restorations which may later require soft tissue augmentation. The possibility of connective tissue adhesion to this material is an exciting prospect in the field of periorestorative dentistry.
Objectives: The objective of this study was to utilize a chitosan-based nanoparticle system as the delivery carrier for glutamic acid, a model for encapsulated biomolecules to visualize the in vitro release and accumulation of the encapsulated glutamic acid from chitosan nanoparticle (CNP) systems.
Methods: CNP was synthesized via ionic gelation routes utilizing tripolyphosphate (TPP) as a cross-linker. In order to track glutamic acid release, the glutamic acid was fluorescently-labeled with fluorescein isothiocyanate prior encapsulation into CNP.
Results: Light Scattering data concluded the successful formation of small-sized and mono-dispersed CNP at a specific volume ratio of chitosan to TPP. Encapsulation of glutamic acid as a model cargo into CNP led to an increase in particle size to >100 nm. The synthesized CNP exhibited spherical shape under Electron Microscopy. The formation of CNP was reflected by the reduction in free amine groups of chitosan following ionic crosslinking reactions. The encapsulation of glutamic acid was further confirmed by Fourier Transform Infrared (FTIR) analysis. Cell viability assay showed 70% cell viability at the maximum concentration of 0.5 mg/mL CS and 0.7 mg/mL TPP used, indicating the low inherent toxicity property of this system. In vitro release study using fluorescently-tagged glutamic acids demonstrated the release and accumulation of the encapsulated glutamic acids at 6 hours post treatment. A significant accumulation was observed at 24 hours and 48 hours later. Flow cytometry data demonstrated a gradual increase in intracellular fluorescence signal from 30 minutes to 48 hours post treatment with fluorescently-labeled glutamic acids encapsulated CNP.
Conclusion: These results therefore suggested the potential of CNP system towards enhancing the intracellular delivery and release of the encapsulated glutamic acids. This CNP system thus may serves as a potential candidate vector capable to improve the therapeutic efficacy for drugs and biomolecules in medical as well as pharmaceutical applications through the enhanced intracellular release and accumulation of the encapsulated cargo.