This study evaluated the potential toxicity, genotoxicity, and cellular uptake of various sizes and concentrations of chitosan (CS) nanoparticles cultured with normal human dental pulp cells. Normal human dental pulp cells (hDPCs) were derived from human dental pulp tissues and cultured with (50–67) nm and (318–350) nm CS-nanoparticles in concentrations of 0.1 mg/mL, 0.5 mg/mL, 1 mg/mL, 2 mg/mL, and 4 mg/mL as study groups and 0 mg/mL as a control group for time intervals of 16 hours, 24 hours, 3 days, 7 days and 14 days. Attachment efficiency and proliferation rate were assessed by measuring the optical density of crystal violet-stained cells. Cell viability was determined by the activity of mitochondrial dehydrogenase enzymes. Genotoxicity was assessed using the cytokinesis-block micronucleus method and by measuring the fluorescent intensity of phosphorylated H2AX nuclear foci. Cellular uptake was determined by tagging chitosan nanoparticles with FITC stain and then measuring the fluorescence intensity of
FITC-tagged chitosan nanoparticles using a spectrophotometer. Statistical analysis was performed using chi-square, one-way ANOVA, and post-hoc Tukey tests.
All concentrations of the (50–67) nm group significantly reduced attachment efficiency in comparison with control (P< 0.01) and with (318–350) nm group (p<0.01). Proliferation rate and cell viability were significantly reduced in cells exposed to various concentrations of (50-67) nm chitosan when compared to (318-350) nm group (P<0.05) and control group (P<0.05). For both size groups, higher concentrations significantly showed lower proliferation rate and cell viability when compared to lower concentration (P< 0.01). CS-nanoparticles were able to internalize hDPCs and significantly induced micronuclei, nuclear buds, and pH2AX at concentrations of 0.5 mg/mL and 2 mg/mL as compared to 0.1 mg/mL (P<0.01) and control groups (P< 0.01). At both the 0.5 mg/mL and 2 mg/mL concentrations, (50–67) nm chitosan significantly induced higher proportions of micronuclei (P= 0.001), nuclear buds (P= 0.009), and pH2AX nuclear foci (P= 0.00004) compared to (318–350) nm chitosan. In conclusion, CS-nanoparticles at sizes (50–67) nm and (318–350) nm at a concentration of (0.5–4) mg/mL internalized hDPCs and exhibited cytotoxic and genotoxic effects in dose-dependent and size-associated manners.
| Identifer | oai:union.ndltd.org:bu.edu/oai:open.bu.edu:2144/42925 |
| Date | 19 August 2021 |
| Creators | Alhomrany, Rami Mohammed |
| Contributors | Chou, Laisheng |
| Source Sets | Boston University |
| Language | en_US |
| Detected Language | English |
| Type | Thesis/Dissertation |
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