الفهرس 
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Abstract
Loss of pulp vitality results in a tooth more susceptible to structural, esthetic failure and secondary infection. Thus, the preservation of pulp vitality is considered an important challenge in order to maintain its function. The regenerative endodontic technology aims at preserving and regenerating the vitality and health of the dental pulp and restoring the physiologically functional dentition.
The aim of the present study is to evaluate the effect of Neo MTA and hydroxyapatite nanoparticulates on odontogenic differentiation and proliferation of human dental pulp stem cells.
The materials used in this study were Neo Mineral Trioxide Aggregate (NeoMTA 2) (Avalon Biomed, Texas, USA), prepared in nanoform at (Nanogate, Cairo, Egypt) and Nanohydroxyapatite (NHAP) (Nanogate, Cairo, Egypt). Human dental pulp tissues were obtained from three impacted third molars. Teeth were collected from three healthy patients aged from 18–20 years after written informed consent. Extraction of teeth was performed under sterile conditions in the department of Oral Surgery Department at Ain Shams University clinics under the approval of its ethics committee.
MSCs were isolated from dental pulp tissue using enzyme digestion method by the following protocol:
• The dental pulp tissue was minced into small pieces in Petri dish containing PBS (pH 7.4) and antibiotics.
• The tissue was digested into collagenase type I and dispase solution with continuous agitation for 5 minutes at room temperature.
• Tissue clumps are passed through a cell strainer to obtain single cell suspension.
• Cells were cultured in a flask 75 cm2, in Gibco Dulbecco’s Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) and 1% of penicillin G sodium (10.000 UI), streptomycin (10 mg) and amphotericin B (25 μg) (PSA). Flask was incubated for 48 hours at 37 °C in an atmosphere of 5% CO2. The media was changed every 24 hours. When the cells reached 70% confluence, the cells were harvested and passaged. Cells from passages 3rd passage were used.
HDPSCs were tested for differentiation and proliferation after application of previously prepared materials.
• group 1: HDPSCs cultured in previously prepared Nano Neomta (9.92 µg/ml).
• group 2: HDPSCs cultured in previously prepared Nano-hydroxyapatite (NHAP 10 µg/mL)
• group 3: Positive control group (OM-PC): HDPSCs cultured in odontogenic differentiation medium (OM).
• group 4: Negative control group (DMEM-NC): HDPSCs cultured in DMEM media.
Plates were incubated at 37 °C and 5% CO2 for 72 hours, each experiment was carried out in triplicate for each group and analyzed in three independent experiments.
Cell Differentiation tested by:
- Alkaline phosphatase assay
- Immunofluorescence staining
Cell Proliferation tested by:
- Trypan blue assay
- MTT assay