Cell & Stem Cell Research

Human dental pulp represents a suitable alternative source of stem cells for the purpose of cell-based therapies in regenerative medicine. Spinal cord injuries at the cervical (neck) level result in full or partial tetraplegia ,low heart rate, low blood pressure, problems in regulating body temperature, and breathing dysfunction. We aim to recommend the best cell population and the best cell transplant criteria to treat cervical spinal cord injury. In this study we successfully isolate CD 24 positive cells from human dental apical papilla from impacted third molar of patient female 15 years using two different techniques enzymatic digestion using Collagenase 3 mg/ml / Dispase 4 mg/ml enzymes also with explant outgrowth method. Characterization done by using flowcytometry cell sorting and we confirm the viability of the sorted cells using trypan blue stain. Interestingly, despite the sorted cells was express

a limited present in the cell population only 5.9% but after culturing in Proliferation Media (PM): DMEM/F12 supplemented with 10% FBS, 100 μM L-ascorbic acid 2-phosphate, 2 mM L-glutamine, 100 units/ml penicillin, 100 mg/ml streptomycin, they give a high proliferation rate. The teeth of the study opened using Hand-held dental pulp isolator device Patent (WO 2016162041 A9). Based on the role of ENTPD1 gene in development of central nervous system during embryogenesis, hydrolyze ATP and other nucleotides to regulate purinergic neurotransmission, anti-inflammatory effect TNF-α-induced cytotoxicity and cell death also its high expression in dental derived stem cells so in the next steps we will compare between seven sub- populations CD24, CD39 (ENTPD1), CD90, CD73, CD105, Nestin and SSEA-3 positive cells from different dental sources as apical papilla, dental follicle and dental pulp of

human tooth. Moreover we will compare between whether cell transplant or whole tissue transplant of the dental pulp in spinal cord injury in sprague dawely rats is more effective. Regarding tissue transplant we will compare between graphene nano-scaffold with and without hydrogel, carbon nano-tubes and high or low porosity PCL Poly (ε-caprolactone) scaffolds and test the Adhesion, Cytotoxicity, Proliferation and neuron-differentiation capacities of the cells or the dental pulp tissue.