IADR Abstract Archives
Dental Origin Stem Cells in Plant Scaffolds
Objectives: To observe the biocompatibility of different human cells on various 3D plant derived scaffolds
Methods: 5 mm thick sheets were obtained from apple, leek, green onion, green garlic and they were carved into circles with a diameter of 16mm. The samples were treated with 0.1% SDS solution and spun at low speed for 72h. Following the decellularization protocol, the scaffolds were incubated in 100mM CaCl2, spun for another 24h to remove any surfactant residue and sterilized in 70% ethanol overnight. Then they were incubated for 24h with a solution of DMEM and 10% FBS. Meanwhile, PerioDontal Ligament Stem Cells (PDLSC), Stem Cells from Apical Papilla (SCAP), Hertwig's Epithelial Root Sheath (HERS), Follicular Stem Cells(FSC), Dental Pulp Stem Cells(DPSC), Bone Marrow Stem Cells(BMC), Stem cells from Human Exfolliated Deciduous teeth (SHED) were isolated from routinely extracted human third molars, using laboratory routine methods (1,2). Cells between 3rd and 5th passage were seeded in the scaffolds and cultivated for 4 weeks. After that the samples were fixed and treated with DAPI for InCell observation.
Results: The 3D plant derived scaffolds are similar to the structure and the size of human cells. All the decellularized scaffolds were found to be supportive for the survival and proliferation of PDLSC, SCAP, HERS, FSC, DPSC, BMC. Fluorescent staining revealed clusters of cell nuclei forming 3D structures on the surface and inside the scaffolds. The plant derived scaffolds did not show any significant toxicity.
Conclusions: Natural 3D scaffolds are a great cellular microenvironment for human culture growth. The decellularized plant tissues supported the cell adhesion, proliferation and functionality of PDLSC, SCAP, HERS, FSC, DPSC, BMC, SHED. Additionally, there was no cytotoxicity exhibited, therefore proving they are a promising substitute of synthetic scaffolds and can be used for many in vivo and in vitro tissue engineering purposes.
Methods: 5 mm thick sheets were obtained from apple, leek, green onion, green garlic and they were carved into circles with a diameter of 16mm. The samples were treated with 0.1% SDS solution and spun at low speed for 72h. Following the decellularization protocol, the scaffolds were incubated in 100mM CaCl2, spun for another 24h to remove any surfactant residue and sterilized in 70% ethanol overnight. Then they were incubated for 24h with a solution of DMEM and 10% FBS. Meanwhile, PerioDontal Ligament Stem Cells (PDLSC), Stem Cells from Apical Papilla (SCAP), Hertwig's Epithelial Root Sheath (HERS), Follicular Stem Cells(FSC), Dental Pulp Stem Cells(DPSC), Bone Marrow Stem Cells(BMC), Stem cells from Human Exfolliated Deciduous teeth (SHED) were isolated from routinely extracted human third molars, using laboratory routine methods (1,2). Cells between 3rd and 5th passage were seeded in the scaffolds and cultivated for 4 weeks. After that the samples were fixed and treated with DAPI for InCell observation.
Results: The 3D plant derived scaffolds are similar to the structure and the size of human cells. All the decellularized scaffolds were found to be supportive for the survival and proliferation of PDLSC, SCAP, HERS, FSC, DPSC, BMC. Fluorescent staining revealed clusters of cell nuclei forming 3D structures on the surface and inside the scaffolds. The plant derived scaffolds did not show any significant toxicity.
Conclusions: Natural 3D scaffolds are a great cellular microenvironment for human culture growth. The decellularized plant tissues supported the cell adhesion, proliferation and functionality of PDLSC, SCAP, HERS, FSC, DPSC, BMC, SHED. Additionally, there was no cytotoxicity exhibited, therefore proving they are a promising substitute of synthetic scaffolds and can be used for many in vivo and in vitro tissue engineering purposes.