Evaluation of bone regeneration potential of dental follicle stem cells for treatment of craniofacial defects

Abstract

Stem cell-based tissue regeneration offers potential for treatment of craniofacial bone defects. The dental follicle (DF), a loose connective tissue surrounding unerupted tooth, has been shown to contain progenitor/stem cells. Dental follicle stem cells (DFSCs) possess strong osteogenesis capability, which makes them suitable for repairing skeletal defects. The objective of this study was to evaluate bone regeneration capability of DFSCs loaded into polycaprolactone (PCL) scaffold for treatment of craniofacial defects. DFSCs were isolated from the first mandibular molars of postnatal Sprague Dawley rats and seeded into PCL scaffold. Cell attachment and cell viability on the scaffold were examined using scanning electron microscopy (SEM) and Alamar blue reduction assay. For in vivo transplantation, critical-size defects (CSD) were created on the skulls of 5 month-old immunocompetent rats, and the cell–scaffold constructs were transplanted into the defects. Skulls were collected at 4 and 8 weeks post-transplantation, and bone regeneration in the defects was evaluated with micro-CT and histological analysis. SEM and Alamar blue assay demonstrated attachment and proliferation of DFSCs in the PCL scaffold. Bone regeneration was observed in the defects treated with DFSC transplantation, but not in the controls without DFSC transplant. Transplanting DFSC-PCL with or without osteogenic induction prior to transplantation achieved approximately 50% bone regeneration at 8 weeks. Formation of woven bone was observed in the DFSC-PCL treatment group. Similar results were seen when osteogenic-induced DFSC-PCL was transplanted to the CSD. This study demonstrated that transplantation of DFSCs seeded into PCL scaffolds can be used to repair craniofacial defects.

Biodegradable zinc oxide composite scaffolds promote osteochondral differentiation of mesenchymal stem cells

“…ZnO was combined with PCL which allowed for the slow release of zinc. PCL has been investigated widely for tissue engineering cartilage and bone (Rezai‐Rad et al, ; Wise, Yarin, Megaridis, & Cho, ; Xue et al, ). With the addition of ZnO to PCL, the chondrogenic differentiation was enhanced where cells on the low percentage of ZnO demonstrated increased GAG and collagen type II production and gene expression of chondrogenic markers.…”

Purmorphamine increased adhesion, proliferation and expression of osteoblast phenotype markers of human dental pulp stem cells cultured on beta-tricalcium phosphate

“…Cells were resuspend in medium containing DMEM (Life Technologies, Carlsbad, CA, United States), 20% fetal bovine serum (FBS) (Invitrogen, Carlsbad, CA, United States) + 1% Penicillin-Streptomycin 10,000 u/ml (Life Technologies, Carlsbad, CA, United States). This medium has been used previously as a growth medium for dental stem cells [23]. Cell suspension was immediately plated in a T-25 flask and placed at 37 C and 5% CO 2 , and non-adherent cells were removed by medium change a day after.…”

Expression of odontogenic ameloblast-associated protein in the dental follicle and its role in osteogenic differentiation of dental follicle stem cells

“…Another study showed that ODAM cooperates with RUNX2 to modulate mineralization (Lee et al, 2010). Studies in our lab also revealed that rat DFSCs cultured in osteogenic medium dramatically and significantly increase the expression of osteogenic marker genes, osteocalcin and bone sialoprotein (Rezai Rad et al, 2015), suggesting that DFSCs can differentiate into osteoblasts under the proper conditions. In this study, we mimicked the increase of ODAM expression by transfecting the plasmid containing rat ODAM coding sequence into human and rat DFSCs, and found that increase of ODAM expression enhanced osteogenesis of the DFSCs at an early stage of osteo-induction when the transfected DFSCs were cultured in osteogenesis medium.…”