The influence of electrospun fibre scaffold orientation and nano-hydroxyapatite content on the development of tooth bud stem cells in vitro.

In stem cell-based dental tissue engineering, the goal is to create tooth-like structures using scaffold materials to guide the dental stem cells. In this study, the effect of fiber alignment and hydroxyapatite content in biodegradable electrospun PLGA scaffolds have been investigated. Fiber orientation of the scaffolds was random or aligned in bundles. For scaffolds with prefabricated orientation, scaffolds were fabricated from PLGA polymer solution containing 0, 10 or 20 % nano-hydroxyapatite. The scaffolds were seeded with porcine cells isolated from tooth buds (dental mesenchymal, dental epithelial, and mixed dental mesenchymal/epithelial cells). Samples were collected at 1, 3 and 6 weeks. Analyses were performed for cell proliferation, ALP activity, and cell morphology. Fiber alignment showed an effect on cell orientation in the first week after cell seeding, but had no long-term effect on cell alignment or organized calcified matrix deposition once the cells reach confluency. Scaffold porosity was sufficient to allow migration of mesenchymal cells. Hydroxyapatite incorporation did not have a positive effect on cell proliferation, especially of epithelial cells, but seemed to promote differentiation. Concluding, scaffold architecture is important to mesenchymal cell morphology, but has no long-term effect on cell alignment or organized ECM deposition. nHA incorporation does have an effect on cell proliferation, differentiation and ECM production, and should be regarded as a bioactive component of dental bioengineered scaffolds.

Discrete phosphorylated retinoblastoma protein isoform expression in mouse tooth development.

Retinoblastoma protein (pRb) phosphorylation plays a central role in mediating cell cycle G1/S stage transition, together with E2F transcription factors. The binding of pRb to E2F is thought to be controlled by the sequential and cumulative phosphorylation of pRb at various amino acids. In addition to well characterized roles as a tumor suppressor, pRb has more recently been implicated in osteoprogenitor and other types of stem cell maintenance, proliferation and differentiation, thereby influencing the morphogenesis of developing organs. In this study, we present data characterizing the expression of pRb and three phosphorylated pRb (ppRb) isoforms-ppRbS780, ppRbS795, ppRbS807/811-in developmentally staged mouse molar and incisor teeth. Our results reveal distinct developmental expression patterns for individual ppRb isoforms in dental epithelial and dental mesenchymal cell differentiation, suggesting discrete functions in tooth development.