There are multiple causes of tooth loss can be caused by multiple reasons, including caries, periodontal disease, trauma and genetic disease. The dental pulp has important functions to sustain teeth, by providing nutrient and oxygen supply, innervation and immune response. Our objective was to determine whether incorporating extracellular matrix derived from decellularized pig tooth buds (tECM) into silk scaffolds would enhance dental pulp derived dental mesenchymal stem cell (DMSC) differentiation. Nine different types of silk scaffolds were fabricated: 1) 6% silk; 2) 6% silk+tECM; 3) 6% silk+collagen; 4) 3% silk, 4g salt; 5) 3% silk, 4g salt + tECM; 6) 3% silk, 4g salt + collagen; 7) 3% silk, 2g salt; 8) 3% silk, 2g salt + tECM; and 9) 3% silk, 2g salt + collagen. Unseeded silk scaffolds were used as controls. Three replicates were used for each experimental condition and controls. Porcine DMSCs (pDMSCs) at passage 2 were seeded into silk scaffolds (3×105 cells/scaffold, ~3×103cells/mm3) and cultured in vitro in osteogenic media for 24 hours, or for 2 and 4 weeks. Histological analyses of paraffin embedded and sectioned constructs revealed pDMSC attachment and proliferation after two and four-week in vitro culture. Immunofluorescent (IF) analysis showed robust dentin sialoprotein (DSP) expression in all pDMSC seeded silk scaffolds. DSP expression was enhanced in scaffolds containing tECM. Statistical analysis demonstrated significant differences in DSP expression in scaffolds with versus without tECM (P≤0.01). Silk scaffolds supported the attachment, proliferation and differentiation of seeded pDMSCs. Furthermore, our research is the first study to demonstrate that tECM can enhance the pDMSC differentiation, suggesting that tECM incorporated scaffolds show promise for dental pulp and/or whole tooth regeneration. Studies of in vivo implanted constructs are being performed to further investigate the effect of tECM on DMSC differentiation and mineralized tissue formation.
|Advisor:||Yelick, Pamela C.|
|Commitee:||Pagni, Sarah, Trotman, Carroll Ann, Zhang, Weibo|
|School:||Tufts University School of Dental Medicine|
|School Location:||United States -- Massachusetts|
|Source:||MAI 57/05M(E), Masters Abstracts International|
|Keywords:||Decellularized tooth bud scaffolds, Dental stem cells, Pulp regeneration, Silk scaffolds|
Copyright in each Dissertation and Thesis is retained by the author. All Rights Reserved
The supplemental file or files you are about to download were provided to ProQuest by the author as part of a
dissertation or thesis. The supplemental files are provided "AS IS" without warranty. ProQuest is not responsible for the
content, format or impact on the supplemental file(s) on our system. in some cases, the file type may be unknown or
may be a .exe file. We recommend caution as you open such files.
Copyright of the original materials contained in the supplemental file is retained by the author and your access to the
supplemental files is subject to the ProQuest Terms and Conditions of use.
Depending on the size of the file(s) you are downloading, the system may take some time to download them. Please be