Histomorphometric Evaluation of Bone Regeneration Induced by Biodegradable Scaffolds as Carriers for Dental Pulp Stem Cells in a Rat Model of Calvarial "Critical Size" DefectSusanna Annibali1*, Roberta Quaranta2#, Antonio Scarano3#, Andrea Pilloni1#, Andrea Cicconetti1#, Maria Paola Cristalli4#, Diana Bellavia2* and Livia Ottolenghi1#
- *Corresponding Authors:
- Susanna Annibali
Associate Professor of Oral Surgery
School of Dentistry Director of Postgraduate
Program in Oral Surgery, Department of Oral
and Maxillofacial Sciences Sapienza University of Rome, Italy
Tel: +39 06 499766511
Fax: +39 06 44230811
E-mail: [email protected]
- Diana Bellavia
Associate Professor of Pathology
Department of Molecular Medicine
Sapienza University of Rome, Italy
Tel: (+39 06) 49255674
E-mail: [email protected]
Received date November 17, 2015; Accepted date January 08, 2016; Published date January 15, 2016
Citation: Annibali S, Quaranta R, Scarano A, Pilloni A, Cicconetti A, et al. (2016) Histomorphometric Evaluation of Bone Regeneration Induced by Biodegradable Scaffolds as Carriers for Dental Pulp Stem Cells in a Rat Model of Calvarial "Critical Size" Defect. J Stem Cell Res Ther 6:322. doi:10.4172/2157-7633.1000322
Copyright: © 2016 Annibali S, et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Objective: The aim of this study was to test specific stem cells that could enhance bone formation in combination with specific scaffolds.
Methods: Dental Pulp Stem Cells (DPSCs) were seeded with Granular Deproteinized Bovine Bone (GDPB) or Beta-Tricalcium Phosphate (ß-TCP) in a rat model of calvarial "critical size" defect. DPSCs were isolated from permanent human teeth, obtained and characterized using specific stem cells markers (Nanog and Oct-4) by real time-PCR and immunofluorescence. Cells were differentiated for 10-15 days towards the osteoblastic phenotype with 100μM L-ascorbic acid, added every day in culture medium and 20 vol. percentage of FBS in α-MEM medium. Osteogenic commitment was evaluated with real time-PCR by measuring the expression of specific markers (osteonectin and runx2). When a sufficient cell number was obtained, DPSCs were trypsinized, washed in culture medium and seeded onto the GDPB and ß-TCP scaffold sat a density of 0.5-1×106 cells/scaffold. Two bilateral critical-size circular defects (5 mm diameter; 1 mm thickness) were created from the parietal bone of the 8 athymic T-cell deficient nude rats. One cranial defect for each rat was filled with the scaffold alone and the other defect with the scaffold seeded with stem cells. After 12 weeks post-surgery animals were euthanized and histomorphometric analysis was performed. Differences between groups were analyzed by one-way analysis of variance (ANOVA) followed by Fisher's Protected Least Significant Difference (PLSD) post-hoc test. A p-value <0.05 was considered statistically significant.
Results: GDPB group presented higher percentage of lamellar bone than that of GDPB/DPSC, ß-TCP alone had lower levels as compared to ß-TCP/DPSC. The addition of stem cells significantly increased woven bone formation in both scaffold-based implants, although still higher in GDPB based implants
Conclusion: Our findings indicate that GDPB and ß-TCP used as scaffold to induce bone regeneration may benefit from adding DPSC to tissue-engineered constructs.