Enhanced Dentin-Like Mineralized Tissue Formation by AdShh-Transfected Human Dental Pulp Cells and Porous Calcium Phosphate Cement

Abstract

The aim of the present study was to investigate the effect of Sonic hedgehog (Shh) on human dental pulp cells (hDPCs) and the potential of complexes with Shh gene modified hDPCs and porous calcium phosphate cement (CPC) for mineralized tissue formation. hDPCs were cultured and transfected with adenoviral mediated human Shh gene (AdShh). Overexpression of Shh and cell proliferation was tested by real-time PCR analysis, western blotting analysis, and MTT analysis, respectively. The odontoblastic differentiation was assessed by alkaline phosphatase (ALP) activity and real-time PCR analysis on markers of Patched-1 (Ptc-1), Smoothened (Smo), Gli 1, Gli 2, Gli 3, osteocalcin (OCN), dentin matrix protein-1 (DMP-1), and dentin sialophosphoprotein (DSPP). Finally, AdShh-transfected hDPCs were combined with porous CPC and placed subcutaneously in nude mice for 8 and 12 weeks, while AdEGFP-transfected and untransfected hDPCs were treated as control groups. Results indicated that Shh could promote proliferation and odontoblastic differentiation of hDPCs, while Shh/Gli 1 signaling pathway played a key role in this process. Importantly, more mineralized tissue formation was observed in combination with AdShh transfected hDPCs and porous CPC, moreover, the mineralized tissue exhibited dentin-like features such as structures similar to dentin-pulp complex and the positive staining for DSPP protein similar to the tooth tissue. These results suggested that the constructs with AdShh-transfected hDPCs and porous CPC might be a better alternative for dental tissue regeneration.

Sonic Hedgehog Promotes Cementoblastic Differentiation via Activating the BMP Pathways

“…In addition, blockage of the SHH signaling pathway attenuated rh-SHH induced growth, cementoblastic differentiation, and in vitro angiogenesis. Similarly, Ad-SHHinduced odontoblastic differentiation has been reported to be inhibited by Cy in HDPCs [29]. Our results suggest that rh-SHH upregulates Ptch1 and Gli1 in an autocrine manner, resulting in cell proliferation, cementoblastic differentiation, and in vitro angiogenesis.…”

“…Exogenous supplementation of recombinant SHH protein has been reported to increase in vitro osteoblastic differentiation of pluripotent MSCs [24], osteoblastic MC3T3-E1 cells [25], bone marrow-derived MSCs [26], preosteoblastic cell line KS483s [27], and primary osteoblasts [28]. Moreover, the overexpression of SHH via an adenoviral-mediated human SHH vector (AdSHH) was able to promote proliferation and odontoblastic differentiation of human dental pulp cells (HDPCs) [29]. However, SHH administration did not change rates of cell proliferation or calcification, but enhanced ALP activity and enamel matrix protein expression in a mouse ameloblast-lineage cell line (ALC) [30].…”

The Effects of Platelet-Derived Growth Factor-BB on Human Dental Pulp Stem Cells Mediated Dentin-Pulp Complex Regeneration

“…Materials and Methods are shown in the Supporting Information (References: [27][28][29][30][31][32][33].…”

Stem Cells and Calcium Phosphate Cement Scaffolds for Bone Regeneration

“…The seeding of hUCMSCs onto the CPC scaffold at approximately 2,654 cells/mm 2 yielded a high percentage of live cells, fast proliferation, and high bone marker expression and mineralization (Zhou et al, 2011). Besides hBMSCs and hUCMSCs, stem cells derived from dental tissues, such as dental pulp stem cells (DPSCs) and deciduous teeth stem cells (DTSCs) with the property to form a calcified matrix, also showed promising results for the regeneration of mineralized tissues in combination with CPCs (Xia et al, 2013). hucMsc AnD hbMsc sEEDing On cPc fOr bOnE rEgEnErAtiOn in vivo Strategies for the acceleration of bone regeneration via CPC include the addition of bioactive agents such as osteocalcin, O-phospho-L -serine (Vater et al, 2010), and bioactive glass (Yu et al, 2013).…”