Characterization of human periodontal ligament cells cultured on three-dimensional biphasic calcium phosphate scaffolds in the presence and absence of L-ascorbic acid, dexamethasone and β-glycerophosphate in vitro

An,Shaofeng; Gao,Yan; Ling,Junqi

doi:10.3892/etm.2015.2706

Characterization of human periodontal ligament cells cultured on three-dimensional biphasic calcium phosphate scaffolds in the presence and absence of L-ascorbic acid, dexamethasone and β-glycerophosphate in vitro

Authors:
- Shaofeng An
- Yan Gao
- Junqi Ling
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Affiliations: Department of Operative Dentistry and Endodontics, Guanghua School of Stomatology, Hospital of Stomatology, Sun Yat‑sen University; Guangdong Provincial Key Laboratory of Stomatology, Guangzhou, Guangdong 510055, P.R. China
Published online on: August 24, 2015 https://doi.org/10.3892/etm.2015.2706
Pages: 1387-1393
Copyright: © An et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of this study was to evaluate the effect of porous biphasic calcium phosphate (BCP) scaffolds on the proliferation and osteoblastic differentiation of human periodontal ligament cells (hPDLCs) in the presence and absence of osteogenic inducer (L‑ascorbic acid, dexamethasone and β‑glycerophosphate). The cell growth within the scaffolds in the absence of osteogenic inducers was studied by cell counting kit‑8 (CCK‑8) assay and scanning electron microscopy (SEM). Alkaline phosphatase (ALP) activity and osteoblastic differentiation markers of hPDLCs in BCP scaffolds were examined in the presence and absence of osteogenic inducers. The cell number of hPDLCs in the BCP scaffolds was less than that of hPDLCs cultured in microplates (control). SEM images showed that cells successfully adhered to the BCP scaffolds and spread amongst the pores; they also produced abundant extracellular cell matrix. In the presence and absence of osteogenic inducers, the ALP activity of hPDLCs within BCP scaffolds was suppressed in varying degrees at all time‑points. In the absence of osteogenic inducers, hPDLCs in BCP scaffolds express significant higher levels of osteopontin (OPN) mRNA than the control, and there were no significant differences for Runx2 and osteocalcin (OCN) mRNA levels compared with those cultured in microplates. In the presence of osteogenic inducers, Runx2 expression levels were significantly higher than those in control. OPN and OCN mRNA levels were downregulated slightly. Three‑dimensional porous BCP scaffolds are able to stimulate the osteoblastic differentiation of hPDLCs in the presence and absence of osteogenic inducer and may be capable of supporting hPDLC-mediated bone formation.

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