Effects of 3-dimensional Bioprinting Alginate/Gelatin Hydrogel Scaffold Extract on Proliferation and Differentiation of Human Dental Pulp Stem Cells

Haiyue Yu; Xiaoyi Zhang; Wenjing Song; Ting Pan; He Wang; Tingting Ning; Qin Wei; Hockin H K Xu; Buling Wu; Dandan Ma

doi:10.1016/j.joen.2019.03.004

Effects of 3-dimensional Bioprinting Alginate/Gelatin Hydrogel Scaffold Extract on Proliferation and Differentiation of Human Dental Pulp Stem Cells

J Endod. 2019 Jun;45(6):706-715. doi: 10.1016/j.joen.2019.03.004. Epub 2019 May 2.

Authors

Haiyue Yu¹, Xiaoyi Zhang², Wenjing Song³, Ting Pan⁴, He Wang², Tingting Ning², Qin Wei⁵, Hockin H K Xu⁶, Buling Wu⁷, Dandan Ma⁸

Affiliations

¹ Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou, China; College of Stomatology, Southern Medical University, Guangzhou, China; Department of Operative and Preventive Dentistry, Charité-Universitätsmedizin Berlin, Berlin, Germany.
² Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou, China; College of Stomatology, Southern Medical University, Guangzhou, China.
³ School of Materials Science and Engineering, South China University of Technology, Guangzhou, China; National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou, China; Key Laboratory of Biomedical Materials and Engineering of the Ministry of Education, South China University of Technology, Guangzhou, China.
⁴ School of Materials Science and Engineering, South China University of Technology, Guangzhou, China; National Engineering Research Center for Tissue Restoration and Reconstruction, Guangzhou, China.
⁵ Department of Operative Dentistry and Endodontics, Guanghua School of Stomatology, Sun Yat-sen University, Guangzhou, China.
⁶ Department of Advanced Oral Sciences and Therapeutics, University of Maryland School of Medicine, Baltimore, Maryland; Center for Stem Cell Biology and Regenerative Medicine, University of Maryland School of Medicine, Baltimore, Maryland; Department of Mechanical Engineering, University of Maryland, Baltimore, Maryland.
⁷ Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou, China; College of Stomatology, Southern Medical University, Guangzhou, China. Electronic address: bulingwu1958@163.com.
⁸ Department of Stomatology, Nanfang Hospital, Southern Medical University, Guangzhou, China; College of Stomatology, Southern Medical University, Guangzhou, China. Electronic address: mdd@smu.edu.cn.

PMID: 31056297
DOI: 10.1016/j.joen.2019.03.004

Abstract

Introduction: Alginate/gelatin hydrogel (Alg-Gel) scaffold has been applied in tissue engineering, but the research on its application in dental tissues regeneration is still lacking. We investigated the effect of this scaffold on human dental pulp stem cells (hDPSCs).

Methods: hDPSCs were cultured in both Alg-Gel and 3D-printed Alg-Gel scaffolds. Cell growth and adhesion were compared using fluorescein isothiocyanate-phalloidin staining and scanning electron microscopic micrographs. Changes in the proliferation in hDPSCs cultured in the complete culture medium containing aqueous extracts of the Alg-Gel or 3D-printed Alg-Gel scaffolds were examined using Cell Counting Kit-8 assay and flow cytometry analysis. Cells were cultured in the mineralization medium containing aqueous extracts of the Alg-Gel or 3D-printed Alg-Gel scaffolds for 7 or 14 days, and the differentiation of cells was shown by alizarin red S staining and alkaline phosphatase staining. The messenger RNA and protein expression of mineralization-related genes were detected with real-time polymerase chain reaction and Western blotting. Elemental analysis was used to test the material extract composition.

Results: More cells were grown and adhered to the 3D-printed Alg-Gel scaffolds than the Alg-Gel scaffolds. The aqueous extracts of 3D-printed scaffolds can promote cell proliferation, and compared with Alg-Gel scaffolds, the extracts of 3D-printed scaffolds were more effective. Compared with the negative control group, 3D-printed Alg-Gel scaffold and Alg-Gel scaffold aqueous extracts promoted osteogenic/odontoblastic differentiation of hDPSCs with the enhanced formation of bone-like nodules and the alkaline phosphatase staining. The expression of mineralization-related genes was also up-regulated. 3D-printed scaffold aqueous extract contained more calcium and phosphorus ions than the Alg-Gel scaffold.

Conclusions: These findings suggest that compared with the Alg-Gel scaffold, 3D-printed Alg-Gel is more suitable for the growth of hDPSCs, and the scaffold extracts can better promote cell proliferation and differentiation.

Keywords: 3D bioprinting; alginate/gelatin; cell adhesion; cell mineralization; cell proliferation; human dental pulp stem cells; scaffold aqueous extracts.

MeSH terms

Alginates*
Bioprinting*
Cell Differentiation*
Cell Proliferation
Cells, Cultured
Dental Pulp*
Gelatin*
Humans
Hydrogels
Osteogenesis
Plant Extracts
Stem Cells
Tissue Scaffolds*

Substances

Alginates
Hydrogels
Plant Extracts
Gelatin