Osteogenic differentiation of human dental papilla mesenchymal cells

doi:10.1016/j.bbrc.2006.02.101

Biochemical and Biophysical Research Communications

Volume 342, Issue 4, 21 April 2006, Pages 1257-1262

https://doi.org/10.1016/j.bbrc.2006.02.101 Get rights and content

Abstract

We isolated dental papilla from impacted human molar and proliferated adherent fibroblastic cells after collagenase treatment of the papilla. The cells were negative for hematopoietic markers but positive for CD29, CD44, CD90, CD105, and CD166. When the cells were further cultured in the presence of β-glycerophosphate, ascorbic acid, and dexamethasone for 14 days, mineralized areas together with osteogenic differentiation evidenced by high alkaline phosphatase activity and osteocalcin contents were observed. The differentiation was confirmed at both protein and gene expression levels. The cells can also be cryopreserved and, after thawing, could show in vivo bone-forming capability. These results indicate that mesenchymal type cells localize in dental papilla and that the cells can be culture expanded/utilized for bone tissue engineering.

Section snippets

Materials and methods

Cell preparation and culture. During orthodontic treatments (extraction of impacted third molars), we obtained molars from patients under sufficient informed consent in accordance with the Ethics Committee of the National Institute of Advanced Industrial Science and Technology (AIST). We first isolated the dental papilla tissue from the molars of patients aged 12–16 years. About 0.4 g of the tissue was digested in a solution consisting of phosphate-buffered saline (PBS), 4 mg/ml collagenase (Wako

Cell surface analysis

Dental papilla tissue was digested by collagenase and used for the primary culture. During the culture, non-adherent floating cells and debris were removed and adherent cells became almost confluent after about 14 days. The shape of the cells was fibroblastic (Fig. 1A). Immunophenotypic analyses of cell surface antigens by FACS showed that the cells were negative for hematopoietic markers (CD14, CD34, and CD45) but positive for markers present in mesenchymal cells (CD29, CD44, CD90, CD105, and

Discussion

Tooth germ consists of the dental papilla, the dental follicle, and the enamel organ. As shown in the results, we confirmed the expressions of bone-forming osteoblastic markers of in vitro cultured cells derived from the dental papilla at both the protein and gene expression levels (Fig. 2). We also demonstrated the noticeable new bone-forming capability of the cultured cells after in vivo implantation (Fig. 3). Significantly, the cells used for in vivo implantation were frozen stocked

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Osteogenic differentiation of human dental papilla mesenchymal cells

Abstract

Section snippets

Materials and methods

Cell surface analysis

Discussion

Trends. Mol. Med.

Biomaterials

Biochem. Biophys. Res. Commun.

Biomaterials

Lancet

Bone

Stromal stem cells: marrow-derived osteogenic precursors

Ciba Found. Symp.

Mesenchymal stem cells

J. Orthop. Res.

Multilineage potential of adult human mesenchymal stem cells

Science

Stem cell technology and bioceramics: from cell to gene engineering

J. Biomed. Mater. Res.

Tissue-engineered bone regeneration

Nat. Biotechnol.

Cultured autologous human cells for hard tissue regeneration: preparation and characterization of mesenchymal stem cells from bone marrow

Artif. Organs

Repair of large bone defects with the use of autologous bone marrow stromal cells

N. Engl. J. Med.

In vitro bone formation by rat marrow cell culture

J. Biomed. Mater. Res.

Osteogenic differentiation of human bone marrow-derived mesenchymal cells cultured on alumina ceramics

Artif. Organs