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Eur J Dent. 2018 Oct-Dec;12(4):566-573. doi: 10.4103/ejd.ejd_261_18.

Bone alkaline phosphatase and osteocalcin expression of rat's Gingival mesenchymal stem cells cultured in platelet-rich fibrin for bone remodeling (in vitro study).

Author information

1
Department of Orthodontic, Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia.
2
Doctoral Student of Medical Science, Faculty of Medicine, Universitas Airlangga, Surabaya, Indonesia.
3
Stem Cell Research and Development Center, Universitas Airlangga, Surabaya, Indonesia.
4
Department of Oral Medicine Faculty of Dental Medicine, Universitas Airlangga, Surabaya, Indonesia.
5
Department of Biochemistry and Molecular, Biochemistry Biomolecular Laboratory, Faculty of Medicine, Universitas Brawijaya, Surabaya, Indonesia.
6
Department of Microbiology, Virology and Immunology Laboratory, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, Indonesia.

Abstract

Objective:

The aim of this study was to analyze the osteogenic differentiation of rat GMSCs cultured in PRF for bone remodeling.

Materials and Methods:

GMSCs were isolated from the lower gingival tissue of four healthy, 250 g, 1-month old, male rats (Rattus norvegicus) cut into small fragments, cultured for 2 weeks, and subsequently passaged every 4-5 days. GMSCs isolated in passage 3 were characterized by CD34, CD45, CD44, CD73, CD90, and CD105 using fluorescein isothiocyanate immunocytochemistry (ICC) examination. GMSCs in passage 3-5 cultured in five M24 plates (N = 108; n = 6/group) for 7, 14, and 21 days with three different mediums as follows: Control (-) group: α-Modified Eagle Medium; Control (+) group: High-dose glucose Dulbecco's Modified Eagle's Medium (DMEM-HG) + osteogenic medium; and treatment group: DMEM-HG + osteogenic medium + PRF. GMSCs were osteogenic differentiation cultured in vitro in three different mediums by bone alkaline phosphatase (BALP) and osteocalcin (OSC) marker using ICC monoclonal antibody.

Statistical Analysis Used:

The one-way analysis of variance was performed (P < 0.05) based on Shapiro-Wilk and Levene's tests (P > 0.05).

Results:

GMSCs were shown to present + CD44, +CD73, +CD90, +CD105 and - CD34, - and CD45 expression as MSCs markers. The treatment group showed the highest BALP expression (16.00 ± 1.732) on day 7, while OSC expression (13.67 ± 2.309) on day 21 showed the statistically significant difference between groups (P < 0.05).

Conclusion:

GMSCs cultured in PRF demonstrated potential osteogenic differentiation ability capable of accelerating in vitro bone remodeling by enhancing BALP and OSC expression.

KEYWORDS:

Bone alkaline phosphatase; gingival mesenchymal stem cells; osteocalcin; osteogenic differentiation; platelet-rich fibrin

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