Effect of Emdogain on proliferation and migration of different periodontal tissue–associated cells

doi:10.1016/j.tripleo.2010.01.007

Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology

Volume 109, Issue 6, June 2010, Pages 924-931

https://doi.org/10.1016/j.tripleo.2010.01.007 Get rights and content

Objectives

Although Emdogain is widely used as a gel in periodontal therapy, the exact mechanisms underlying its regenerative ability still need to be further investigated. Therefore, we tested in vitro the effect of the product Emdogain on proliferation, viability, and migration of various human cell types of periodontium.

Study design

Proliferation and viability of alveolar osteoblasts (AOBs), epithelial cell line HSC-2, and human umbilical vein endothelial cells (HUVECs) were measured using [³H]-thymidine uptake and 3,4,5-dimethylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT)-assay, respectively. Cell migration was investigated in microchemotaxis chamber.

Results

The proliferation and viability of AOB, HSC-2, and HUVECs were significantly stimulated by Emdogain (12.5-250 μg/mL) in direct relationship with the amount of product present in the cell culture medium. Cell migration was stimulated in AOB and HUVECs depending on Emdogain amount. In contrast, in HSC-2 cells the migration was stimulated only by less than 50 μg/mL of Emdogain, whereas at higher amounts this stimulating effect was either diminished or absent.

Conclusion

Emdogain stimulates proliferation, viability, and migration of AOB, HSC-2, and HUVECs in vitro. This biological versatility of Emdogain could correspond to an essential mechanism underlying its ability to promote periodontal regeneration.

Section snippets

Cell culture

Commercially available cell lines of human oral epithelial cells HSC-2 (Health Science Research Resources Bank, Sennan, Japan) and human umbilical vein endothelial cells (HUVECs, Technoclone, Vienna, Austria) were used in the present study. Alveolar osteoblasts (AOBs) were isolated from alveolar bone, which was taken from periodontally healthy patients undergoing routine third-molar extraction. The study was performed in accordance with permission from the local ethics committee of the Medical

Cell proliferation and viability

The effect of Emdogain on the proliferation of AOB, HSC-2, and HUVEC cells measured by [³H]-thymidine uptake is shown in Fig. 1. As can be seen, the proliferation rate in all 3 cell types was significantly increased with increased amount of Emdogain in the cell culture medium. Interestingly, the stimulating effect in endothelial cells (HUVECs) was somewhat lower than that in osteoblasts and epithelial cells.

The effect of Emdogain on the viability of AOB, HSC-2, and HUVEC cells measured by

Discussion

Periodontal disease is one of the most common oral diseases, which induces destruction of tooth-supporting bone structure and soft tissue around the tooth and may result in tooth loss. Successful treatment of periodontal disease and regeneration of periodontal tissue are the main tasks of periodontal therapy. Although Emdogain is widely used in periodontal therapy, the mechanisms of action underlying its ability to promote regeneration of periodontal tissues still need to be further

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The apoptosis assay was performed in triplicate. Cell migration was assessed in a 48-well microchemotaxis chamber (Neuroprobe, Gaithersburg, MD, USA) on a polycarbonate filter with 8 μm pore size as described previously [23]. The chamber consisted of acrylic top and bottom plates, each containing 48 matched wells.
Enamel matrix derivative (EMD) is an effective biomaterial for periodontal tissue regeneration and might stimulate angiogenesis. In order to clarify mechanisms underlying its biological activity, we separated two EMD fractions with different molecular weight protein components and investigated their effects on human umbilical vein endothelial cells (HUVECs) in vitro.
Fraction Low-Molecular Weight (LMW) included proteins with a molecular weight (M.W.) < 8 kDa. Fraction LMW-depleted included proteins with M.W. > 8 kDa and lower than approximately 55 kDa. The effect of EMD fractions on proliferation/viability, apoptosis, migration and expression of angiopoetin-2 (ang-2), von Willebrand factor (vWF), E-selectin, intracellular adhesion molecules 1 (ICAM-1), vascular endothelial growth factor (VEGF) receptors Flt-1 and KDR was investigated.
The proliferation/viability of HUVECs was inhibited by both LMW and LMW-depleted at concentrations 100 μg/ml, whereas EMD slightly increased cell proliferation/viability. The expression of all investigated proteins was up-regulated by EMD. However, differences in the effect of EMD fractions on the protein expression were observed. The effect of LMW-depleted on the expression of ICAM-1 and E-selectin was markedly higher compared to LMW. In contrast, the expression of vWF and VEGF receptors Flt-1 and KDR was primarily affected LMW than by LMW depleted. The expression of ang-2 was not influenced by LMW and LMW-depleted. HUVECs migration was stimulated more strongly by LMW than by EMD and LMW-depleted.
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In addition, the effect of EMD on the osteoblast proliferation seems to depend on the cell lineage. In particular, EMD stimulates proliferation of primary human osteoblasts,15,25 human Sam-1 osteoblast cells,26 human alveolar osteoblast,27 and MC3T3 mouse osteoblast cell line,14,28 but does not influence that of MG-63 cells.29 Our data further showed that EMD increase the ALP activity and production of osteocalcin (Figs. 2 and 3), which are the early and the late osteoblast differentiation markers, respectively.30,31
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Osteoblast-like MG-63 cells were seeded on coarse-grit–blasted and acid-etched surface Ti implant disks and stimulated with various EMD concentrations. Cell proliferation/viability, alkaline phosphatase activity, osteocalcin production, and expression levels of osteoprotegerin (OPG) and receptor activator of nuclear factor κB ligand (RANKL) were determined.
EMD inhibited the proliferation/viability of MG-63 cells. Furthermore, EMD significantly increased the alkaline phosphatase activity and osteocalcin production in MG-63 cells grown on Ti surfaces. Finally, EMD enhanced mRNA expression level of OPG and did not influence that of RANKL.
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EndodontologyEffect of Emdogain on proliferation and migration of different periodontal tissue–associated cells

Objectives

Study design

Results

Conclusion

Section snippets

Cell culture

Cell proliferation and viability

Discussion

Biomaterials

Oral Surg Oral Med Oral Pathol Oral Radiol Endod

Oral Surg Oral Med Oral Pathol Oral Radiol Endod

Dent Mater

Oral Surg Oral Med Oral Pathol Oral Radiol Endod

Arch Oral Biol

J Chromatogr

Enamel matrix, cementum development and regeneration

J Clin Periodontol

The use of enamel matrix derivative in the treatment of periodontal defects: a literature review and meta-analysis

Crit Rev Oral Biol Med

Ten-year results following treatment of intra-bony defects with enamel matrix proteins and guided tissue regeneration

J Clin Periodontol

Porcine fetal enamel matrix derivative stimulates proliferation but not differentiation of pre-osteoblastic 2T9 cells, inhibits proliferation and stimulates differentiation of osteoblast-like MG63 cells, and increases proliferation and differentiation of normal human osteoblast NHOst cells

J Periodontol

Biological mediators and periodontal regeneration: a review of enamel matrix proteins at the cellular and molecular levels

J Clin Periodontol

Effect of heat treatment on bioactivities of enamel matrix derivatives in human periodontal ligament (HPDL) cells

J Periodontal Res

Osteoprotegerin and receptor activator of nuclear factor-kappa B ligand modulation by enamel matrix derivative in human alveolar osteoblasts

J Periodontol

Involvement of FGF-2 in the action of Emdogain on normal human osteoblastic activity

Oral Dis

In vitro assessment of motility and proliferation of human osteogenic cells on different isolated extracellular matrix components compared with enamel matrix derivative by continuous single-cell observation

Clin Oral Implants Res

Endodontology
Effect of Emdogain on proliferation and migration of different periodontal tissue–associated cells