Effects of enamel matrix derivative and titanium on the proliferation and differentiation of osteoblasts.
10.5051/jkape.2003.33.3.359
- Author:
Sang Hyun PARK
1
;
In Kyeong LEE
;
Seung Min YANG
;
Seung Yun SHIN
;
Yong Moo LEE
;
Young KU
;
In Chul RHYU
;
Chong Pyoung CHUNG
;
Soo Boo HAN
;
Sang Mook CHOI
Author Information
1. Department of Periodontology, College of Dentistry, Seoul National University, Korea.
- Publication Type:Original Article
- Keywords:
enamel matrix derivative(EMD);
osteoblast;
titanium;
RT-PCR
- MeSH:
Humans
- From:The Journal of the Korean Academy of Periodontology
2003;33(3):359-372
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
Among objectives of periodontal therapy, the principal one is the morphological and functional reconstruction of lost periodontal supporting tissues. This includes de novo formation of connective tissue attachment and the regrowth of alveolar bone. The use of enamel matrix derivative(EMD) may be a suitable means of regeneration new periodontal attachment in the infrabony defects. Implant used to replace lost tooth but, implantitis occurred after installation. The purpose of this study was to investigate the effects of EMD on differentiation and growth of osteoblast in titanium disc. Twentyfive millimeter diameter and 1mm thick Ti disc which was coated 25, 50, 100, 200microgram/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/ml of rhBMP-2 as positive control group, and no coat as negative control group. A human osteosarcoma cell line Saos-2 was cultured in Ti disc and cell proliferation and Alkaline phosphatase (ALP) activity were measured at 1 and 6 days. PCR was performed at 2 and 8 hours. Semi-quantitative RT-PCR for mRNA expressions of various osteoblastic differentiation markers - type I collagen, ALP, osteopontin, and bone sialoprotein - were performed at appropriate concentrations based upon the results of MTT and ALP assay. Cultured cell-disc complexes were prepared for scanning electron microscopy (SEM) at 2 hour. Data were analyzed using Mann-Whitney and repeated- measures 1-way analysis of variance(SPSS software version 10,SPSS, Chicago, IL). After culture, there was more osteoblast in EMD100microgram/ml than in EMD50, 200microgram/ml on day 6. There was significant difference in experimental and positive control group compared control group, as times go by(1 and 6 days). Alkaline phosphatase activity was different significantly in EMD100, 200microgram/ml and BMP100, 200ng/ml on day 6. The results of reverse transcriptase-polymerase chain reaction (RT-PCR) showed that expression of mRNA for ALPase, collagen type I, osteopontin, bone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMD200microgram/ml subgroup and BMP100ng/ml subgroup. The results of this study suggest that application of enamel matrix derivative on osteoblast attached to titanium surface facilitate the expression of bone specific protein and the differentiation and growth of osteoblast.
