생분해성 Calcium Metaphosphate골이식재의 골조직재생효과에 관한 연구

채한승,이용무,양승민,천성수,김석영,구영,정종평,한수부,최상묵,류인철,Chae, Han-seung,Lee, Yong-Moo,Yang, Seung-Min,Chun, Sung-Soo,Kim, Suk-Young,Ku, Young,Choung, Chong-Pyoung,Han, Soo-Boo,Choi, Sang-Mook,Rhyu, In-Chul 대한치주과학회 2003 Journal of Periodontal & Implant Science Vol.33 No.1

Periodontal regeneration therapy with bone-substituting materials has gained favorable clinical efficacy by enhancing osseous regeneration in periodontal bony defect. As bone-substituting materials, bone powder, calcium phosphate ceramic, modified forms of hydroxyapatite, and hard tissue replacement polymer have demonstrated their periodontal bony regenerative potency. Bone-substituting materials should fulfill several requirements such as biocompatibility, osteogenecity, malleability, biodegradability. The purpose of this study was to investigate biocompatibility, osteo-conduction capacity and biodegradability of $Na_2O$, $K_2O$ added calcium metaphosphate(CMP). Beta CMP was obtained by thermal treatment of anhydrous $Ca_2(H_2PO_4)_2$. $Na_2O$ and $K_2O$ were added to CMP. The change of weight of pure CMP, $Na_2O$-CMP, and $K_2O$-CMP in Tris-buffer solution and simulated body fluid for 30 days was measured. Twenty four Newzealand white rabbits were used in negative control, positive control(Bio-Oss), pure CMP group, 5% $Na_2$-CMP group, 10% $Na_2O$-CMP goup, and 5% $K_2O$-CMP group. In each group, graft materials were placed in right and left parietal bone defects(diameter 10mm) of rabbit. The animals were sacrificed at 3 months and 6 months after implantation of the graft materials. Degree of biodegradability of $K_2O$ or $Na_2O$ added CMP was greater than that of pure CMP in experimental condition. All experimental sites were healed with no clinical evidence of inflammatory response to all CMP implants. Histologic observations revealed that all CMP grafts were very biocompatible and osseous conductive, and that in $K_2O$-CMP or $Na_2O$-CMP implanted sites, there was biodegradable pattern, and that in site of new bone formation, there was no significant difference between all CMP group and DPBB(Bio-Oss) group. From this result, it was suggested that all experimental CMP group graft materials were able to use as an available bone substitution.

The Effect of Ethanolic Extracts Mixture from Magnoliae cortex and Carthami semen on the' Progression of Experimental Periodontitis in Beagle Dogs

설양조,배규현,이용무,구영,류인철,한수부,배기환,최상묵,정종평,Seol, Yang-Jo,Bae, Kyoo-Hyun,Lee, Yong-Moo,Ku, Young,Rhyu, In-Chul,Han, Soo-Boo,Bea, Ki-Hwan,Choi, Sang-Mook,Chung, Chong-Pyoung The Korean Academy of Periodontoloy 2001 Journal of Periodontal & Implant Science Vol.31 No.4

후박추출물은 치주병인균에는 항균효과가 있고, 치은섬유아세포에는 세포독성이 적은 것으로 알려져 있으며 홍아씨 추출물은 골아세포를 활성화시키는 효과가 있어 골조직재생에 효과가 있는 것으로 알려져 있다. 이 실험의 목적은 비글견에 인위적으로 치주염을 유도하고 후박추출물과 홍화씨추출물의 혼합물을 투여함으로써, 이 두 가지 혼합물이 치주염 진행에 예방효과가 잇는지, 또, 치료효과가 있는지 알아보는 것이다. 후박과 홍화씨를 구입하여 에탄올에서 추출하여 정제하고 진공상태에서 건조시켜 농축하였다. 후박추출물과 홍화씨 추출물을 1:5의 질량비로 혼합하고 250㎎씩 캡슐에 넣었다. 비글견의 상하악 소구치와 하악 제1대구치를 8주 동안 철사와 견사로 결찰하여 치주염을 유발하였다. 8주 후에, 실험군 비글견에게는 미리 후박추출물과 홍화씨추출물 혼합물을 매 12시간마다 하루에 1,500㎎ 씩 12주 동안 투약하였다. 대조군 비글견에는 투약을 하지 않았다. 투약을 시작한 후에 치은지수, 부착수준을 매 4주마다 측정하였다. 매 4주마다 치주낭에서 페이퍼포인트를 이용하여 혐기성세균과 호기성세균을 채취, 배양하여 각각의 집락 수를 세었다. 그 결과 실험군의 치은지수는 8주, 12주에서 대조군보다 유의하게 낮았고(p<0.05)), 임상적으로도 상당한 치은염의 개선을 보였다. 실험군의 혐기성세균 집락수는 감소하여 4, 8, 및 12주에서 대조군에 비해 유의하게 낮았고(p<0.001), 호기성 세균 집락수는 4, 8, 및 12주에서 대조군에 비해 유의하게 낮았다.(p<0.05)이 실험 결과, 후박 추출물과 홍화씨 추출물 혼합물이 치주질환의 예방 및 치료에 이용되어질 수 있을 것으로 생각된다.

칫솔 강모 형태에 관한 연구

한승민,양승민,이용무,구영,류인철,정종평,최상묵,한수부,Han, Seung-Min,Yang, Seung-Min,Lee, Young-Moo,Ku, Young,Rhyu, In-Chul,Chung, Chong-Pyoung,Choi, Sang-Mook,Han, Soo-Boo 대한치주과학회 2002 Journal of Periodontal & Implant Science Vol.32 No.4

The aim of this study is to investigate the end of filaments of the different toothbrushes in the market through the stereomicroscope and to evaluate the % of rounded-end filaments considered to be acceptable. 9 brands, total 11 type toothbrushes were tested. 2 toothbrushes of each type which is marked as rounded-end filaments were tested. The toothbrushes which are not marked as rounded-end filaments were excluded. The domestic as well as foreign toothbrushes which are familiar to consumers were tested. 2 tufts of each toothbrushes were cut and examined by stereomicroscope using $40{\times}$ magnification. The procedure was carried out with blind-technique, and the digital photographs were taken. Besides the % of rounded-end filaments, total tufts number, material of the tuft, stiffness, and other special characteristics were recorded. By the classification of Silverstone and Featherstone, rounded-end filaments were examined and counted. The results shows that there are different range of rounded-end filaments according to the toothbrush types(17.7%-91.2%). Atman toothbrush has the most rounded-end filaments(91.2%) among the observed toothbrushes, and the Advantage Plus(Ora1-B) has the next(86.75%). E-Clean #411 has the least(17.70%) and E-Clean #410 of the same brand has also low % rounded-end filaments(20.60%). While G.U.M #409(Butler) has 67.90% rounded-end filaments, G.U.M #471 of the Same brand has comparative low 41.83% rounded-end filaments. 4 types of total 11 have the rounded-end filaments over 80%, however other 4 types have under even 50%. Considering that the correct brushing habit with a toothbrush which has rounded-end filaments can protect the gingival injury and tooth abrasion, it is thought that we dentists need to give the correct information about toothbrush to the patients

미노클린 첨부제의 지치주위염에 대한 유효성 및 안전성 평가를 위한 위약대조 이중 맹검 제3상 비교임상시험

설양조,고영경,이용무,구영,류인철,한수부,최상묵,정종평,Seol, Yang-Jo,Ko, Young-Kyung,Lee, Yong-Moo,Ku, Young,Rhyu, In-Cheul,Han, Soo-Boo,Choi, Sang-Mook,Chung, Chong-Pyoung 대한치주과학회 2000 Journal of Periodontal & Implant Science Vol.30 No.3

This clinical study was designed to determine the clinical and microbiological outcomes and safety of using minocycline loaded polycaprolactone strip for pericoronitis patients. 64 patients showing symptoms and signs of pericoronitis were enrolled according to the inclusion criteria in this double blind study. They were randomly assigned to two groups. 32 patients comprised control group and they received only polycaprolactone films in pericoronal spaces, and another 32 patients comprised experimental group and they received polycaprolactone films loaded with 30% minocycline. Informed consent was obtained from all the participants before beginning the study. At the initial visit, gingival index(GI), papillary bleeding index(PBI), amount of gingival crevicular fluid(GCF) were recorded, and microbiological sampling was done. Then, loaded or unloaded polycaprolactone film was inserted into the pericoronal spaces. No drug was prescribed excepting this film. After one week, clinical and microbiological exam was repeated. Presence of any side effects or inconveniences were checked. Chi-square test and t-test was performed to compare outcomes. At baseline, there were no significant differences in all the criteria between experimental group and control group. Experimental group showed significant improvement compared with control group both in GI(p<0.01) and PBI(p<0.01). The amount of GCF of the experimental group was significantly decreased compared with the control group(p<0.01) and baseline(p<0.01). In microbiological study, percentage of motile rod was prominently decreased in the experimental group. Also, aerobic(p<0.001), anaerobic(p<0.001) and black pigmented(p<0.01) bacteria were significantly decreased from the baseline. Furthermore, no side effects or inconveniences was reported in the experimental group. From this study, it was concluded that insertion of polycaprolactone film with 30% minocycline into the pericoronal spaces would be effective and safe treatment for pericoronitis.

Biological Activities of Calcium Polyphosphate

설양조,이재일,이용무,임윤탁,김석영,구영,류인철,함병도,한수부,최상묵,정종평,Seol, Yang-Jo,Lee, Jae-Il,Lee, Yong-Moo,Lim, Yoon-Tak,Kim, Seok-Young,Ku, Young,Rhyu, In-Chul,Hahm, Byung-Do,Han, Soo-Boo,Choi, Sang-Mook,Chung, Chong-Pyoung The Korean Academy of Periodontoloy 2000 Journal of Periodontal & Implant Science Vol.30 No.2

이 연구의 목적은 다공성의 CPP 내부에 쥐의 장골의 골수에서 유래된 세포를 접종하고 3차 원적으로 배양하여 CPP가 골 형성을 위한 조직공학의 지지체로 적용가능한가를 연구하는 것과 Calcium PolyPhosphate(CPP)의 돌연변이 유발성을 검사하는 것이다. 무수 ($Ca(H_2PO_4)$)를 condensation하여 무결정의 ($Ca(PO_3)$)를 얻고 이를 용융하고 냉각시킨 후 분쇄하여 Calcium polyphosphate(CPP) powder를 얻었다. 다공성의 CPP는 5% $SiO_2$를 첨가하여 sponge 형태로 $450-550{\mu}m$ 소공의 크기를 가지는 것과(CPP-45ppi) $200-300{\mu}m$의 소공의 크기를 가지는 것(CCP-60ppi) 2가지로 제작하였다. 각각의 CPP matrices는 $5mm{\times}5mm{\times}1mm$의 블록 형태로 만들었다. 체중 100g 내외의 백서에서 장골(femur, tibia)을 채취하여 백서의 장골 골수 세포를 분리하여 배양한 후 24well에 CPP block을 넣고 CPP block 당 $10^5$개의 배양한 세포를 접종하였다. 배양 1, 7, 14, 및 21 일째에 각 well에서 trypsin EDTA를 이용하여 2회 반복하여 cell을 분리하였고, 원심분리한 후 hemacytometer로 측정하였다. 또, 45ppi와 60ppi, 그 리 고 Tissue Culture Polystyrene(control group)에 접종, 배양된 세포들의 염기성 인산분해효소활성도를 배양 7, 14, 및 21 일째에 각각 측정하였다. 각 기간별로 배양된 세포-CPP 혼합체내에서 세포의 부착 및 증식과 형성된 조직의 3차원적 형태를 관찰하기 위하여 주사전자현미경하에서의 관찰하였다. CPP의 돌연변이 유발성 검사 (mutagenicity test)를 위해 hypoxanthine-guanine phosphoribosyl transferase(HPRT) assay를 하였다. NIH3T3 cell line과 CHO-K1 cell line으로 각각 $1000{\mu}g/m{\ell}$, $100{\mu}g/m{\ell}$, $10{\mu}g/m{\ell}$ 그리고 $1{\mu}g/m{\ell}$의 CPP 농도에서 측정하였다. 통계적 분석을 위해서 모든 측정은 각군당 4개체 이상 시험하였고, 각 측정값은 평균값${\pm}$표준편차로 나타내었다. 각 군간의 통계적 유의성 검정을 위해서 Analysis of variance(ANOVA)를 이용하였고 Tukey의 방법으로 사후분석을 실시하였다. 제작된 CPP matrices 소공들이 서로간에 연결이 잘 되어있는 형태였다. 두 가지로 제조된 CPP(45ppi와 60ppi) 모두에서 세포의 부착이 잘 일어났고, 부착된 세포의 분열도 잘 일어났다. 2 가지의 CPP 모두에서 7, 14, 21일째의 세포 수는 1일째에 비해 유의성 있게 증가하였다(P<0.01). 3차원적 구조인 Calcium PolyPhosphate에서 배양한 세포는 24well dish(tissue culture polystyrene)에서 평면적으로 배양한 대조군의 세포에서 보다 염기성 인산분해효소 (Alkaline Phosphatase)를 유의성 있게 높게 나타냈다. 주사전자현미경에서 세포-CPP 혼합체를 관찰한 결과, CPP block에 세포들이 잘부착되어 있었고, 시간이 지남에 따라 세포가 여러 층을 형성하면서 뭉치는 현상을 보였다. 또, HPRT assay 결과 , Calcium PolyPhosphate는 돌연변이 유발성을 보이지 않았다. 이상의 결과로 볼 때 CPP에는 세포부착이 잘 일어나고, 지지체 상에서 세포의 분열도 활발하게 일어나므로 골조직을 위한 조직공학의 우수한 지지체가 될 수 있을 것으로 사료된다.

혈소판유래성장인자를 함유한 Chitosan/Calcium Metaphosphate의 골조직재생효과에 관한 연구

이승열,설양조,이용무,이주연,이승진,김석영,구영,류인철,한수부,최상묵,정종평,Lee, Seung-Yeol,Seol, Yang-Jo,Lee, Yong-Moo,Lee, Ju-Yeon,Lee, Seung-Jin,Kim, Suk-Young,Ku, Young,Rhyu, In-Chul,Han, Soo-Boo,Choi, Sang-Mook,Chung, Chong-Pyou 대한치주과학회 2001 Journal of Periodontal & Implant Science Vol.31 No.1

Chitosan is biodegradable natural polymer that has been demonstrated its ability to improve wound healing, and calcium metaphosphate(CMP) is a unique class of phosphate minerals having a polymeric structure. In this study, chitosan/CMP and platelet derived growth factor(PDGF-BB) loaded chitosan/CMP sponges were developed, and the effect of the sponges on bone regeneration and their possibility as scaffolds for bone formation by three-dimensional osteoblast culture were examined. PDGF-BB loaded chitosan/CMP sponges were prepared by freeze-drying of a mixture of chitosan solution and CMP powder, and soaking in a PDGF-BB solution. Fabricated sponge retained its 3-dimensional porous structure with $100-200\;{\mu}m$ pores. The release kinetics of PDGF-BB loaded onto the sponge were measured in vitro with $^{125}I-labeled$ PDGF-BB. In order to examine their possibility as scaffolds for bone formation, fetal rat calvarial osteoblastic cells were isolated, cultured, and seeded into the sponges. The cell-sponge constructs were cultured for 28 days. Cell proliferation, alkaline phosphatase activity were measured at 1, 7, 14 and 28 days, and histologic examination was performed. In order to examine the effect on the healing of bone defect, the sponges were implanted into rat calvarial defects. Rats were sacrificed 2 and 4 weeks after implantation and histologic and histomorphometrical examination were performed. An effective therapeutic concentration of PDGF-BB following a high initial burst release was maintained throughout the examination period. PDGF-BB loaded chitosan/CMP sponges supported the proliferation of seeded osteoblastic cells as well as their differentiation as indicated by high alkaline phosphatase activities. Histologic findings indicated that seeded osteoblastic cells well attached to sponge matrices and proliferated in a multi-layer fashion. In the experiments of implantation in rat calvarial defects, histologic and histomorphometric examination revealed that chitosan/CMP sponge promoted osseous healing as compared to controls. PDGF-BB loaded chitosan/CMP sponge further echanced bone regeneration. These results suggested that PDGF-BB loaded chitosan/CMP sponge was a feasable scaffolding material to grow osteoblast in a three-dimentional structure for transplantation into a site for bone regeneration.

구연산과 테트라싸이클린으로 처리한 치근면에서 rhBMP-2가 치주인대섬유아세포와 골아세포의 활성에 미치는 영향

심정민,한수부,설양조,이용무,김경화,계승범,최상묵,정종평,Shim, Jung-Min,Han, Soo-Boo,Seol, Yang-Jo,Lee, Yong-Moo,Kim, Kyeong-Hwa,Kye, Seung-Beom,Choi, Sang-Mook,Chung, Chong-Pyoung 대한치주과학회 2001 Journal of Periodontal & Implant Science Vol.31 No.1

The goal of Periodontal treatment is predictable periodontal regeneration. But until now, many products including GTR materials and growth factors are beyond of complete regeneration. BMP can induce ectopic bone formation when implanted into sites such as rat muscle and can greatly enhance healing of bony defects when applied exogenously. BMP can promote periodontal regeneration by their ability to stimulate new bone and new cementum formation. But little is known about optimal conditions required for the application. Root conditioning is used for bioacive root change so altered root surface provides a substrate that promotes chemotaxis, migration and attachment of peridontal cells encouraging connective attachment to the denuded root surface. The aim of this study is to investigate whether the acid conditioning change effect of rhBMP-2 on human periodontal ligament cell and osteoblast cell line. 288 periodontally involved root dentin slices are divided into 6 groups, each 48, 1)control, 2)treated with BMP, 3)treated with citric acid 4)treated with citric acid+BMP 5)treated with tetracycline 6)treated with TC+BMP. Each group was devided half, so 12 root dentin slices were seeded with periodontal ligament cells and 12 were seeded with osteoblasts. At day 2 and 7, cell number, protein assay, ALP activitiy was measured. To investigate morphology of cultured cells, SEM was employed. Statistical analysis was performed with SPSS 8.0 either t-test or ANOVA test. The results are ; Protein assay and cell number was slightly decreased in CA+BMP group compared to Ca group but it was not statistically significant and ALP activity was much more increased in CA+BMP group compared to CA group so there was no statistically significance between BMP and CA+BMP group and statistically significant compared to control group. Cell number and protein assay was slightly increased in TC group and ALP activity was much less the BMP group and CA group. Cell number and protein and ALP activity was not much increased in TC+BMP group. TC group and TC+BMP group showed cell morphology change in SEM. This results suggested that application of root surface with citric acid before BMP treatment might give better result in periodontal regeneration.

Ames test 결과와 QSAR를 이용한 변이원성예측치와의 비교

양숙영(Sook Young Yang),맹승희(Seung Hee Maeng),이종윤(Jong Yun Lee),이용묵(Yang Mook Lee),정호근(Ho Keun Chung),정해원(Hai Won Chung),유일재(Il Je Yu) 한국환경성돌연변이발암원학회 2000 한국환경성돌연변이·발암원학회지 Vol.20 No.1

Recently there is increasing interest in the use of structure activity relationships for predicting the biological activity of chemicals. The reasons for the interest include the decrease cost and time per chemical as compared with animal or cell system for identifying toxicological effects of chemicals and the reduction in the use of animals for toxicological testing. This study is to test the validity of the mutagenicity data generated from QSAR (Quantitative Structure Activity Relationship) program. Thirty chemicals, which had been evaluated by Ames test during 1997-1999, were assessed with TOPKAT QSAR mutagenicity prediction module. Among 30 chemicals experimented, 28 were negative and 2 were positive for Ames test. On the contrary, 23 chemicals showed the high confidence level indicating high prediction rate in mutagenicity evaluation, and 7 chemicals showed the low to moderate confidence level indicating low prediction in mutagenicity evaluation. Overall mutagenicity prediction rate was 77% (23/30). The prediction rates for non-mutagenic chemicals were 79% (22/28) and mutagenic chemicals were 50% (1/2). QSAR could be a useful tool in providing toxicological data for newly introduced chemicals or in furnishing data for MSDS or in determining the dose in toxicity testing for chemicals with no known toxicological data.

Antiinflammatory Effects of Tetracycline-loaded Biodegradable Membranes in Experimental Periodontitis of Beagle Dogs

전형식,설양조,박윤정,이용무,구영,류인철,이승진,한수부,최상묵,권수경,정종평,Jun, Hyung-Sik,Seol, Yang-Jo,Park, Yoon-Jeong,Lee, Yong-Moo,Ku, Young,Rhyu, In-Cheul,Lee, Seung-Jin,Han, Soo-Boo,Choi, Sang-Mook,Kwon, Soo-Kyoung,Chung, Chon The Korean Academy of Periodontoloy 2000 Journal of Periodontal & Implant Science Vol.30 No.3

조직유도재생술 과정에 사용된 차폐막의 술 후 오염 혹은 시술부위의 감염으로 조직재생유도가 제대로 이루어지지 않는 경우가 많이 있다. 테트라싸이클린은 넓은 범위의 치주 원인균에 효과적이고, 중성구 교원분해효소를 억제 함으로써 결합조직파괴를 억제하기 때문에 전신적 혹은 국소적으로 치주질환 치료에 널리 이용되어져왔다. 이번 연구의 목적은 비글견에 실험적으로 치주염을 유발시킨 후 테트라싸이클린 함유 차폐막을 이용하여 조직유도재생술을 하고서 테트라싸이클린의 유리반응을 관찰하는 것과 테트라싸이클린에 의한 항염, 항균효과를 알아보는 것이다. 실험 2-3개월 전에 비글견의 구강에서 치조골 결손부를 형성하여 치주염을 유발시켰다. 결손부 형성 2-3개월 후 실험군으로 테트라싸이클린 함유 차폐막으로 조직유도재생술을 하였고, 대조군으로 테트라 싸이클린이 함유되지 않은 차폐막으로 조직유도재생술을 하였고, 음성대조군으로 치은판막술만을 하였다. 시술 전과 시술 후 1, 2, 4주 간격으로 치은지수, 치태지수, 치은열구액의 양 및 혐기성 세균과 호기성 세균의 군락수를 측정하였다. 술 후, 1, 3, 5, 7, 및 14일 간격으로 유리된 테트라싸이클린의 농도를 측정하였다. 테트라싸이클린 함유 차폐막은 임상적 치은지수는 술 후 2, 4주째, 치태지수는 1, 2, 4주째 유의하게 감소하였다.(p<0.05) 1주에서 혐기성 및 호기성 세균집락수는 테트라싸이클린 함유 차폐막을 사용한 군이 약물이 함유되지 않은 차폐막보다 유의성있게 적었다.(p<0.001) 테트라싸이클린 함유 차폐막은 처음 1일은 높은 농도로 유리되었고 그 후 1주일 동안 일정하게 MIC 이상으로 유리되었다. 하지만 14일째는 유리가 관찰되지 않았다. 본 연구 결과, 테트라싸이클린 함유 생분해성 차폐막은 1주이상 약제가 MIC 이상 유리되었고, 유리된 테트라싸이클린에 의한 항염, 항균작용이 있어 치주조직 재생유도술시에 초기 치유가 잘 진행되도록 하는 효과가 있을 것이다.

법랑기질유도체를 도포한 타이태늄 표면에서 조골세포의 증식 및 분화

박상현,이인경,양승민,신승윤,이용무,구영,류인철,정종평,한수부,최상묵,Park, Sang-Hyun,Lee, In-Kyeong,Yang, Seung-Min,Shin, Seung-Yun,Lee, Yong-Moo,Ku, Young,Rhyu, In-Chul,Chung, Chong-Pyoung,Han, Soo-Boo,Choi, Sang-Mook 대한치주과학회 2003 Journal of Periodontal & Implant Science Vol.33 No.3

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, 200${\mu}g$/ml of EMD(Emdogain(R)) used as experimental group, 25, 50, 100, 200ng/d 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 EMD100${\mu}g$/ml than in EMD50, 200${\mu}g$/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, 200${\mu}g$/ml and BMP100, 200${\mu}g$/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. hone sialoprotein and BMP-2 was detected at 2 hour and 8 hour in EMI 200${\mu}g$/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.