• Title/Summary/Keyword: Alkaline phosphatase

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The effect of admixture of vitamin $D_3$ and dexamethasone on the activity of osteoblastic cells (Vitamin $D_3$와 Dexamethasone의 복합 투여가 골모세포에 미치는 영향에 관한 연구)

  • Lim, Na-Won;Park, Young Joo;Kim, Sang-Cheol
    • The korean journal of orthodontics
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    • v.29 no.3 s.74
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    • pp.383-397
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    • 1999
  • Bone is a dynamic tissue which is constantly remodelled by subsequent cycles of bone resorption and formation. Glucocorticoid and vitamine $D_3$ are known as regulating substances in bone metabolism. In vitro experiments using bone tissue, it was suggested that glucocorticoid inhibits bone resorption, whereas the effect of glucocorticoid on bone formation are complex- increasing or decreasing effect. The active form of vitamin $D_3$, 1,25-dihydroxycholecalciferol[1.25-$(OH)_2D_3$], has been reported to stimulate osteoblastic activities including the production of ALP, type I collagen, and osteoclacin. The purpose of this study was to evaluate the effect of admixture of vitamin $D_3$ and dexamethasone, one of glucocorticoids, on osteoblastic cell line(MC3T3-E1). Alkaline phosphatase(ALP) and MTT assay were conducted in the cultivated cells with 1, 10, 100nM/ml of 1,25-$(OH)_2D_3$ and/or 10nM/ml, 100nM/ml, $1{\mu}M/ml$ of dexamethasone. The observed results were as follows. 1. The activity of osteoblastic cells with $1{\mu}M/ml$ of dexamethasone was significantly increased at 1-day cultivation with comparison to control group, but was decreased afterwards. But the activity of ALP was greatest in $1{\mu}M/ml$ of dexamethasone and increased with time lapsed. 2. The activity of osteoblastic cells with vitamin $D_3$ was significantly increased dose-dependently at 1-day cultivation, but was significantly decreased in l00nM/.ml at 2-day cultivation, and was a little increased again at 3-day cultivation. The activity of ALP was increased in 10nM/ml or 100nM/ml at 2-day or 3-day cultivation, and was greatest in 100nM/ml at 3-day cultivation. 3. In case of admixture of dexamethasone and vitamin $D_3$, the cellular activity was decreased in any concentration of vitamin $D_3$ at 2-day cultivation, but was increased again at 3-day cultivation, which was greater than that in control or dexamethasone only group. The activity of ALP was decreased at 1-day cultivation, but was increased in the admixture of 10nM/ml or 100nM/ml of dexamethasone with 100nM/ml of vitamin $D_3$ at 2-day cultivation, and was again decreased at 3-day cultivation.

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Early histological change in hard tissue from orthodontic force placed on microscrews in ovariectomized rats (난소 적출 백서에 식립된 마이크로스크류에 교정력 부여 시 나타난 초기 경조직 변화에 관한 연구)

  • Lee, Dea-Seung;Chang, Moon-Jung;Lee, Jin-Woo
    • The korean journal of orthodontics
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    • v.36 no.2 s.115
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    • pp.103-113
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    • 2006
  • Most elderly women experience a decrease in their bone density due to a deficiency of calcium intake, ovariectomy, or menopause. This study evaluated the usability of the microscrew as a skeletal anchorage system in these orthodontic treatment cases, using rats as a research group. The 4 month old sprague-dawley species rats were divided into two groups, the OS (Ovariectomy Screw), and the SS (Sham operation Screw) group. In both the OS and SS groups, microscrews were implanted into the palatal bone between the upper molar teeth and two upper incisors were retracted using NETE coil spring with 75 g of force. After 3days, the again after 7 days, 7 rats in each group were sacrificed. Three days before they were sacrificed, Alizarin red S was intraperitoneally injected, and their maxillary bone, tibia and blood from their hearts were taken. The components of the extracted blood were biochemically analyzed and non-decalcified grinding resin sections for maxillary bone and tibia were made. The sections were examined with a polarization microscope, and fluorescent microscope. Smaller concentrations of Ca and P, the inorganic substances closely related to bone density, were found in the extracted blood of the OS group. Both OS and SS groups showed a possibility of bone remodeling with a high concentration of ALP after 7 days. An increase in bone density on the tension and compression sides of the microscrew and the tension side of the tooth for both OS and SS groups was confirmed with a polarization microscope. However, the bone density of the pressure side of the tooth and apical side was decreased. More deposits of Alizarin red S in the bone after 7 days rather than 3 days seen with a fluorescent microscope suggested the existence of new bone formation.

Antioxidaitve and Differentiation Effects of Artemisia capillaris T. Extract on Hydrogen Peroxide-induced Oxidative Damage of MC3T3-E1 Osteoblast Cells (사철쑥(Artemisia capillaris T.) 추출물의 항산화 활성 및 H2O2로 산화적 스트레스를 유도한 조골세포의 활성과 분화에 미치는 영향)

  • Seo, Jee-Eun;Hwang, Eun-Sun;Kim, Gun-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.11
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    • pp.1532-1536
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    • 2011
  • In this study, the antioxidative activity of Artemisia capillaris T. extract on the proliferation and differentiation of MC3T3-E1 cells under $H_2O_2$-induced oxidative stress was investigated in order to determine its protective effect against oxidative stress as well as its availability as an antioxidant material related to treatment of bone diseases. As a result, the total polyphenol content of A. capillaris extract was 90.10 mg/g, whereas the flavonoid content was 4.45 mg/g. A. capillaris extract increased proliferation of MC3T3-E1 cells under $H_2O_2$-induced oxidative stress, and also increased the proliferation of differentiated osteoblast cells under oxidative stress. In addition, two differentiation markers, alkaline phosphatase activity and mineralization level, in A. capillaris extract tended to increase. These results indicate that A. capillaris extract suppresses the damage to osteoblasts caused by oxidative stress, which demonstrates its availability as an antioxidant material for preventing bone diseases.

Effects of Tea Fungus/Kombucha Beverage on Lipid Metabolism and Protein Level in Diabetic Female Rats (발효홍차 음료가 당뇨 암쥐의 지질대사 및 단백질농도에 미치는 영향)

  • 노민희;고진복
    • Journal of Life Science
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    • v.12 no.6
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    • pp.661-668
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    • 2002
  • We investigated the effect of tea fungus/kombucha beverage(TF) on the body weights, the blood glucose levels, lipid and protein concentrations, and enzyme activities in diabetic female rats. Sprague-Dawley rats were fed drinking water supplemented with 20% or 40% TF groups, respectively for 7 weeks. The female rats (mean weight 155.5$\pm$9.3 g) were assigned to one control and three diabetic groups. Diabetic groups were divided into diabetic control (TF free water), 20% or 40% TFD groups (20% or 40% TF in water) according to the levels of TF supplementation. Diabetes was experimentally induced by intraperitonially administration of streptozotocin in citrate buffer(pH 4.3) after 2 weeks feeding of four experimental water. Animals were sacrificed at the 5 weeks of diabetic state. The diabetic groups showed significantly decrease of body weight(6.8-7.5 g) compared with the control group(48.3 g). The hepatic, kidney and pancreatic weights of 20% or 40% TFD groups were not significantly different with D-control group. The fasting serum glucose level were higher in all diabetic groups than that of the control group. The concentrations of serum triglyceride in 40% TFD group and serum LDL-cholesterol in 20% TFD group were significantly decreased compared with the D-control group. The concentrations of serum total cholesterol and HDL-cholesterol, HDL-cholesterol/total cholesterol ratio, and atherogenic index in 20% or 40% TFD groups were similar to those in D-control group. The concentrations of hepatic triglyceride in 20% or 40% TFD groups were significantly decreased compared with the D-control group, but the concentrations of hepatic cholesterol and phospholipid were similar to all diabetic groups. The concentrations of serum and hepatic total protein, serum albumin, and the activities of GOT, GPT and LDH in the serum were the same levels of all diabetic groups.

The Clinical Aspects of Acute Septic Arthritis and Acute Osteomyelitis in Children (소아기 급성 화농성 관절염과 급성 골수염의 임상적 고찰)

  • Kim, Jin-Man;Lee, So-Yeon;Kim, Young-Ho;Shin, Eon-Woo;Oh, Phil-Soo;Kim, Kwang-Nam
    • Pediatric Infection and Vaccine
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    • v.12 no.2
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    • pp.149-156
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    • 2005
  • Purpose : Acute septic arthritis and acute osteomyelitis are not rare diseases in pediatric population. But when the diagnosis is delayed or inappropriate treatments are given, permanent disabilities of joint or bone can be followed. We analysed clinical manifestations, laboratory findings, X-ray findings, causative microorganisms and antibiotic susceptibility results of the two diseases in children. Methods : During January 1992 and May 2002, we conducted a retrospective study of 103 children who were diagnosed as acute septic arthritis and acute osteomyelitis. We selected out 34 children who had positive culture results in the blood or involved sites. Results : 19 cases were diagnosed as acute septic arthritis and 15 cases were acute osteomyelitis. These diseases were most common in preschool children and next in neonates. Hip joints and tibia were the most common sites in each disease. X-ray findings showed abnormalities in 6 cases(36%) of acute septic arthritis and 7 cases(50%) of acute osteomyelitis on admission. The most common microorganism isolated from the involved sites was Staphylococcus aureus; 12 out of 14 cases in acute septic arthritis and 6 out of 13 cases in acute osteomyelitis. Conclusion : It is difficult to make a clear initial diagnosis of the two diseases. We could not find any differences between these two diseases on clinical manifestations such as fever, swelling, tenderness and limitation of movements in joint and bone. The most common microorganism was Staphylococcus aureus.

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Dynamic Study on the Hepatobiliary Diseases with Combination of $^{131}I$-Rose bengal and $^{198}Au$-Colloid Scintiphotography (간담도질환(肝膽道疾患)의 Scintiphotography 상(像)에 있어서 $^{131}I$-Rose bengal 및 $^{198}Au$-colloid의 섭취(攝取)와 배설(排泄)에 관(關)한 역동학적(力動學的) 연구(硏究))

  • Rhee, Yong-Kook
    • The Korean Journal of Nuclear Medicine
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    • v.5 no.1
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    • pp.49-64
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    • 1971
  • The radioactive $^{131}I$-rose bengal serial scintiphotography was performed in 62 patients with the hepatobiliary diseases and in 20 normal subjects. This approach permitted visualization of the hepatic uptake of $^{131}I$-rose bengal from the circulation and its excretion into the biliary trees and the intestines. In some of these patients, gallbladder function was examined, using eggs as a gallbladder constrictor. The time of maximum hepatic uptake was well correlated to the conventional biochemical liver function tests. In addition to $^{131}I$-rose bengal scintiphotography, $^{198}Au$-colloid scintiphotography was also performed to make comparison of these two tests. The results obtained were as follows: 1. In normal subjects, the maximum hepatic uptake of $^{131}I$-rose bengal occurred at $23{\pm}2.9$ minutes, the initial hepatic excretion at $34{\pm}5.1$ minutes, the visualization of the gallbladder at $29{\pm}5.7$ minutes and the intestinal visualization at $54{\pm}25.8$ minutes. The radioactivity in the gallbladder decreased to $10.7{\pm}5.0%$ one hour after the ingestion of eggs. 2. In the patients with cirrhosis of the liver, there was a delayed and decreased hepatic uptake. The maximun hepatic upake occurred at $43{\pm}12.9$ minutes. The differences in the results of uptake between the cirrhotic and the normal group were statistically significant. The initial hepatic excretion occurred at $60{\pm}18.5$ minutes and had tendency of delaying compared with the normal controls. The gallbladder was visualized in 13 of 16 cases (81%) and its visualization occurred at $49{\pm}14.6$ minutes with a tendency to be delayed compared with the normal controls. The intestinal visualization occurred at $63{\pm}15.8$ minutes and its delaying tendency was somewhat more prominent. 3. In patients with hepatitis, the maximum hepatic uptake occurred at $59{\pm}21.4$ minutes and was significantly delayed. The initial hepatic excretion occurred at $82{\pm}34.3$ minutes and the results revealed a delaying tendency. The gallbladder was visualized in 15 of 20 cases (75%) at $57{\pm}18.7$ minutes, which was significantly delayed. The Intestinal visualization was noted in all cases with marked delay. 4. In patients with obstructive jaundice, the maximum hepatic uptake was noted at $83{\pm}14.7$ minutes, showing the most significant delay. The hepatic excretion into biliary trees and intestines was not entirely noted in all cases except the only one case with gallbladder visualization. 5. In patients with cholelithiasis, the maximum hepatic upake and the initial hepatic excretion were slightly delayed with mean times of $39{\pm}11.2\;and\;48{\pm}17.1$ minutes respectively. The visualization of the gallbladder was demonstrated in 10 of 17 cases (59%) and occurred at $52{\pm}25.6$ minutes with a slight delay. The intestinal visualization occurred at $67{\pm}47.7$ minutes and was slightly delayed. $^{131}I$-rose bengal in the gallbladder remained high, $49.3{\pm}21.3%$, which suggested quantitatively decreased power of gallbladder constriction. 6. The time of the maximum hepetic uptake was correlated well to BSP retention and serum alkaline phosphatase ativity. However, the maximum hepatic uptake had no definite correlation with serum albumin, serum globulin, TTT, serum cholesterol, SGPT or SGOT. 7. In the diagnosis of the hepatobiliary diseases with jaundice, $^{131}I$-rose bengel serial scintiphotography has proved to be more useful than $^{198}Au$-colloid scintiphotography. With these results, it could be justified that $^{131}I$-rose bengal scintiphotography is an excellent diagnostic aid for dynamic hepatobiliary function studies in the clinical practice.

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Assessment of the permanent canine bone support after secondary bone graft In UCLP patients (편측성 순구개열 환자에서 이차 골이식후 맹출된 영구 견치의 치조골 지지도에 관한 연구)

  • Park, Ki-Tae
    • The korean journal of orthodontics
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    • v.31 no.6 s.89
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    • pp.601-610
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    • 2001
  • The purpose of this retrospective study was to evaluate the level oi alveolar bone support of the erupted Permanent canine through the reconstructed cleft region compared to the contralateral canine on the non-cleft side. This study was limited to children with complete unilateral cleft lip and palate who underwent secondary alveolar iliac bone gvaft and the apices of the erupted canine roots were closed at the time of evaluation. With these criteria the study included 21 children whose average age at the time of bone graft reconstruction was 9.8 years, with a minimum of 12.4 years of age at the time of the evaluation. The study was limited to the use of iliac cancellous bone as the autograft material for reconstruction of the alveolar cleft. Cranial bone graft and other autogenous bone sources were excluded. The periapical radiographs were used to evaluate alveolar bone level of each canine. The percentages of root supported by the bone were established by dividing the amount of root covered with the bone by the anatomic root length. The canine oi the non-cleft side was used as an internal control and the canine on the cleft side was used as an experimental. There was a statistically significant difference in the alveolar bone support ratio between the control ($92.9\%$) and experimental canines ($8.7\%$). An average of $95\%$ level of alveolar bone support was achieved for the experimental canine in comparison to the control canine. Neither the presence of lateral incisor, nor the stage of root development of the canine at the time of the bone graft appeared to have affected the alveolar bone support ratio of the canine after the secondary bone graft.

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Effect of RGD peptide coating of implant titanium surface on human mesenchymal stem cell response (양극산화 티타늄 표면에 서로 다른 RGD 펩타이드 코팅 방법이 인간간엽줄기세포 반응에 미치는 영향)

  • Kim, Min-Su;Jeong, Chang-Mo;Jeon, Young-Chan;Ryu, Jae-Jun;Huh, Jung-Bo;Yun, Mi-Jung
    • The Journal of Korean Academy of Prosthodontics
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    • v.49 no.3
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    • pp.245-253
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    • 2011
  • Purpose: The aim of this in vitro study was to estimate surface characteristic after peptide coating and investigate biological response of human mesenchymal stem cell to anodized titanium discs coated with RGD peptide by physical adhesion and chemical fixation. Materials and methods: Fluorescence isothiocyanate (FITC) modified RGD-peptide was coated on the anodized titanium discs (diameter 12 mm, height 3 mm) using two methods. One was physical adhesion method and the other was chemical fixation method. Physical adhesion was performed by dip and dry procedure, chemical fixation was performed by covalent bond via silanization. In this study, human mesenchymal stem cell was used for experiments. The experiments consisted of surface characteristic evaluation after peptide coating, analysis about cell adhesion, proliferation, differentiation, and mineralization. Obtained data are statistically treated using Kruskal-Wallis test and Bonferroni test was performed as post hoc test (P=.05). Results: The evaluation of FE-SEM images revealed no diffenrence at micro-surfaces between each groups. Total coating dose was higher at physical adhesion experimental group than at chemical fixation experimental group. In cell adhesion and proliferation, RGD peptide coating did not show a statistical significance compared with control group (P>.05). In cell differentiation and mineralization, physical adhesion method displayed significantly increased levels compared with control group and chemical fixation method (P<.05). Conclusion: RGD peptide coating seems to enhance osseointegration by effects on the response of human mesenchymal stem cell. Especially physical adhesion method showed more effective than chemical fixation method on response of human mesenchymal stem cell.

EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN PERIOSTEAL-DERIVED CELLS (배양된 인간 골막기원세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Lee, Sung-Gyoon;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.28 no.6
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    • pp.511-519
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    • 2006
  • Autogenous bone grafts have been considered the gold standard for maxillofacial bony defects. However, this procedure could entail a complicated surgical procedure as well as potential donor site morbidity. Possibly the best solution for bone-defect regeneration is a tissue engineering approach, i.e. the use of a combination of a suitable scaffold with osteogenic cells. A major source of osteogenic cells is the bone marrow. Bone marrow-derived mesenchymal stem cells are multipotent and have the ability to differentiate into osteoblastic, chondrocytic, and adipocytic lineage cells. However, the isolation of cells from bone marrow has someproblems when used in clinical setting. Bone marrow aspiration is sometimes potentially more invasive and painful procedure and carries of a risk of morbidity and infection. A minimally invasive, easily accessible alternative would be cells derived from periosteum. The periosteum also contains multipotent cells that have the potential to differentiate into osteoblasts and chondrocytes. In the present study, we evaluated the osteogenic activity and mineralization of cultured human periosteal-derived cells. Periosteal explants were harvested from mandibule during surgical extraction of lower impacted third molar. The periosteal cells were cultured in the osteogenic inductive medium consisting of DMEM supplemented with 10% fetal calf serum, 50g/ml L-ascorbic acid 2-phosphate, 10 nmol dexamethasone and 10 mM -glycerophosphate for 42 days. Periosteal-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 14 of culture period, then decreased in intensity during the culture period. ALP mRNA expression increased up to day 14 with a decrease thereafter. Osteocalcin mRNA expression appeared at day 7 in culture, after that its expression continuously increased in a time-dependent manner up to the entire duration of culture. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. In conclusion, our study showed that cultured human periosteal-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix. As the periosteal-derived cells, easily harvested from intraoral procedure such as surgical extraction of impacted third molar, has the excellent potential of osteogenic capacity, tissue-engineered bone using periosteal-derived cells could be the best choice in reconstruction of maxillofacial bony defects.

EVALUATION OF OSTEOGENIC ACTIVITY AND MINERALIZATION OF CULTURED HUMAN DENTAL PAPILLA-DERIVED CELLS (배양된 치유두 유래세포의 조골활성 및 골기질 형성의 평가)

  • Park, Bong-Wook;Byun, June-Ho;Choi, Mun-Jeoung;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.29 no.4
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    • pp.279-288
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    • 2007
  • In the present study, we focused on stem cells in the dental papilla of the tooth germ. The tooth germ, sometimes called the tooth bud, is the primordial structure from which a tooth is formed. The tooth germ consists of the enamel organ, the dental papilla, and the dental follicle. The dental papilla lies below a cellular aggregation of the enamel organ. Mesenchymal cells within the dental papilla are responsible for formation of dentin and pulp of a tooth. Tooth germ disappears as a tooth is formed, but that of a third molar stays in the jawbone of a human until the age of 10 to 16, because third molars grow slowly. Impacted third molar tooth germs from young adults are sometimes extracted for orthodontic treatment. In the present study, we evaluated the osteogenic activity and mineralization of cultured human dental papilla-derived cells. Dental papillas were harvested from mandible during surgical extraction of lower impacted third molar from 3 patients aged 13-15 years. After passage 3, the dental papilla-derived cells were trypsinized and subsequently suspended in the osteogenic induction DMEM medium supplemented with 10% fetal bovine serum, 50 g/ml L-ascorbic acid 2-phosphate, 10 nM dexamethasone and 10 mM -glycerophosphate at a density of $1\;{\times}10^6\;cells/dish$ in a 100-mm culture dish. The dental papilla-derived cells were then cultured for 6 weeks and the medium was changes every 3 days during the incubation period. Dental papilla-derived cells showed positive alkaline phosphatase (ALP) staining during 42 days of culture period. The formation of ALP stain showed its maximal manifestation at day 7 of culture period, then decreased in intensity during the culture period. ALP mRNA level was largely elevated at 1 weeks and gradually decreased with culture time. Osteocalcin mRNA expression appeared at day 14 in culture, after that its expression continuously increased in a time-dependent manner up to day 28. The expression remained constant thereafter. Runx2 expression appeared at day 7 with no detection thereafter. Von Kossa-positive mineralization nodules were first present at day 14 in culture followed by an increased number of positive nodules during the entire duration of the culture period. Osteocalcin secretion was detectable in the culture medium from 1 week. The secretion of osteocalcin from dental papilla-derived cells into the medium greatly increased after 3 weeks although it showed a shallow increase by then. In conclusion, our study showed that cultured human dental papilla-derived cells differentiated into active osteoblastic cells that were involved in synthesis of bone matrix and the subsequent mineralization of the matrix.