• Title/Summary/Keyword: alkaline phosphatase.

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Analysis of ceramide metabolites in differentiating epidermal keratinocytes treated with calcium or vitamin C

  • Kim, Ju-Young;Yun, Hye-Jeong;Cho, Yun-Hi
    • Nutrition Research and Practice
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    • v.5 no.5
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    • pp.396-403
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    • 2011
  • Ceramides (Cer) comprise the major constituent of sphingolipids in the epidermis and are known to play diverse roles in the outermost layers of the skin including water retention and provision of a physical barrier. In addition, they can be hydrolyzed into free sphingoid bases such as $C_{18}$ sphingosine (SO) and $C_{18}$ sphinganine (SA) or can be further metabolized to $C_{18}$ So-1-phosphate (S1P) and $C_{18}$ Sa-1-phosphate (Sa1P) in keratinocytes. The significance of ceramide metabolites emerged from studies reporting altered levels of SO and SA in skin disorders and the role of S1P and Sa1P as signaling lipids. However, the overall metabolism of sphingoid bases and their phosphates during keratinocyte differentiation remains not fully understood. Therefore, in this study, we analyzed these Cer metabolites in the process of keratinocyte differentiation. Three distinct keratinocyte differentiation stages were prepared using 0.07 mM calcium (Ca$^{2+}$) (proliferation stage), 1.2 mM Ca$^{2+}$ (early differentiation stage) in serum-free medium, or serum-containing medium with vitamin C (50 ${\mu}L$/mL) (late differentiation stage). Serum-containing medium was also used to determine whether vitamin C increases the concentrations of sphingoid bases and their phosphates. The production of sphingoid bases and their phosphates after hydrolysis by alkaline phosphatase was determined using high-performance liquid chromatography. Compared to cells treated with 0.07 mM Ca$^{2+}$, levels of SO, SA, S1P, and SA1P were not altered after treatment with 1.2 mM Ca$^{2+}$. However, in keratinocytes cultured in serum-containing medium with vitamin C, levels of SO, SA, S1P, and SA1P were dramatically higher than those in 0.07- and l.2-mM Ca$^{2+}$-treated cells; however, compared to serum-containing medium alone, vitamin C did not significantly enhance their production. Taken together, we demonstrate that late differentiation induced by vitamin C and serum was accompanied by dramatic increases in the concentration of sphingoid bases and their phosphates, although vitamin C alone had no effect on their production.

Antioxidant and hepatoprotective effects of Korean ginseng extract GS-KG9 in a D-galactosamine-induced liver damage animal model

  • Jo, Yun Ho;Lee, Hwan;Oh, Myeong Hwan;Lee, Gyeong Hee;Lee, You Jin;Lee, Ji Sun;Kim, Min Jung;Kim, Won Yong;Kim, Jin Seong;Yoo, Dae Seok;Cho, Sang Won;Cha, Seon Woo;Pyo, Mi Kyung
    • Nutrition Research and Practice
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    • v.14 no.4
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    • pp.334-351
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    • 2020
  • BACKGROUND/OBJECTIVES: This study was designed to investigate the improvement effect of white ginseng extract (GS-KG9) on D-galactosamine (Ga1N)-induced oxidative stress and liver injury. SUBJECTS/METHODS: Sixty Sprague-Dawley rats were divided into 6 groups. Rats were orally administrated with GS-KG9 (300, 500, or 700 mg/kg) or silymarin (25 mg/kg) for 2 weeks. The rats of the GS-KG9- and silymarin-treated groups and a control group were then intraperitoneally injected Ga1N at a concentration of 650 mg/kg for 4 days. To investigate the protective effect of GS-KG9 against GalN-induced liver injury, blood liver function indicators, anti-oxidative stress indicators, and histopathological features were analyzed. RESULTS: Serum biochemical analysis indicated that GS-KG9 ameliorated the elevation of aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) in GalN-treated rats. The hepatoprotective effects of GS-KG9 involved enhancing components of the hepatic antioxidant defense system, including glutathione, glutathione peroxidase (GPx), superoxide dismutase (SOD), and catalase (CAT). In addition, GS-KG9 treatment inhibited reactive oxygen species (ROS) production induced by GalN treatment in hepatocytes and significantly increased the expression levels of nuclear factor erythroid-2-related factor 2 (Nrf2) and heme oxygenase-1 (HO-1) proteins, which are antioxidant proteins. In particular, by histological analyses bases on hematoxylin and eosin, Masson's trichrome, α-smooth muscle actin, and transforming growth factor-β1 staining, we determined that the administration of 500 mg/kg GS-KG9 inhibited hepatic inflammation and fibrosis due to the excessive accumulation of collagen. CONCLUSIONS: These findings demonstrate that GS-KG9 improves GalN-induced liver inflammation, necrosis, and fibrosis by attenuating oxidative stress. Therefore, GS-KG9 may be considered a useful candidate in the development of a natural preventive agent against liver injury.

Effects of Calcium Supplementations on Mineral Metabolism during Pregnancy with Calcium-Deficient Young Adult Rats (가임기 동안 칼슘섭취 부족 흰쥐에서 임신기의 칼슘보충 수준이 무기질대사에 미치는 영향)

  • 이연숙;김은애;박미나
    • Journal of Nutrition and Health
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    • v.36 no.5
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    • pp.459-469
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    • 2003
  • We examined the effects of calcium intake levels on mineral metabolism during pregnancy using calcium-deficient young adult rats. Five week-old female Sprague-Dawley rats were fed normal Ca (NCa, 0.5%) and low Ca (LCa, 0.15%) diets for five weeks (pre-pregnancy). The low Ca intake group was then divided into three groups and fed low Ca (0.15%), normal Ca (0.5%) and high Ca (1.5%) diets for 3 weeks (pregnancy). All of the rats were mated with normal male rats. The control group was fed a consistently normal Ca (0.5%) diet during pre-pregnancy and pregnancy. On the day after delivery, dams and their pups were sacrificed. We measured total protein, albumin, alkaline phosphatase and mineral content in serum and weight, length, breaking force, ash and mineral content in the femur and lumbar (L2-L4) of the dams. Whole body mineral content was measured in the pups. There was no difference in weight gain and food intake among the groups. Serum total protein and albumin were in the normal range but a little lower during pregnancy. High Ca supplementation decreased serum Mg and Fe during pregnancy. Weight, ash and Ca of the femur and lumbar significantly decreased in rats fed a chronically low Ca diet during pre-pregnancy and pregnancy. Calcium supplementation levels were above normal during pregnancy and increased the bone weight and breaking force of rats fed the low Ca diet during pre-pregnancy. However, Ca supplementation did not increase the ash and Ca contents in the bones. High Ca supplementation during pregnancy significantly decreased Mg in the bones and increased Ca and P in the kidneys. Ash content of pups from dams fed the chronically low Ca diet decreased but there was no difference in whole body Ca among the groups. Mg and Fe in the whole body decreased in pups from dams fed the high Ca diet. Pregnancy performance was reduced in dams fed the low Ca diet. These results suggest that above normal Ca supplementation levels during pregnancy restored maternal bone status to some extent in rats fed the chronically low Ca diet. The same could not be said for mineral content. Also, high Ca supplementation during pregnancy may deteriorate mineral composition in bones and other tissues. Therefore, more detailed research is needed to facilitate sound recommendations on appropriate calcium intake during pregnancy. (Korean J Nutrition 36(5): 459∼469, 2003)

Nutrition Survey in a Korean Mountainous Farming Area (산간지(山間地) 농촌주민(農村住民)의 영양실태조사(營養實態調査))

  • Ham, Jung-Rea;Kim, Hyong-Soo;Lee, Ki-Yull;Kim, Young-Hoo
    • Journal of Nutrition and Health
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    • v.6 no.3
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    • pp.37-45
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    • 1973
  • The main character of the Korean diet has been found to be low in protein both quantity and quality and high in carbohydrate. The purpose of this survey was to study the amount of salt intake related to the dietary pattern in Korea. The nutrition survey was conducted in a mountainous farming area located in Auhchun-ri, Gaebuk-myon, Changsoo-gun, Chunbuk Province, February 14-19 in 1973 (7 days). The precise weighing method was used in evaluating the kinds of foods and nutrients intake for 24 households during a three day period. The physical examinations were performed by a doctor on 120 persons and a detailed biochemical test on both blood and urine was made on 42 persons over 40 years old. The results obtained are summarized as follows: (1) Average nutrients intake of an adult per day: calorie intake was 2,446 Cal and its components-protein(61.1g) was 10 percent, fat(12.9g) was 5 percent and carbohydrate(521g) contributed 85 percent of the total calories. Other nutrients-calcium (443mg), thiamine(1.09mg), riboflavin (0.90mg), niacin (14.4mg) and vitamin C (63.2mg) were lower than the recommended daily allowance but vitamin A(2,083 I.U.), iron(11mg) and phosphorous(998mg) were slightly higher than that. (2) To evaluate the nutritional deficiences, clinical examinations were conducted. Angular stomatitis was present in 16.7 percent of those examiners. No edema was found. The rate of osteoarthritis, hepatomegaly diseases appeared in 20 percent of the total subjects and the symptoms appeared highest among those Iron 50 to 59 years old. (3) The following chemical components of blood serum were analyzed and found to be within the normal range: glucose, blood urea nitrogen, uric acid, total protein, albumin, globulin, bilirubin, total cholesterol, inorganic phosphate, alkaline phosphatase, sodium, potassium, chloride, calcium and lecithine dehydrase. One case of each of the following were found: hyperglycemia, hypocholesterolemia, renal problem, hypoproteinaemia and diabetes mellitus, and two persons were classified as showing hypoglycemia and hyponaturemia. (4) The sodium content in urine was 199.6 mEq/L, potassium content was 24.6 mEq/L. The sugar, pH and specific gravity in the urine was shown to be normal.

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Effect of Ethanol Extract of Yukmijiwhang-Whan on Trabecular Bone Area in OVX Rats (육미지황환(六味地黃丸) 에탄올 추출물이 난소제거 흰쥐의 경골 소주골에 미치는 영향)

  • Kim, Chung-Sook;Ha, Hye-Kyung;Lee, Je-Hyun;Song, Kye-Yong;Kim, Hye-Jin;Shin, Sun-Mi
    • Korean Journal of Oriental Medicine
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    • v.6 no.1
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    • pp.123-132
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    • 2000
  • Bone is continuously remodeled during adult life with the resorption of old bone by osteoclasts and its subsequent replacement by osteoblast. Bone homeostasis is maintained by a balance between activities of osteoblasts and osteoclasts, but an imbalance between resorption and formation results in bone diseases including osteoporosis. Osteoblasts line up on the bone surface, especially regions of new bone formation, lay down bone matrix (osteoid) in orderly lamellae and induce its mineralization. Thus, the increased activity of osteoblasts is helpful to treat and prevent osteoporosis. In this study, we examined whether 80% EtOH extract of yukmijiwhang-whan is capable of affecting osteoblast proliferation using human osteoblast-like cell line, MG-63 and Saos-2. In an in vivo experiment, extract of yukmijiwhang-whan was administered for 9 weeks to ovariectomized (OVX) rats. At necropsy, uterus weights were measured, and trabecular bone areas (TBAS) of tibia and the sixth lumbar vertebra were measured by bone histomorphology. The maximum cell proliferation of MG-63 caused by extract of yukmijiwhang-whan at $5\;{\times}\;10^{-6}\;mg/ml$ was approximately 115% compared with control. In Saos-2, cell proliferation was approximately 145% of control at $5\;{\times}\;10^{-4}\;mg/ml$ and maximum alkaline phosphatase (ALP) activity was approximately 143% of control at $5\;{\times}\;10^{-5}\;mg/ml$. In animal study, however, the tibia and lumbar TBAS of the yukmijiwhang-whan group did not increased than the OVX control group. In conclusion, the 80% EtOH extract of yukmijiwhang-whan increased proliferation of osteoblasts but did not prevent bone loss in OVX rats.

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Downregulation of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in human keratinocytes by melanogenic inhibitors

  • Ahn, Kwang-Seok;Lee, Jinseon;Kim, Yeong-Shik
    • Proceedings of the SCSK Conference
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    • 2003.09a
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    • pp.780-803
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    • 2003
  • Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB ($\textrm{NF}_{-{\kappa}}\textrm{B}$) activators [e.g. tumor necrosis factor-$\alpha$, interleukin-1, lipopolysaccharides, and ultraviolet light] leads to phosphorylation and degradation of the inhibitory protein, $\textrm{I}_{{\kappa}}\textrm{B}$. Liberated $\textrm{NF}_{-{\kappa}}\textrm{B}$ is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. In order to demonstrate the possible role of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of $\textrm{NF}_{-{\kappa}}\textrm{B}$ induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with $\textrm{pNF}_{-{\kappa}}\textrm{B}$-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the $\textrm{NF}_{-{\kappa}}\textrm{B}$ activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation was measured with the SEAP reporter gene assay using a fluorescence detection method. Of the Mis tested, kojic acid ($IC_{50}$/ = 60 $\mu$M) was found to be the most potent inhibitor of UVB-upregulating $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in transfectant HaCaT cells, which is followed by niacinamide ($IC_{50}$/= 540 $\mu$M). Pretreatment of the transfectant HaCaT cells with the Mis, especially kojic acid and niacinamide, effectively lowered $\textrm{NF}_{-{\kappa}}\textrm{B}$ binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. These observations suggest that Mis working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte.

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Anticancer and Immunopotentiating Activities of Crude Polysaccharides from Pleurotus nebrodensis on Mouse Sarcoma 180

  • Cha, Youn Jeong;Alam, Nuhu;Lee, Jae Seong;Lee, Kyung Rim;Shim, Mi Ja;Lee, Min Woong;Kim, Hye Young;Shin, Pyung Gyun;Cheong, Jong Chun;Yoo, Young Bok;Lee, Tae Soo
    • Mycobiology
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    • v.40 no.4
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    • pp.236-243
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    • 2012
  • Pleurotus nebrodensis is an edible and commercially available mushroom in Korea. This study was conducted in order to evaluate the anticancer and immunopotentiating activities of crude polysaccharides, extracted in methanol, neutral saline, and hot water (hereafter referred to as Fr. MeOH, Fr. NaCl, and Fr. HW, respectively) from the fruiting bodies of P. nebrodensis. ${\beta}$-Glucan and protein contents in Fr. MeOH, Fr. NaCl, and Fr. HW extracts of P. nebrodensis ranged from 23.79~36.63 g/100 g and 4.45~6.12 g/100 g, respectively. Crude polysaccharides were not cytotoxic against sarcoma 180, HT-29, NIH3T3, and RAW 264.7 cell lines at a range of $10{\sim}2,000{\mu}g/mL$. Intraperitoneal injection with crude polysaccharides resulted in a life prolongation effect of 11.76~27.06% in mice previously inoculated with sarcoma 180. Treatment with Fr. NaCl resulted in an increase in the numbers of spleen cells by 1.49 fold at the concentration of $50{\mu}g/mL$, compared with control. Fr. HW improved the immuno-potentiating activity of B lymphocytes through an increase in alkaline phosphatase activity by 1.65 fold, compared with control at $200{\mu}g/mL$. Maximum production of nitric oxide ($14.3{\mu}M$) was recorded in the Fr. NaCl fraction at $200{\mu}g/mL$. Production of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interleukin-$1{\beta}$ (IL-$1{\beta}$), and interleukin-6 (IL-6) was significantly higher, compared to control, and IL-6 production was highest, in contrast to TNF-${\alpha}$, IL-$1{\beta}$, and positive control, concanavalin at the tested concentration of the various fractions. Results of the current study suggest that polysaccharides extracted from P. nebrodensis have a strong anticancer effect and may be useful as an ingredient of biopharmaceutical products for treatment of cancer.

Prognosis and Surgical Treatment of the Urethra Embedding Leiomyosarcoma in a Dog (개에서 요도를 포매한 평활근육종의 수술적 처치 및 예후)

  • Kim, Ji-Hyun;Lee, Jun-Am;Kim, Ill-Hwa;Jang, Dong-Woo;Kang, Hyun-Gu
    • Journal of Veterinary Clinics
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    • v.31 no.4
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    • pp.307-312
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    • 2014
  • A 16-year-old female Shih-Tzu, weighing 5 kg, presented with clinical symptoms of abdominal distension and urinary and fecal incontinence. Abdominal palpation detected a large mass. According to the radiographic findings, the bladder had been moved to the umbilicus by the mass and a large abdominal mass was confirmed in the lower abdominal area. Ultrasonography indentified a large heterogeneous mass with heterogeneous parenchyma and a focal anechoic area in the lower abdominal area. The complete blood count abnormalities suggested thrombocytosis and mild neutrophilia, and the serum chemistry indicated an elevated alkaline phosphatase value. During laparotomy, a firm mass that measured $10.5{\times}9.6cm$ was found between the uterine cervix and urinary bladder. The urethra was embedded in the mass. A diagnosis of leiomyosarcoma was established based on histopathology and histochemistry. One week after surgery, urinary retention symptoms that did not appear to be related to mechanical obstruction presented suddenly, but they did not respond to several drug treatments, thus long-term conservative therapy was adopted. The urinary symptoms disappeared on day 27 and the patient started to void large quantities of urine in a smooth and frequent manner. This case report describes the serial changes in the patient's status and the response after surgical remove of the urethra embedding leiomyosarcoma.

Antioxidant and anti-fatigue effects of abalone (Haliotis discus hannai) composites containing natural plants (전복 복합물의 항산화능 및 피로개선 효과)

  • Lee, Soo-Jung;Oh, Soo-Jeong;Kang, Min-Jung;Shin, Jung-Hye;Kang, Shin-Kwon
    • Food Science and Preservation
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    • v.22 no.4
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    • pp.598-606
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    • 2015
  • To develop a functional drink using abalone, two abalone composites (APM-1, APM-2) were prepared by mixing the abalone and natural plants (Lycii fructus and Rubus coreanus Miq.). Their antioxidant and anti-fatigue effects were evaluated using rats running on a treadmill after 4 weeks supplementation of the abalone composites. Experimental groups were divided into four groups including normal (non-exercised group), control (exercised group), one dose per day (EAPM-1), and three doses per day (EAPM-2) with exercise by running. Antioxidant activities, and total phenols and flavonoids contents of APM-2 were significantly higher than those of APM-1 (p<0.05). Total lipid content in serum of EAPM-2 was significantly lower than that of control group (p<0.05). Lactate dehydrogenase activity and blood urea nitrogen content of EAPM-1 and EAPM-2 groups were significantly lower than that of control group. There were no significant difference in aspartate aminotransferase activity among control, EAPM-1 and EAPM-2 groups. In the meanwhile, alanine aminotransferase and alkaline phosphatase activities were significantly lower than that of control group. In serum and liver tissues of EAPM-1 and EAPM-2 groups, lipid peroxide contents significantly decreased compared to control group. DPPH radical scavenging activities in liver tissues of EAPM-1 and EAPM-2 groups were significantly higher than those of control group. Therefore, abalone composites were effective for the alleviation of oxidative stress caused by treadmill running, which was dependent on antioxidant activity and phenolic compounds content.

EFFECT OF DEXAMETHASONE CONCENTRATIONS ON OSTEOGENIC ACTIVITY OF CULTURED HUMAN PERIOSTEAL-DERIVED CELLS (배양된 인간 골막기원세포의 조골활성에 대한 덱사메타손 농도의 효과)

  • Kim, Jong-Ryoul;Park, Bong-Wook;Lee, Chang-Il;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Byun, June-Ho
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.31 no.4
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    • pp.287-293
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    • 2009
  • Long-term treatment with glucocorticoid leads to the development of osteoporosis and osteonecrosis. In contrast to the marked inhibitory effect of pharmacological doses of glucocorticoids on bone formation, the relationship between physiological concentrations of glucocorticoids and osteoprogenitor cell proliferation and phenotypes has not been elucidated yet. In addition, the effects of dexamethasone treatment on the proliferation and osteoblastic differentiation of osteoprogenitor cells are also controversial. The purpose of this study was to examine the effects of dexamethasone on the proliferation and osteoblastic differentiation of periosteal-derived cells. Periosteal-derived cells were obtained from mandibular periosteums and introduced into the cell culture. After passage 3, the cells were further cultured for 21 days in the osteogenic induction medium with different dexamethasone concentrations of 0, 10, and 100 nM. The proliferation and osteoblastic phenotypes of periosteal-derived cells were promoted in dexamethasone-treated cells than in untreated cells. Among the dexamethasone-treated cells, cell proliferation was slightly greater in 10 nM dexamethasone-treated cells than in 100 nM dexamethasone-treated cells. Histochemical staining and the bioactivity of alkaline phosphatase (ALP) were higher in 100 nM dexamethasone-treated cells than in 10 nM dexamethasone-treated cells. Similarly, von Kossa-positive mineralization nodules and calcium content were also more evident in 100 nM dexamethasone-treated cells than in 10 nM dexamethasone-treated cells. These results suggest that dexamethasone enhances the in vitro osteoblastic differentiation of periosteal-derived cells. The present study also demonstrates that higher dexamethasone concentrations reduce the in vitro proliferation of periosteal-derived cells.