• Journal of Society of Preventive Korean Medicine
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• v.11 no.2
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• pp.159-174
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• 2007

Introduction : The experiments was undertaken to evaluate the effects of herbal medicine, Sibjeondaebotang, Yugmijihwangtang, in pregnant rats and fetuses. Female Sprague-Dawley rats were orally administered with the Sibjeondaebotang, Yugmijihwangtang at dose of 5mg/kg/day for 20 days. Pregnant rats were sacrificed at 20th day of gestation, and observed internal and reproductive organs. Approximately live fetuses in the 20th day of gestation were randomly selected and fixed in 95% ethanol. To observe skeletal malformations, fetuses were stained with alcian blue and alizarin red S. Effect on pregnant rats : Maternal body weight of Sibjeondaebotang, Yugmijihwangtang treated group has a tendency to increase compared to that of control group. There were no significant difference in internal and reproductive organs. There were no significant changes between two groups in blood chemistry and hematological values. There were no significant changes in number of corpus luteum, implantation, live fetuses and implantation rate, delivery rate, and sex ratio. But Yugmijihwangtang administered group showed higher early resorption rate than the control group(p<0.05). Also Sibjeondaebotang administered group showed higher late resorption rate than the control group(p<0.05). From the sex ratio, number of females, bigger than number of males in the control group, and more males than females in Yugmijihwangtang administered group(p<0.05).

• Journal of Physiology & Pathology in Korean Medicine
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• v.22 no.4
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• pp.802-809
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• 2008

Gamijangsing-tang (GMJST) has been used for treatment of bone formation in traditional korean medicine. The purpose of this study is to examine effects of GMJST on bone metabolism. The effects on the osteoblasts were determined by measuring (1) cell proliferation, (2) alkaline phosphatase (ALP) activity, (3) osteoprotegerin (OPG) secretion. (4) The morphologic changes of cells were observed by light microscopy and electron microscopy. Mineralization of calcium was determined by quantitative alizarin red-S assay and mineralization of phosphate was observed by von kossa staining. The morphologic changes of mineralization on the cells were observed by transmission electron microscopy (TEM). The effects on the osteoclast were investigated by tartrate-resistant acid phosphatase (TRAP) staining. Following results were obtained: Celluar activity of osteoblastic cells (MG-63) was significantly increased in 10-5 of dilution of GMJST. ALP and OPG activity of osteoblastic cells were increased in GMJST than normal MG-63 cell. Mineralization of osteoblastic cells were increased in GMJST than normal MG-63 cell. The activity of osteoclast cells (RAW 264.7) was significantly decreased in GMJST than normal MG-63 cell. From the results, GMJST stimulated the proliferation and mineralization of bone-forming osteoblast and inhibited by bone- lysis osteoclast.

• Korean Journal of Veterinary Research
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• v.33 no.1
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• pp.7-21
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• 1993

The morphogenesis of the stifle joint in Korean cattle embryos and fetuses was observed by radiography, alizarin red S stain and light microscopy. Fourty-eight(48) embryos and fetuses, ranging from 11mm to 160mm in crown-rump length (C-R L), were used for this study. The experimental samples were divided into twelve separate groups according to their C-R L. The first to the nineth group ranged from 11-100mm in C-R L, spaced at 10mm intervals. The tenth to the twelfth group ranged in C-R L from 101mm to 160mm, and were spaced at 20mm intervals. The results were as follows; 1. The first appearance of the interzone between the femur and tibia was formed in the second experimental group. The patellar ligament and cruciate ligament appeared in the third group and the patella, menisci and synovial cavity appeared in the fourth group. 2. The lateral and medial menisci were first obseved in the fourth group, and these structures showed crescent shape in the fifth group of the fetuses. 3. The first appearance of the femoro-tibial joint cavity was in the third group. The fifth group contained the first appearance of the patello-femoral cavity. Also, the femoro-tibial cavity was divided into two cavities by the anterior and posterior cruciate ligament.

• Korean Journal of Materials Research
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• v.17 no.12
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• pp.669-675
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• 2007

Hydroxyapatite (HAp) and biphasic calcium phosphate (BCP) nano powders were synthesized using the microwave-assisted synthesis process dependent on pH and microwave irradiation time. The average size of a powder was less than 100 nm in diameter. Through in-vitro cytotoxicity tests by an extract dilution method, the HAp and BCP nano powders have shown to be cytocompatible for L-929 fibroblast cells, osteoblastlike MG-63 cells and osteoclast-like Raw 264.7 cells. The activation of osteoblast was estimated by alkaline phosphatase (ALP) activity. When the HAp and BCP were treated to MG-63 cells, alkaline phosphatase activities increased on day 3, compared with those of the untreated cells. Also, the collagen fibers increased when the HAp and BCP powders suspension were treated to MG-63 cells, compared to those of the untreated cells. Quantitative alizarin red S mineralization assays showed a trend toward increasing mineralization in osteoblast cultured with powder suspension. In conclusion, hydroxyapatite and biphasic calcium phosphate appeared to be a bone graft substitute material with optimal biocompatibility and could be further applied to clinical use as an artificial bone graft substitute.

• Nutrition Research and Practice
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• v.10 no.2
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• pp.148-153
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• 2016

BACKGROUND/OBJECTIVES: Bone formation and bone resorption continuously occur in bone tissue to prevent the accumulation of old bone, this being called bone remodeling. Osteoblasts especially play a crucial role in bone formation through the differentiation and proliferation. Therefore, in this study, we investigated the effects of Scytosiphon lomentaria extract (SLE) on osteoblastic proliferation and differentiation in MC3T3-E1 cells. MATERIALS/METHODS: A cell proliferation assay, alkaline phosphatase (ALP) activity assay, alizarin red staining and protein expression analysis of osteoblastic genes were carried out to assess the osteoblastic proliferation and differentiation. RESULTS: The results indicated that treatment of SLE promoted the proliferation of MC3T3-E1 cells and improved ALP activity. And, SLE treatment significantly promoted mineralized nodule formation compared with control. In addition, cells treated with SLE significantly upregulated protein expression of ALP, type 1 collagen, bone morphogenetic protein 2, runt-related transcription factor 2, osterix, and osteoprotegerin. CONCLUSIONS: The results demonstrate that SLE promote differentiation inducement and proliferation of osteoblasts and, therefore may help to elucidate the transcriptional mechanism of bone formation and possibly lead to the development of bone-forming drugs.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.35 no.1
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• pp.7-12
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• 2009

Purpose: Gelatin-hydroxyapatite nanocomposite is similar to inorganic nanostructure of bone. To make a scaffold with osteoinductivity, bone marrow derived stem cells from rabbit femur were impinged into the nanocomposite. This vitro study was to test osteogenic differentiation of the stem cells in the nanocomposite, which was made by authors. Material & Methods: Gel-HA nanocomposite with 10g of HA, 3 g of Gel has been made by co-precipitation process. Bone marrow was obtained from femur of New Zealand White rabbits and osteogenic differentiation was induced by culturing of the BMSCs in an osteogenic medium. The BMSCs were seeded into the Gel-HA nanocomposite scaffold using a stirring seeding method. The scaffolds with the cells were examined by scanning electron microscopy (SEM), colorimetry assay, biochemical assay with alkaline phosphatase (ALP) diagnostic kit, osteocalcin ELISA kit. Results: Gel-HA nanocomposite scaffolds were fabricated with relatively homogenous microscale pores ($20-40{\mu}m$). The BMSCs were obtained from bone marrow of rabbit femurs and confirmed with flow cytometry, Alizarin red staining. Attachment and proliferation of BMSCs in Gel-HA nanocomposite scaffold could be identified by SEM, ALP activity and osteocalcin content of BMSCs. Conclusion: The Gel-HA nanocomposite scaffold with micropores could be fabricated and could support BMSCs seeding, osteogenic differentiation.

• Journal of Periodontal and Implant Science
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• v.42 no.4
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• pp.113-118
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• 2012

Purpose: The aim of this study was to investigate the effects of synthetic fibronectin (FN) fragments, including fibrin binding sites from amino-terminal FN fragments containing type I repeats 1 to 5, on osteoblast-like cell activity. Methods: Oligopeptides ranging from 9 to 20 amino acids, designated FF1, FF3, and FF5, were synthesized by a solid-phase peptide synthesizing system, and we investigated the effects of these peptides on cell attachment and extent of mineralization using confocal microscopy, 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays, and Alizarin red S staining. Results: FF3 and FF5 peptides increased the number of attached human osteoblastic cells, and FF3 administration led to prominent cell spreading. Mineralization was increased in FF3 and FF5 compared to FF1 and the untreated control. Conclusions: Taken together, it can be concluded that the fibrin-binding oligopeptides FF3 and FF5 enhanced cell attachment and mineralization on osteoblast-like cells. These results indicate that FF3 and FF5 have the potential to increase osteoblast-like cell activity. Performing an in vivo study may provide further possibilities for surface modification of biomimetic peptides to enhance osteogenesis, thus improving the regeneration of destroyed alveolar bone.

• Journal of Periodontal and Implant Science
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• v.50 no.5
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• pp.291-302
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• 2020

Purpose: The objective of this study was to investigate whether phelligridin D could reduce glucose-induced oxidative stress, attenuate the resulting inflammatory response, and restore the function of human periodontal ligament cells (HPDLCs). Methods: Primary HPDLCs were isolated from healthy human teeth and cultured. To investigate the effect of phelligridin D on glucose-induced oxidative stress, HPDLCs were treated with phelligridin D, various concentrations of glucose, and glucose oxidase. Glucose-induced oxidative stress, inflammatory molecules, osteoblast differentiation, and mineralization of the HPDLCs were measured by hydrogen peroxide (H₂O₂) generation, cellular viability, alkaline phosphatase (ALP) activity, alizarin red staining, and western blot analyses. Results: Glucose-induced oxidative stress led to increased production of H₂O₂, with negative impacts on cellular viability, ALP activity, and calcium deposition in HPDLCs. Furthermore, HPDLCs under glucose-induced oxidative stress showed induction of inflammatory molecules (intercellular adhesion molecule-1, vascular cell adhesion protein-1, tumor necrosis factor-alpha, interleukin-1-beta) and disturbances of osteogenic differentiation (bone morphogenetic protein-2, and -7, runt-related transcription factor-2), cementogenesis (cementum protein-1), and autophagy-related molecules (autophagy related 5, light chain 3 I/II, beclin-1). Phelligridin D restored all these molecules and maintained the function of HPDLCs even under glucose-induced oxidative stress. Conclusions: This study suggests that phelligridin D reduces the inflammation that results from glucose-induced oxidative stress and restores the function of HPDLCs (e.g., osteoblast differentiation) by upregulating autophagy.

• Journal of Periodontal and Implant Science
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• v.33 no.1
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• pp.49-60
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• 2003

Several growth factors and polypeptides are not commonly yet used for regenerators of bone tissue or alveolar bone because of the insufficiency of studies on their side effects, genetic engineering for mass production and stability for clinical application. Recently, many herbal medicines, which have advantage of less side effects and possibility of long-term use, have been studied for their capacity and effects of anti-bacterial, antiinflammatory and regenerative potential of periodontal tissues. Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have been traditionally used as medicines for treatment of bone disease in Eastern medicine. The objective of the present study is to examine the ability of alkaline phosphatase (ALP) activity of human fetal osteoblast (hFOB1) when several natural medicines were supplemented. hFOB1 were cultured with Dulbecuo's Modified Eagle's Medium Nutrient Mixture F-12 HAM ( DMEM/F-12 1:1 Mixture, Sigma, USA) and negative control, dexamethasone (positive control), and each natural medicines for 3 days. And then ALP activity was measured by spectrophotometer for enzyme activity and Alizarin red S staining for morphometry. Among the natural medicines of this study, Morindae Radix, Cibotium Barometz (L.) and Cistanchis Herba induced higher activity of ALP synthesis than negative controls in all experimental group. Albizziae Cortex showed mild increases than negative control group. According to measurement of positively stained area, all of the natural medicines of this study increased compared to negative control. Especially, Cibotium Barometz (L.) and Cistanchis Herba showed statistical significance compared to negative control (p<0.05). These results indicate that Morindae Radix, Cibotium Barometz (L.), Albizziae Cortex, Cistandhis Herba have an inducing ability of ALP synthesis on osteoblast.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.36 no.1
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• pp.7-15
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• 2010

Complex human tissues harbor stem cells and precursor cells, which are responsible for tissue development or repair. Recently, dental tissues such as dental pulp, periodontal ligament (PDL), dental follicle have been identified as easily accessible sources of undifferentiated cells. These tissues contain mesenchymal stem cells that can be differentiate into bone, cartilage, fat or muscle by exposing them to specific growth conditions. In this study, the authors procured the stem cell from pulp, PDL, and dental follicle and differentiate them into osteoblast and examine the bone induction capacity. Dental pulp stem cell (DPSC), periodontal ligament stem cell (PDLSC), and dental follicle precursor cell (DFPC) were obtained from human 3rd molar and cultured. Each cell was analyzed for presence of stem cell by fluorescence activated cell sorter (FACs) against CD44, CD105 and CD34, CD45. Each stem cell was cultured, expanded and grown in an osteogenic culture medium to allow formation of a layer of extracellular bone matrix. Osteogenic pathway was checked by alizarin red staining, alkaline phosphatase (ALP) activity test and RT-PCR for ALP and osteocalcin (OCN) gene expression. According to results from FACs, mesenchymal stem cell existed in pulp, PDL, and dental follicle. As culturing with bone differentiation medium, stem cells were differentiated to osteoblast like cell. Compare with stem cell from pulp, PDL and dental follicle-originated stem cell has more osteogenic effect and it was assumed that the character of donor cell was able to affect on differential potency of stem cell. From this article, we are able to verify the pulp, PDL, and dental follicle from extracted tooth, and these can be a source of osteoblast and stem cell for tissue engineering.