• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• v.35 no.5
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• pp.294-298
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• 2009

Purpose: The development of a microvascularization is important for the homeostasis of normal bone. Vascular endothelial growth factor (VEGF) is one of the most important factors in vessel formation. The purpose of this study was to examine VEGF-related autocrine growth in periosteal-derived cells. Materials and methods: Periosteal-derived cells were obtained from mandibular periosteums and introduced into the cell culture. After passage 3, the periosteal-derived cells were further cultured for 21 days in an osteogenic inductive culture medium containing dexamethasone, ascorbic acid, and $\beta$-glycerophosphate. Results: The expression of four VEGF isoforms and VEGFRs was observed in periosteal-derived cells. Treatment with cultures with VEGFR-1 and VEGFR-2 Kinase Inhibitor inhibited osteoblastic differentiation and alkaline phosphatase (ALP) activity of periosteal-derived cells. In addition, exogenous VEGF treatment increased calcium content in the periosteal-derived cells. Conclusion: These results suggest that VEGF might act as an autocrine growth molecule during osteoblastic differentiation of cultured human periosteal-derived cells.

• The Journal of Dong Guk Oriental Medicine
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• v.1
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• pp.237-249
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• 1992

In order to study tile effects of moxibustion on renal failure induced by $HgCl_2$ in rats aminal experiments were carry out. The moxibustion was treat into Sin-soo locus(VII23), We-soo locus(VII21) and Sin-soo + We-soo loci these loci were correspond to human body. Experimental group were divided into 4 groups : lontrol group, moxibustion groups treat on Sin-Soo and We-Soo, Sin-Soo + We-Soo group. The obtained results were as follows : 1. The concentration of serum sodium were inreased in all the groups, compared with those of the control group. Sin-Soo, We-Soo, Sin-Soo + We-Soo loci showed increased on the 1st and 3rd days. All three experimental groups showed a significant increased on 3rd day. 2. The concentration of serum potassium were decreased in all the group, compared with those of the control group. Sin-Soo, We-Soo showed a significant decrease on 3rd day. Sin-Soo + We-Soo showed a little decrease but do not significant decrease. 3. The levels of serum total protein were increased in all the group, compare with those of the control group. Sin-Soo, We-Soo show a significant increased or 3rd day. Sin-Soo + We-Soo do not showed a significant increased. 4. The levels of serum albumin were decreased in all the groups, compared with those of the control group. Sin-Soo, Sin-Soo + We-Soo showed a significant decrease on 3rd day. We-Soo does not showed a significant decrease. 5. The concentration of serum blood urea nitrogen were decreased in all the groups, compared with those of the control group. Sin-Soo, We-Soo, Sin-Soo + We-Soo showed a significant decrease on 3rd day. 6. The levels of serum alkaline phosphatase were decreased in all the group, compared with those of the control groups. Sin-Soo, We-Soo showed a significant decrease on 3rd day Sin-Soo + We-Soo not a significant decrease. In conclusion, the result of this study seemed to effect on the treatment for renal failure induced by $HgCl_2$.

• Journal of Periodontal and Implant Science
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• v.46 no.4
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• pp.220-233
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• 2016

Purpose: The aim of this study was to present new a model that allows the study of the bone healing process, with an emphasis on the biological behavior of different graft-to-host interfaces. A standardized "over-inlay" surgical technique combined with a differential histomorphometric analysis is presented in order to optimize the use of critical-size calvarial defects in pre-clinical testing. Methods: Critical-size defects were created into the parietal bone of 8 male Wistar rats. Deproteinized bovine bone (DBBM) blocks were inserted into the defects, so that part of the block was included within the calvarial thickness and part exceeded the calvarial height (an "over-inlay" graft). All animals were sacrificed at 1 or 3 months. Histomorphometric and immunohistochemical evaluation was carried out within distinct regions of interest (ROIs): the areas adjacent to the native bone (BA), the periosteal area (PA) and the central area (CA). Results: The animals healed without complications. Differential morphometry allowed the examination of the tissue composition within distinct regions: the BA presented consistent amounts of new bone formation (NB), which increased over time ($24.53%{\pm}1.26%$ at 1 month; $37.73%{\pm}0.39%$ at 3 months), thus suggesting that this area makes a substantial contribution toward NB. The PA was mainly composed of fibrous tissue ($71.16%{\pm}8.06%$ and $78.30%{\pm}2.67%$, respectively), while the CA showed high amounts of DBBM at both time points ($78.30%{\pm}2.67%$ and $74.68%{\pm}1.07%$, respectively), demonstrating a slow remodeling process. Blood vessels revealed a progressive migration from the interface with native bone toward the central area of the graft. Osterix-positive cells observed at 1 month within the PA suggested that the periosteum was a source of osteoprogenitor elements. Alkaline phosphatase data on matrix deposition confirmed this observation. Conclusions: The present model allowed for a standardized investigation of distinct graft-to-host interfaces both at vertically augmented and inlay-augmented sites, thus possibly limiting the number of animals required for pre-clinical investigations.

• Journal of Periodontal and Implant Science
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• v.41 no.5
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• pp.227-233
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• 2011

Purpose: The aim of this study was to compare osteoblast behavior on zirconia and titanium under conditions cultured with bone morphogenetic protein-2. Methods: MC3T3-E1 cells were cultured on sandblasted zirconia and sandblasted/etched titanium discs. At 24 hours after seeding MC3T3-E1, the demineralized bone matrix (DBM) gel alone and the DBM gel with bone morphogenetic protein-2 (BMP-2) were added to the culture medium. The surface topography was examined by confocal laser scanning microscopy. Cellular proliferation was measured at 1, 4, and 7 days after gel loading. Alkaline phosphatase activity was measured at 7 days after gel loading. The mRNA expression of ALPase, bone sialoprotein, type I collagen, runt-related transcription factor 2 (Runx-2), osteocalcin, and osterix were evaluated by real-time polymerase chain reaction at 4 days and 7 days. Results: At 1, 4, and 7 days after loading the DBM gel alone and the DBM gel with BMP-2, cellular proliferation on the zirconia and titanium discs was similar and that of the groups cultured with the DBM gel alone and the DBM gel with BMP-2 was not significantly different, except for titanium with BMP-2 gel. ALPase activity was higher in the cells cultured with BMP-2 than in the other groups, but there was no difference between the zirconia and titanium. In ALPase, bone sialoprotein, osteocalcin, Runx-2 and osterix gene expression, that of cells on zirconia or titanium with BMP-2 gel was much more highly increased than titanium without gel at day 7. The gene expression level of cells cultured on zirconia with BMP-2 was higher than that on titanium with BMP-2 at day 7. Conclusions: The data in this study demonstrate that the osteoblastic cell attachment and proliferation of zirconia were comparable to those of titanium. With the stimulation of BMP-2, zirconia has a more pronounced effect on the proliferation and differentiation of the osteoblastic cells compared with titanium.

• Journal of Periodontal and Implant Science
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• v.39 no.sup2
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• pp.279-286
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• 2009

Purpose: Gene therapy (ex vivo) has recently been used as a means of delivering bone morphogenetic proteins (BMPs) to sites of tissue regeneration. In the present study, we investigated the effect of co-transduction of adenoviruses expressing BMP-2 and BMP-7 on osteogenesisof C2C12 cells in vitro. Methods: A replication-defective human adenovirus 5 (Ad5) containing a cDNA for BMPs in the E1 region of the virus (Ad5BMP-2 and Ad5BMP-7) was constructed by in vivo homologous recombination. Functional activity of Ad5BMP-2 and Ad5BMP-7 were evaluated in mouse stromal cells (W20-17cells). C2C12 cells are transduced with various MOI (multiplicity of infection) of Ad5BMP-2 and Ad5BMP-7 to assess most effective and stable titer. Based on this result, C2C12 cells were transduced with Ad5BMP-2 and Ad5BMP-7 alone or by combination. BMPs expression, alkaline phosphatase (ALPase) activity, cell proliferation, and mineralization were assessed. Results: Ad5BMP-2 and Ad5BMP-7 are successfully transduced to W20-17 cells, and secreted BMPs stimulated cell differentiation. Also, C2C12 cells transduced with Ad5BMPs showed expression of BMPs and increased ALPaseactivity. In all groups, cell proliferation was observed over times. At 7days, cells co-transduced with Ad5BMP-2 and Ad5BMP-7 showed lower proliferation than the others. C2C12 cells co-transduced with Ad5BMP-2 and Ad5BMP-7 had greater ALPaseactivity than that would be predicted if effect of individual Ad5BMPs were additive. Little mineralized nodule formation was detected in cells transduced with individual Ad5BMPs. In contrast, Ad5BMP-2 and Ad5BMP-7 combination stimulated mineralization after culturing for 10 days in mineralizing medium. Conclusions: Present study demonstrated that adenoviruses expressing BMPs gene successfully produced BMPs protein and these BMPs stimulated cells to be differentiated into osteoblastic cells. In addition, the osteogenic activity of Ad5BMPs can be synergistically increased by co-transduction of cells with Ad5BMP-2 and Ad5BMP-7.

• Journal of the Korean Society of Food Science and Nutrition
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• v.42 no.2
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• pp.195-202
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• 2013

This study was carried out to identify the relationship of biochemical bone turnover markers, bone mineral density (BMD), and general characteristics in Korean women. One hundred eighty healthy women, 20 to 50 years of age, living in Gwangju and Chonnam participated. Serum bone-specific alkaline phosphatase (BAP) and osteocalcin (OC) were used as bone formation markers and N-telopeptide of type 1 collagen (NTx) was used as a bone resorption maker to evaluate the state of bone turnover. T-scores were measured to evaluate BMD. We analyzed general characteristics, including age, menopause status, osteoporosis history, alcohol consumption, physical activity level, and degree of obesity (BMI, percent of fat). The BAP level significantly decreased in the group of twenty-year olds and increased in the non-alcohol consuming group, the group with a family history of osteoporosis, the menopause group, and the obese group (p<0.05). The OC level was lower in the group of twenty-year olds and increased in the non-alcohol intake group and the menopause group (p<0.05). BMD significantly decreased in the obese group (p<0.05). In conclusion, BAP and OC were affected by age, alcohol consumption, osteoporosis history, menopause status, and obesity. BMD was affected by obesity degree. These results suggest that the management of alcohol consumption and obesity are important for maintaining bone status during aging in Korean women.

• Journal of Periodontal and Implant Science
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• v.34 no.2
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• pp.269-279
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• 2004

백악질 세포의 분리 및 배양방법을 확립하고, 이를 이용하여 백악질 세포의 형질특성을 알아보고자 하였다. 교정목적으로 발거된 소구치를 이용하여, 치은섬유아세포, 치주인대 세포 및 백악질 유래세포를 분리, 배양하였다. 백악질 유래 세포 배양시에는 백악질을 절제한 후 Collagenase P를 이용하여 백악질 유래 세포 외의 다른 세포의 개제를 배제하였고, 기질을 분해하여 세포의 분리 및 배양이 용이하도록 하였다. 분리 및 배양시기의 세포의 형태를 광화현미정을 이용하여 관찰하였다. 조골세포의 특성을 가지는 SaOs-2 세포를 대조군으로 이용하여 분리 및 배양된 세포군들을 동일한 조건으로 배양하였다. 3일 및 7일째에 세포증식도를 측정하였고 7일째에 ALPase 효소 활성도를 측정하였다. 각 세포의 형질 특성을 알아보기 위해 RT-PCR을 실시하여 조골세포 분화 표식자와 연관된 osteopontin(OPN), Alkaline phosphatase(ALP), type I collagen(COL-I), Bone sialoprotein(BSP), BMP-2 및 osteocalcin(OC)의 발현을 비교 관찰하였다. 백악질 유래 세포의 분리 및 배양을 시도한 5명의 치아 중에서 3명의 치아에서 세포군을 배양해 낼 수 있었다. 배양한 백악질 유래 세포는 섬유아세포와 유사한 형태와 증식을 보였다. ALPase 효소 활성도 검사 결과 백악질 유래 세포는 SaOs-2 세포보다 낮은 활성도를 나타내었으며, 배양된 세포의 RT-PCR 결과 백악질 유래 세포군에서는 ALPase의 발현이 나타나지 않았고, 다른 조골세포 표식자의 발현도 낮게 나타났다. 이는 백악질 유래 세포가 조골세포 및 다른 대조군의 세포와는 다른 형질 특성을 가지고 있다는 것을 시사한다. 이상의 관찰결과로 사람의 백악질 유래 세포롤 백악질의 절제 및 효소처리 방법으로 효과적인 분리 및 배양이 가능하며, 이는 향후 백악질 세포의 형질 특성 및 백악질 형성의 분자적 기전을 파악하는 중요한 연구자료로 활용 될 수 있을 것으로 사료된다.

• Journal of Periodontal and Implant Science
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• v.33 no.1
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• pp.79-89
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• 2003

치주질환은 세균감염에 의해 치조골이 파괴되는 염증성질환으로서 치아상실의 주된 원인이다. Treponema denticola는 성인성 치주염의 병소에서 자주 발견되는 세균으로서 부착능 및 단백분해효소생성능과 같은 독성 인자가 밝혀져 치주조직 파괴에 있어서 중요성이 강조되어 왔다. 골개조는 조골세포의 골형성및 파골세포에 의한 골흡수의 균형에 의하여 유지되며 치주염시 야기되는 치조골파괴는 조골세포 및 파골세포 기능의 불균형에 의하여 야기되는 것으로 설명되고 있다. 골세포에 대한 영향으로서 T. denticola는 파골세포의 형성을 촉진시키는 것으로 보고되었으나 조골세포에 대한 영향은 아직 밝혀져 있지 않다. 따라서 본 연구에서는 T. denticola가 골형성에 미치는 영향을 알아보고자 마우스의 두개골세포로부터 조골세포를 분리한 후 T. denticola분쇄액으로 처리하여 본 세균이 조골세포의 alkaline phosphatase(ALPase) 활성, 석회화결절 형성 및 Prostaglandin $E_2\;(PGE_2)$ 생성에 미치는 영향을 평가하였다. ALPase활성은 p-nitrophenylphosphate분해능, 석회화결절형성은 Von Kossa 염색법, 그리고 PGE2의 농도는 효소면역측정법으로 측정하였다. T. denticola분쇄액 (2.5 ug/ml)은 마우스 두개골세포의 ALPase활성을 억제하였으며 석회화결절의 형성을 감소시켰다. 또한 동일한 농도의 균분쇄액은 마우스 두개골세포의 $PGE_2$ 생산을 증가시켰다. 균분쇄액과 prostaglandin의 합성억제제인 indomethacin으로 세포를 동시에 처리한 경우 T .denticola분쇄액에 의한 $PGE_2$의 생산은 감소되었으나, ALPase의 활성억제에는 변화가 없었다. 균분쇄액을 열처리하여 마우스 두개골세포에 처리하였을 때에도 ALPase의 활성이 억제되는 것에는 변함이 없었다. 이러한 결과는 T. denticola의 구성성분 중 열에 안정한 물질이 prostaglandin과 무관한 경로를 통해 조골세포의 분화를 억제함을 시사하며 이와 같은 T. denticola에 의한 골형성억제가 치주염시 야기되는 치조골 파괴에 관여할 수 있을 것으로 생각된다.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.27 no.1
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• pp.1-8
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• 2005

This study was aimed to characterize osteogenic potential of rat bone marrow stromal cells (BMSC) isolated with standard flushing method and investigate the plasticity of transdifferentiation between osteoblastic and adipocytic lineage of cultured BMSC. Unlike aspiration method in human, rat bone marrow was extracted by means of irrigation with culture media that elevates the possibility of co-extraction of committed osteoprogenitor, or preosteoblast or other progenitor cells of several types present inside bone marrow. The cultured stromal cells showed high ALP activity which is representative marker of osteoblast without any treatment. Osteogenic inducers such as Dex and BMP-2 were examined for the evaluation of their effect on osteogenic and adipocytic differentiation of stromal cells, because they function as osteoinductive agent in stromal cells, but simultaneously induce adipogenic differentiation. Osteogenic differentiation was evaluated by measuring alkaline phosphatase activity or mRNA expression of osteoblast markers such as osteopontin, bone sialoprotein, collagen type I and CbfaI, and in vitro matrix mineralization by von Kossa staining. Oil red staining method was used to detect adipocyte and adipocytic marker, aP2 and $PPAR{\gamma}2$ expression was examined using RT-PCR. It can be supposed that irrigation procedure resulted in high portion of already differentiation-committed osteoprogenitor cell showing elevated ALP activity and strong mineralization only under the supplement of $100{\mu}M$ ascorbic 2-phosphate and 10mM ${\beta}$-glycerophosphate without any treatment of osteogenic inducers such as Dex and BMP-2. Dex and BMP-2 seemed to transdifferentiate osteoprogenitor cells having high ALP activity into adipocytes temporarily, but continuous treatment redifferentiated into osteoblast and developed in vitro matrix mineralization. This property must be considered either in tissue engineering for bone regeneration, or in research of characterization of osteogenic differentiation, with rat BMSC isolated by the standard irrigation method.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.35 no.1
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• pp.31-37
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• 2013

Purpose: Maxilla and mandible have different patterns of cortical and trabecular bone and different bone mineral densities, even though both are components of the jaw bone. However, cellular differences between maxilla- and mandible derived osteoblasts (OBs) have rarely been studied. We hypothesize that maxilla- and mandible-derived OBs show different responses to $17{\beta}$-estradiol (E2), which is one of the critical factors for bone formation. This study compares skeletal site-specific cell responses between maxilla- and mandible-derived human OBs to E2. Methods: Maxilla- and mandible-derived OBs derived from an identical donor were separately isolated from a total of five normal healthy subjects aged 18~44 years old, cultured with a treatment of 100 nM estrogen. The responses between maxilla- and mandible-derived OBs to E2 were evaluated and compared using cell proliferation, alkaline phosphatase (ALP) activity and gene expression of osteoprotegerin (OPG), ALP, insulin-like growth factor-1 (IGF-1), and estrogen receptor ${\alpha}$ ($ER{\alpha}$). Results: E2 did not have any distinct effects on the proliferation of both types of OBs. Mandible-derived OBs exhibited higher ALP activity than maxilla-derived OBs in the non-treated condition, which was common in all tested individuals. ALP activities of both types of OBs showed a minor increasing tendency with the treatment of E2, even though there was no statistical significance in some specimens. The gene expression of OB by E2 was diverse, depending on the individuals. There was increased expression of OPG, IGF-1, or $ER{\alpha}$ gene in the part of subjects, which was more repeated in maxilla-derived OBs. In particular, OPG or ALP induction by E appeared less frequently in mandible-derived OBs. Conclusion: Current results revealed that E2 affects maxilla- and mandible-derived OBs into facilitating the osteogenic process despite individual differences. Mandible-derived OBs are less sensitive to bone-forming gene expression by E2.