• Journal of Periodontal and Implant Science
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• 제44권4호
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• pp.178-183
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• 2014

Purpose: The aim of this study was to assess the possible paracrine effect of bone morphogeneticprotein-2 (BMP-2) at the experimental site on the adjacent control site for validating a rabbit calvarial defect model as a means of verifying the effect of BMP-2. Methods: Sixteen rabbits were divided into two groups (n=8 in each) according to whether or not BMP-2 would be used. Two circular defects (8 mm in diameter) were created side by side, 2 mm apart, in the calvarium of all of the rabbits. In each animal, one of the defects was grafted with either BMP-2-loaded carrier or carrier material alone. The control defects adjacent to these grafted defects, designated CB (the nongrafted defect adjacent BMP-2-loaded carrier-grafted defect) and CC (the nongrafted defect adjacent to carrier only-grafted defect), respectively, were the focus of this study, and were filled only with a blood clot in all of the animals. Histologic observation and histomorphometric analysis were performed at 2 and 8 weeks (n=4 animals per point in time) after surgery. Results: There was no noteworthy difference in the healing pattern, and no statistically significant differences in histomorphometric parameters such as the defect closure, new bone area, or total augmented area between the CC and CB groups. Conclusions: The results of this study suggest that rabbit calvarial defects separated by a distance of 2 mm are suitable for evaluating the effects of BMP-2 and the control defect can be regarded not to be affected by BMP-2 applied defect.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제34권4호
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• pp.419-427
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• 2008

The present study aimed to investigate the osteogenic potentials of differentiated osteoblast-like cells (DOCs) induced from bone marrow-derived mesenchymal stem cells (MSCs) on ${\beta}-tricalcium$ phosphate (${\beta}-TCP$) with recombinant human bone morphogenetic protein (rhBMP-2) in vitro. Osteoblast differentiation was induced in confluent cultures by adding 100 nM dexamethasone, 10 mM ${\beta}$-glycerophosphate, 50 mM L-ascorbic acid. The Alizarin red S staining and reverse transcriptase-polymerase chain reaction (RT-PCR) were perfomed to examine the mRNA expression of alkaline phosphatase (ALP), bone sialoprotein (BSP), osteocalcin (OCN), receptor activator for nuclear factor ${\kappa}B$ ligand (RANKL), runt-related transcription factor 2 (RUNX2), collagen-Ⅰ (COL-Ⅰ). There were no significant differences in the osteogenic potentials of DOCs induced from MSCs on ${\beta}-TCP(+/-)$. According to the incubation period, there were significant increasing of Alizadin red S staining in the induction 3 weeks. The mRNA expression of ALP, RUNX2, and RANKL were higher in DOCs/${\beta}-TCP(-)$ than DOCs/${\beta}-TCP(+)$. According to rhBMP-2 concentrations, the mRNA expression of BSP was significantly increased in DOCs/${\beta}-TCP(+)$ compared to that of DOCs/${\beta}-TCP(-)$ on rhBMP 10 ng/ml. Our study presented the ${\beta}-TCP$ will have the possibility that calcium phosphate directly affect the osteoblastic differentiation of the bone marrowderived MSCs.

• Journal of Periodontal and Implant Science
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• 제30권4호
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• pp.757-779
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• 2000

The aim of our study is to achieve complete periodontal tissue regeneration by the application of BMP and resorbable membrane. Three beagle dogs aged over one and half years and weighed 14 to 16 kg were used in this study. Mandibular 1st, 2nd premolars were extracted bilaterally. Horizontal furcation defects were induced around 3rd, 4th premolars bilaterally. BMP-4 were applied in the right side with resorbable membranes and only resorbable membranes were applied in the left side respectively. Each animal was sacrificed at 2, 4, and 8weeks, after regenerative surgery. Specimens were prepared with Hematoxylin-Eosin stain and Goldner's modified Masson Trichrome stain for light microscopic evaluation. The results were as follows: 1. At 2 weeks after regenerative surgery, downgrowth of junctional epithelium was observed both in the membraneapplied site and BMP-4-and-membrane-applied site. 2. At 4 weeks after regenerative surgery, resorbable membranes were completely resolved, therefore would not prevent downgrowth of junctional epithelium. New bone formation, new cementum formation and Sharpey's fiber were observed in BMP-4-andmembrane-applied site. 3. At 8 weeks after regenerative surgery, downgrowth of junctional epithelium was observed in the membrane-applied site. But, new cementum formation was observed in the same site. The extensive regeneration of new bone, new cementum and remarkable formation of Shapey's fiber were showed in BMP-4-and-membrane-applied site. 4. Resorbable membranes were resolved via the cell-mediated processes. 5. Periodontal tissue regeneration were better achieved in the BMP-4-andmembrane-applied site than in the membrane-applied site. Within the above results, BMP-4 may have the strong capability to form the new bone and resorbable membrane may be able to prevent the bony ankylosis. However, resolution rate of resorbable membrane may not be enough to protect rapid epithelial downgrowth for ideal periodontal regeneration. In conclusion, I suggest BMP-4 may have the strong possibility to be utilized in the clinical periodontal treat-

• BMB Reports
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• 제45권4호
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• pp.247-252
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• 2012

Although previous studies have demonstrated that BMP9 is highly capable of inducing osteogenic differentiation of mesenchymal stem cells, the molecular mechanism involved remains to be fully elucidated. In this study, we showed that BMP9 simultaneously promotes the activation of Smad1/5/8, p38 and ERK1/2 in C3H10T1/2 cells. Knockdown of Smad4 with RNA interference reduced nuclear translocation of Smad1/5/8, and disrupted BMP9-induced osteogenic differentiation. BMP9-induced osteogenic differentiation was blocked by p38 inhibitor SB203580, whereas enhanced by ERK1/2 inhibitor PD98059. SB203580 decreased BMP9-activated Smads singling, and yet PD98059 stimulated Smads singling in C3H10T1/2 cells. The effects of inhibitor were reproduced with adenovirus expressing siRNA targeted p38 and ERK1/2, respectively. Taken together, our findings revealed that Smads, p38 and ERK1/2 are involved in BMP9-induced osteogenic differentiation. Also, it is noteworthy that p38 and ERK1/2 may play opposing regulatory roles in mediating BMP9-induced osteogenic differentiation of C3H10T1/2 cells.

• 한방재활의학과학회지
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• 제30권4호
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• pp.17-30
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• 2020

Objectives The purpose of this study is to evaluate the healing effect of Yukmijihwang-tang (YM) on femur fractured mice. Methods Mice were randomly divided into 6 groups: normal, control, positive control, YM with low, medium, high dosage each. All groups were prepared with femur fracture and treated diffrently. In order to measure bone regeneration effects, we analysed the levels of cyclooxygenase-2 (COX2), bone morphogenetic protein-2 (BMP2), collagen type II alpha 1 chain (Col2a1), Sox9, runt-related transcription factor 2 (Runx2), and osterix genes expressed in bone. For morphological analysis, muscles were removed and femur was observed with naked eye. Results COX2 gene expression in bone marrow significantly decreased. BMP2 gene expression significantly increased. Col2a1 gene expression significantly increased. Sox9 gene expression increased as well. Runx2 gene expression in bone marrow increased, but there was no statistical significance. Osterix gene expression significantly increased. Union of the fracture site progressed more in YM group compared to the control group. The fracture union score was significantly decreased in YM group compared to the control group. Conclusions YM showed anti-inflammatory effect, promoted bone regeneration by stimulating the bone regeneration factor. In conclusion, YM can help fracture healing and it well be applied clinically to patients with fracture.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제39권4호
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• pp.150-155
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• 2013

Objectives: The objective of this study is to determine the adequate concentration and to evaluate the osteogenic potential of simvastatin in human maxillary sinus membrane-derived stem cells (hSMSC). Materials and Methods: Mesenchymal stem cells derived from the human maxillary sinus membrane were treated with various concentrations of simvastatin. The adequate concentration of simvastatin for osteogenic induction was determined using bone morphogenetic protein (BMP-2). The efficacy of osteogenic differentiation of simavastatin was verified using osteocalcin mRNA, and the mineralization efficacy of hSMSCs and simvastatin treatment was compared with alkaline phosphatase and von Kossa staining. Results: Expression of BMP-2 mRNA and protein was observed after three days and was dependent on the concentration of simvastatin. Expression of osteocalcin mRNA was observed after three days in the $1.0{\mu}M$ simvastatin-treated group. Mineralization was observed after three days in the simvastatin-treated group. Conclusion: These results suggest that simvastatin induces the osteogenic potential of mesenchymal stem cells derived from the human maxillary sinus membrane mucosa.

• Maxillofacial Plastic and Reconstructive Surgery
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• 제35권2호
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• pp.73-81
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• 2013

Purpose: Smad4 is a central mediator for transforming growth factor-${\beta}$/bone morphogenetic protein ($TGF-{\beta}/BMP$) signals, which are involved in regulating cranial neural crest cell formation, migration, proliferation, and fate determination. Accumulated evidences indicate that $TGF-{\beta}/BMP$ signaling plays key roles in the early tooth morphogenesis. However, their roles in the late tooth formation, such as cellular differentiation and matrix formation are not clearly understood. The objective of this study is to understand the roles of Smad4 in vivo during enamel and dentin formation through tissue-specific inactivation of Smad4. Methods: We generated and analyzed mice with dental epithelium-specific inactivation of the Smad4 gene (K14-Cre:$Smad4^{fl/fl}$) and dental mesenchyme-specific inactivation of Smad4 gene (Osr2Ires-Cre:$Smad4^{fl/fl}$). Results: In the tooth germs of K14-Cre:$Smad4^{fl/fl}$, ameloblast differentiation was not detectable in inner enamel epithelial cells, however, dentin-like structure was formed in dental mesenchymal cells. In the tooth germs of Osr2Ires-Cre:$Smad4^{fl/fl}$ mice, ameloblasts were normally differentiated from inner enamel epithelial cells. Interestingly, we found that bone-like structures, with cellular inclusion, were formed in the dentin region of Osr2Ires-Cre:$Smad4^{fl/fl}$ mice. Conclusion: Taken together, our study demonstrates that Smad4 plays a crucial role in regulating ameloblast and odontoblast differentiation, as well as in regulating epithelial-mesenchymal interactions during tooth development.

• 한국콘텐츠학회논문지
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• 제10권3호
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• pp.196-203
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• 2010

본 연구는 BMP-4의 발현을 통해 골절 후 골유합에 대한 미세전류의 효과를 관찰하였다. 실험동물은 체중 2.5~3 kg내외의 6개월 령 뉴질랜드 웅성 토끼 24마리를 사용하였으며 경골 골절 후 미세전류를 적용한 실험군과 비적용군인 대조군으로 나누었고, 시간경과에 따른 변화를 관찰하기 위하여 3일, 7일, 14일 및 28일군으로 나누어 BMP-4에 대한 면역조직화학적 염색을 실행하여 다음과 같은 결론을 얻었다. BMP-4의 발현은 미세전류를 적용한 실험군과 자연치유군인 대조군 모두 시간이 경과함에 따라 증가하다가 감소되었다. 그러나 골절 7일 후 까지 동일 시점에서 실험군이 대조군에 비해 더욱 강한 면역양성 반응을 보였다. 특히 경골 골절 7일 후 대조군은 하버씨계의 동심원과 간질층판을 중심으로 중등도의 갈색의 면역양성반응을 보인데 반해 실험군의 경우 바깥층판을 포함하여 매우 강한 갈색의 면역양성반응을 보였다. 위의 결과로 보면 골절 후 미세전류를 적용할 때 치유과정 초기에 골형성단백질인 BMP-4의 발현을 증가시켜 골절 치유를 촉진시킴을 알 수 있었다.

• Molecules and Cells
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• 제37권3호
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• pp.220-225
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• 2014

Suppression of bone morphogenetic protein (BMP) signaling induces neural induction in the ectoderm of developing embryos. BMP signaling inhibits neural induction via the expression of various neural suppressors. Previous research has demonstrated that the ectopic expression of dominant negative BMP receptors (DNBR) reduces the expression of target genes down-stream of BMP and leads to neural induction. Additionally, gain-of-function experiments have shown that BMP downstream target genes such as MSX1, GATA1b and Vent are involved in the suppression of neural induction. For example, the Vent1/2 genes are involved in the suppression of Geminin and Sox3 expression in the neural ectodermal region of embryos. In this paper, we investigated whether PV.1, a BMP downstream target gene, negatively regulates the expression of FoxD5b, which plays a role in maintaining a neural progenitor population. A promoter assay and a cyclohexamide experiment demonstrated that PV.1 negatively regulates FoxD5b expression.

• Journal of Periodontal and Implant Science
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• 제41권4호
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• pp.167-175
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• 2011

Purpose: Periodontal ligament (PDL) cell differentiation into osteoblasts is important in bone formation. Bone formation is a complex biological process and involves several tightly regulated gene expression patterns of bone-related proteins. The expression patterns of bone related proteins are regulated in a temporal manner both in vivo and in vitro. The aim of this study was to observe the gene expression profile in PDL cell proliferation, differentiation, and mineralization in vitro. Methods: PDL cells were grown until confluence, which were then designated as day 0, and nodule formation was induced by the addition of 50 ${\mu}g$/mL ascorbic acid, 10 mM ${\beta}$-glycerophosphate, and 100 nM dexamethasone to the medium. The dishes were stained with Alizarin Red S on days 1, 7, 14, and 21. Real-time polymerase chain reaction was performed for the detection of various genes on days 0, 1, 7, 14, and 21. Results: On day 0 with a confluent monolayer, in the active proliferative stage, c-myc gene expression was observed at its maximal level. On day 7 with a multilayer, alkaline phosphatase, bone morphogenetic protein (BMP)-2, and BMP-4 gene expression had increased and this was followed by maximal expression of osteocalcin on day 14 with the initiation of nodule mineralization. In relationship to apoptosis, c-fos gene expression peaked on day 21 and was characterized by the post-mineralization stage. Here, various genes were regulated in a temporal manner during PDL fibroblast proliferation, extracellular matrix maturation, and mineralization. The gene expression pattern was similar. Conclusions: We can speculate that the gene expression pattern occurs during PDL cell proliferation, differentiation, and mineralization. On the basis of these results, it might be possible to understand the various factors that influence PDL cell proliferation, extracellular matrix maturation, and mineralization with regard to gene expression patterns.