• 대한치과의사협회지
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• 제46권8호
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• pp.494-504
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• 2008

Osseointegration is a result of bone formation and bone regeneration process, which take place at the interface between bone and implant and biologic determinants such as cytokine, growth factors, bone matrix proteins play an important role in osseointegration. The purpose of this study is to compare the expressoin of TGF-$\beta$, IGF-I, BMP2, BMP4 during osseointegration. We designed an experimental group which was inserted with a RBM surface titanium implants and machined surface, and compared with a control group which had a simple bone cavity and normal bone. Titanium implants were placed into tibia of 8 rabbits. We compared the expression of TGF-$\beta$, IGF-I, BMP2, BMP4 using RT_PCR (reverse transcriptase chain reaction)analysis in day 3,7,14 and 28 of implant insertion. According to the results, growth factors of experimental groups were more expressed than control groups. Among experimental groups, expression of TGF-$\beta$, IGF-I, BMP4 of BMP group had tedency to increase more at 14th, 28th days than Machined surface group. Therefore, our results suggest that TGF-$\beta$, IGF-I, BMP4 are expressed within the bone around the implant and more increased around rough surface implants while osseointegration occurs after dental implant insertion.

• BMB Reports
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• 제45권9호
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• pp.509-514
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• 2012

We have found that the previously uncharacterized bone morphogenetic protein-9 (BMP9) is one of the most osteogenic factors. However, it is unclear if BMP9 cross-talks with $TGF{\beta}1$ during osteogenic differentiation. Using the recombinant BMP9 adenovirus, we find that low concentration of rh$TGF{\beta}1$ synergistically induces alkaline phosphatase activity in BMP9-transduced C3H10T1/2 cells and produces more pronounced matrix mineralization. However, higher concentrations of $TGF{\beta}1$ inhibit BMP9-induced osteogenic activity. Real-time PCR and Western blotting indicate that BMP9 in combination with low dose of $TGF{\beta}1$ potentiates the expression of later osteogenic markers osteopontin, osteocalcin and collagen type 1 (COL1a2), while higher concentrations of $TGF{\beta}1$ decrease the expression of osteopontin and osteocalcin but not COL1a2. Cell cycle analysis reveals that $TGF{\beta}1$ inhibits C3H10T1/2 proliferation in BMP9-induced osteogenesis and restricts the cells in $G_0/G_1$ phase. Our findings strongly suggest that $TGF{\beta}1$ may exert a biphasic effect on BMP9-induced osteogenic differentiation of mesenchymal stem cells.

• 한국가금학회지
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• 제44권3호
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• pp.211-223
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• 2017

Transforming growth factor beta ($TGF-{\beta}$) signaling pathways are involved in the regulation of proliferation, differentiation, immunity, survival, and apoptosis of many cells. The aim of this study was to investigate the differential expression of $TGF-{\beta}$-related genes, and their interactions and regulators in the spleen of two genetically disparate chicken lines (Marek's disease resistant line 6.3 and Marek's disease-susceptible line 7.2) induced with necrotic enteritis (NE) by Eimeria maxima and Clostridium perfringens infection. By using high-throughput RNA-sequencing, we investigated 76 $TGF-{\beta}$-related genes that were significantly and differentially expressed in the spleens of the chickens. Approximately 20 $TGF-{\beta}$ pathway genes were further verified by qRT-PCR, and the results were consistent with our RNA sequencing data. All 76 identified genes were analyzed through Gene Ontology and mapped onto the KEGG chicken $TGF-{\beta}$ pathway. Our results demonstrated that several key genes, including $TGF-{\beta}$1-3, bone morphogenetic proteins (BMP)1-7, inhibitor of differentiation (ID) proteins ID1-3, SMAD1-9, and Jun, showed a markedly differential expression between the two chicken lines, relative to their respective controls. We then further predicted 24 known miRNAs that targeted BMP7 mRNA from 139 known miRNAs in the two chicken lines. Among these, six miRNAs were measured by qRT-PCR. In conclusion, this study is the first to analyze most of the genes, interactions, and regulators of the $TGF-{\beta}$ pathway in the innate immune responses of NE afflicted chickens.

• Journal of Chest Surgery
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• 제36권2호
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• pp.86-90
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• 2003

폐 형성에 활성적인 또는 억제적인 기능을 갖고 있다고 알려져 있는 FGF-7 성장인자, BMP수용체 II, 그리고 TGF-$\beta$ 수용체 II 유전자의 비정상 발현이 폐기포 생성에 관여하는지를 각각의 단일성 클론 항체를 사용하여 수술로 절제된 자연기흉 환자의 폐기포 조직들을 면역조직염색 방법으로 염색하여 관찰하였다. 대상 및 방법: 재발성 또는 지속성 기흉으로 흉강경 또는 개흉술로 폐기포 절제술을 실시한 환자들을 대상으로 하였다. 총 31명의 환자로 15세에서 39세까지 연령분포를 보였으며 남자 30명, 여자 1명이었다. 폐기포 절제는 비디오흉강경이나 소절개개흠술을 통하여 폐기포벽에 손상을 가하지 않게 주의하면서 비디오흉강경용 스태플러(Endo GIA stapler)를 이용하여 절제하였으며 가능한 원형을 유지하여 신선한 상태로 포르마린에 고정하여 면역조직화학적 연구를 위한 표본을 만들었다. 폐기포 조직 슬라이드를 단일클론성 항 TGF-$\beta$ 수용체 II, BMP수용체 II 그리고 FGF-7인자 항체를 이용하여 면역조직학적 염색방법으로 관찰하였다. 결과: 전체 환자 31명중 TGF-$\beta$ 수용체 II항체에 양성 반응을 나타낸 환자수는 모두 24명이었다. 이들 중에는 18명이 강한 양성 반응을 보였고, 6명이 약한 양성 반응을 보였다 면역조직화학적 염색 결과를 고배율 현미경으로 살펴보면, TGF-$\beta$ 수용체 II의 염색이 기흉과 정상 폐조직 경계 부위에서 측히 강하게 염색됨이 관찰되었다. 이에 반하여, BMP수용체 II 그리고 FGF-7인자의 항체를 이용한 면역조직학적 염색 결과는 모든 환자의 조직들에서 음성으로 관찰되었다. 결론: 폐 조직이 형성될 때, 억제유전자의 역할을 담당하고 있다고 알려진 TGF-$\beta$ 수용체 II의 발현이 증가되면서 폐기포가 생성될 수 있다는 가능성을 제시하였다. 이번 결론은 면역조직학적 염색 실험 결과만으로 밝혀진 사실임으로 좀 더 체계적인 분자생물학적 인 연구가 요구된다.

• Asian-Australasian Journal of Animal Sciences
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• 제32권4호
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• pp.477-484
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• 2019

Objective: We aimed to observe hair follicle (HF) development in the dorsal skin and elucidate the expression patterns of genes and proteins related to skin and HF development in Rex rabbits from birth to 8 weeks of age. Methods: Whole-skin samples were obtained from the backs of Rex rabbits at 0, 2, 4, 6, and 8 weeks of age, the morphological development of primary and secondary HFs was observed, and the gene transcript levels of insulin-like growth factor-I (IGF-I), epidermal growth factor (EGF), bone morphogenetic protein 2 (BMP2), transforming growth factor ${\beta}-1$, 2, and 3 ($TGF{\beta}-1$, $TGF{\beta}-2$, and $TGF{\beta}-3$) were examined using quantitative real-time polymerase chain reaction (PCR). Additionally, Wnt family member 10b (Wnt10b) and ${\beta}$-Catenin gene and protein expression were examined by quantitative real-time PCR and western blot, respectively. Results: The results showed significant changes in the differentiation of primary and secondary HFs in Rex rabbits during their first 8 weeks of life. The IGF-I, EGF, $TGF{\beta}-2$, and $TGF{\beta}-3$ transcript levels in the rabbits were significantly lower at 2 weeks of age than at birth and gradually increased thereafter, while the BMP2 and $TGF{\beta}-1$ transcript levels at 2 weeks of age were significantly higher than those at birth and gradually decreased thereafter. ${\beta}$-Catenin gene expression was also significantly affected by age, while the Wnt10b transcript level was not. However, the Wnt10b and ${\beta}$-catenin protein expression levels were the lowest at 2 and 4 weeks of age. Conclusion: Our data showed that a series of changes in HFs in dorsal skin occurred during the first 8 weeks. Many genes, such as IGF-I, EGF, BMP2, $TGF{\beta}-1$, $TGF{\beta}-2$, $TGF{\beta}-3$, and ${\beta}$-Catenin, participated in this process, and the related proteins Wnt10b and ${\beta}$-Catenin in skin were also affected by age.

• BMB Reports
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• 제38권1호
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• pp.9-16
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• 2005

Transforming Growth Factor (TGF)-$\beta$ family, including TGF-$\beta$, bone morphorgenic protein (BMP), and activn, plays an important role in essential cellular functions such as proliferation, differentiation, apoptosis, tissue remodeling, angiognesis, immune responses, and cell adhesions. TGF-$\beta$ predominantly transmits the signals through serine/threonine receptor kinases and cytoplasmic proteins called Smads. Since the discovery of TGF-$\beta$ in the early 1980s, the dysregulation of TGF-$\beta$/Smad signaling has been implicated in the pathogenesis of human diseases. Among signal transducers in TGF-$\beta$/Smad signaling, inhibitory Smads (I-Smads), Smad6 and Smad7, act as major negative regulators forming autoinhibitory feedback loops and mediate the cross-talking with other signaling pathways. Expressions of I-Smads are mainly regulated on the transcriptional levels and post-translational protein degradations and their intracellular levels are tightly controlled to maintain the homeostatic balances. However, abnormal levels of I-Smads in the pathological conditions elicit the altered TGF-$\beta$ signaling in cells, eventually causing TGF-$\beta$-related human diseases. Thus, exploring the molecular mechanisms about the regulations of I-Smads may provide the therapeutic clues for human diseases induced by the abnormal TGF-$\beta$ signaling.

• Clinical and Experimental Reproductive Medicine
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• 제34권4호
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• pp.239-252
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• 2007

목 적: 사람의 HAD와 HUC를 심근세포로 분화 유도하고자 하였다. 연구방법: 사람의 HAD와 HUC를 분리하여 5-azacytidine을 24시간 처리하고 여러 가지 BMP와 FGF을 첨가하여 배양하였다. 또한 HUC은 BMP와 FGF와 함께 activin A 또는 TGF-$\beta$1 또는 Wnt inhibitor를 첨가하여 배양한 후 심근세포 특이 유전자의 발현을 조사하였다. 결 과: HAD를 5-azacytidine 처리하고 기본배양액에서 4주 동안 배양하였을 때 TnT 유전자가 새로이 발현하였으며 Cmlc1과 kv4.3의 발현 양이 증가하였다. 5-azacytidine 처리 후에 BMP-4와 함께 FGF-4 (B4/F4) 또는 FGF-8 (B4/F8)을 첨가하여 배양하였을 때는 $\beta$-MHC 유전자 발현이 새로이 유도되었으며, Cmlc1, TnT, TnI 그리고 Kv4.3 유전자 발현 양이 더 많이 증가하였다. HUC은 5-azacytidine 및 BMP와 FGF 처리에 의해 유전자 발현 변화가 없었다. 그러나 BMP와 FGF와 함께 activin A 또는 TGF-$\beta$1을 첨가하여 배양하였을 때, BMP-2와 FGF-8 (B2/F8)을 첨가하여 배양한 세포에서 $\beta$-MHC 발현이 새로이 유도되었으며 $\alpha$-CA, TnT 그리고 Kv4.3 유전자의 발현이 증가하였다. 또한 BMP와 FGF와 함께 Wnt inhibitor를 처리하여 1주 동안 배양하였을 때 Cinlc1 유전자 발현이 새로이 유도되었으며 $\alpah$-CA, TnT, TnI 그리고 Kv4.3의 발현이 증가되었다. 결 론: HAD는 BMP와 FGF 처리에 의해 심근세포 특이 유전자의 발현증가를 유도할 수 있었으며 HUC는 BMP와 FGF와 함께 activin A 또는 TGF-$\beta$1 또는 Wnt inhibitor를 처리함으로써 심근세포 특이 유전자의 발현증가를 유도할 수 있었다. 따라서 HAD와 HUC는 심장질환 치료를 목적으로 하는 세포 치료에 이용될 수 있을 것으로 사료된다.

• 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.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
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• 제33권4호
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• pp.288-296
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• 2007

Distraction osteogenesis(DO) is a technique of lengthning bone including soft tissue by gradual separation of surgically divided bone surfaces. Distraction osteogenesis combination with a compression stimulation(DO-CO) was a new technique by authors to enhance new bone quality and to shorten the consolidation period. The purpose of this study was to compare DO with DO combined with compression force in efficiency by evaluating the expression of TGF-${\beta}1$, osteonectin and BMP-4 on bone regenerate in rabbit mandible. Fourty two rabbits were used for this experiment. On the control group, the distraction was carried out at the rate of 1 mm per day to obtain the amount of 8 mm distraction for 8 days. On the experimental group, the distraction was carried out at the rate of 1 mm per day for 10 days, 3 days-latency period, and then the compression was carried out as counter direction 1 mm per day for 2 days. After 0 day, 5 days, 13 days, 20 days, 27 days, 34 days and 41 days, three rabbits on each group were sacrificed and the distracted portion of mandible were cut and treated for RT-PCR observation. The level of expression of TGF-${\beta}1$ and osteonectin were shown more and longer expression in the experimental group than in the control group. The expression of BMP-4 was maintained with high level during the entire experimental period in both groups. These findings suggested that DO with compression stimulation could be a favorable technique for obtaining a good new bone quality.

• BMB Reports
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• 제46권2호
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• pp.107-112
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• 2013

Although BMP6 is highly capable of inducing osteogenic differentiation of mesenchymal progenitor cells (MPCs), the molecular mechanism involved remains to be fully elucidated. Using dominant negative (dn) mutant form of type I and type II $TGF{\beta}$ receptors, we demonstrated that three dn-type I receptors (dnALK2, dnALK3, dnALK6), and three dn-type II receptors (dnBMPRII, dnActRII, dnActRIIB), effectively diminished BMP6-induced osteogenic differentiation of MPCs. These findings suggested that ALK2, ALK3, ALK6, BMPRII, ActRII and ActRIIB are essential for BMP6-induced osteogenic differentiation of MPCs. However, MPCs in this study do not express ActRIIB. Moreover, RNA interference of ALK2, ALK3, ALK6, BMPRII and ActRII inhibited BMP6-induced osteogenic differentiation in MPCs. Our results strongly suggested that BMP6-induced osteogenic differentiation of MPCs is mediated by its functional $TGF{\beta}$ receptors including ALK2, ALK3, ALK6, BMPRII, and ActRII.