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- 주제어 : Osteogenic differentiation (검색결과 176 건)
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BMP-2와 GDF-5의 Rat 골수 유래 중간엽 줄기세포의 골 분화 효과 비교
권영삼,유정준,전흥재,김희중 한국조직공학과 재생의학회 2009 조직공학과 재생의학 Vol.6 No.13
Recently, it has been reported that growth and differentiation factor-5 (GDF-5) regulates and promotes osteoblastic and osteogenic differentiation. We here compared effects of GDF-5 and BMP-2 on the proliferation and osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBM-MSCs). Cell proliferation was significantly increased in BMP-2 and GDF-5 treated group compared to those in control group. The ALP activities of osteogenic differentiation group, BMP-2 treated group, and GDF-5 treated group was higher than in control group. The ALP activity of GDF-5 group was similar with that of BMP-2 group. When the Ca activity was compared in all groups, the Ca level of GDF-5 group was highest and was similar with that of BMP-2 group. As a result of Von Kossa staining and RT-PCR, the calcium deposition and osteogenic gene expression were increased in the GDF-5 group than in the osteogenic differentiation group. In the present study, GDF-5 appears to be an effective stimulator of proliferation and osteogenic differentiation of rBM-MSCs. To this regard, GDF-5 may be a useful regulator for promoting the bone regeneration and formation by inducing rBM-MSCs proliferation and osteogenic differentiation. Recently, it has been reported that growth and differentiation factor-5 (GDF-5) regulates and promotes osteoblastic and osteogenic differentiation. We here compared effects of GDF-5 and BMP-2 on the proliferation and osteogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBM-MSCs). Cell proliferation was significantly increased in BMP-2 and GDF-5 treated group compared to those in control group. The ALP activities of osteogenic differentiation group, BMP-2 treated group, and GDF-5 treated group was higher than in control group. The ALP activity of GDF-5 group was similar with that of BMP-2 group. When the Ca activity was compared in all groups, the Ca level of GDF-5 group was highest and was similar with that of BMP-2 group. As a result of Von Kossa staining and RT-PCR, the calcium deposition and osteogenic gene expression were increased in the GDF-5 group than in the osteogenic differentiation group. In the present study, GDF-5 appears to be an effective stimulator of proliferation and osteogenic differentiation of rBM-MSCs. To this regard, GDF-5 may be a useful regulator for promoting the bone regeneration and formation by inducing rBM-MSCs proliferation and osteogenic differentiation.
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Kim, Ji Hye,Yoon, Sei Mee,Song, Sun U.,Park, Sang Gyu,Kim, Won-Serk,Park, In Guk,Lee, Jinu,Sung, Jong-Hyuk MDPI 2016 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.17 No.9
<P>Hypoxia has diverse stimulatory effects on human adipose-derived stem cells (ASCs). In the present study, we investigated whether hypoxic culture conditions (2% O<SUB>2</SUB>) suppress spontaneous mineralization and osteogenic differentiation of ASCs. We also investigated signaling pathways and molecular mechanisms involved in this process. We found that hypoxia suppressed spontaneous mineralization and osteogenic differentiation of ASCs, and up-regulated mRNA and protein expression of Insulin-like growth factor binding proteins (IGFBPs) in ASCs. Although treatment with recombinant IGFBPs did not affect osteogenic differentiation of ASCs, siRNA-mediated inhibition of IGFBP3 attenuated hypoxia-suppressed osteogenic differentiation of ASCs. In contrast, overexpression of IGFBP3 via lentiviral vectors inhibited ASC osteogenic differentiation. These results indicate that hypoxia suppresses spontaneous mineralization and osteogenic differentiation of ASCs via intracellular IGFBP3 up-regulation. We determined that reactive oxygen species (ROS) generation followed by activation of the MAPK and PI3K/Akt pathways play pivotal roles in IGFBP3 expression under hypoxia. For example, ROS scavengers and inhibitors for MAPK and PI3K/Akt pathways attenuated the hypoxia-induced IGFBP3 expression. Inhibition of Elk1 and NF-κB through siRNA transfection also led to down-regulation of IGFBP3 mRNA expression. We next addressed the proliferative potential of ASCs with overexpressed IGFBP3, but IGFBP3 overexpression reduced the proliferation of ASCs. In addition, hypoxia reduced the osteogenic differentiation of bone marrow-derived clonal mesenchymal stem cells. Collectively, our results indicate that hypoxia suppresses the osteogenic differentiation of mesenchymal stem cells via IGFBP3 up-regulation.</P>
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( Jung Bok Lee ),( Sung Min Jeong ),( Kyoung Jun Kim ),( Dong Hyun Cho ),( Il Keun Kwon ),( In Chan Yoon ),( Kui Won Choi ),( Jun Kyo Francis Suh ),( Jae Hong Park ),( Yong Duk Park ),( Jong Hyuk Chun 한국조직공학·재생의학회 2009 조직공학과 재생의학 Vol.6 No.1
Dexamethasone(DEX), a synthetic steroidal anti-inflammatory drug, is an efficient and dependable drug that induces osteogenic differentiation. The aim of this study is to fabricate the DEX loaded PLGA nanofibers by electrospinning method. Adipose-derived stem cells(ADSCs) were seeded into the nanofiber and the sustained release of DEX from PLGA nanofiber scaffolds promoted their osteogenic differentiation. The properties of DEX loaded PLGA nanofiber scaffold were characterized by scanning electron microscopy(SEM) and the release kinetics of DEX from PLGA nanofibers in vitro(1 h to 14 days) was evaluated by high performed liquid chromatography(HPLC). To evaluate the cellular response of the ADSCs seeded onto DEX loaded PLGA nanofiber, we performed F-actin, cytotoxicity, alkaline phosphatase activity, alizarin red S, von Kossa staining and immunocytochemistry assays for osteogenic differentiation. DEX loaded PLGA nanofiber scaffold was observed to have sustained release in vitro during experimental periods. The cytotoxicity test of the DEX loaded PLGA nanofiber scaffold indicated there was almost no-toxic effects in regards to proliferation and differentiation of adiposederived stem cells(ADSCs) as compared with a control. Alkaline phosphatase activity and alizarin red S were more significantly increased after 14days with increased DEX concentration of PLGA nanofiber scaffold than with PLGA nanofiber only. Also, von Kossa staining results confirmed a larger area of calcium deposition with increased DEX concentration of PLGA nanofiber scaffold. We observed that osteocalcin was also increased with increasing DEX concentration. These results demonstrate that DEX acts as an osteogenic inductive factor, and increased DEX concentration promoted more osteogenic differentiation of ADSCs. In conclusion, DEX loaded PLGA nanofiber would be valuable tool for bone tissue regeneration.
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Jin Jeon,이민석,Hee Seok Yang 한국생체재료학회 2018 생체재료학회지 Vol.22 No.1
Background: The extracellular matrix (ECM) can directly or indirectly influence on regulation of cell functions such as cell adhesion, migration, proliferation and differentiation. The cell derived ECM (CD-ECM) is a useful in vitro model for studying the comprehensive functions of CD-ECM because it maintains a native-like structure and composition. In this study, the CD-ECM is obtained and a test is carried out to determine the effectiveness of several combinations of decellularized methods. These methods were used to regulate the optimal ECM compositions to be induced by osteogenic differentiation using primary isolated osteoblasts. Result: We investigated the effect of osteoblasts re-seeded onto normal osteoblast ECM under the growth medium (GM-ECM) and the osteogenic differentiation medium (OD-ECM). The osteoblasts were then cultured statically for 1, 2, and 4 weeks in a growth medium or differentiation medium. Before osteoblast culture, we performed immunostaining with filamentous actin and nuclei, and then performed DNA quantification. After each culture period, the osteogenic differentiation of the osteoblasts re-seeded on the OD-ECMs was enhanced osteogenic differentiation which confirmed by alkaline phosphatase staining and quantification, Alizarin Red S staining and quantification, and von Kossa staining. The OD-ECM-4 W group showed more effective osteogenic differentiation than GM-ECM and OD-ECM-2 W. Conclusions: The OD-ECM-4 W has a better capacity in a microenvironment that supports osteogenic differentiation on the GM-ECM and OD-ECM-2 W. The ECM substrate has a wide range of applications as cell culture system or direct differentiation of stem cell and excellent potential as cell-based tissue repair in orthopedic tissue engineering.
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안국영,Sung Eun Kim,윤태훈,최인성,Daewon Park,최성욱 한국생체재료학회 2021 생체재료학회지 Vol.25 No.4
Background: Alendronate (Alen) is promising material used for bone-targeted drug delivery due to its high bone affinity and therapeutic effects on bone diseases. In addition, Alen can enhance the osteogenic differentiation of osteoblastic cell. Recently, nanodiamonds (NDs) with hardness, non-toxicity, and excellent biocompatibility are employed as promising materials for carrier systems and osteogenic differentiation. Therefore, we prepared Alenconjugated NDs (Alen-NDs) and evaluated their osteogenic differentiation performances. Methods: Alen-NDs were synthesized using DMTMM as a coupling reagent. Morphological change of Mouse calvaria-derived preosteoblast (MC3T3-E1) treated with Alen-NDs was observed using the confocal microscope. The osteogenic differentiation was confirmed by cell proliferation, alkaline phosphatase (ALP), calcium deposition, and real-time polymerase chain reaction assay. Results: Alen-NDs were prepared to evaluate their effect on the proliferation and differentiation of osteoblastic MC3T3-E1 cells. The Alen-NDs had a size of about 100 nm, and no cytotoxicity at less than 100 μg/mL of concentration. The treatment of NDs and Alen-NDs reduced the proliferation rate of MC3T3-E1 cells without cell death. Confocal microscopy images confirmed that the treatment of NDs and Alen-NDs changed the cellular morphology from a fibroblastic shape to a cuboidal shape. Flow cytometry, alkaline phosphatase (ALP) activity, calcium deposition, and real-time polymerase chain reaction (RT-PCR) confirmed the higher differentiation of MC3T3-E1 cells treated by Alen-NDs, compared to the groups treated by osteogenic medium and NDs. The higher concentration of Alen-ND treated in MC3T3-E1 resulted in a higher differentiation level. Conclusions: Alen-NDs can be used as potential therapeutic agents for osteoporosis treatment by inducing osteogenic differentiation.
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Cho, Hyun Hwa,Park, Hyung Taek,Kim, Yeon Jeong,Bae, Yong Chan,Suh, Kuen Taek,Jung, Jin Sup Wiley Subscription Services, Inc., A Wiley Company 2005 Journal of cellular biochemistry Vol.96 No.3
<P>Valproic acid (VPA) has been used as an anticonvulsant agent for the treatment of epilepsy, as well as a mood stabilizer for the treatment of bipolar disorder, for several decades. The mechanism of action for these effects remains to be elucidated and is most likely multifactorial. Recently, VPA has been reported to inhibit histone deacetylase (HDAC) and HDAC has been reported to play roles in differentiation of mammalian cells. In this study, the effects of HDAC inhibitors on differentiation and proliferation of human adipose tissue-derived stromal cells (hADSC) and bone marrow stromal cells (hBMSC) were determined. VPA increased osteogenic differentiation in a dose dependent manner. The pretreatment of VPA before induction of differentiation also showed stimulatory effects on osteogenic differentiation of hMSC. Trichostatin A (TSA), another HDAC inhibitor, also increased osteogenic differentiation, whereas valpromide (VPM), a structural analog of VPA which does not possess HDAC inhibitory effects, did not show any effect on osteogenic differentiation on hADSC. RT-PCR and Real-time PCR analysis revealed that VPA treatment increased osterix, osteopontin, BMP-2, and Runx2 expression. The addition of noggin inhibited VPA-induced potentiation of osteogenic differentiation. VPA inhibited proliferation of hADSC and hBMSC. Our results suggest that VPA enhance osteogenic differentiation, probably due to inhibition of HDAC, and could be useful for in vivo bone engineering using hMSC. © 2005 Wiley-Liss, Inc.</P>
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강경신,홍정민,강조아,이종원,정영훈,조동우 생화학분자생물학회 2013 Experimental and molecular medicine Vol.45 No.1
Many studies have reported that an electromagnetic field can promote osteogenic differentiation of mesenchymal stem cells. However, experimental results have differed depending on the experimental and environmental conditions. Optimization of electromagnetic field conditions in a single, identified system can compensate for these differences. Here we demonstrated that specific electromagnetic field conditions (that is, frequency and magnetic flux density) significantly regulate osteogenic differentiation of adipose-derived stem cells (ASCs) in vitro. Before inducing osteogenic differentiation, we determined ASC stemness and confirmed that the electromagnetic field was uniform at the solenoid coil center. Then, we selected positive (30/45 Hz, 1 mT) and negative (7.5 Hz, 1 mT) osteogenic differentiation conditions by quantifying alkaline phosphate (ALP)mRNA expression. Osteogenic marker (for example, runt-related transcription factor 2) expression was higher in the 30/45 Hz condition and lower in the 7.5 Hz condition as compared with the nonstimulated group. Both positive and negative regulation of ALP activity and mineralized nodule formation supported these responses. Our data indicate that the effects of the electromagnetic fields on osteogenic differentiation differ depending on the electromagnetic field conditions. This study provides a framework for future work on controlling stem cell differentiation.
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The effect of extracellular matrix for osteogenic differentiation
전진,양희석 한국공업화학회 2018 한국공업화학회 연구논문 초록집 Vol.2018 No.0
The extracellular matrix (ECM) can influence on regulation of cell functions such as cell adhesion, migration, proliferation, and differentiation. The cell derived ECM (CD-ECM) is a useful in vitro model for studying the comprehensive functions of CD-ECM because it maintains a native-like structure and composition. In this study, the CD-ECM is obtained and a test is carried out to determine the effectiveness of several combinations of decellularized methods. We investigated the effect of osteoblasts re-seeded onto normal osteoblast ECM under the growth medium ECM (GM-ECM) and the osteogenic differentiation medium (OD-ECM). The osteoblasts were then cultured for 1, 2, and 4 weeks in a growth medium or differentiation medium. After each culture period, the osteogenic differentiation of the osteoblasts re-seeded on the OD-ECMs was enhanced osteogenic differentiation. The OD-ECM-4W group showed more effective osteogenic differentiation than GM-ECM and OD-ECM-2W.
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Ting Chen,Yan Zhou,Wen-Song Tan 한국생물공학회 2009 Biotechnology and Bioprocess Engineering Vol.14 No.6
A functional relationship between the growth and the progression of events associated with osteogenic differentiation of human amniotic mesenchymal stem cells (hAMSCs) has been a fundamental question, which remains unclear. This study is aimed at investigating the effects of low temperature and lactate individually, and in combination on the growth and osteogenic differentiation of hAMSCs. It was shown that the growth of hAMSCs in growth medium was inhibited by both low-cultivation temperature and lactate. By extending culture period at low temperature, cell growth declined gradually, while the ALP expression and calcium deposition increased progressively. However, the growth of hAMSCs induced in osteogenic medium at 37oC was markedly enhanced by additional lactate. The ALP expression and calcium deposition, on the contrary, were significantly depressed. Furthermore, the synergistic actions of long-term low temperature and lactate resulted in more intense inhibition on both cell growth and osteogenic differentiation. Therefore, these findings may imply the co-contribution of the culture environment on the selective manipulation on the growth capacity and osteogenic differentiation potential of hAMSCs A functional relationship between the growth and the progression of events associated with osteogenic differentiation of human amniotic mesenchymal stem cells (hAMSCs) has been a fundamental question, which remains unclear. This study is aimed at investigating the effects of low temperature and lactate individually, and in combination on the growth and osteogenic differentiation of hAMSCs. It was shown that the growth of hAMSCs in growth medium was inhibited by both low-cultivation temperature and lactate. By extending culture period at low temperature, cell growth declined gradually, while the ALP expression and calcium deposition increased progressively. However, the growth of hAMSCs induced in osteogenic medium at 37oC was markedly enhanced by additional lactate. The ALP expression and calcium deposition, on the contrary, were significantly depressed. Furthermore, the synergistic actions of long-term low temperature and lactate resulted in more intense inhibition on both cell growth and osteogenic differentiation. Therefore, these findings may imply the co-contribution of the culture environment on the selective manipulation on the growth capacity and osteogenic differentiation potential of hAMSCs
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김기주,Moon Seop Choi,Jong Won Rhie,Jin Hyung Shim 한국조직공학과 재생의학회 2019 조직공학과 재생의학 Vol.16 No.4
BACKGROUND: Silica particles (SPs) induce cell proliferation and osteogenic differentiation. We reported that SPs in the scaffold induced early stage osteogenic differentiation. METHODS: A polycaprolactone (PCL) scaffold was fabricated with a 10 wt% SPs. The surface of PCL scaffold was coated with a 10 lg/mL collagen solution. Next, the scaffold was conjugated with 2 lM SPs, 2 lg/mL bone morphogenetic protein 2 (BMP2), or 2 lM BMP2-conjugated SPs (BCSPs). Green fluorescent protein-coupled BMP2 was applied to fabricate the scaffold. The fluorescence intensity was analyzed by confocal microscopy. The mRNA levels of the early osteogenic differentiation marker, alkaline phosphatase (ALP), were analyzed by real-time quantitative polymerase chain reaction. Levels of BMP2, RUNX2, ERK1/2, and AKT were assessed by western blotting. RESULTS: ALP mRNA levels were significantly higher in the BCSP-conjugated scaffold than in the other scaffolds. In the early stage of osteogenic differentiation, the protein levels of BMP2, RUNX2, ERK1/2, and AKT in cells were significantly higher in the BCSP-conjugated scaffold than in other scaffolds. Thus, the BCSP composite scaffold induced rapid osteogenic differentiation. CONCLUSION: These results suggest that BCSP composite can be used to promote early stage osteogenic differentiation and show promise as a material for use in scaffolds for bone regeneration.
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