• 제목/요약/키워드: Bone morphogenetic protein 9

검색결과 38건 처리시간 0.019초

Rabbit Model for in vivo Study of Intervertebral Disc Degeneration and Regeneration

  • Kong, Min-Ho;Do, Duc-H.;Miyazaki, Masashi;Wei, Feng;Yoon, Sung-Hwan;Wang, Jeffrey C.
    • Journal of Korean Neurosurgical Society
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    • 제44권5호
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    • pp.327-333
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    • 2008
  • Objective: The purpose of this study is to verify the usefulness of the rabbit model for disc degeneration study. Materials: The L1-L2, L2-L3, L3-L4. or L4-L5 lumbar intervertebral disc (IVD) of 9 mature male New Zealand White rabbits were injured by inserting a 16-gauge needle to a depth of 5 mm in the left anterolateral annulus fibrosus while leaving L5-L6 IVD uninjured. Three other rabbits also received intradiscal injections of rabbit disc cells transfected with adenovirus and bone morphogenetic protein-2 (ad-BMP-2) at L4-L5 in addition to injury by 16-gauge needle at the L1-L2 level. Using digitized radiographs, measurements of IVD height were made and analyzed by using the disc height index (DHI). Magnetic resonance imaging (MRI) scans of the injured discs, injected discs, and uninjured L5-L6 discs were performed at 15 weeks post surgery and compared with preoperative MRI scans. Results: All twelve rabbits showed consistent results of disc degeneration within 15 weeks following annular puncture. DHIs of injured discs were significantly lower than that of the uninjured L5-L6 discs (p<0.05). The mean value of disc degeneration grade of injured discs was significantly higher than that of uninjured discs (p<0.05). The injection of disc cell transfected with ad-BMP-2 did not induce disc regeneration at 15 weeks after injection. Conclusion: This study showed that the injured disc had a significant change in DHI on simple lateral radiograph and disc degeneration grade on MRI scans within 15 weeks in all rabbits. Rabbit annular puncture model can be useful as a disc degeneration model in vivo.

Nox4-Mediated Cell Signaling Regulates Differentiation and Survival of Neural Crest Stem Cells

  • Lee, Ji-Eun;Cho, Kyu Eun;Lee, Kyung Eun;Kim, Jaesang;Bae, Yun Soo
    • Molecules and Cells
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    • 제37권12호
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    • pp.907-911
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    • 2014
  • The function of reactive oxygen species (ROS) as second messengers in cell differentiation has been demonstrated only for a limited number of cell types. Here, we used a well-established protocol for BMP2-induced neuronal differentiation of neural crest stem cells (NCSCs) to examine the function of BMP2-induced ROS during the process. We first show that BMP2 indeed induces ROS generation in NCSCs and that blocking ROS generation by pretreatment of cells with diphenyleneiodonium (DPI) as NADPH oxidase (Nox) inhibitor inhibits neuronal differentiation. Among the ROS-generating Nox isozymes, only Nox4 was expressed at a detectable level in NCSCs. Nox4 appears to be critical for survival of NCSCs at least in vitro as down-regulation by RNA interference led to apoptotic response from NCSCs. Interestingly, development of neural crest-derived peripheral neural structures in Nox4-/- mouse appears to be grossly normal, although Nox4-/- embryos were born at a sub-Mendelian ratio and showed delayed over-all development. Specifically, cranial and dorsal root ganglia, derived from NCSCs, were clearly present in Nox4-/- embryo at embryonic days (E) 9.5 and 10.5. These results suggest that Nox4-mediated ROS generation likely plays important role in fate determination and differentiation of NCSCs, but other Nox isozymes play redundant function during embryogenesis.

A Study on Effect of Carrying FecB Gene on Body Weight in Garole and Garole×Malpura Sheep

  • Kolte, A.P.;Mishra, A.K.;Kumar, S.;Arora, A.L.;Singh, V.K.
    • Asian-Australasian Journal of Animal Sciences
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    • 제18권10호
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    • pp.1379-1382
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    • 2005
  • High prolificacy in Garole sheep is due to existence of FecB mutation in an autosomal gene, bone morphogenetic protein receptor. The mutation enhances ovulation rate and in turn litter size in Garole sheep. Garole sires were crossed with non-prolific Malpura ewes with the aim to introduce prolificacy into Garole${\times}$Malpura (G${\times}$M) crosses through FecB introgression programme. In the present study, the effect of carrying booroola allele on litter size and live body weight was analyzed. The average litter size at birth was found to be 1.87 and 1.48 in the Garole and the G${\times}$M crosses, respectively. At weaning, 6-month, 9-month and 12-month of age, body weights were not affected by the presence of booroola allele (p>0.05); however, a significant effect (p<0.05) was found on body weight at birth in G${\times}$M crosses. In Garole sheep, no significant effect of FecB was observed on live weights in any age group. The interaction between the genetic group and the FecB genotype was also found to be non-significant.

Biphasic effects of TGFβ1 on BMP9-induced osteogenic differentiation of mesenchymal stem cells

  • Li, Rui-Dong;Deng, Zhong-Liang;Hu, Ning;Liang, Xi;Liu, Bo;Luo, Jin-Yong;Chen, Liang;Yin, Liangjun;Luo, Xiaoji;Shui, Wei;He, Tong-Chuan;Huang, Wei
    • 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.

Activation of JNKs is essential for BMP9-induced osteogenic differentiation of mesenchymal stem cells

  • Zhao, Yan-Fang;Xu, Jing;Wang, Wen-Juan;Wang, Jin;He, Juan-Wen;Li, Li;Dong, Qian;Xiao, Yan;Duan, Xing-Lian;Yang, Xue;Liang, Yi-Wen;Song, Tao;Tang, Min;Zhao, Dan;Luo, Jin-Yong
    • BMB Reports
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    • 제46권8호
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    • pp.422-427
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    • 2013
  • Although BMP9 is highly capable of promoting osteogenic differentiation of mesenchymal stem cell (MSCs), the molecular mechanism involved remains to be fully elucidated. Here, we explore the possible involvement and detail role of JNKs (c-Jun N-terminal kinases) in BMP9-induced osteogenic differentiation of MSCs. It was found that BMP9 stimulated the activation of JNKs in MSCs. BMP9-induced osteogenic differentiation of MSCs was dramatically inhibited by JNKs inhibitor SP600125. Moreover, BMP9-activated Smads signaling was decreased by SP600125 treatment in MSCs. The effects of inhibitor are reproduced with adenoviruses expressing siRNA targeted JNKs. Taken together, our results revealed that JNKs was activated in BMP9-induced osteogenic differentiation of MSCs. What is most noteworthy, however, is that inhibition of JNKs activity resulted in reduction of BMP9-induced osteogenic differentiation of MSCs, implying that activation of JNKs is essential for BMP9 osteoinductive activity.

생체골의 열전도성 및 열처리된 골의 염전력 변화에 대한 실험적 연구 (Experimental Studies on Heat Conductivity of Human Bone and Torsional Strength of Pasteurized Porcine Tibia)

  • 박일형;김신근;신동규;인주철
    • 대한골관절종양학회지
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    • 제1권1호
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    • pp.7-16
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    • 1995
  • 인체골 4 시편(specimen)과 돼지 경골 25쌍을 이용하여, 생체골의 열전도성 측정과 열처리후 열처리온도와 시간에 따른 골의 염전력을 실험한 결과를 요약하면 다음과 같다. 인체골에 있어서 골수강을 제거하지 않고 $60^{\circ}C$의 항온식염수에서 열처리하면, 골심부의 온도가 $20^{\circ}C$에서 $58^{\circ}C$에 도달하는데 소요된 시간은 경골근위부가 32분 50초, 대퇴골 원위부가 30.분, $80^{\circ}C$ 항온조에서는 경골근위부가 12분 50초, 대퇴골 원위부가 11분 10초 소요되었다. 돼지 경골간부의 피질골내부(endosteum)에 열전대를 부착하고 뼈 양끝을 밀봉하여 같은 실험을 행한 결과 $50^{\circ}C$까지는 시간에 비례해서 일정한 비율로 온도상승이 이루어 졌으며, $20^{\circ}C$에서 $58^{\circ}C$에 이르는 시간이 $60^{\circ}C$ 항온조에서는 7분, $70^{\circ}C$에서는 3분 30초, $80^{\circ}C$에서는 2분이었다. 따라서 임상에서 골수강을 제거 후 장골의 간부(shaft) 만을 항온조에 달굴때는 골구강내에도 데워진 심염수로 가득차게 되므로 상기 시간의 절반이 못되는 짧은 시간내에 피질골의 내부가 $58^{\circ}C$에 이르리라고 판단되었다. 골수강을 소파하지 않은 돼지 경골을 각각 4쌍씩 우측만을 $60^{\circ}C$ 35분, $80^{\circ}C$ 15분 열처리한 후 실험군의 최대염전력은 대조군과 비교할 때 +7.0%, -5.1%, -3.2%, -4.2%의 변화가 있었고, $80^{\circ}C$ 15분 열처리후는 -4.3%, -3.8%, -1.4%(1예는 실험 오류로 제외됨)의 변화가 있었다. 골수강을 완전 제거한 되재 경골을 각각 4쌍씩 우측만을 $60^{\circ}C$, $70^{\circ}C$, $80^{\circ}C$에서 15분 열처리 후 실험군의 최대염전력은 대조군과 비교할 때 -3.4%, -4.2%, -0.7%, +2.7%의 변화가 있었고, $70^{\circ}C$ 15분 열처리후는 -2.8%, -3.9%, -2.1%(1예는 실험 오류로 제외됨)의 변화가 있었으며, $80^{\circ}C$ 15분 열처리후는 +5.2%, -4.4%, -2.9%, -0.3%의 염전력 변화가 있었다. 그러므로 골수강을 제거하지 않고 $80^{\circ}C$ 35분, $60^{\circ}C$ 15분 열처리 하거나, 골수강을 완전소파 후 $60^{\circ}C$ 15분, $70^{\circ}C$ 15분, $80^{\circ}C$ 15분 열처리해서는 각군사이에 염전력의 유의한 차이는없었다. 이상의 결과로 돼지 경골의 경우 $60^{\circ}C$ 항온에서는 35분까지, $80^{\circ}C$이하의 항온에서는 15분까지 열처리하여도 골강도에는 거의 영향이 없는 것으로 나타났다.

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BMP4 처리에 의한 인간 배아줄기세포 유래 KDR 양성 중배엽성 세포군의 분화 양상 조사 (Identification and Characterization of a KDR-positive Mesoderm Population Derived from Human Embryonic Stem Cells Post BMP4 Treatment)

  • 김정모;손온주;조윤정;이재호;정형민
    • Reproductive and Developmental Biology
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    • 제35권1호
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    • pp.9-15
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    • 2011
  • The functional cardiovascular system is comprised of distinct mesoderm-derived lineages including endothelial cells, vascular smooth muscle cells and other mesenchymal cells. Recent studies in the human embryonic stem cell differentiation model have provided evidence indicating that these cell lineages are developed from the common progenitors such as hemangioblasts and cardiovascular progenitor cells. Also, the studies have suggested that these progenitors have a common primordial progenitor, which expresses KDR (human Flk-1, also known as VEGFR2, CD309). We demonstrate here that sustained activation of BMP4 (bone morphogenetic protein 4) in hESC line, CHA15 hESC results in $KDR^+$ mesoderm specific differentiation. To determine whether the $KDR^+$ population derived from hESCs enhances potential to differentiate along multipotential mesodermal lineages than undifferentiated hESCs, we analyzed the development of the mesodermal cell types in human embryonic stem cell differentiation cultures. In embryoid body (EB) differentiation culture conditions, we identified an increased expression of $KDR^+$ population from BMP4-stimulated hESC-derived EBs. After induction with additional growth factors, the $KDR^+$ population sorted from hESCs-derived EBs displays mesenchymal, endothelial and vascular smooth muscle potential in matrix-coated monolayer culture systems. The populations plated in monolayer cultures expressed increased levels of related markers and exhibit a stable/homologous phenotype in culture terms. In conclusion, we demonstrate that the $KDR^+$ population is stably isolated from CHA15 hESC-derived EBs using BMP4 and growth factors, and sorted $KDR^+$ population can be utilized to generate multipotential mesodermal progenitors in vitro, which can be further differentiated into cardiovascular specific cells.

The antioxidant icariin protects porcine oocytes from age-related damage in vitro

  • Yoon, Jae-Wook;Lee, Seung-Eun;Park, Yun-Gwi;Kim, Won-Jae;Park, Hyo-Jin;Park, Chan-Oh;Kim, So-Hee;Oh, Seung-Hwan;Lee, Do-Geon;Pyeon, Da-Bin;Kim, Eun-Young;Park, Se-Pill
    • Animal Bioscience
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    • 제34권4호
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    • pp.546-557
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    • 2021
  • Objective: If fertilization does not occur within a specific period, the quality of unfertilized oocytes in the oviduct (in vivo aging) or in culture (in vitro aging) will deteriorate over time. Icariin (ICA), found in all species of Epimedium herbs, has strong antioxidant activity, and is thought to exert anti-aging effects in vitro. We asked whether ICA protects oocytes against age-related changes in vitro. Methods: We analyzed the reactive oxygen species (ROS) levels and expression of antioxidant, maternal, and estrogen receptor genes, and along with spindle morphology, and the developmental competence and quality of embryos in the presence and absence of ICA. Results: Treatment with 5 μM ICA (ICA-5) led to a significant reduction in ROS activity, but increased mRNA expression of glutathione and antioxidant genes (superoxide dismutase 1 [SOD1], SOD2, peroxiredoxin 5, and nuclear factor erythroid 2-like 2), during aging in vitro. In addition, ICA-5 prevented defects in spindle formation and chromosomal alignment, and increased mRNA expression of cytoplasmic maturation factor genes (bone morphogenetic protein 15, cyclin B1, MOS proto-oncogene, serine/threonine kinase, and growth differentiation factor-9). It also prevented apoptosis, increased mRNA expression of antiapoptotic genes (BCL2-like 1 and baculoviral IAP repeat-containing 5), and reduced mRNA expression of pro-apoptotic genes (BCL2 antagonist/killer 1 and activation of caspase-3). Although the maturation and cleavage rates were similar in all groups, the total cell number per blastocyst and the percentage of apoptotic cells at the blastocyst stage were higher and lower, respectively, in the control and ICA-5 groups than in the aging group. Conclusion: ICA protects oocytes against damage during aging in vitro; therefore, it can be used to improve assisted reproductive technologies.