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Exp Ther Med：转染人BMP-2和EGFP重组腺病毒可增强兔骨髓间充质干细胞的成骨作用

2018-11-08 MedSci MedSci原创

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骨髓间充质干细胞（BMSCs）是骨组织工程中最重要的种子细胞。本研究旨在探讨兔BMSCs转染人BMP-2和EGFP重组腺病毒后在成骨过程中的潜力。分离兔BMSCs，通过流式细胞仪检测表面干细胞标志物，包括CD29、CD44和CD45。采用逆转录-定量聚合酶链反应和Western blot分析检测BMSCs中BMP-2 mRNA和蛋白的表达。成骨培养基诱导后，使用ALP活性试剂盒，免疫组织化学染色和

骨髓间充质干细胞（BMSCs）是骨组织工程中最重要的种子细胞。本研究旨在探讨兔BMSCs转染人BMP-2和EGFP重组腺病毒后在成骨过程中的潜力。

分离兔BMSCs，通过流式细胞仪检测表面干细胞标志物，包括CD29、CD44和CD45。采用逆转录-定量聚合酶链反应和Western blot分析检测BMSCs中BMP-2 mRNA和蛋白的表达。成骨培养基诱导后，使用ALP活性试剂盒，免疫组织化学染色和茜素红S染色，分别检测第7天的碱性磷酸酶（ALK）活性，第14天的I型胶原和第21天的钙结节。通过蛋白质印迹分析检测与Wnt信号传导通路相关的蛋白质的表达水平。

结果显示，CD29，CD44和CD45的阳性率分别为62.92±1.99,93.55±0.99和0.21±0.12％。在Ad-BMP-2/EGFP转染的BMSCs中BMP-2 mRNA和蛋白的表达显著上调。此外，Ad-BMP-2/EGFP诱导ALP活性，促进兔BMSCs中I型胶原和钙结节形成的产生。在Ad-hBMP-2/EGFP转染的BMSCs中，β-连环蛋白、细胞周期蛋白D1、Runx2和c-myc的水平上调，而GSK3β的水平显著降低。结果还表明，Ad-hBMP-2/EGFP过表达BMP-2，通过刺激Wnt信号通路，促进β-catenin的积累和GSK3β的抑制，提高了兔BMSCs的成骨分化能力。预期Ad-hBMP-2/EGFP转染的兔BMSCs是骨组织工程中的良好种子细胞。

原始出处：

Wu CC, Wang F, Enhancement of osteogenesis of rabbit bone marrow derived mesenchymal stem cells by transfection of human BMP-2 and EGFP recombinant adenovirus via Wnt signaling pathway. Exp Ther Med. 2018 Nov;16(5):4030-4036. doi: 10.3892/etm.2018.6735. Epub 2018 Sep 13.

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    2019-08-11 12498f17m73暂无昵称

    #BMP-2#

    0

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    2019-02-01 HNYYM

    #BMP#

    0

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    2018-11-19 ms7514935887898259

    #重组腺病毒#

    0

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    2018-12-12 h.y200607_23674418

    #间充质干细#

    0

  5. [GetPortalCommentsPageByObjectIdResponse(id=1642663, encodeId=2eec164266362, content=<a href='/topic/show?id=814c35e6e2' target=_blank style='color:#2F92EE;'>#BMP-2#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=49, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3576, encryptionId=814c35e6e2, topicName=BMP-2)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=120c22999408, createdName=12498f17m73暂无昵称, createdTime=Sun Aug 11 02:20:00 CST 2019, time=2019-08-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1634054, encodeId=0bed16340545d, content=<a href='/topic/show?id=596635e549' target=_blank style='color:#2F92EE;'>#BMP#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=33, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3575, encryptionId=596635e549, topicName=BMP)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=d95322064897, createdName=HNYYM, createdTime=Fri Feb 01 20:20:00 CST 2019, time=2019-02-01, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1687223, encodeId=e998168e22366, content=<a href='/topic/show?id=6a5b9604106' target=_blank style='color:#2F92EE;'>#重组腺病毒#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=46, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=96041, encryptionId=6a5b9604106, topicName=重组腺病毒)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=0ddd28353190, createdName=ms7514935887898259, createdTime=Mon Nov 19 15:20:00 CST 2018, time=2018-11-19, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1815406, encodeId=11a7181540634, content=<a href='/topic/show?id=ae359e231d4' target=_blank style='color:#2F92EE;'>#间充质干细#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=40, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=97231, encryptionId=ae359e231d4, topicName=间充质干细)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4b632500090, createdName=h.y200607_23674418, createdTime=Wed Dec 12 00:20:00 CST 2018, time=2018-12-12, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1705489, encodeId=c5cc1e05489de, content=<a href='/topic/show?id=31f9102015b3' target=_blank style='color:#2F92EE;'>#骨髓间充质干细胞#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=36, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=102015, encryptionId=31f9102015b3, topicName=骨髓间充质干细胞)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=723a30986413, createdName=ying_wu, createdTime=Mon Mar 18 03:20:00 CST 2019, time=2019-03-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1351895, encodeId=5e081351895db, content=<a href='/topic/show?id=e95954093cd' target=_blank style='color:#2F92EE;'>#成骨#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54093, encryptionId=e95954093cd, topicName=成骨)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=zhaojie88, createdTime=Sat Nov 10 10:20:00 CST 2018, time=2018-11-10, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1609301, encodeId=91d1160930126, content=<a href='/topic/show?id=eb6b1145939' target=_blank style='color:#2F92EE;'>#Med#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=11459, encryptionId=eb6b1145939, topicName=Med)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=hb2008ye, createdTime=Sat Nov 10 10:20:00 CST 2018, time=2018-11-10, status=1, ipAttribution=)]
    2019-03-18 ying_wu

    #骨髓间充质干细胞#

    0

  6. [GetPortalCommentsPageByObjectIdResponse(id=1642663, encodeId=2eec164266362, content=<a href='/topic/show?id=814c35e6e2' target=_blank style='color:#2F92EE;'>#BMP-2#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=49, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3576, encryptionId=814c35e6e2, topicName=BMP-2)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=120c22999408, createdName=12498f17m73暂无昵称, createdTime=Sun Aug 11 02:20:00 CST 2019, time=2019-08-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1634054, encodeId=0bed16340545d, content=<a href='/topic/show?id=596635e549' target=_blank style='color:#2F92EE;'>#BMP#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=33, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3575, encryptionId=596635e549, topicName=BMP)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=d95322064897, createdName=HNYYM, createdTime=Fri Feb 01 20:20:00 CST 2019, time=2019-02-01, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1687223, encodeId=e998168e22366, content=<a href='/topic/show?id=6a5b9604106' target=_blank style='color:#2F92EE;'>#重组腺病毒#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=46, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=96041, encryptionId=6a5b9604106, topicName=重组腺病毒)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=0ddd28353190, createdName=ms7514935887898259, createdTime=Mon Nov 19 15:20:00 CST 2018, time=2018-11-19, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1815406, encodeId=11a7181540634, content=<a href='/topic/show?id=ae359e231d4' target=_blank style='color:#2F92EE;'>#间充质干细#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=40, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=97231, encryptionId=ae359e231d4, topicName=间充质干细)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4b632500090, createdName=h.y200607_23674418, createdTime=Wed Dec 12 00:20:00 CST 2018, time=2018-12-12, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1705489, encodeId=c5cc1e05489de, content=<a href='/topic/show?id=31f9102015b3' target=_blank style='color:#2F92EE;'>#骨髓间充质干细胞#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=36, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=102015, encryptionId=31f9102015b3, topicName=骨髓间充质干细胞)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=723a30986413, createdName=ying_wu, createdTime=Mon Mar 18 03:20:00 CST 2019, time=2019-03-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1351895, encodeId=5e081351895db, content=<a href='/topic/show?id=e95954093cd' target=_blank style='color:#2F92EE;'>#成骨#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54093, encryptionId=e95954093cd, topicName=成骨)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=zhaojie88, createdTime=Sat Nov 10 10:20:00 CST 2018, time=2018-11-10, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1609301, encodeId=91d1160930126, content=<a href='/topic/show?id=eb6b1145939' target=_blank style='color:#2F92EE;'>#Med#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=11459, encryptionId=eb6b1145939, topicName=Med)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=hb2008ye, createdTime=Sat Nov 10 10:20:00 CST 2018, time=2018-11-10, status=1, ipAttribution=)]
    2018-11-10 zhaojie88

    #成骨#

    0

  7. [GetPortalCommentsPageByObjectIdResponse(id=1642663, encodeId=2eec164266362, content=<a href='/topic/show?id=814c35e6e2' target=_blank style='color:#2F92EE;'>#BMP-2#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=49, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3576, encryptionId=814c35e6e2, topicName=BMP-2)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=120c22999408, createdName=12498f17m73暂无昵称, createdTime=Sun Aug 11 02:20:00 CST 2019, time=2019-08-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1634054, encodeId=0bed16340545d, content=<a href='/topic/show?id=596635e549' target=_blank style='color:#2F92EE;'>#BMP#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=33, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=3575, encryptionId=596635e549, topicName=BMP)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=d95322064897, createdName=HNYYM, createdTime=Fri Feb 01 20:20:00 CST 2019, time=2019-02-01, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1687223, encodeId=e998168e22366, content=<a href='/topic/show?id=6a5b9604106' target=_blank style='color:#2F92EE;'>#重组腺病毒#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=46, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=96041, encryptionId=6a5b9604106, topicName=重组腺病毒)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=0ddd28353190, createdName=ms7514935887898259, createdTime=Mon Nov 19 15:20:00 CST 2018, time=2018-11-19, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1815406, encodeId=11a7181540634, content=<a href='/topic/show?id=ae359e231d4' target=_blank style='color:#2F92EE;'>#间充质干细#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=40, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=97231, encryptionId=ae359e231d4, topicName=间充质干细)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4b632500090, createdName=h.y200607_23674418, createdTime=Wed Dec 12 00:20:00 CST 2018, time=2018-12-12, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1705489, encodeId=c5cc1e05489de, content=<a href='/topic/show?id=31f9102015b3' target=_blank style='color:#2F92EE;'>#骨髓间充质干细胞#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=36, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=102015, encryptionId=31f9102015b3, topicName=骨髓间充质干细胞)], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=723a30986413, createdName=ying_wu, createdTime=Mon Mar 18 03:20:00 CST 2019, time=2019-03-18, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1351895, encodeId=5e081351895db, content=<a href='/topic/show?id=e95954093cd' target=_blank style='color:#2F92EE;'>#成骨#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=24, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=54093, encryptionId=e95954093cd, topicName=成骨)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=zhaojie88, createdTime=Sat Nov 10 10:20:00 CST 2018, time=2018-11-10, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=1609301, encodeId=91d1160930126, content=<a href='/topic/show?id=eb6b1145939' target=_blank style='color:#2F92EE;'>#Med#</a>, beContent=null, objectType=article, channel=null, level=null, likeNumber=28, replyNumber=0, topicName=null, topicId=null, topicList=[TopicDto(id=11459, encryptionId=eb6b1145939, topicName=Med)], attachment=null, authenticateStatus=null, createdAvatar=, createdBy=4c37265, createdName=hb2008ye, createdTime=Sat Nov 10 10:20:00 CST 2018, time=2018-11-10, status=1, ipAttribution=)]
    2018-11-10 hb2008ye

    #Med#

    0