SCI 干细胞国际 1687 - 9678 1687 - 966 x Hindawi 10.1155 / 2018/2791632 2791632 研究文章 Alginate-Microencapsulated脂肪Tissue-Derived关节内注射间充质干细胞治疗骨关节炎的兔子 http://orcid.org/0000 - 0003 - 2644 - 6521 崔 Seongjae 1 金 Jun-Hyung 1 哈 Jeongho 1 宋 Bo-Ing 2 荣格 Yun陈 2 李 Geun-Shik 1 吸引 Heung-Myong 1 http://orcid.org/0000 - 0002 - 2419 - 1775 康 Byung-Jae 1 常 彭 1 兽医学院兽医科学研究所 Kangwon国立大学 春川24341 韩国 kangwon.ac.kr 2 KPC 光州12773 韩国 kpclab.co.kr 2018年 26 6 2018年 2018年 14 12 2017年 17 04 2018年 20. 05年 2018年 26 6 2018年 2018年 版权©2018 Seongjae Choi et al。 这是一个开放的文章在知识共享归属许可下发布的,它允许无限制的使用,分布和繁殖在任何媒介,提供最初的工作是正确的引用。

我们调查的影响alginate-microencapsulated脂肪tissue-derived关节内注射间充质干细胞在骨关节炎(OA)(对asc)发展的兔子模型前交叉韧带横断(ACLT)。我们在成熟诱导OA双边ACLT新西兰白兔。抑制关节被分为四组根据关节内注射材料。藻酸盐微和对asc microencapsulated使用振动喷嘴技术准备。ACLT两周后,兔子获得连续三周0.9%的氯化钠,关节内注射海藻酸微磁,对asc,或对asc microencapsulated,到每个关节。ACLT 9周后,我们实施安乐死收集的兔子和双边股骨髁部宏观、组织学和免疫组织化学分析。宏观评价使用修改后的OA研究学会国际(OARSI)得分和总软骨损伤分数显示股骨髁软骨退化是microencapsulated-ASC组相对较低。的组织学分析股外侧髁部表明chondroprotective有对asc microencapsulated效果显著。免疫组织化学,MMP-13关节软骨损伤后的表达相对较低的microencapsulated-ASC-treated抑制关节。在实验办公自动化的发展,仅对asc相比,关节内注射对asc microencapsulated显著降低OA的过程和程度。 科技部、ICT和未来规划 nrf - 2015 r1c1a1a01051759 韩国国家研究基金会 1。介绍<gydF4y2Ba/title> <p>退化性关节疾病或骨关节炎(OA)的膝盖是最常见的关节炎和降低生活质量,引起疼痛,僵硬,身体残疾(<gydF4y2Baxref ref-type="bibr" rid="B1"> 1<gydF4y2Ba/xref>]。OA的特点是进步的软骨的恶化和破坏由于受损的细胞外基质合成和/或分解代谢的平衡(<gydF4y2Baxref ref-type="bibr" rid="B2"> 2<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B3"> 3<gydF4y2Ba/xref>]。关节软骨的自我修复能力较低(<gydF4y2Baxref ref-type="bibr" rid="B4"> 4<gydF4y2Ba/xref>]。它是具有挑战性的增强透明软骨组织的再生(<gydF4y2Baxref ref-type="bibr" rid="B5"> 5<gydF4y2Ba/xref>]。有许多临床治疗OA管理(<gydF4y2Baxref ref-type="bibr" rid="B6"> 6<gydF4y2Ba/xref>]。这些治疗包括非甾体类抗炎药、富含血小板血浆,止痛剂,透明质酸,mevastatin,间充质干细胞(msc) [<gydF4y2Baxref ref-type="bibr" rid="B7"> 7<gydF4y2Ba/xref>- - - - - -<gydF4y2Baxref ref-type="bibr" rid="B11"> 11<gydF4y2Ba/xref>]。尽管许多治疗与msc已经发达,脂肪tissue-derived间充质干细胞(对asc)有几个优势。在人类中,关节内注射对asc改善功能和减轻疼痛和膝关节软骨缺损<gydF4y2Baxref ref-type="bibr" rid="B11"> 11<gydF4y2Ba/xref>]。此外,对asc可以更容易地培养和获得的大大大数量不那么咄咄逼人的方法msc可以(<gydF4y2Baxref ref-type="bibr" rid="B12"> 12<gydF4y2Ba/xref>- - - - - -<gydF4y2Baxref ref-type="bibr" rid="B14"> 14<gydF4y2Ba/xref>]。<gydF4y2Ba/p> <p>全身或局部干细胞疗法代表一个治疗OA的成长领域,导致关节软骨的修复(<gydF4y2Baxref ref-type="bibr" rid="B15"> 15<gydF4y2Ba/xref>]。尽管如此,研究日益显示可怜的可行性和较低的存活率移植干细胞的流感传染的网站(<gydF4y2Baxref ref-type="bibr" rid="B16"> 16<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B17"> 17<gydF4y2Ba/xref>]。因此最近的研究集中在提高细胞生存能力在当地网站受到疾病和治疗涉及msc的旁分泌作用[<gydF4y2Baxref ref-type="bibr" rid="B18"> 18<gydF4y2Ba/xref>- - - - - -<gydF4y2Baxref ref-type="bibr" rid="B21"> 21<gydF4y2Ba/xref>]。大多数msc治疗的影响被认为行为以旁分泌的方式通过促进血管生成,组织再生和生产可溶性抗炎因子(<gydF4y2Baxref ref-type="bibr" rid="B16"> 16<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B22"> 22<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B23"> 23<gydF4y2Ba/xref>]。Alginate-microencapsulated细胞提供一个机械屏障作为人工细胞外基质,增加细胞的生存能力和允许阻止cell-produced生长因子和抗炎因子的释放到周围受伤组织(<gydF4y2Baxref ref-type="bibr" rid="B24"> 24<gydF4y2Ba/xref>]。<gydF4y2Ba/p> <p>我们调查的影响周期对asc alginate-microencapsulated在OA关节内注射兔前交叉韧带横断模型(ACLT)。我们提出对asc microencapsulated将减少对asc OA进展更有效地比独自一人。<gydF4y2Ba/p> </sec> <sec id="sec2"> <title>2。材料和方法<gydF4y2Ba/title> <sec id="sec2.1"> <title>2.1。动物<gydF4y2Ba/title> <p>十五岁成年新西兰白兔(3.0 - -3.5公斤,25 - 30周大,男)与闭合骺被用于这项研究。十一个兔子被分配到OA模型和四个兔子allogeneic-ASC收割。动物保健和研究协议是审查和批准的机构动物保健和使用委员会Kangwon国立大学(kw - 170315 - 1)。<gydF4y2Ba/p> </sec> <sec id="sec2.2"> <title>2.2。兔子ACLT模型<gydF4y2Ba/title> <p>十一个兔子anesthetised tiletamine-zolazepam通过肌肉注射(20毫克/公斤;Zoletil Virbac,首尔,韩国)和甲苯噻嗪盐酸盐(5毫克/公斤;Rompun拜耳韩国,汉城,韩国)。静脉注射ketoprofen(3毫克/公斤;韩国首尔的SCD制药)给出先发制人的镇痛。静脉注射头孢唑啉(22毫克/公斤;庄Kun Dang制药,首尔,韩国)管理(预防感染)时的手术和术后3 d每隔24小时。兔子麻醉时,电剪刀用来剪了头发覆盖每一个膝关节,和无菌手术获得皮下脂肪组织进行。外侧髌腱移位后,两国后肢的前交叉韧带切断完全没有。11手术刀叶片。缝合后,10个颅画运动引发了联合刺激,ACLT完全证实。 The rabbits were closely monitored for infection and other complications and were allowed to rest in a cage without immobilisation.</p> </sec> <sec id="sec2.3"> <title>2.3。ASC隔离和文化<gydF4y2Ba/title> <p>兔子对asc孤立的胶原酶灌注技术根据先前描述的方法(<gydF4y2Baxref ref-type="bibr" rid="B22"> 22<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B25"> 25<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B26"> 26<gydF4y2Ba/xref>]。约5克脂肪收集四个兔子颈后皮下脂肪组织的混合,用磷酸盐(PBS)。脂肪组织是切成条10分钟孵化中胶原酶我(热费希尔科学、沃尔瑟姆,妈,美国)6-well板。胶原酶溶解在PBS所以它的浓度是0.1%,25毫升,是用来消化脂肪组织在37°C水浴中90分钟。混合物动摇了每10分钟消化期间。反应完成后,25毫升high-glucose杜尔贝科修改鹰的介质(美国DMEM,表达载体,卡尔斯巴德,CA)被添加到中和胶原酶的活动。最终的解决方案是透过100<我t一个l我c> μ<gydF4y2Ba/italic>m细胞的过滤器。滤液离心机在1500 g×6分钟25°C,和上层的移除。接下来,对asc的颗粒被播种在细胞培养中菜肴和培养在标准条件下high-glucose DMEM补充10%的胎牛血清(美国的边后卫,表达载体,卡尔斯巴德,CA)和1%的antibiotic-antimycotic解决方案(热费希尔科学、沃尔瑟姆,妈,美国)。媒介取代每隔48 h,直到细胞汇合的。融合后的细胞达到90%,他们收获0.25%胰蛋白酶乙二胺四乙酸(EDTA);Welgene Gyeongsan-si、韩国)和存储在液态氮或亚文化。当细胞密度~ 2.5×10<年代up>4<gydF4y2Ba/sup>细胞/厘米<年代up>2<gydF4y2Ba/sup>,使对asc。使用低温贮藏对asc到通道5。<gydF4y2Ba/p> </sec> <sec id="sec2.4"> <title>2.4。制备海藻酸微<gydF4y2Ba/title> <p>1.2%的海藻酸钠溶液作为聚合物。无菌过滤等张1.8%的海藻酸钠溶液(Buchi Labortechnik AG) Flawil,瑞士)浓度与无菌生理盐水稀释至1.2%。1.2%的海藻酸钠溶液制备了注射器。藻酸盐微振动喷嘴产生的技术通过Buchi Encapsulator b - 395 Pro (Buchi Labortechnik AG) Flawil,瑞士)参数之前报道的制备海藻酸微适合通过23-gauge针:喷嘴大小,120<我t一个l我c> μ<gydF4y2Ba/italic>m;1000赫兹频率;流量,23毫升/分钟;和电极电位,1000 V (<gydF4y2Baxref ref-type="bibr" rid="B21"> 21<gydF4y2Ba/xref>]。胶凝溶液制备氯化钙在100毫米。海藻酸滴胶凝在100毫米氯化钙30分钟。完成后的海藻酸微过滤40<我t一个l我c> μ<gydF4y2Ba/italic>在PBS m细胞过滤器和清洗。的形态和大小海藻酸微磁光显微镜下进行分析。<gydF4y2Ba/p> </sec> <sec id="sec2.5"> <title>2.5。准备对asc Alginate-Microencapsulated<gydF4y2Ba/title> <p>藻朊酸盐解决方案包含对asc是由混合1.2%的海藻酸钠溶液的浓度对asc和10<年代up>7<gydF4y2Ba/sup>细胞/毫升。这种悬架受到微型胶囊然后过滤优化工艺参数。接下来,对asc microencapsulated洗1 x PBS,和微胶囊的大小和形状对asc包含被亮场显微镜分析。<gydF4y2Ba/p> </sec> <sec id="sec2.6"> <title>2.6。这项研究计划<gydF4y2Ba/title> <p>兔子每星期3星期注射一种材料根据一个实验组,ACLT在2周后开始。ACLT 9周后,所有的兔子都实施安乐死,膝关节样本收集和评估(图<gydF4y2Baxref rid="fig1a" ref-type="fig"> 1(一)<gydF4y2Ba/xref>)。<gydF4y2Ba/p> <fig-group id="fig1"> <label>图1<gydF4y2Ba/label> <p>研究方案与材料内注入根据实验团体和显微镜图像。(一)研究方案内注入的材料根据实验组。(b)的光学显微镜图像对asc纺锤状形态。(c)的光学显微镜图像海藻酸微没有细胞。(d) Light-microscopic外观microencapsulated植入的时候也用300 - 400细胞在每一个400 - 500<我t一个l我c> μ<gydF4y2Ba/italic>m胶囊。规模<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M1"> <mml:mi mathvariant="normal"> b<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> r<gydF4y2Ba/mml:mi> <mml:mo> =<gydF4y2Ba/mml:mo> <mml:mn> 500年<gydF4y2Ba/mml:mn> <mml:mtext> </mml:mtext> <mml:mi> μ<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 米<gydF4y2Ba/mml:mi> </mml:math> </inline-formula>。<gydF4y2Ba/p> <fig id="fig1a"> <label>(一)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.001a"></graphic> </fig> <fig id="fig1b"> <label>(b)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.001b"></graphic> </fig> <fig id="fig1c"> <label>(c)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.001c"></graphic> </fig> <fig id="fig1d"> <label>(d)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.001d"></graphic> </fig> </fig-group> </sec> <sec id="sec2.7"> <title>2.7。关节内注射的材料<gydF4y2Ba/title> <p>内注入任何材料之前,每只兔子anesthetised的肌内注射10毫克/公斤tiletamine-zolazepam (Zoletil 50, Virbac,首尔,韩国)和5毫克/公斤甲苯噻嗪盐酸盐(Rompun拜耳韩国,汉城,韩国)。1毫升塑料注射器满载着0.5毫升的0.9%氯化钠的材料之一,有或没有细胞悬液;这个注射器,21-gauge皮下注射针连接。<gydF4y2Ba/p> <p>匹配配对分析,11个兔子(22膝盖)。ACLT在2、3和4周后,在五兔子,被注入到左侧膝关节也和对asc microencapsulated注入右膝关节,通过内侧的方法。在六个兔子,0.9%生理盐水注入左侧膝关节,和藻酸盐微注入右膝关节,通过内侧的方法。每只兔子举行了5分钟,以便滑膜注射材料的附件。<gydF4y2Ba/p> </sec> <sec id="sec2.8"> <title>2.8。宏观分析<gydF4y2Ba/title> <p>十一个兔子ACLT安乐死在9周后,通过静脉注射氯化钾过量。股骨髁部都是精心收集,以避免任何损害软骨表面。关节的关节表面沾墨汁(美国美国MasterTech,洛迪,CA)对宏观检验和研究关于颤和侵蚀<gydF4y2Baxref ref-type="bibr" rid="B27"> 27<gydF4y2Ba/xref>]。软骨表面涂上墨汁两次,我们每次冲洗与PBS软骨;墨水应用3分钟后清洗。宏观照片拍摄使用索尼数码相机(索尼,东京,日本)。<gydF4y2Ba/p> <p>宏观OA分数计算加法内侧和外侧髁分数和通过修改版本的骨关节炎研究学会国际(OARSI)评分系统(补充表<gydF4y2Baxref ref-type="sec" rid="supplementary-material-1"> 1<gydF4y2Ba/xref>)[<gydF4y2Baxref ref-type="bibr" rid="B28"> 28<gydF4y2Ba/xref>]。<gydF4y2Ba/p> <p>确定软骨损伤总分(tcd)、股骨髁部分级基于油墨保留;量化的缺陷区域在股骨髁部进行ImageJ (NIH,贝塞斯达,医学博士,美国):图像分析软件。计算的核心是通过添加软骨损伤分数(CDSs)的内侧和外侧股骨髁部。这些信用违约掉期的计算是通过乘以每个髁关节软骨受损面积的百分比的墨水retention-based年级(1级,完整的表面:表面正常的外观,而不是保留印度墨水;二年级,颤:表面保留墨汁作为细长的斑点或浅灰色或黑色斑点;三年级,侵蚀:暴露软骨损失)。信用违约掉期范围从0到300(0 =没有软骨损伤;300 =完全暴露软骨下骨)。TCDSs通过以下方程计算(<gydF4y2Baxref ref-type="bibr" rid="B29"> 29日<gydF4y2Ba/xref>]:<d我年代p-formula> <mml:math 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</mml:mtd> </mml:mlabeledtr> <mml:mtr> <mml:mtd> <mml:mi mathvariant="normal"> T<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> C<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> D<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 年代<gydF4y2Ba/mml:mi> <mml:mo> =<gydF4y2Ba/mml:mo> <mml:msub> <mml:mrow> <mml:mi mathvariant="normal"> C<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> D<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 年代<gydF4y2Ba/mml:mi> </mml:mrow> <mml:mrow> <mml:mi mathvariant="normal"> 米<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> e<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> d<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 我<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> l<gydF4y2Ba/mml:mi> <mml:mtext> </mml:mtext> <mml:mi mathvariant="normal"> f<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> e<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 米<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> o<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> r<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> l<gydF4y2Ba/mml:mi> <mml:mtext> </mml:mtext> <mml:mi mathvariant="normal"> c<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> o<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> n<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> d<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> y<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> l<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> e<gydF4y2Ba/mml:mi> </mml:mrow> </mml:msub> <mml:mo> +<gydF4y2Ba/mml:mo> <mml:msub> <mml:mrow> <mml:mi mathvariant="normal"> C<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> D<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 年代<gydF4y2Ba/mml:mi> </mml:mrow> <mml:mrow> <mml:mi mathvariant="normal"> l<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> t<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> e<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> r<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> l<gydF4y2Ba/mml:mi> <mml:mtext> </mml:mtext> <mml:mi mathvariant="normal"> f<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> e<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 米<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> o<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> r<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> l<gydF4y2Ba/mml:mi> <mml:mtext> </mml:mtext> <mml:mi mathvariant="normal"> c<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> o<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> n<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> d<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> o<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> y<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> l<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> e<gydF4y2Ba/mml:mi> </mml:mrow> </mml:msub> <mml:mo> 。<gydF4y2Ba/mml:mo> </mml:mtd> </mml:mtr> </mml:mtable> </mml:math> </disp-formula></p> <p>宏观OA分数和tcd测定两个兽医,其中一个是被注射材料。<gydF4y2Ba/p> </sec> <sec id="sec2.9"> <title>2.9。组织学分析<gydF4y2Ba/title> <p>组织学检查,远端股骨软骨都固定在10%中性缓冲福尔马林溶液(热费希尔科学、沃尔瑟姆,妈,美国)后宏观分析。标本在4% EDTA脱钙的解决方案和嵌入式石蜡。石蜡包埋的部分被切割成厚度为4<我t一个l我c> μ<gydF4y2Ba/italic>在旁矢状面的平面上。部分是通过退化最严重的区域。所有的样品被安装到幻灯片和苏木精和伊红染色一般病理观察。此外,所有的样品都沾Safranin-O和复染色与快速绿色OARSI分数可以确定。一组织病理学部分髁是独立评估在光学显微镜(日本东京奥林巴斯光学有限公司)由两个兽医那些失明组分布。关节软骨的变化进行评估根据OARSI兔组织评估指南(<gydF4y2Baxref ref-type="bibr" rid="B28"> 28<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B30"> 30.<gydF4y2Ba/xref>]。OA分数是一个指数的等级和舞台在显微切片(<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M3"> <mml:mi mathvariant="normal"> 年代<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> c<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> o<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> r<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> e<gydF4y2Ba/mml:mi> <mml:mo> =<gydF4y2Ba/mml:mo> <mml:mi mathvariant="normal"> g<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> r<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> d<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> e<gydF4y2Ba/mml:mi> <mml:mo> ×<gydF4y2Ba/mml:mo> <mml:mi mathvariant="normal"> 年代<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> t<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> g<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> e<gydF4y2Ba/mml:mi> </mml:math> </inline-formula>)。六年级代表一个级别:表面完好无损,不连续,垂直裂缝,侵蚀、剥蚀或变形。舞台被分配的基础上的水平程度涉及底层OA软骨表面缺陷等级。第一阶段代表参与不到10%,第二阶段意味着10 - 25%的参与,参与第三阶段表示25 - 50%,第四阶段代表超过50%的参与。<gydF4y2Ba/p> </sec> <sec id="sec2.10"> <title>2.10。免疫组织化学(包含IHC)分析<gydF4y2Ba/title> <p>从每个块,串行部分被削减到4<我t一个l我c> μ<gydF4y2Ba/italic>米厚度和改善附件安装到gelatin-coated幻灯片。幻灯片是deparaffinised二甲苯和下洗乙醇(100%,95%)和PBS。关节软骨包含IHC分析手动执行通过鼠标反MMP-13单克隆抗体(1:20;美国马热费希尔科学、沃尔瑟姆)。对于抗原检索,部分被孵化,然后在5分钟的高压釜处理在95°C使用0.1%柠檬酸钠缓冲(pH值6.0)。之后,冲洗三次PBS和幻灯片被孵化的peroxidase-conjugated聚合物DAKO真正想象/合兔/鼠标/装备(真正的设想检测系统K5007, DAKO,哥本哈根,丹麦)30分钟,再冲洗三次与PBS和孵化DABb工具包的5分钟,根据制造商的指示。最后,核与苏木精复染色30年代,法蓝在自来水下,乙醇洗,cover-slipped。进行半定量的分析的软骨外侧股骨髁部。免疫反应性的细胞和正常细胞在关节面区域计算细胞每平方毫米。包含IHC价值观,MMP-13阳性细胞的比例,确定蛋白表达一个完整的评估,最高分数的100%。 The assay was conducted by two blinded investigators [<xref ref-type="bibr" rid="B31"> 31日<gydF4y2Ba/xref>]。<gydF4y2Ba/p> </sec> <sec id="sec2.11"> <title>2.11。统计分析<gydF4y2Ba/title> <p>执行这个分析棱镜5.00 (GraphPad软件公司,圣地亚哥,美国)。并给出了结果<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M4"> <mml:mi mathvariant="normal"> 米<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> e<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> n<gydF4y2Ba/mml:mi> <mml:mo> ±<gydF4y2Ba/mml:mo> <mml:mi mathvariant="normal"> 年代<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> t<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> n<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> d<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> r<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> d<gydF4y2Ba/mml:mi> </mml:math> </inline-formula>偏差(SD)。统计学意义是克鲁斯卡尔-沃利斯检验评估的其次是Mann-Whitney事后测试(比较组)。数据与<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M5"> <mml:mi> P<gydF4y2Ba/mml:mi> <mml:mo> <<gydF4y2Ba/mml:mo> <mml:mn> 0.05<gydF4y2Ba/mml:mn> </mml:math> </inline-formula>为所有的测试被认为是重要的。<gydF4y2Ba/p> </sec> </sec> <sec id="sec3"> <title>3所示。结果<gydF4y2Ba/title> <sec id="sec3.1"> <title>3.1。形态对asc的确认、藻酸盐微和对asc Alginate-Microencapsulated<gydF4y2Ba/title> <p>使用光学显微镜,我们确认对asc纺锤状(图<gydF4y2Baxref rid="fig1b" ref-type="fig"> 1 (b)<gydF4y2Ba/xref>)。藻酸盐微和微胶囊含有对asc也证实了光学显微镜主要正常的球形。我们确定在藻酸盐微磁珠直径差异对asc和微胶囊。海藻酸盐微是250 - 300<我t一个l我c> μ<gydF4y2Ba/italic>米直径,包含对asc的微胶囊是400 - 500<我t一个l我c> μ<gydF4y2Ba/italic>米直径。每个微胶囊包含对asc包括300 - 400细胞,含有细胞的数量不同的珠直径(数据显示<gydF4y2Baxref rid="fig1c" ref-type="fig"> 1 (c)<gydF4y2Ba/xref>和<gydF4y2Baxref rid="fig1d" ref-type="fig"> 1 (d)<gydF4y2Ba/xref>)。<gydF4y2Ba/p> </sec> <sec id="sec3.2"> <title>3.2。宏观分析<gydF4y2Ba/title> <p>总值目视检查后,所有抑制关节显示完整的前交叉韧带横断和严重侵蚀内侧和外侧股骨髁部或裂缝。有更广泛的病变比横向侧内侧髁部。双边侧的等级是“侵蚀”控制和海藻酸microbead组。同时,内侧髁部的品位是侵蚀,侧侧的等级是“裂缝”ASC组和microencapsulated-ASC组。此外,该地区的受损软骨microencapsulated-ASC组似乎比其他组小得多。图<gydF4y2Baxref rid="fig2a" ref-type="fig"> 2(一个)<gydF4y2Ba/xref>提出了每组髁典型的例子,沾满了墨汁。显著降低宏观OA得分观察microencapsulated-ASC组(13.5±0.94)和ASC组(16.9±2.86)与对照组(21.58±2.11)。此外,宏观OA microencapsulated-ASC组的得分显著低于ASC组。然而,没有明显差异之间的控制和海藻酸microbead组(20.33±4.48)(图<gydF4y2Baxref rid="fig2b" ref-type="fig"> 2 (b)<gydF4y2Ba/xref>)。显著降低tcd观察microencapsulated-ASC组相对于对照组(26.6±5.59)(50.17±15.89),海藻酸microbead组(71.33±23.99)和ASC组(41±3.08)(图<gydF4y2Baxref rid="fig2c" ref-type="fig"> 2 (c)<gydF4y2Ba/xref>)。较低的宏观OA microencapsulated-ASC组分数和tcd显示显著减少损害软骨表面。<gydF4y2Ba/p> <fig-group id="fig2"> <label>图2<gydF4y2Ba/label> <p>的宏观分析股骨髁部ACLT 9周后。(a)一个代表性样本的髁(从每组)沾墨汁来识别任何颤和侵蚀。(b)的宏观OA得分。(c)浴室。<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M6"> <mml:msup> <mml:mrow></mml:mrow> <mml:mrow> <mml:mo> ∗<gydF4y2Ba/mml:mo> </mml:mrow> </mml:msup> </mml:math> </inline-formula>从对照组显著差异(<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M7"> <mml:mi> P<gydF4y2Ba/mml:mi> <mml:mo> <<gydF4y2Ba/mml:mo> <mml:mn> 0.05<gydF4y2Ba/mml:mn> </mml:math> </inline-formula>)。<年代up>#<gydF4y2Ba/sup>海藻酸的显著差异microbead集团(<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M8"> <mml:mi> P<gydF4y2Ba/mml:mi> <mml:mo> <<gydF4y2Ba/mml:mo> <mml:mn> 0.05<gydF4y2Ba/mml:mn> </mml:math> </inline-formula>)。<年代up>__<gydF4y2Ba/sup>ASC组的显著差异(<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M9"> <mml:mi> P<gydF4y2Ba/mml:mi> <mml:mo> <<gydF4y2Ba/mml:mo> <mml:mn> 0.05<gydF4y2Ba/mml:mn> </mml:math> </inline-formula>)。<gydF4y2Ba/p> <fig id="fig2a"> <label>(一)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.002a"></graphic> </fig> <fig id="fig2b"> <label>(b)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.002b"></graphic> </fig> <fig id="fig2c"> <label>(c)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.002c"></graphic> </fig> </fig-group> </sec> <sec id="sec3.3"> <title>3.3。组织学分析<gydF4y2Ba/title> <p>这个评价抑制关节显示更具体的横向侧(图的差异<gydF4y2Baxref rid="fig3a" ref-type="fig"> 3(一个)<gydF4y2Ba/xref>)。内侧髁软骨表面显示大多是剥蚀或变形,和组之间没有明显差异。在横向侧,OARSI OA得分没有显著差异之间的对照组(15.88±4.50),另一组除了microencapsulated-ASC组(4.35±3.04)(图<gydF4y2Baxref rid="fig3b" ref-type="fig"> 3 (b)<gydF4y2Ba/xref>)。在对照组和海藻酸microbead组(12.96±9.14),外侧关节面显示剥蚀或变形。虽然没有达到统计学意义,更有利的分数的关节面观察microencapsulated-ASC组比ASC组(7.3±5.34)。<gydF4y2Ba/p> <fig-group id="fig3"> <label>图3<gydF4y2Ba/label> <p>的组织学分析股骨外侧髁部。(a)一个代表性样本的外侧髁每组沾Safranin-O和复染色快速绿色。(b) OA的OARSI得分。<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M10"> <mml:msup> <mml:mrow></mml:mrow> <mml:mrow> <mml:mo> ∗<gydF4y2Ba/mml:mo> </mml:mrow> </mml:msup> </mml:math> </inline-formula>从对照组显著差异(<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M11"> <mml:mi> P<gydF4y2Ba/mml:mi> <mml:mo> <<gydF4y2Ba/mml:mo> <mml:mn> 0.05<gydF4y2Ba/mml:mn> </mml:math> </inline-formula>)。规模<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M12"> <mml:mi mathvariant="normal"> b<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> r<gydF4y2Ba/mml:mi> <mml:mo> =<gydF4y2Ba/mml:mo> <mml:mn> 200年<gydF4y2Ba/mml:mn> <mml:mtext> </mml:mtext> <mml:mi> μ<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 米<gydF4y2Ba/mml:mi> </mml:math> </inline-formula>。<gydF4y2Ba/p> <fig id="fig3a"> <label>(一)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.003a"></graphic> </fig> <fig id="fig3b"> <label>(b)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.003b"></graphic> </fig> </fig-group> </sec> <sec id="sec3.4"> <title>3.4。包含IHC分析<gydF4y2Ba/title> <p>的表达MMP-13被包含IHC染色法半定量的分析,使用ACLT样品准备在9周后。包含IHC分析抑制外侧髁关节显示更具体的差异(图<gydF4y2Baxref rid="fig4a" ref-type="fig"> 4(一)<gydF4y2Ba/xref>)。<gydF4y2Ba/p> <fig-group id="fig4"> <label>图4<gydF4y2Ba/label> <p>包含IHC分析MMP-13软骨。从每组(a)一个代表性样本评估MMP-13股骨外侧髁。(b) MMP-13-positive细胞的比例。<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M13"> <mml:msup> <mml:mrow></mml:mrow> <mml:mrow> <mml:mo> ∗<gydF4y2Ba/mml:mo> </mml:mrow> </mml:msup> </mml:math> </inline-formula>从对照组显著差异(<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M14"> <mml:mi> P<gydF4y2Ba/mml:mi> <mml:mo> <<gydF4y2Ba/mml:mo> <mml:mn> 0.05<gydF4y2Ba/mml:mn> </mml:math> </inline-formula>)。<年代up>#<gydF4y2Ba/sup>海藻酸的显著差异microbead集团(<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M15"> <mml:mi> P<gydF4y2Ba/mml:mi> <mml:mo> <<gydF4y2Ba/mml:mo> <mml:mn> 0.05<gydF4y2Ba/mml:mn> </mml:math> </inline-formula>)。<年代up>__<gydF4y2Ba/sup>ASC组的显著差异(<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M16"> <mml:mi> P<gydF4y2Ba/mml:mi> <mml:mo> <<gydF4y2Ba/mml:mo> <mml:mn> 0.05<gydF4y2Ba/mml:mn> </mml:math> </inline-formula>)。规模<我nl我ne-formula> <mml:math xmlns:mml="http://www.w3.org/1998/Math/MathML" id="M17"> <mml:mi mathvariant="normal"> b<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 一个<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> r<gydF4y2Ba/mml:mi> <mml:mo> =<gydF4y2Ba/mml:mo> <mml:mn> One hundred.<gydF4y2Ba/mml:mn> <mml:mtext> </mml:mtext> <mml:mi> μ<gydF4y2Ba/mml:mi> <mml:mi mathvariant="normal"> 米<gydF4y2Ba/mml:mi> </mml:math> </inline-formula>。<gydF4y2Ba/p> <fig id="fig4a"> <label>(一)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.004a"></graphic> </fig> <fig id="fig4b"> <label>(b)<gydF4y2Ba/label> <graphic xlink:href="//www.newsama.com/downloads/journals/sci/2018/2791632.fig.004b"></graphic> </fig> </fig-group> <p>MMP-13-positive细胞的比例显著低于microencapsulated-ASC组(17.2±2.77)比其他组织,包括ASC组(22.6±2.30)。对照组没有显著区别(31±5.14)和海藻酸microbead组(38.17±8.73)(图<gydF4y2Baxref rid="fig4b" ref-type="fig"> 4 (b)<gydF4y2Ba/xref>)。<gydF4y2Ba/p> </sec> </sec> <sec id="sec4"> <title>4所示。讨论<gydF4y2Ba/title> <p>许多策略防止软骨变性和OA已经开发和临床测试。msc nonimmunogenic有效性和强力的免疫抑制活性的报道,和同种异体的msc可以安全使用在OA免疫抑制药物(<gydF4y2Baxref ref-type="bibr" rid="B32"> 32<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B33"> 33<gydF4y2Ba/xref>]。以增强其效果,改善MSC可行性,并延长MSC在严酷的生存微环境,MSC可以结合传统的治疗,如注射透明质酸(<gydF4y2Baxref ref-type="bibr" rid="B34"> 34<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B35"> 35<gydF4y2Ba/xref>]。最近,据报道干细胞消失后第七天关节内注射;定期注射更有效。然而,这种方法需要许多细胞和重复注射(<gydF4y2Baxref ref-type="bibr" rid="B36"> 36<gydF4y2Ba/xref>- - - - - -<gydF4y2Baxref ref-type="bibr" rid="B38"> 38<gydF4y2Ba/xref>]。如果细胞生存能力增加,有可能进一步提高治疗效果,同时减少MSC注射的频率和大小,和这种方法可能更划算。当脚手架和干细胞应用于骨缺损,可以增加治疗效果增加msc的分化潜能(<gydF4y2Baxref ref-type="bibr" rid="B39"> 39<gydF4y2Ba/xref>]。另一方面,这种支架没有被雇佣在关节和应用具有侵略性。最近,据报道,微型胶囊的细胞海藻酸的小说,是一种无创检查方法联合注射[的增加细胞的生存能力<gydF4y2Baxref ref-type="bibr" rid="B21"> 21<gydF4y2Ba/xref>]。增加细胞的生存能力,最重要的是保护细胞注入宿主的免疫反应。保护的方法之一是封装细胞半透系统内(<gydF4y2Baxref ref-type="bibr" rid="B40"> 40<gydF4y2Ba/xref>藻朊酸盐等)。海藻酸是一种天然生物聚合物从褐藻中提取,是生物相容性,相对无毒,廉价的(<gydF4y2Baxref ref-type="bibr" rid="B24"> 24<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B41"> 41<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B42"> 42<gydF4y2Ba/xref>]。<gydF4y2Ba/p> <p>最近报告了对asc发挥旁分泌作用,间接刺激分泌细胞因子和生长因子等生物活性的因素<gydF4y2Baxref ref-type="bibr" rid="B31"> 31日<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B43"> 43<gydF4y2Ba/xref>]。三维gel-sphere脚手架的海藻酸微是一种多孔结构,允许扩散的氧气和营养物质和代谢产物的运输<gydF4y2Baxref ref-type="bibr" rid="B24"> 24<gydF4y2Ba/xref>]。这些海藻酸微磁结构特点可能促进对asc的旁分泌活动中使用的干细胞治疗。<gydF4y2Ba/p> <p>在这项研究中,我们进行了三个定期注射,因为ACLT 2周后,治疗OA。它已经证明了退行性变化可以在4周后观察到早期OA ACLT [<gydF4y2Baxref ref-type="bibr" rid="B44"> 44<gydF4y2Ba/xref>]。在急性OA,我们认为干细胞注射2,3,4周后ACLT不会有直接治疗效果(例如,软骨再生),因为会没有足够先进的关节损伤。不过,如果渐进损伤的关节软骨发生自ACLT 4周后,将有效地抑制对asc OA进展通过旁分泌作用而不是通过关节软骨再生。在其他的研究中,发现了细胞注射到关节生存~ 7 d [<gydF4y2Baxref ref-type="bibr" rid="B36"> 36<gydF4y2Ba/xref>- - - - - -<gydF4y2Baxref ref-type="bibr" rid="B38"> 38<gydF4y2Ba/xref>]。尽管目前的研究没有实验确定关节对asc microencapsulated存活多久,据报道其他皮下注射microencapsulated细胞表现出更多的可行性<我t一个l我c> 在活的有机体内<gydF4y2Ba/italic>比细胞直接注入没有微型胶囊(<gydF4y2Baxref ref-type="bibr" rid="B18"> 18<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B21"> 21<gydF4y2Ba/xref>]。先前的研究的基础上,我们认为海藻酸微型胶囊可能延长生存的干细胞治疗和提高OA关节功效以延长旁分泌活动。<gydF4y2Ba/p> <p>宏观分析ACLT 9周后,因为所有的迹象期间在场的OA ACLT[8 - 12周后<gydF4y2Baxref ref-type="bibr" rid="B44"> 44<gydF4y2Ba/xref>]。观察胫骨平台时,OA轻度或中度病变在这个领域。这一现象发生在ACLT模型兔子,同意一项研究Mero et al。<gydF4y2Baxref ref-type="bibr" rid="B45"> 45<gydF4y2Ba/xref>]。鉴于轻度或中度病变,胫骨高原似乎是一个不合理的指标来评估软骨退化。此外,组间没有差异的总评价。此外,急性病变对股骨髁ACLT具有最大的影响,一些研究也表明,股骨髁软骨退化的是一个很好的指标(<gydF4y2Baxref ref-type="bibr" rid="B44"> 44<gydF4y2Ba/xref>]。由于这个原因,我们专注于股骨髁上病变。<gydF4y2Ba/p> <p>抑制关节中经常使用OA模型。载荷分布和步态力学抑制关节因物种而异。不像其他动物,兔子会被加载到膝关节的横向方面(<gydF4y2Baxref ref-type="bibr" rid="B46"> 46<gydF4y2Ba/xref>]。轻微的关节炎的早期变化ACLT后第4周开始出现的时候,和严重的软骨变性首先发生在股骨外侧髁,其次是股骨内侧髁和半月板(<gydF4y2Baxref ref-type="bibr" rid="B45"> 45<gydF4y2Ba/xref>]。因此,膝关节外侧隔间的兔子是确认的网站早期软骨的变化(<gydF4y2Baxref ref-type="bibr" rid="B47"> 47<gydF4y2Ba/xref>]。在外侧股骨髁软骨变性发生早期,软骨损伤microencapsulated-ASC组显著低于其他组。尽管如此,我们严重评估了半月板,损害已经发生。此外,半月板时评估的评分方法亚当斯et al。<gydF4y2Baxref ref-type="bibr" rid="B48"> 48<gydF4y2Ba/xref>),实验组之间没有显著差异。这一发现表明,早期对asc microencapsulated管理局可能预防或延迟外侧股骨髁软骨损伤,这损害代表早期关节病变。相比之下,对asc microencapsulated并没有阻止或减缓股骨内侧髁和半月板损伤,损伤后发达在OA进展。预计对asc microencapsulated将独自生存超过对asc,但是早期的重复政府不会阻止软骨损伤后长时间管理。因此,额外的管理4周后或缩短收获时间可以防止损伤半月板和股骨内侧髁。<gydF4y2Ba/p> <p>墨汁染色了,有一个优势,可以更直观地确认。对照组没有显著区别,海藻酸microbead集团,这意味着海藻酸微不承担额外antiarthritis效果。我们解释之间的不同结果microencapsulated-ASC组和ASC组作为延迟关节炎发展的象征。各种研究表明,海藻酸微型胶囊能增加干细胞的可行性和延长治疗性细胞因子的分泌<gydF4y2Baxref ref-type="bibr" rid="B49"> 49<gydF4y2Ba/xref>]。因此,宏观评价表明,软骨变性microencapsulated-ASC组显著降低。相比之下,组织学评估显示无显著差异的内侧髁缺陷在所有组中,因为损害的程度。此外,microencapsulated-ASC集团在侧关节表现出显著差异只有在与对照组进行比较。海藻酸盐microbead集团体现大变化的结果。尽管如此,由于海藻酸微生物相容性,这些结果似乎没有由于海藻酸微磁的特性。变化的结果可以认为是由于样本量不足和实验诱导OA的模型的局限性。如果收获时间略早,我们可以观察到OA进展的巨大差异。进一步研究注入时机和频率在急性和慢性OA应该表示对asc microencapsulated如何更有效地应用。<gydF4y2Ba/p> <p>透明软骨损伤的程度可以通过包含IHC染色评估关于upregulation MMP-13,证明在关节软骨基质降解酶(<gydF4y2Baxref ref-type="bibr" rid="B50"> 50<gydF4y2Ba/xref>]。之前报道,透明软骨矩阵由II型胶原和蛋白多糖高分子aggrecan。II型胶原蛋白被OA时,基质降解酶包括金属蛋白酶- 1、MMP-8, MMP-13调节(<gydF4y2Baxref ref-type="bibr" rid="B51"> 51<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B52"> 52<gydF4y2Ba/xref>]。<gydF4y2Ba/p> <p>在这项研究中,确认对asc microencapsulated的影响,我们认为只有通过包含IHC MMP-13表达分析。MMP-13-positive细胞比例显著低于microencapsulated-ASC组相比,所有其他组织。这些结果表明,微型胶囊可能会增加对asc施加的旁分泌作用。化验额外的酶和因素,如collagenase-generated乳沟neoepitope II型胶原蛋白,这被认为是一个敏感的OA生物标志物(<gydF4y2Baxref ref-type="bibr" rid="B53"> 53<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B54"> 54<gydF4y2Ba/xref>),确认可能影响对asc的微型胶囊。<gydF4y2Ba/p> <p>在人类中,治疗关节炎的药物被发现的影响取决于剂量的干细胞注入。高剂量组显示临床、放射学和关节镜的结果比那些来自更有利的组织接受较低或中等剂量(<gydF4y2Baxref ref-type="bibr" rid="B11"> 11<gydF4y2Ba/xref>]。这些数据表明,足够数量的msc的流感传染的网站应该送到最好的结果。细胞剂量的重要性已经被几位作者提出<gydF4y2Baxref ref-type="bibr" rid="B55"> 55<gydF4y2Ba/xref>- - - - - -<gydF4y2Baxref ref-type="bibr" rid="B57"> 57<gydF4y2Ba/xref>]。一些报道称,注入10<年代up>7<gydF4y2Ba/sup>msc导致完全愈合的伤疤在老鼠<gydF4y2Baxref ref-type="bibr" rid="B55"> 55<gydF4y2Ba/xref>),而另一些已经证明,应用msc的数量不足产生低劣的结果(<gydF4y2Baxref ref-type="bibr" rid="B58"> 58<gydF4y2Ba/xref>]。预计海藻酸微型胶囊会增加关节的干细胞的可行性。在重复注射,microencapsulated-ASC组存活细胞的数量应该高于ASC组。这种生存能力可能对asc可能意味着对asc microencapsulated更有效治疗关节炎。<gydF4y2Ba/p> <p>本研究的局限性如下:有必要研究长期软骨保护作用,因为它反映了短期的早期OA软骨保护活动。还有一个需要长期研究潜在的并发症。此外,alginate-microencapsulated细胞的生存能力已经被调查,和体内研究表明,注射alginate-microencapsulated细胞比自由细胞的生存和保留<gydF4y2Baxref ref-type="bibr" rid="B21"> 21<gydF4y2Ba/xref>]。然而,因为目前的研究没有探讨海藻酸的降解动力学微在关节内,有必要研究微胶囊在关节退化时间评估效果在临床应用前的联合。最后,许多类型的codelivery策略目前正在开发和讨论(<gydF4y2Baxref ref-type="bibr" rid="B59"> 59<gydF4y2Ba/xref>,<gydF4y2Baxref ref-type="bibr" rid="B60"> 60<gydF4y2Ba/xref>]。因此,有必要证明与其他codelivery策略相比,藻酸盐微确保更高的生存能力和保留的细胞。<gydF4y2Ba/p> </sec> <sec id="sec5"> <title>5。结论<gydF4y2Ba/title> <p>对asc Microencapsulated放缓OA的恶化和减少它的程度,更对asc比自由。Alginate-based微型胶囊可能增加ASC膝关节内的可行性。据我们所知,本研究首次采用alginate-based脚手架改善ASC-based治疗关节炎的功效。<gydF4y2Ba/p> </sec> <back> <sec> 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