调节骨髓间充质干细胞向皮肤创面迁移过程中的血小板衍生生长因子

背景：血小板衍生生长因子具有创伤修复作用,对其研究大部分集中于骨组织的修复,在皮肤创伤愈合中的修复作用研究较少。 目的：观察血小板衍生生长因子在皮肤创伤愈合过程中调控骨髓间充质干细胞向创面迁移促进创面愈合的作用。 方法：培养大鼠骨髓间充质干细胞,免疫荧光检测细胞表面标记物CD34、CD44。选取健康雄性SD大鼠30只,随机分为5组,各组大鼠尾静脉注射PKH26标记的骨髓间充质干细胞。注射1周后,于大鼠背部正中线划长为3 cm的切口,制备皮肤创伤模型。造模后即刻于皮肤创伤处多点注射不同质量浓度血小板衍生生长因子干预药物,对照组注射等体积的生理盐水。注射14 d后留取皮肤创面组织进行相关指标检测。 结果与结论：荧光显微镜下观察血小板衍生生长因子能够剂量依赖性诱导骨髓间充质干细胞向皮肤创伤组织处迁移和聚集,进而促进皮肤创伤修复。Masson染色结果显示,随着血小板衍生生长因子干预质量浓度的增加,创面炎性细胞浸润减轻、胶原纤维数量不断增多。Western blot检测结果显示,血小板衍生生长因子能够抑制皮肤创伤组织基质金属蛋白酶1的表达,促进基质金属蛋白酶组织抑制因子1的表达,抑制胶原降解,发挥间接促愈合作用。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

基质细胞衍生因子1α预处理大鼠骨髓间充质干细胞的迁移

背景：骨髓间充质干细胞移植过程中只有少量细胞能定向迁移到损伤组织,因此如何提高细胞定向迁移的数量是干细胞移植的关键因素。 目的：观察基质细胞衍生因子1α预处理对大鼠骨髓间充质干细胞定向迁移的影响。 方法：体外分离培养骨髓间充质干细胞,予以传代。使用Transwell体外迁移体系,观察不同浓度的基质细胞衍生因子1α趋化骨髓间充质干细胞定向迁移,选取最佳浓度值趋化细胞。在基质细胞衍生因子1α预处理、CXCR 4型受体阻断剂AMD3100和Akt通路阻断剂LY294002的干预下观察骨髓间充质干细胞的趋化迁移情况,检测基质细胞衍生因子1α预处理对骨髓间充质干细胞中CXCR 4型受体 mRNA、蛋白水平及AKT磷酸化的影响。 结果与结论：0.2 mg/L的基质细胞衍生因子1α孵育细胞10 h具有最佳趋化细胞迁移效应。骨髓间充质干细胞的定向趋化迁移作用随着基质细胞衍生因子1α预处理细胞浓度的增加而增强,预处理时间为6 h;而AMD3100和LY294002可阻断基质细胞衍生因子1α预处理的促迁移作用;基质细胞衍生因子1α预处理可上调骨髓间充质干细胞CXCR 4型受体的表达并促进Akt蛋白磷酸化。提示基质细胞衍生因子1α预处理可通过增加骨髓间充质干细胞表面的CXCR 4型受体数量,从而增强基质细胞衍生因子1α/CXCR4型受体介导的骨髓间充质干细胞定向迁移,该预处理效应可能与Akt信号途径有关。     

山羊骨髓间充质干细胞向纤维软骨分化的形态学改变

背景：前期初步研究发现,碱性成纤维细胞生长因子可诱导骨髓间充质干细胞向颞下颌关节盘细胞方向分化,且碱性成纤维细胞生长因子 10 μg/L诱导组合成胶原量明显高于5 μg/L诱导组。目的：观察经不同浓度碱性成纤维细胞生长因子诱导后的骨髓间充质干细胞的超微结构变化。方法：原代分离培养山羊骨髓间充质干细胞,选P3,P4细胞,用5,10 μg/L碱性成纤维细胞生长因子诱导骨髓间充质干细胞,以未加碱性成纤维细胞生长因子培养的骨髓间充质干细胞做对照。倒置相差显微镜观察细胞生长状况,用第7,14,21天的细胞爬片行蕃红O、天狼猩红和Ⅰ型胶原免疫组织化学染色,并观察第21天细胞的超微结构。结果与结论：经不同浓度碱性成纤维细胞生长因子诱导后,骨髓间充质干细胞可向颞下颌关节盘成纤维细胞样细胞形态分化,10 μg/L组细胞更像关节盘成纤维细胞样细胞。提示骨髓间充质干细胞有向颞下颌关节盘细胞方向分化的形态学基础。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

基质细胞衍生因子1预处理抑制大鼠骨髓间充质干细胞的凋亡

背景：研究发现基质细胞衍生因子1除参与趋化干细胞定向迁移途径,还具有抗凋亡作用。 目的：观察基质细胞衍生因子1预处理后对骨髓间充质干细胞凋亡的影响。 方法：以不同浓度H₂O₂诱导大鼠骨髓间充质干细胞凋亡,取最适宜浓度100 μmol/L用于实验。不同质量浓度基质细胞衍生因子1干预100 μmol/L H₂O₂诱导后的大鼠骨髓间充质干细胞,选择0.2 mg/L最佳保护质量浓度用于实验。取第3代大鼠骨髓间充质干细胞,随机分组：正常对照组不进行任何处理;损伤组在培养液中加入H₂O₂作用24 h;基质细胞衍生因子1预处理组于H₂O₂损伤细胞前6 h加入基质细胞衍生因子1;基质细胞衍生因子1+AMD3100(基质细胞衍生因子1受体CXCR4的阻断剂)组于H₂O₂细胞损伤前6 h加入基质细胞衍生因子1与AMD3100共孵。 结果与结论：H2O2能体外模拟缺血缺氧环境诱导骨髓间充质干细胞凋亡,且作用呈剂量依赖性。与损伤组比较,加入基质细胞衍生因子1预处理后细胞凋亡明显减轻(P < 0.01),细胞E2F6基因表达增强(P < 0.05),E2F1基因表达减少(P < 0.05),线粒体细胞色素C转位减少(P < 0.05),Caspase-3活性降低(P < 0.05),AMD3100可阻断基质细胞衍生因子1对骨髓间充质干细胞的保护作用。提示基质细胞衍生因子1可能通过增强E2F6基因,负性调控E2F1基因抑制线粒体损伤导致的骨髓间充质干细胞凋亡。     

前列腺素E1对大鼠骨髓间充质干细胞增殖的影响

背景：目前还缺乏前列腺素E1对骨髓间充质干细胞增殖影响的研究。 目的：观察前列腺素E1与骨髓间充质干细胞共培养对细胞增殖作用的影响。 方法：体外培养SD大鼠骨髓间充质干细胞。取第3 代细胞利用流式细胞仪鉴定细胞表面表型。以每孔1×104数量接种于96 孔板,将前列腺素E1分别以0(对照组),1,2,5,10,20,40,100 μg/L的浓度作用骨髓间充质干细胞72 h,以CCK-8法检测细胞增殖情况,观察最佳增殖浓度10%的前列腺素E1在不同时间(24,48,72 h)对骨髓间充质干细胞增殖的影响。 结果与结论：P3代骨髓间充质干细胞表面阳性率CD11b/c为4.93%,CD29为99.93%,CD45为0.1%,CD90为99.58%,符合骨髓间充质干细胞特征。质量浓度为1,2,5,10,20,40,100 μg/L的前列腺素E1均可以促进骨髓间充质干细胞在体外的增殖,以10 μg/L的作用最强。10 μg/L前列腺素E1在作用48 h后能显著促进大鼠骨髓间充质干细胞增殖(P < 0.05),其作用呈一定程度的时间依赖性,72 h达最高。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

长期培养对基质细胞衍生因子1/CXCR4介导间充质干细胞迁移的影响**◆

背景：人外源性的间充质干细胞能特异性地向损伤部位迁移,参与多种组织的损伤修复,然而其定向迁移的机制尚不清楚。 目的：观察基质细胞衍生因子1对骨髓间充质干细胞定向迁移的影响。 方法：体外培养不同个体的骨髓间充质干细胞,培养至第10代,检测细胞衰老状态。RT-PCR和细胞免疫荧光检测骨髓间充质干细胞CXCR4受体表达情况;体外迁移体系(Transwell)检测基质细胞衍生因子1对骨髓间充质干细胞迁移的影响。 结果与结论：骨髓间充质干细胞在体外长期增殖后生长逐渐缓慢,在第6,7代衰老细胞增多,其CXCR4受体表达减少。基质细胞衍生因子1对骨髓间充质干细胞的趋化作用呈剂量依赖性。 关键词：趋化作用;骨髓间充质干细胞;细胞衰老;基质细胞衍生因子1;CXCR4受体 doi:10.3969/j.issn.1673-8225.2012.01.013      

软骨损伤后基质细胞衍生因子1的表达

背景：研究表明基质细胞衍生因子1对骨髓间充质干细胞的迁移、聚集有影响。 目的：观察软骨损伤后不同时间损伤修复区组织基质细胞衍生因子1的表达以及与骨髓间充质干细胞迁移的关系。 方法：建立兔软骨损伤模型,分别于建模后2,5,7,10,14,28 d取损伤灶及边缘区组织,检测基质细胞衍生因子1表达和体外细胞迁移实验,观察基质细胞衍生因子1对骨髓间充质干细胞、软骨细胞的迁移影响。 结果与结论：基质细胞衍生因子1的表达呈现出时间变化的趋势,软骨损伤后第7天达到高峰(P < 0.05)。体内移植的骨髓间充质干细胞主要聚集在软骨损伤灶周围,但在封闭CXC趋化因子受体4后,此聚集现象逐渐减弱(P < 0.05)。结果证实,局部组织中基质细胞衍生因子1的表达在软骨损伤早期明显升高,对骨髓间充质干细胞向软骨损伤修复区迁移有重要作用。     

转化生长因子β1对骨髓干细胞分化类髓核细胞的影响

背景：转化生长因子β1是骨髓间充质干细胞分化为类髓核细胞的首选生长因子,适当浓度能诱导骨髓间充质干细胞的增殖和分化。 目的：观察不同质量浓度转化生长因子β1对大鼠骨髓间充质干细胞体外向类髓核细胞分化的影响,优化骨髓间充质干细胞体外培养条件。 方法：取成年大鼠股骨骨髓,体外培养、纯化。取第3代成年大鼠骨髓间充质干细胞,实验组用加入不同质量浓度转化生长因子β1(0,1,10,20 μg/L)的HG-DMEM无血清培养液诱导3,7,14,21 d;对照组普通状态下用含体积分数为10%胎牛血清的HG-DMEM培养液自然分化。 结果与结论：10 μg/L转化生长因子β1诱导液诱导的骨髓间充质干细胞蛋白聚糖与Ⅱ型胶原蛋白水平明显高于其他实验组和对照组(P < 0.01)。各实验组14 d时蛋白聚糖的表达均较3,7,21 d时高(P < 0.01)。对照组各时间点蛋白聚糖Aggrecan及Ⅱ型胶原表达均呈阴性。提示10 μg/L的转化生长因子β1能提高大鼠骨髓间充质干细胞体外诱导分化类髓核的数量,从而使骨髓间充质干细胞发挥更高的治疗效率。     

芍药苷对骨髓间充质干细胞增殖的影响

背景：研究显示芍药苷具有补血及治疗自身免疫性疾病的功效,骨髓间充质干细胞对机体的造血及免疫功能也起着重要的作用,但芍药苷对骨髓间充质干细胞的增殖及细胞因子的分泌和表达有何影响报道较少。 目的：探讨芍药苷对人骨髓间充质干细胞增殖及白细胞介素6表达的影响。 方法：采用密度梯度离心法和贴壁培养法体外分离培养人骨髓间充质干细胞,用流式细胞术和成脂及成骨诱导法鉴定人骨髓间充质干细胞生物学特性,MTT法检测不同浓度芍药苷对人骨髓间充质干细胞增殖的影响,ELISA 法测定芍药苷干预人骨髓间充质干细胞后培养上清液中白细胞介素6的分泌水平,RT-PCR 检测芍药苷干预后白细胞介素6 mRNA的表达情况。 结果与结论：成功分离出骨髓间充质干细胞,具有成骨、成脂分化潜能。与对照组相比,芍药苷浓度为2 μmol/L和10 μmol/L可明显促进骨髓间充质干细胞增殖。10 μmol/L芍药苷干预骨髓间充质干细胞后,G₀/G₁期细胞比例显著降低,S期细胞比例显著升高。10 μmol/L芍药苷干预组骨髓间充质干细胞白细胞介素6的分泌和mRNA表达均显著高于对照组(P < 0.01)。由此得出,一定浓度的芍药苷可促进骨髓间充质干细胞增殖,并提高骨髓间充质干细胞分泌白细胞介素6水平和基因表达。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

小鼠骨髓间充质干细胞体外成肌分化中基质金属蛋白酶1的作用

背景：小鼠骨髓间充质干细胞体外成肌诱导分化效率较低。 目的：观察基质金属蛋白酶1在体外对小鼠骨髓间充质干细胞成肌分化的作用。 方法：利用差速贴壁法体外分离培养小鼠骨髓间充质干细胞并进行鉴定后,按照不同基质金属蛋白酶1药物处理浓度将小鼠骨髓间充质干细胞分成4组,即10 μg/L组、1 μg/L组、0.1 μg/L组、对照组,给予相应处理后,Real-time定量PCR检测MyoD、Desmin mRNA表达,Western Blotting检测Desmin蛋白表达。 结果与结论：利用差速贴壁法分离培养的小鼠骨髓间充质干细胞形态较一致,表面分子检测示CD29⁺、Sca-1⁺、CD45^-、CD34^-,并具有多向分化潜能;经基质金属蛋白酶1处理后,Real-time定量PCR检测示Desmin、MyoD mRNA表达上调,Western Blotting检测示Desmin蛋白表达上调,并且以上各成肌相关基因、蛋白表达增高程度与基质金属蛋白酶1具有浓度依赖性。提示基质金属蛋白酶1在体外可促进骨髓间充质干细胞向肌肉细胞分化。     

骨髓间充质干细胞分泌基质细胞衍生因子1保护心肌细胞

背景：有研究显示表达CXCR4的干细胞能够沿着基质细胞衍生因子1的浓度梯度迁移到心肌梗死部位再生心肌和血管而改善心脏的功能。 目的：探索间充质干细胞通过其分泌的基质细胞衍生因子1对心肌细胞的保护作用。 方法：收集培养2 d的间充质干细胞条件培养基。在缺氧条件,利用基质细胞衍生因子1受体CXCR4阻断剂AMD3100或PI3-K/Akt途径阻断剂LY294002预处理H9C2细胞后,利用AnnexinV/PI双标法流式细胞术分析间充质干细胞条件培养基作用下H9C2细胞凋亡的变化;Western blotting分析H9C2细胞磷酸化Akt蛋白的表达;RT-PCR分析间充质干细胞基质细胞衍生因子1的表达。 结果与结论：RT-PCR结果显示间充质干细胞表达基质细胞衍生因子1,Western blotting结果显示间充质干细胞条件培养基增加了H9C2细胞磷酸化Akt蛋白的水平。AnnexinV/PI分析发现间充质干细胞条件培养基明显降低了H9C2细胞缺氧复氧后的凋亡,且这种抗凋亡作用能被CXCR4阻断剂AMD3100或PI3-K/Akt途径阻断剂LY294002所阻断。说明间充质干细胞通过其分泌的基质细胞衍生因子1通过激活PI3-K/Akt途径保护H9C2细胞,增加H9C2细胞的幸存能力。     

骨髓间充质干细胞定向趋化及骨修复中基质细胞衍生因子1的作用

背景：骨折愈合与聚集在骨折端的骨髓间充质干细胞的数量及功能密切相关,基质细胞衍生因子1可增强骨髓间充质干细胞的趋化功能。 目的：采用携带绿色荧光蛋白转基因骨髓间充质干细胞的骨髓嵌合体小鼠,制作左胫骨骨折模型,观察基质细胞衍生因子1对骨髓间充质细胞定向迁移的影响及其骨折修复中的作用,并初步探讨其作用机制。 方法：利用密度梯度离心法从携带绿色荧光蛋白转基因小鼠C57BL的骨髓内分离培养出携带绿色荧光蛋白的骨髓间充质干细胞;将经X射线照射后携带绿色荧光蛋白转基因的骨髓间充质干细胞与雄性小鼠的骨髓非贴壁细胞联合移植,最后建立起稳定的骨髓嵌合型小鼠模型,再建立左胫骨骨折模型。建模后分别用基质细胞衍生因子1和基质细胞衍生因子1抗体干预,并设置对照组。 结果与结论：建模后第1,3,7,14天各相应时相点骨折端的骨髓间充质干细胞数量,建模后第14,21天各相应时相点的骨痂量,建模后第28天抗折力,均为基质细胞衍生因子1组>对照组>基质细胞衍生因子1抗体组(P < 0.05);在建模后第28天基质细胞衍生因子1组骨痂量减少(P < 0.05),骨小梁融合成片,部分骨髓腔再通,对照组和基质细胞衍生因子1抗体组髓腔未通。证实,基质细胞衍生因子1 可促进骨髓间充质干细胞向骨折端迁移,具有促进骨折愈合的作用。     

Contribution of bone marrow-derived cells to skin: collagen deposition and wound repair

The bone marrow provides inflammatory cells and endothelial progenitor cells to healing cutaneous wounds. To further explore the bone marrow contribution to skin and healing wounds, we used a chimeric mouse model in which the bone marrow from enhanced green fluorescent protein (EGFP) transgenic mice is transplanted into normal C57BL mice. We found that normal skin is a target organ for bone marrow-derived cells from both the hematopoietic and the mesenchymal stem cell pool. We present evidence that the bone marrow contribution to normal skin and the healing cutaneous wound is substantially greater than the previously recognized CD45+ subpopulation, where 15%-20% of the spindle-shaped dermal fibroblasts were bone marrow-derived (EGFP+). Furthermore, the bone marrow-derived cells were able to contract a collagen matrix and transcribe both collagen types I and III, whereas the skin-resident cells transcribed only collagen type I. Whereas endothelial progenitor cells were found early during the wound repair process, bone marrow-derived endothelial cells were not seen after epithelialization was complete. Our data show that wound healing involves local cutaneous cells for reconstituting the epidermis but distant bone marrow-derived cells and the adjacent uninjured dermal mesenchymal cells for reconstituting the dermal fibroblast population.     

碱性成纤维细胞生长因子及胰岛素样生长因子影响骨髓间充质干细胞增殖及胶原合成

BACKGROUND: How to control the orderly formation of collage in skin repair and scarring process is worthy of attention. OBJECTIVE: To investigate the effect of basic fibroblast growth factor (bFGF) combined with insulin-like growth factor 1 (IGF-1) on the proliferation and collagen synthesis of rat bone marrow mesenchymal stem cells in vitro. METHODS: Rat bone marrow mesenchymal stem cells were isolated and cultured to induce adipogenic differentiation assessed by oil red O staining and osteogenic differentiation identified by alizarin red staining in vitro. Passage 3 cells were cultured in the medium containing bFGF, IGF-1, combination of them or the control fluid, respectively. MTT assay was used to detect cell proliferation at 12, 24, 48, 72 and 96 hours of culture. The expression of type I collagen and type III collagen were detected by RT-PCR and western blot after 10 days of incubation. RESULTS AND CONCLUSION: Compared with the control group, bFGF or IGF-1 alone significantly promoted the proliferation of bone marrow mesenchymal stem cells, and inhibited the expression of type I collagen and type III collagen. After combined use of bFGF and IGF-1, the proliferation of bone marrow mesenchymal stem cells was improved more significantly, and the expression of type I collagen and type III collagen returned to normal levels. These findings indicate that the combination of IGF-1 and bFGF can promote proliferation of bone marrow mesenchymal stem cells and restrain the expression of type I collagen and type III collagen, which may be helpful for control and repair of scar formation during wound healing.      

内源性骨髓间充质干细胞与骨折微环境中的相关趋化因子

背景：研究表明,骨髓间充质干细胞定向迁移依赖于损伤局部表达趋化因子与细胞表面相应受体的相互作用。然而,哪些趋化因子在介导骨髓间充质干细胞向骨折部位定向迁移过程中起关键作用,目前尚不清楚。 目的：对内源性骨髓间充质干细胞进行示踪,观察其在骨修复中的作用;检测并筛选出骨折微环境中表达量高且与骨髓间充质干细胞趋化规律相关的因子。 方法：建立C57BL/6小鼠荧光/普通骨髓嵌合体,利用嵌合体动物模型制造胫骨骨折模型,在不同时相点检测骨折部位组织绿色荧光蛋白阳性细胞占所有细胞的比例、来源于骨髓间充质干细胞的成骨细胞占全部成骨细胞的比例,判断来源于骨髓的骨髓间充质干细胞在骨折修复中的作用;利用免疫组化等方法检测骨折部位不同时相点趋化因子的蛋白表达水平。 结果与结论：骨折局部绿色荧光蛋白阳性细胞占所有细胞比例在术后1,5,14 d分别为(3.011±0.911)%,(9.031±0.145)%,(12.064±0.145)%;来源于骨髓间充质干细胞的成骨细胞占全部成骨细胞的50%以上;骨折后各时相点微环境中基质细胞衍生因子1、集落刺激因子、肝细胞生长因子、单核细胞趋化蛋白1、间质金属蛋白酶9有不同程度的表达,粒细胞集落刺激因子无明显表达。基质细胞衍生因子1的表达量相对最高。经相关分析认为：基质细胞衍生因子1在骨折微环境中的表达与骨髓间充质干细胞趋化规律具有相关关系。结果表明骨髓内的骨髓间充质干细胞参与骨折修复并在其中起到了重要作用;基质细胞衍生因子1促进骨髓间充质干细胞的定向趋化并参与骨组织的修复。     

Enhanced wound healing by topical administration of mesenchymal stem cells transfected with stromal cell-derived factor-1

The objective of this study was to investigate the ability of mesenchymal stem cells (MSC) genetically engineered with stromal cell-derived factor-1 (SDF-1) to heal skin wounds. When transfected with SDF-1 plasmid DNA, MSC which were isolated from the bone marrow of rats, secreted SDF-1 for 7 days. In vitro cell migration assay revealed that the SDF-1-engineered MSC (SDF-MSC) enhanced the migration of MSC and dermal fibroblasts to a significantly greater extent than MSC. The SDF-MSC secreted vascular endothelial growth factor, hepatocyte growth factor, and interleukin 6 at a significantly high level. A skin defect model of rats was prepared and MSC and SDF-MSC were applied to the wound to evaluate wound healing in terms of wound size and histological examinations. The wound size decreased significantly faster with SDF-MSC treatment than with MSC and PBS treatments. The length of the neoepithelium and the number of blood vessels newly formed were significantly larger. A cell-tracing experiment with fluorescently labeled cells demonstrated that the percent survival of SDF-MSC in the tissue treated was significantly high compared with that of MSC. It was concluded that SDF-1 genetic engineering is a promising way to promote the wound healing activity of MSC for a skin defect.     

小分子药物Kartogenin诱导骨髓间充质干细胞定向分化中基质金属蛋白酶2的表达

BACKGROUND:Kartogenin can induce chondrogenic differentiation of mesenchymal stem cells as reported in in vitro experiments. The discovery of Kartogenin finds a novel path to cartilage repair, and it is expected to develop into a new drug to treat osteoarthritis. OBJECTIVE:To observe the inductive role of Kartogenin in the process of human bone marrow mesenchymal stem cells differentiating into chondrocytes in vitro. METHODS:In vitro cultured human bone marrow mesenchymal stem cells were grown to the logarithmic phase, and then divided into control group (0 μmol/L Kartogenin), 1 μmol/L Kartogenin group, and 10 μmol/L Kartogenin group. After 72 hours of culture, cell proliferation and differentiation were observed microscopically. Matrix metalloproteinase 2 and type II collagen levels in the cell supernatant were detected by enzyme linked immunosorbent assay and immunofluorescence staining, respectively. RESULTS AND CONCLUSION:Under the microscope, Kartogenin was shown to significantly promote the proliferation and differentiation of human bone marrow mesenchymal stem cells. With the increase of Kartogenin concentrations, the level of type II collagen was increased, while the level of matrix metalloproteinase 2 was decreased. These findings indicate that Kartogenin can induce human bone marrow mesenchymal stem cells to differentiate into chondrocytes, and with the increase of Kartogenin concentration, destruction of the cartilage extracellular matrix may be inhibited.     

骨形态发生蛋白2抑制骨髓间充质干细胞成脂分化的优选浓度

BACKGROUND:Bone morphogenetic protein 2 cannot only promote osteogenesis, but also at different concentration ranges regulate the differentiation of bone marrow mesenchymal stem cells into adipocytes. Therefore, it is very important for the prevention and treatment of osteoporosis, if the range of concentrations of bone morphogenetic protein 2 can be found to regulate the differentiation of bone marrow mesenchymal stem cells into adipocytes, after the induction of bone marrow mesenchymal stem cells. OBJECTIVE:To determine the optimal concentration of bone morphogenetic protein 2 to inhibit the differentiation of bone marrow mesenchymal stem cells into adipocytes. METHODS:Bone marrow samples were obtained from the femur and tibia of rats. Primary cultured bone marrow mesenchymal stem cells were obtained by adherent culture method in vitro. Flow cytometry was used to identify the surface antigens of passage 2 bone marrow mesenchymal stem cells. Cells at passages 2 and 3 were taken and cultured in adipogenic induction medium for 3 days followed by the addition of 0.5, 5, 50, 100, 200 μg/L bone morphogenetic protein 2, or culture medium of bone marrow mesenchymal stem cells (control group), respectively. RESULTS AND CONCLUSION: Bone marrow mesenchymal stem cells cultured were identified by the flow cytometry. Oil red O staining results showed that the increasing number of lipid droplets was shown in bone marrow mesenchymal stem cells induced by bone morphogenetic protein 2 under the increasing concentration of 0.5 to 50 μg/L; while the number of lipid droplets in bone marrow mesenchymal stem cells decreased after 17 days of induction with bone morphogenetic protein 2 under the concentration of 100 to 200 μg/L. These findings indicate that 100 μg/L bone morphogenetic protein 2 can inhibit adipogenic differentiation of bone marrow mesenchymal stem cell, so as to reduce the formation of adipocytes in rats. Therefore, 100 μg/L bone morphogenetic protein 2 is better to inhibit adipogenic differentiation of bone marrow mesenchymal stem cells.