摘要：背景:小分子药物kartogenin是Smad4/Smad5通路激活剂,拥有极强的促骨分化能力,可促进骨髓间充质干细胞定向分化为成骨细胞,但其药物效果较为局限。目的:设计并研制kartogenin/Pluronic F127胶束,观察其对骨髓间充质干细胞定向成骨分化的作用。方法:将小分子药物kartogenin溶于二甲基亚砜中,然后与Pluronic F127溶液混合均匀,通过真空旋转蒸发仪去除有机溶剂购形成一干燥的药脂膜,加入PBS制备为kartogenin/Pluronic F127胶束。取对数生长期的大鼠骨髓间充质干细胞,分4组处理,分别加入常规细胞培养液(空白组)、含Pluronic F127胶束溶液的细胞培养液(Pluronic F127组)、含kartogenin的细胞培养液(药物组)、含kartogenin/Pluronic F127胶束溶液的细胞培养液(实验组),其中药物组与实验组中药物浓度均为1μmol/L,Pluronic F127组与实验组胶束浓度一致。检测细胞增殖、凋亡及成骨相关蛋白表达。结果与结论:①透射电镜显示kartogenin/Pluronic F127胶束呈现出不规则的球状形态,粒径分布较为均匀,粒径范围为32.6-118.9 nm,多集中在77.3 nm;②MTT检测显示,随着培养时间的延长,4组细胞存活率下降,但均保持在90%以上,4组间细胞生存率比较差异无显著性意义(P>0.05);③处理24 h后的流式细胞仪检测显示,4组间的细胞凋亡率比较差异无显著性意义(P>0.05);④处理24 h后的Western blot检测显示,药物组、实验组的骨桥蛋白、碱性磷酸酶、骨形态发生蛋白2及基质金属蛋白酶2表达均高于Pluronic F127胶束组、空白组(P<0.05),实验组的碱性磷酸酶、骨形态发生蛋白2及基质金属蛋白酶2表达高于药物组(P<0.05);⑤结果表明,kartogenin/Pluronic F127胶束可促进骨髓间充干细胞的成骨分化。

摘要：Abstract Meniscus injury is a common disease in *** it was not treated in time,it leads to osteoarthritis which brings unbearable pain and heavy economic burden to the *** present,meniscectomy and meniscus suture are widely used in the treatment for meniscus ***,It is not ideal for poor self-healing ability of *** recruitment of endogenous stem cells is an attractive option for wounded meniscus *** reduced high-mobility group box 1 protein(HMGB1)can accelerate the regeneration of multiple tissues by endogenous stem cell activation,migration and ***(KGN)has shown to induce the chondrogenesis of the stem ***,no study has explored such effects of HMGB1 and KGN in wounded meniscus ***,in order to improve the regeneration of meniscus,we intend to use a novel bioactive microsphere which was developed by combining fully reduced high mobility group box 1(frHMGB1)and kartogenin(KGN)with alginate gel which slowly release high concentrations of HMGB1 and KGN to activate rat bone marrow stem cells(BMSCs)and promote cell *** results showed that this HMGB1–KGN microsphere released and kept high concentrations of HMGB1 and KGN in the wound area for more than 2 *** vitro experimental results showed that the HMGB1–KGN microsphere can promote cell proliferation via recruiting rat bone marrow stem cells(BMSCs)and activating the BMSCs from G_(0) to G_(Alert) stage as evidenced by cell migration testing and 5-bromo-2′-deoxyuridine(BrdU)incorporation *** vivo results indicated that this HMGB-KGN microsphere can recruit GFP-labeled BMSCs from tail vein to wounded meniscus and induce these GFP-labeled BMSCs to differentiate into *** results demonstrated that the HMGB1–KGN-containing bioactive microsphere induced cell migration in vitro and recruited the cells to wound area to promote wounded rat meniscus healing in vivo.