摘要： 目的：通过制备基质细胞衍生因子（SDF1-α）的Pluronic F-127复合凝胶材料,观察这种复合材料在体外对兔骨髓间充质干细胞（BMSCs）的矿化及成血管作用和复合凝胶材料在兔上颌窦外提升术中的成骨及成血管效果。方法：将兔骨髓间充质干细胞、50ng/ml SDF1-α和质量百分比浓度0.1%的Pluronic F-127复合培养，MTT法检测其细胞毒性，碱性磷酸酶活性测定及茜素红染色。将18只新西兰大白兔双侧进行上颌窦外提升术，双侧植入不同的复合材料（共3种）并进行对照。材料分别为：β-磷酸三钙（β-TCP），生理盐水及SDF-1α复合Pluronic F-127凝胶。于术后4、8、12周处死，X线片观察双侧上颌窦骨质形成情况，行HE染色、CD31+、CD34+、BMP-2及VEGF免疫组织化学染色观察成骨及成血管作用，并进行统计学分析。结果：体外实验采用MTT法证实了0.1% w/w Pluronic F-127无细胞毒性（P>0.05）。BMSCs-SDF1α复合凝胶碱性磷酸酶染色下，细胞质可见大量碱性磷酸酶染色沉淀，其余3组未见沉淀。BMSCs-SDF1α复合凝胶、BMSCs-单纯凝胶、单纯BMSCs在成骨诱导培养条件下均产生矿化结节，非成骨诱导组未见矿化结节形成。体内实验，X线片结果显示BMSCs-SDF1-α复合凝胶及β-TCP充填侧均有阻射影，生理盐水组未见明显阻射影；HE染色结果显示BMSCs-SDF1-α复合凝胶成骨效果与β-TCP成骨效果明显，生理盐水组未见骨质形成，统计学分析表明BMSCs-SDF1-α复合凝胶成血管作用较β-TCP及生理盐水组明显增加。结论：SDF-1α复合Pluronic F-127凝胶能够成功诱导BMSCs分化为成骨细胞，无细胞毒性，在体外及兔上颌窦内成骨、成血管效果明显。

关键词: 基质细胞衍生因子SDF1-α, Pluronic F-127水凝胶, 骨髓间充质干细胞, 成骨诱导分化, 上颌窦外提升术

Abstract: Objective: The aim of this study was to analyze  the effect of mineralization and vascularization of a new composite material: stromal cell-derived factor (SDF1-α) loaded on Pluronic F-127 which is created by ourselves. Methods: We observed the effect of this composite material by two-part of experiment. One underwent in vitro, to assess the effect of the composite material on the BMSCs. The other, in vivo test, we placed the new composite material inside the rabbit’s maxillary sinus for bone augmentation. First, the  BMSCs were cultured with 50 ng/ml SDF1-α and 0.1% w/w Pluronic F-127. Using the thiazolyl blue tetrazolium bromide (MTT) assay testing the biocompatibility, alkaline phosphatase activity assay(AKP) for testing the substance whether the cell can vascularization, and alizarin red staining(ARS) for testing the ability of mineralization. Eighteen New Zealand white rabbits were obtained and bilateral sinus lifting surgery, were performed. Three different composites were used for augmentation:Group A,β- tricalcium phosphateβ-TCP, (Ca3(PO4)2); Group B, SDF-1α composite Pluronic F-127 gel, and Group C, normal saline. 18 New Zealand white rabbits were sacrificed at 4-, 8-, and 12-weeks,  X-ray observation of bilateral maxillary bone formation, HE staining for histological examination, CD31, CD34, BMP-2 and VEGF immunohistochemical staining and statistical analysis were evaluated. Results: MTT assay confirmed the 0.1% w/w Pluronic F-127 has no effect on cell proliferation (P>0.05). BMSCs-SDF1α composite gel under alkaline phosphatase staining revealed a lot of precipitation, and the remaining three groups have no precipitation. BMSCs-SDF1α composite gel, BMSCs-Pluronic F-127, pure BMSCs cultured in osteogenic conditions, both produced mineralized nodules, no mineralization nodule formation for non-osteogenic group. In vivo experiments, X-ray film showed that both BMSCs-SDF1-α composite gel and β-TCP filling side have the radiopacity images, however, normal saline group doesn’t. HE staining showed osteogenic effect of BMSCs-SDF1α composite gel and β-TCP were significant, but no bone formation in saline group. In the meantime, BMSCs-SDF1α composite gel had promoted significant vascularization differentiation compared to β-TCP and physiological saline group. Conclusion: Our findings demonstrate that the SDF-1α composite Pluronic F-127 gels cultured with BMSCs exhibited significant osteogenic potential in rabbit maxillary sinus augmentation.

Key words: stromal cell derived factor SDF1-α, Pluronic F-127 hydrogel, bone marrow mesenchymal stem cells, osteogenic differentiation, sinus augmentation

中图分类号: 

• R782.1