Wnt11 plays an important role in the osteogenesis of human mesenchymal stem cells in a PHA/FN/ALG composite scaffold: possible treatment for infected bone defect.
Wnt11 plays an important role in the osteogenesis of human mesenchymal stem cells in a PHA/FN/ALG composite scaffold: possible treatment for infected bone defect.
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Wnt11 在 PHA/FN/ALG 复合支架中的人间充质干细胞成骨中发挥重要作用：感染性骨缺损的可能治疗方法
DOI：
10.1186/s13287-016-0277-4
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发表时间：
2016-01-27
影响因子：
7.5
通讯作者：
Xu YQ
中科院分区：
文献类型：
--
作者：
Wang H;He XQ;Jin T;Li Y;Fan XY;Wang Y;Xu YQ
由于感染性骨缺损难以治愈，给骨科医生带来了巨大的挑战。以人骨髓间充质干细胞（human mesenchymal stem cells，hMSCs）为基质的组织工程骨（tissue engineering bone），是目前临床上一种很有前途的治疗方案。在先前的研究中，多孔羟基磷灰石/纤维连接蛋白/藻酸盐（PHA/FN/ALG）复合支架作为一种新型支架显示出良好的生物学性能，被认为优于单一材料支架。此外，Wnt 11已被证明在成骨细胞的发育中发挥重要作用，但直到最近，其在感染环境中hMSCs的成骨分化中的作用仍不清楚。本研究采用层层法构建PHA/FN/ALG复合支架。此外，我们还通过慢病毒转导构建了Wnt 11沉默（RNAi）和过表达的hMSCs。通过定量PCR和Western blot分析证实了基因转导效率。以hMSCs和PHA/FN/ALG复合支架构建组织工程骨，植入感染性骨缺损模型，通过定量PCR、大体观察、显微CT和组织学分析评价组织工程骨的成骨能力。所有细胞在支架上的粘附能力和增殖能力相似。组织工程骨植入后，体内有较高水平的全身炎症因子，抗生素治疗后3天明显下降。植入后1 ~ 2个月，成骨相关基因分析、大体观察、显微CT和组织学检查结果一致显示，Wnt 11过表达hMSC组的成骨能力最强，而Wnt 11-RNAi hMSC组的成骨能力较低。而空白对照组和单纯复合支架无细胞植入组均表现出极弱的成骨能力。我们的研究结果表明，Wnt 11基因在hMSCs中起着重要的作用，在感染环境中促进成骨。
Infected bone defect poses a great challenge for orthopedists because it is difficult to cure. Tissue-engineered bone based on the human mesenchymal stem cells (hMSCs), has currently taken a promising treatment protocol in clinical practice. In a previous study, a porous hydroxyapatite/fibronectin/alginate (PHA/FN/ALG) composite scaffold displayed favorable biological properties as a novel scaffold, which was considered better than single-material scaffolds. In addition, Wnt11 has been demonstrated to play an important role in the development of osteoblasts, but until recently, its role in the osteogenic differentiation of hMSCs in infectious environment remained unclear. In this study, we constructed a PHA/FN/ALG composite scaffold with layer-by-layer technology. Furthermore, we also constructed Wnt11-silenced (RNAi) and -overexpressing hMSCs by lentiviral transduction. The gene transduction efficacy was confirmed by quantitative PCR assay and Western blot analysis. Tissue-engineered bone was constructed with hMSCs and PHA/FN/ALG composite scaffolds, and then was implanted into an infected bone defect model for evaluating the osteogenic capacity by quantitative PCR, gross observation, micro-CT and histology analysis. All those cells showed similar adhesion abilities and proliferation capacities in scaffolds. After tissue-engineered bone implantation, there were high levels of systemic inflammatory factors in vivo, which significantly declined three days after antibiotic therapy. One or two months after implantation, the results of osteogenic-related gene analyses, gross observation, micro-CT and histology consistently showed that the Wnt11 over-expression hMSC group displayed the strongest osteogenesis capacity, whereas the Wnt11-RNAi hMSC group displayed inferior osteogenesis capacity, when compared with the other cell-containing groups. However, the blank control group and the only composite scaffold without cell implantation group both showed extremely weak osteogenesis capacity. Our results revealed that the Wnt11 gene plays an important role in hMSCs for enhancing the osteogenesis in an infectious environment.