背景:生物活性物质Biodentine具有良好的抗压强度、粘接强度及较少的微渗漏等诸多优点,已被成功应用于多种临床适应证,然而有关Biodentine是否可促进成骨细胞成骨的研究罕见。目的:研究不同浓度Biodentine对人骨肉瘤细胞MG-63增殖及成...背景:生物活性物质Biodentine具有良好的抗压强度、粘接强度及较少的微渗漏等诸多优点,已被成功应用于多种临床适应证,然而有关Biodentine是否可促进成骨细胞成骨的研究罕见。目的:研究不同浓度Biodentine对人骨肉瘤细胞MG-63增殖及成骨作用的影响。方法:制备不同浓度梯度(1、1/2、1/4、1/8和1/16)的Biodentine浸提液。将人骨肉瘤细胞MG-63分6组培养,分别加入MEM培养液(对照组)及5种浓度的Biodentine浸提液,采用CCK-8法检测培养第1,3,5,7天的细胞增殖,筛选Biodentine浸提液最佳浓度。将人骨肉瘤细胞MG-63分2组培养,分别加入MEM培养液(空白对照组)及最佳浓度的Biodentine材料浸提液(实验组),培养第1,3,5,7天,采用Real-time PCR法检测细胞中成骨因子Runx2 m RNA表达;培养第10,14天行茜素红染色,观察细胞钙化结节形成情况。结果与结论:(1)培养第1,3天时,不同浓度Biodentine材料浸提液组活细胞数量与对照组相比无差异;培养第5天,1浓度材料浸提液组活细胞数量明显低于对照组(P<0.05),1/2、1/4、1/8浓度材料浸提液组活细胞数量与对照组比较无差异,1/16浓度材料浸提液组活细胞数量明显高于对照组(P<0.05),因此实验选择1/16浓度材料浸提液进行后续实验;(2)实验组培养第1,3,5,7天的Runx2 m RNA表达量分别为空白对照组的1.14,5.29,1.08,2.11倍(P均<0.05);(3)培养第10天时,实验组与空白对照组均未见矿化结节;培养第14天,两组均可见矿化结节,实验组的面积更大、颜色更深;(4)结果表明在一定浓度下,Biodentine可促进人骨肉瘤细胞MG-63增殖及成骨活性。展开更多
Background:Mesenchymal stem or stromal cells(MSCs)derived from the induced pluripotent stem cells(iPSCs)have uniform biological activity,which makes the clinical application of MSCs in bone repair possible.Culturing t...Background:Mesenchymal stem or stromal cells(MSCs)derived from the induced pluripotent stem cells(iPSCs)have uniform biological activity,which makes the clinical application of MSCs in bone repair possible.Culturing the iPSC-MSCs onto osteoconductive materials is a promising tissue engineering-based strategy in bone regeneration.The aim of this work was to evaluate the effects of semaphorin 3A(Sema3A)and hypoxia inducible factor 1 subunit alpha(HIF1α)co-overexpression on the survival and osteogenic differentiation of iPSC-MSCs.Methods:Sema3A and HIF1αwere linked together with the three(GGGGS;G,glycine;S,serine)peptide fragment,and their co-expression in iPSC-MSCs was mediated by a lentiviral vector.The fusion protein retained the immune reactivity for both Sema3A and HIF1αas determined with Western blotting.iPSC-MSCs were infected with overexpression lentivirus(oeLenti)as negative control,oeLenti-Sema3A,oeLenti-HIF1αor oeLenti-Sema3A-HIF1αlentiviruses.Results:Sema3A overexpression alone promoted the osteogenic differentiation of iPSC-MSCs(the activity and/or expression of osteoblast markers,such as alkaline phosphatase,osteopontin,and osteocalcin,were upregulated),and suppressed cell survival.The Sema3A-HIF1αfusion protein showed a comparable osteoconductive effect to that of Sema3A without reducing cell survival.We further seeded iPSC-MSCs modified by SemaA-HIF1αoverexpression onto hydroxyapatite(HA)scaffolds,and evaluated their growth and differentiation on this three-dimensional material.Additional data indicated that,as compared to iPSC-MSCs cultured in ordinary two-dimensional dishes,cells cultured in HA scaffolds grew(blank vs.HA scaffolds:0.83 vs.1.39 for survival)and differentiated better(blank vs.HA scaffolds:11.29 vs.16.62 for alkaline phosphatase activity).Conclusion:Modifying iPSC-MSCs with pro-osteogenic(Sema3A)and pro-survival(HIF1α)factors may represent a promising strategy to optimize tissue engineering-based strategy in bone repair.展开更多
Objective: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor,...Objective: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor, type II(BMPR2), E1 A binding protein p300(EP300), transforming growth factor-β2(TGFβ2), and tumor necrosis factor, and alpha-induced protein 3(TNFAIP3) gene expression patterns in B-cell malignancies were studied. Methods: The relative expression levels of BMPR2, EP300, TGFβ2, and TNFAIP3 mRNA in B-lymphoma cell lines, myeloid cell lines, as well as in cells from healthy volunteers, were determined by real-time quantitative reverse transcriptpolymerase chain reaction(qRT-PCR) with SYBR Green Dye. Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) was used as reference. Results: The expression level of TGFβ2 mRNA in B-lymphoma cell lines was significantly higher than those in the cells from the healthy control(P<0.05). However, the expression level of TNFAIP3 mRNA in B-malignant cells was significantly lower than that of the healthy control(P<0.05). The expression levels of BMPR2 and EP300 mRNA showed no significant difference between B-malignant cell lines and the healthy group(P>0.05). In B-lymphoma cell lines, correlation analyses revealed that the expression of BMPR2 and TNFAIP3(r=0.882, P=0.04) had significant positive relation. The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in cell lines from myeloid leukemia were significantly lower than those in the cells from the healthy control(P<0.05). The expression levels of TGFβ2 mRNA showed no significant difference between myeloid leukemia cell lines and the healthy control or B-malignant cell lines(P>0.05). The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in B-lymphoma cells were significantly higher than those of the myeloid leukemia cells(P<0.05).Conclusion: Different expression patterns of BMPR2, EP300, TGFβ2, and TNFAIP3 genes in B-lymphoma cells exist.展开更多
文摘背景:生物活性物质Biodentine具有良好的抗压强度、粘接强度及较少的微渗漏等诸多优点,已被成功应用于多种临床适应证,然而有关Biodentine是否可促进成骨细胞成骨的研究罕见。目的:研究不同浓度Biodentine对人骨肉瘤细胞MG-63增殖及成骨作用的影响。方法:制备不同浓度梯度(1、1/2、1/4、1/8和1/16)的Biodentine浸提液。将人骨肉瘤细胞MG-63分6组培养,分别加入MEM培养液(对照组)及5种浓度的Biodentine浸提液,采用CCK-8法检测培养第1,3,5,7天的细胞增殖,筛选Biodentine浸提液最佳浓度。将人骨肉瘤细胞MG-63分2组培养,分别加入MEM培养液(空白对照组)及最佳浓度的Biodentine材料浸提液(实验组),培养第1,3,5,7天,采用Real-time PCR法检测细胞中成骨因子Runx2 m RNA表达;培养第10,14天行茜素红染色,观察细胞钙化结节形成情况。结果与结论:(1)培养第1,3天时,不同浓度Biodentine材料浸提液组活细胞数量与对照组相比无差异;培养第5天,1浓度材料浸提液组活细胞数量明显低于对照组(P<0.05),1/2、1/4、1/8浓度材料浸提液组活细胞数量与对照组比较无差异,1/16浓度材料浸提液组活细胞数量明显高于对照组(P<0.05),因此实验选择1/16浓度材料浸提液进行后续实验;(2)实验组培养第1,3,5,7天的Runx2 m RNA表达量分别为空白对照组的1.14,5.29,1.08,2.11倍(P均<0.05);(3)培养第10天时,实验组与空白对照组均未见矿化结节;培养第14天,两组均可见矿化结节,实验组的面积更大、颜色更深;(4)结果表明在一定浓度下,Biodentine可促进人骨肉瘤细胞MG-63增殖及成骨活性。
基金This work was supported by a grant from the National Natural Science Foundation of China(No.81601726).
文摘Background:Mesenchymal stem or stromal cells(MSCs)derived from the induced pluripotent stem cells(iPSCs)have uniform biological activity,which makes the clinical application of MSCs in bone repair possible.Culturing the iPSC-MSCs onto osteoconductive materials is a promising tissue engineering-based strategy in bone regeneration.The aim of this work was to evaluate the effects of semaphorin 3A(Sema3A)and hypoxia inducible factor 1 subunit alpha(HIF1α)co-overexpression on the survival and osteogenic differentiation of iPSC-MSCs.Methods:Sema3A and HIF1αwere linked together with the three(GGGGS;G,glycine;S,serine)peptide fragment,and their co-expression in iPSC-MSCs was mediated by a lentiviral vector.The fusion protein retained the immune reactivity for both Sema3A and HIF1αas determined with Western blotting.iPSC-MSCs were infected with overexpression lentivirus(oeLenti)as negative control,oeLenti-Sema3A,oeLenti-HIF1αor oeLenti-Sema3A-HIF1αlentiviruses.Results:Sema3A overexpression alone promoted the osteogenic differentiation of iPSC-MSCs(the activity and/or expression of osteoblast markers,such as alkaline phosphatase,osteopontin,and osteocalcin,were upregulated),and suppressed cell survival.The Sema3A-HIF1αfusion protein showed a comparable osteoconductive effect to that of Sema3A without reducing cell survival.We further seeded iPSC-MSCs modified by SemaA-HIF1αoverexpression onto hydroxyapatite(HA)scaffolds,and evaluated their growth and differentiation on this three-dimensional material.Additional data indicated that,as compared to iPSC-MSCs cultured in ordinary two-dimensional dishes,cells cultured in HA scaffolds grew(blank vs.HA scaffolds:0.83 vs.1.39 for survival)and differentiated better(blank vs.HA scaffolds:11.29 vs.16.62 for alkaline phosphatase activity).Conclusion:Modifying iPSC-MSCs with pro-osteogenic(Sema3A)and pro-survival(HIF1α)factors may represent a promising strategy to optimize tissue engineering-based strategy in bone repair.
基金supported by the Guangdong Province Key Foundation of Science and Technology Program (Grant No.2009B0507000029)the Guangdong Province Science and Technology Program (Grant No.2012B031800474)a grant from the Overseas Chinese Affairs Office of the State Council Key Discipline Construction Fund (Grant No.51205002)
文摘Objective: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor, type II(BMPR2), E1 A binding protein p300(EP300), transforming growth factor-β2(TGFβ2), and tumor necrosis factor, and alpha-induced protein 3(TNFAIP3) gene expression patterns in B-cell malignancies were studied. Methods: The relative expression levels of BMPR2, EP300, TGFβ2, and TNFAIP3 mRNA in B-lymphoma cell lines, myeloid cell lines, as well as in cells from healthy volunteers, were determined by real-time quantitative reverse transcriptpolymerase chain reaction(qRT-PCR) with SYBR Green Dye. Glyceraldehyde-3-phosphate dehydrogenase(GAPDH) was used as reference. Results: The expression level of TGFβ2 mRNA in B-lymphoma cell lines was significantly higher than those in the cells from the healthy control(P<0.05). However, the expression level of TNFAIP3 mRNA in B-malignant cells was significantly lower than that of the healthy control(P<0.05). The expression levels of BMPR2 and EP300 mRNA showed no significant difference between B-malignant cell lines and the healthy group(P>0.05). In B-lymphoma cell lines, correlation analyses revealed that the expression of BMPR2 and TNFAIP3(r=0.882, P=0.04) had significant positive relation. The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in cell lines from myeloid leukemia were significantly lower than those in the cells from the healthy control(P<0.05). The expression levels of TGFβ2 mRNA showed no significant difference between myeloid leukemia cell lines and the healthy control or B-malignant cell lines(P>0.05). The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in B-lymphoma cells were significantly higher than those of the myeloid leukemia cells(P<0.05).Conclusion: Different expression patterns of BMPR2, EP300, TGFβ2, and TNFAIP3 genes in B-lymphoma cells exist.
