[目的]探讨体外负压培养对hBMSCs成骨活性的影响。[方法]取第3代hBMSCs分为实验组和对照组,实验组进行间歇性负压培养,设置压力为17 kPa,30 m in/次,4次/d,干预2周;对照组于普通CO2培养箱中常规培养。倒置显微镜下观察细胞形态,MTT法和...[目的]探讨体外负压培养对hBMSCs成骨活性的影响。[方法]取第3代hBMSCs分为实验组和对照组,实验组进行间歇性负压培养,设置压力为17 kPa,30 m in/次,4次/d,干预2周;对照组于普通CO2培养箱中常规培养。倒置显微镜下观察细胞形态,MTT法和流式细胞仪检测细胞增殖和凋亡,检测ALP活性,茜素红染色观察钙节结形成,免疫组织化学检测Ⅰ型胶原和VEGF的表达。[结果]诱导2周,实验组细胞增殖能力下降,S期细胞百分比为(5.14±1.56)%,较对照组(13.45±3.51)%约下降62.4%,细胞凋亡增加。实验组ALP活性为(15.68±1.97)mU/mg,对照组为(6.34±1.21)mU/mg,两组比较差异有统计学意义(P<0.05);与对照组比较,实验组钙结节形成增多,VEGF表达较对照组显著提高;实验组Ⅰ型胶原表达呈阳性,对照组呈阴性反应。[结论]负压能抑制hBMSCs增殖,促进细胞凋亡,但可以提高细胞成骨活性。展开更多
The bone marrow microenvironment is critical for the maintenance and functionality of stem/progenitor cells,which are essential for bone development and regeneration.However,the composition and potential use of bone m...The bone marrow microenvironment is critical for the maintenance and functionality of stem/progenitor cells,which are essential for bone development and regeneration.However,the composition and potential use of bone marrow interstitial fluid have not been well explored.In this study,we report the role of neonatal bovine bone marrow interstitial fluid(NBIF)in enhancing the bone regeneration capacity of human bone marrow mesenchymal stem cells(hBMSCs).Unlike adult bovine bone marrow interstitial fluid(ABIF),NBIF-fed hBMSCs exhibit enhanced self-renewal and osteogenic potential and bone marrow homing ability,along with transcriptome changes as compared to hBMSCs cultured in standard fetal bovine serum(FBS)supplemented medium.Mass spectrometry analysis reveals that multiple secreted factors associated with tissue repair and bone development are enriched in NBIF compared to FBS and ABIF.The combined use of NBIF-enriched Nerve Growth Factor(NGF),Lactoferrin(LTF),and High Mobility Group Protein B1(HMGB1),together with Insulin-Like Growth Factor 1(IGF1)for culturing hBMSCs in the presence of FBS can enhance osteogenic potential and bone marrow homing ability,mimicking NBIF's effects.These findings highlight the role of interstitial fluid in the bone marrow microenvironment and its potential to optimize stem cell-based therapies.展开更多
To assess a novel cell manipulation technique of tissue engineering with respect to its ability to augment superparamagnetic iron oxide particles （SPIO） labeled mesenchymal stem cells （MSCs） density at a localized...To assess a novel cell manipulation technique of tissue engineering with respect to its ability to augment superparamagnetic iron oxide particles （SPIO） labeled mesenchymal stem cells （MSCs） density at a localized cartilage defect site in an in vitro phantom by applying magnetic force. Meanwhile, non-invasive imaging techniques were use to track SPIO-labeled MSCs by magnetic resonance imaging （MRI）. Human bone marrow MSCs were cultured and labeled with SPIO. Fresh degenerated human osteochondral fragments were obtained during total knee arthroplasty and a cartilage defect was created at the center. Then, the osteochondral fragments were attached to the sidewalls of culture flasks filled with phosphate-buffered saline （PBS） to mimic the human joint cavity. The SPIO-labeled MSCs were injected into the culture flasks in the presence of a 0.57 Tesla （T） magnetic force. Before and 90 min after cell targeting, the specimens underwent T2-weighted turbo spin-echo （SET2WI） sequence of 3.0 T MRI. MRI results were compared with histological findings. Macroscopic observation showed that SPIO-labeled MSCs were steered to the target region of cartilage defect. MRI revealed significant changes in signal intensity （P0.01）. HE staining exibited that a great number of MSCs formed a three-dimensional （3D） cell ＂sheet＂ structure at the chondral defect site. It was concluded that 0.57 T magnetic force permits spatial delivery of magnetically labeled MSCs to the target region in vitro. High-field MRI can serve as an very sensitive non-invasive technique for the visualization of SPIO-labeled MSCs.展开更多
文摘[目的]探讨体外负压培养对hBMSCs成骨活性的影响。[方法]取第3代hBMSCs分为实验组和对照组,实验组进行间歇性负压培养,设置压力为17 kPa,30 m in/次,4次/d,干预2周;对照组于普通CO2培养箱中常规培养。倒置显微镜下观察细胞形态,MTT法和流式细胞仪检测细胞增殖和凋亡,检测ALP活性,茜素红染色观察钙节结形成,免疫组织化学检测Ⅰ型胶原和VEGF的表达。[结果]诱导2周,实验组细胞增殖能力下降,S期细胞百分比为(5.14±1.56)%,较对照组(13.45±3.51)%约下降62.4%,细胞凋亡增加。实验组ALP活性为(15.68±1.97)mU/mg,对照组为(6.34±1.21)mU/mg,两组比较差异有统计学意义(P<0.05);与对照组比较,实验组钙结节形成增多,VEGF表达较对照组显著提高;实验组Ⅰ型胶原表达呈阳性,对照组呈阴性反应。[结论]负压能抑制hBMSCs增殖,促进细胞凋亡,但可以提高细胞成骨活性。
基金financially supported by the Guangzhou National Laboratory(grant#GZNL2025C02022,A.M.#QNPG2317,J.Z.)partially by the National Natural Science Foundation of China(31988101)。
文摘The bone marrow microenvironment is critical for the maintenance and functionality of stem/progenitor cells,which are essential for bone development and regeneration.However,the composition and potential use of bone marrow interstitial fluid have not been well explored.In this study,we report the role of neonatal bovine bone marrow interstitial fluid(NBIF)in enhancing the bone regeneration capacity of human bone marrow mesenchymal stem cells(hBMSCs).Unlike adult bovine bone marrow interstitial fluid(ABIF),NBIF-fed hBMSCs exhibit enhanced self-renewal and osteogenic potential and bone marrow homing ability,along with transcriptome changes as compared to hBMSCs cultured in standard fetal bovine serum(FBS)supplemented medium.Mass spectrometry analysis reveals that multiple secreted factors associated with tissue repair and bone development are enriched in NBIF compared to FBS and ABIF.The combined use of NBIF-enriched Nerve Growth Factor(NGF),Lactoferrin(LTF),and High Mobility Group Protein B1(HMGB1),together with Insulin-Like Growth Factor 1(IGF1)for culturing hBMSCs in the presence of FBS can enhance osteogenic potential and bone marrow homing ability,mimicking NBIF's effects.These findings highlight the role of interstitial fluid in the bone marrow microenvironment and its potential to optimize stem cell-based therapies.
基金supported by a grant from the National Natural Sciences Foundation of China (No. 30870639)
文摘To assess a novel cell manipulation technique of tissue engineering with respect to its ability to augment superparamagnetic iron oxide particles （SPIO） labeled mesenchymal stem cells （MSCs） density at a localized cartilage defect site in an in vitro phantom by applying magnetic force. Meanwhile, non-invasive imaging techniques were use to track SPIO-labeled MSCs by magnetic resonance imaging （MRI）. Human bone marrow MSCs were cultured and labeled with SPIO. Fresh degenerated human osteochondral fragments were obtained during total knee arthroplasty and a cartilage defect was created at the center. Then, the osteochondral fragments were attached to the sidewalls of culture flasks filled with phosphate-buffered saline （PBS） to mimic the human joint cavity. The SPIO-labeled MSCs were injected into the culture flasks in the presence of a 0.57 Tesla （T） magnetic force. Before and 90 min after cell targeting, the specimens underwent T2-weighted turbo spin-echo （SET2WI） sequence of 3.0 T MRI. MRI results were compared with histological findings. Macroscopic observation showed that SPIO-labeled MSCs were steered to the target region of cartilage defect. MRI revealed significant changes in signal intensity （P0.01）. HE staining exibited that a great number of MSCs formed a three-dimensional （3D） cell ＂sheet＂ structure at the chondral defect site. It was concluded that 0.57 T magnetic force permits spatial delivery of magnetically labeled MSCs to the target region in vitro. High-field MRI can serve as an very sensitive non-invasive technique for the visualization of SPIO-labeled MSCs.
