PDF(Pubmed)
Abstract：
UNASSIGNED: Osteoporotic-related fractures remains a significant public health concern, thus imposing substantial burdens on our society. Excessive activation of osteoclastic activity is one of the main contributing factors for osteoporosis-related fractures. While polylactic acid (PLA) is frequently employed as a biodegradable scaffold in tissue engineering, it lacks sufficient biological activity. Microdroplets (MDs) have been explored as an ultrasound-responsive drug delivery method, and mesenchymal stem cell (MSC)-derived exosomes have shown therapeutic effects in diverse preclinical investigations. Thus, this study aimed to develop a novel bioactive hybrid PLA scaffold by integrating MDs-NFATc1-silencing siRNA to target osteoclast formation and MSCs-exosomes (MSC-Exo) to influence osteogenic differentiation (MDs-NFATc1/PLA-Exo).
UNASSIGNED: Human bone marrow-derived mesenchymal stromal cells (hBMSCs) were used for exosome isolation. Transmission electron microscopy (TEM) and confocal laser scanning microscopy were used for exosome and MDs morphological characterization, respectively. The MDs-NFATc1/PLA-Exo scaffold was fabricated through poly(dopamine) and fibrin gel coating. Biocompatibility was assessed using RAW 264.7 macrophages and hBMSCs. Osteoclast formations were examined via TRAP staining. Osteogenic differentiation of hBMSCs and cytokine expression modulation were also investigated.
UNASSIGNED: MSC-Exo exhibited a cup-shaped structure and effective internalization into cells, while MDs displayed a spherical morphology with a well-defined core-shell structure. Following ultrasound stimulation, the internalization study demonstrated efficient delivery of bioactive MDs into recipient cells. Biocompatibility studies indicated no cytotoxicity of MDs-NFATc1/PLA-Exo scaffolds in RAW 264.7 macrophages and hBMSCs. Both MDs-NFATc1/PLA and MDs-NFATc1/PLA-Exo treatments significantly reduced osteoclast differentiation and formation. In addition, our results further indicated MDs-NFATc1/PLA-Exo scaffold significantly enhanced osteogenic differentiation of hBMSCs and modulated cytokine expression.
UNASSIGNED: These findings suggest that the bioactive MDs-NFATc1/PLA-Exo scaffold holds promise as an innovative structure for bone tissue regeneration. By specifically targeting osteoclast formation and promoting osteogenic differentiation, this hybrid scaffold may address key challenges in osteoporosis-related fractures.
UNASSIGNED: Human bone marrow-derived mesenchymal stromal cells (hBMSCs) were used for exosome isolation. Transmission electron microscopy (TEM) and confocal laser scanning microscopy were used for exosome and MDs morphological characterization, respectively. The MDs-NFATc1/PLA-Exo scaffold was fabricated through poly(dopamine) and fibrin gel coating. Biocompatibility was assessed using RAW 264.7 macrophages and hBMSCs. Osteoclast formations were examined via TRAP staining. Osteogenic differentiation of hBMSCs and cytokine expression modulation were also investigated.
UNASSIGNED: MSC-Exo exhibited a cup-shaped structure and effective internalization into cells, while MDs displayed a spherical morphology with a well-defined core-shell structure. Following ultrasound stimulation, the internalization study demonstrated efficient delivery of bioactive MDs into recipient cells. Biocompatibility studies indicated no cytotoxicity of MDs-NFATc1/PLA-Exo scaffolds in RAW 264.7 macrophages and hBMSCs. Both MDs-NFATc1/PLA and MDs-NFATc1/PLA-Exo treatments significantly reduced osteoclast differentiation and formation. In addition, our results further indicated MDs-NFATc1/PLA-Exo scaffold significantly enhanced osteogenic differentiation of hBMSCs and modulated cytokine expression.
UNASSIGNED: These findings suggest that the bioactive MDs-NFATc1/PLA-Exo scaffold holds promise as an innovative structure for bone tissue regeneration. By specifically targeting osteoclast formation and promoting osteogenic differentiation, this hybrid scaffold may address key challenges in osteoporosis-related fractures.
摘要：
■骨质疏松相关骨折仍然是一个重大的公共卫生问题，从而给我们的社会带来了巨大的负担。破骨细胞活性的过度激活是骨质疏松相关骨折的主要促成因素之一。虽然聚乳酸（PLA）经常被用作组织工程中的生物可降解支架，它缺乏足够的生物活性。微滴（MD）已被探索作为一种超声响应的药物递送方法，和间充质干细胞（MSC）衍生的外泌体在不同的临床前研究中显示出治疗效果。因此，这项研究旨在通过整合MDs-NFATc1沉默siRNA来靶向破骨细胞形成和MSC-外泌体（MSC-Exo）以影响成骨分化（MDs-NFATc1/PLA-Exo），开发一种新型的生物活性混合PLA支架。
■人骨髓来源的间充质基质细胞(hBMSC)用于外泌体分离。透射电子显微镜（TEM）和共聚焦激光扫描显微镜用于外泌体和MD的形态表征，分别。MDs-NFATc1/PLA-Exo支架通过聚(多巴胺)和纤维蛋白凝胶涂层制备。使用RAW264.7巨噬细胞和hBMSC评估生物相容性。通过TRAP染色检查破骨细胞形成。还研究了hBMSCs的成骨分化和细胞因子表达调节。
■MSC-Exo表现出杯状结构并有效地内化到细胞中，而MD表现出具有明确定义的核-壳结构的球形形态。超声刺激后，内化研究证明了生物活性MD有效递送到受体细胞中.生物相容性研究表明MDs-NFATc1/PLA-Exo支架在RAW264.7巨噬细胞和hBMSCs中没有细胞毒性。MDs-NFATc1/PLA和MDs-NFATc1/PLA-Exo处理均显着降低了破骨细胞的分化和形成。此外,我们的结果进一步表明MDs-NFATc1/PLA-Exo支架显着增强了hBMSCs的成骨分化并调节了细胞因子的表达。
■这些发现表明，生物活性MD-NFATc1/PLA-Exo支架有望成为骨组织再生的创新结构。通过特异性靶向破骨细胞形成和促进成骨分化，这种混合支架可能解决骨质疏松症相关骨折的关键挑战.
■人骨髓来源的间充质基质细胞(hBMSC)用于外泌体分离。透射电子显微镜（TEM）和共聚焦激光扫描显微镜用于外泌体和MD的形态表征，分别。MDs-NFATc1/PLA-Exo支架通过聚(多巴胺)和纤维蛋白凝胶涂层制备。使用RAW264.7巨噬细胞和hBMSC评估生物相容性。通过TRAP染色检查破骨细胞形成。还研究了hBMSCs的成骨分化和细胞因子表达调节。
■MSC-Exo表现出杯状结构并有效地内化到细胞中，而MD表现出具有明确定义的核-壳结构的球形形态。超声刺激后，内化研究证明了生物活性MD有效递送到受体细胞中.生物相容性研究表明MDs-NFATc1/PLA-Exo支架在RAW264.7巨噬细胞和hBMSCs中没有细胞毒性。MDs-NFATc1/PLA和MDs-NFATc1/PLA-Exo处理均显着降低了破骨细胞的分化和形成。此外,我们的结果进一步表明MDs-NFATc1/PLA-Exo支架显着增强了hBMSCs的成骨分化并调节了细胞因子的表达。
■这些发现表明，生物活性MD-NFATc1/PLA-Exo支架有望成为骨组织再生的创新结构。通过特异性靶向破骨细胞形成和促进成骨分化，这种混合支架可能解决骨质疏松症相关骨折的关键挑战.
