含羟基磷灰石的纳米纤维支架和微流控制备的聚酰胺胺/BMP-2 质粒树枝状复合物在骨组织工程中的应用。
Nanofibrous Scaffolds Containing Hydroxyapatite and Microfluidic-Prepared Polyamidoamin/BMP-2 Plasmid Dendriplexes for Bone Tissue Engineering Applications.
机构信息
Department of Pharmaceutical Biomaterials, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Department of Biotechnology, University of Tehran, Tehran, Iran.
出版信息
Int J Nanomedicine. 2020 Apr 21;15:2633-2646. doi: 10.2147/IJN.S244416. eCollection 2020.
OBJECTIVE
The aim of this study is to fabricate functional scaffolds to gene delivery bone morphogenetic protein-2 (BMP-2) plasmid for bone formation in bone tissue engineering.
METHODS
Dendriplexes (DPs) of generation 4 polyamidoamin (G4-PAMAM)/BMP-2 plasmid were prepared through microfluidic (MF) platform. The physiochemical properties and toxicity of DPs were evaluated by DLS, AFM, FESEM and MTT assay. In order to create a suitable environment for stem cell growth and differentiation, poly-l-lactic acid (PLLA) and poly-l-lactic acid/poly (ethylene oxide) (PLLA/PEO) scaffolds containing hydroxyapatite nanoparticles (HA) and DPs were fabricated by the electrospinning method. The osteogenic potency of the scaffolds on human adipose tissue-derived mesenchymal stem cells (hASCs) was investigated.
RESULTS
The results revealed that tuning the physical properties of DPs by adjusting flow parameters in microfluidic platform can easily improve the cell viability compared to conventional bulk mixing method. Also, the result showed that the presence of HA and DPs in PLLA/PEO scaffold enhanced alkaline phosphatase (ALP) activity and increased the amount of deposited Ca, as well as, related to osteogenesis gen markers.
CONCLUSION
This study indicated that on using the MF platform in preparation of DPs and loading them along with HA in PLLA/PEO scaffold, the osteogenic differentiation of hASCs could be tuned.
目的
本研究旨在构建用于骨组织工程中成骨的基因递释骨形态发生蛋白 2(BMP-2)质粒的功能性支架。
方法
通过微流控(MF)平台制备了第 4 代聚酰胺-胺(G4-PAMAM)/BMP-2 质粒的树枝状聚合物(DPs)。通过 DLS、AFM、FESEM 和 MTT 测定法评估了 DPs 的理化性质和毒性。为了为干细胞的生长和分化创造合适的环境,通过静电纺丝法制备了含有纳米羟基磷灰石(HA)和 DPs 的聚-l-乳酸(PLLA)和聚-l-乳酸/聚(氧化乙烯)(PLLA/PEO)支架。研究了支架对人脂肪组织来源间充质干细胞(hASCs)的成骨能力。
结果
结果表明,通过在微流控平台中调整流动参数来调整 DPs 的物理性质,与传统的批量混合方法相比,可轻松提高细胞活力。此外,结果表明,HA 和 DPs 的存在增强了 PLLA/PEO 支架中的碱性磷酸酶(ALP)活性和钙沉积量,并与成骨基因标志物有关。
结论
本研究表明,在制备 DPs 时使用 MF 平台,并将其与 PLLA/PEO 支架中的 HA 一起加载,可以调节 hASCs 的成骨分化。