Zakhireh Solmaz, Barar Jaleh, Beygi-Khosrowshahi Younes, Barzegari Abolfazl, Omidi Yadollah, Adibkia Khosro
Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran.
Bioimpacts. 2022;12(3):183-193. doi: 10.34172/bi.2021.24. Epub 2021 Dec 18.
The current study, for the first time, suggests nature-made pollen grains (PGs) of L. as a potential candidate for using as scaffolding building blocks with encapsulation capability of bioactive compounds, such as bone morphogenetic protein 4 (BMP4). A modified method using KOH (5%, 25ºC) was developed to produce nonallergic hollow pollen grains (HPGs), confirmed by energy dispersive X-ray (EDX) analysis, field emission scanning electron microscopy (FESEM), and DNA and protein staining techniques. The study was conducted on human adipose-derived mesenchymal stem cells (hAD-MSCs) to investigate the applicability of HPGs as bone scaffolding building blocks. Cytocompability was evaluated by FESEM, MTT assay, and gene expression analysis of apoptotic markers (BAX and BCL2). The osteoconductive potential of HPGs was assessed by alkaline phosphatase (ALP) activity measurement and gene expression analysis of osteogenic markers (RUNX2 and osteocalcin). Findings demonstrated that HPGs can be considered as biocompatible compounds increasing the metabolic activities of the cells. Further, the bioactive nature of HPGs resulted in suitable cellular adhesion properties, required for a potent scaffold. The investigation of apoptotic gene expression indicated a reduced BAX/BCL2 ratio reflecting the protective effect of HPGs on hAD-MSCs. The increased ALP activity and expression of osteogenic genes displayed the osteoconductive property of HPGs. Moreover, the incorporation of BMP4 in HPGs initiated a synergistic effect on osteoblast maturation. Owing to the unique compositional and surface nanotopographical features of the L. HPG, this microscale architecture provides a favorable microenvironment for the bottom-up remodeling of bone.
当前研究首次表明，天然的枸杞花粉粒（PGs）有潜力作为支架构建模块，用于封装生物活性化合物，如骨形态发生蛋白4（BMP4）。开发了一种使用KOH（5%，25ºC）的改良方法来制备无过敏反应的中空花粉粒（HPGs），通过能量色散X射线（EDX）分析、场发射扫描电子显微镜（FESEM）以及DNA和蛋白质染色技术进行了确认。该研究以人脂肪来源的间充质干细胞（hAD-MSCs）为对象，研究HPGs作为骨支架构建模块的适用性。通过FESEM、MTT法以及凋亡标志物（BAX和BCL2）的基因表达分析来评估细胞相容性。通过碱性磷酸酶（ALP）活性测定和成骨标志物（RUNX2和骨钙素）的基因表达分析来评估HPGs的骨传导潜力。研究结果表明，HPGs可被视为具有生物相容性的化合物，能提高细胞的代谢活性。此外，HPGs的生物活性特性使其具有适合作为有效支架所需的细胞黏附特性。对凋亡基因表达的研究表明，BAX/BCL2比值降低，反映了HPGs对hAD-MSCs的保护作用。ALP活性的增加和成骨基因的表达显示了HPGs的骨传导特性。此外，将BMP4掺入HPGs中对成骨细胞成熟产生了协同作用。由于枸杞HPG独特的组成和表面纳米拓扑特征，这种微观结构为骨的自下而上重塑提供了有利的微环境。
