Keywords
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Pollen grain,
Pistacia vera L.
Bottom-up tissue engineering,
Building block,
Bone tissue,
Human adipose-derived
mesenchymal stem cells
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Abstract
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The current study, for the first time, suggests nature-made pollen grains (PGs) of Pistaciavera L. as a potential candidate for using as scaffolding building blocks with encapsulation capability of bioactive compounds, such as bone morphogenetic
protein 4 (BMP4).
Methods: 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 in-vitro 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).
Results: 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.
Conclusion: Owing to the unique compositional and surface nanotopographical features of the
Pistacia vera L. HPG, this microscale architecture provides a favorable microenvironment for the
bottom-up remodeling of bone.
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