Zirconium Oxide Thin Films Obtained by Atomic Layer Deposition Technology Abolish the Anti-Osteogenic Effect Resulting from miR-21 Inhibition in the Pre-Osteoblastic MC3T3 Cell Line
Aleksandra Seweryn , Ariadna Pielok , Krystyna Ławniczak-Jabłońska , Rafał Pietruszka , Klaudia Marcinkowska , Mateusz Sikora , Bartłomiej S. Witkowski , Marek Godlewski , Krzysztof Marycz , Agnieszka Śmieszek
AbstractIntroduction: The development of the field of biomaterials engineering is rapid. Various bioactive coatings are created to improve the biocompatibility of substrates used for bone regeneration, which includes formulation of thin zirconia coatings with pro-osteogenic properties. The aim of this study was to assess the biological properties of ZrO2 thin films grown by Atomic Layer Deposition (ALD) technology (ZrO2 ALD). Methodology: The cytocompatibility of the obtained layers was analysed using the mice pre-osteoblastic cell line (MC3T3) characterized by decreased expression of microRNA 21-5p (miR-21-5p) in order to evaluate the potential pro-osteogenic properties of the coatings. The in vitro experiments were designed to determine the effect of ZrO2 ALD coatings on cell morphology (confocal microscope), proliferative activity (cell cycle analysis) and metabolism, reflected by mitochondrial membrane potential (cytometric-based measurement). Additionally, the influence of layers on the expression of genes associated with cell survival and osteogenesis was studied using RT-qPCR. The following genes were investigated: B-cell lymphoma 2 (Bcl-2), Bcl-2-associated X protein (Bax), p53 and p21, as well as osteogenic markers, i.e. collagen type 1 (Coll-1), osteopontin (Opn), osteocalcin (Ocl) and runt-related transcription factor 2 (Runx-2). The levels of microRNA (miRNA/miR) involved in the regulation of osteogenic genes were determined, including miR-7, miR-21, miR-124 and miR-223. Results: The analysis revealed that the obtained coatings are cytocompatible and may increase the metabolism of pre-osteoblast, which was correlated with increased mitochondrial membrane potential and extensive development of the mitochondrial network. The obtained coatings affected the viability and proliferative status of cells, reducing the population of actively dividing cells. However, in cultures propagated on ZrO2 ALD coatings, the up-regulation of genes essential for bone metabolism was noted. Discussion: The data obtained indicate that ZrO2 coatings created using the ALD method may have pro-osteogenic properties and may improve the metabolism of bone precursor cells. Given the above, further development of ZrO2 ALD layers is essential in terms of their potential clinical application in bone regenerative medicine.
|Journal series||International Journal of Nanomedicine, ISSN 1176-9114, e-ISSN 1178-2013, (N/A 140 pkt)|
|Publication size in sheets||0.75|
|Keywords in English||ALD, Cytocompatibility, Osteoblasts precursors, Pro-osteogenic properties, Zirconia-based coatings|
|ASJC Classification||; ; ; ; ;|
|License||Journal (articles only); published final; ; with publication|
|Score||= 140.0, 14-09-2020, ArticleFromJournal|
|Publication indicators||= 0; = 0; : 2018 = 1.16; : 2018 = 4.471 (2) - 2018=5.029 (5)|
|Citation count*||1 (2020-09-21)|
* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.