Algorithms for the multidimensional microscopy images analysis. Application to study of macrophages-Fe based degradable biomaterials interactions

FAGALI NS; LOZANO PUERTO RM; PÉREZ MACEDA BT; MADRID MA; FERNÁNDEZ LORENZO MA

Oxford

Congreso; 10th Symposium on Biodegradable Metals; 2018

INTRODUCTION: Advantages and disadvantages of the use of pure iron (Fe) and some of its alloys for the manufacture of biodegradable temporary stents have been discussed in the last decades. The evaluation of biocompatibility ?in vivo? (in the area of the implant), is complex and the use of cell cultures is a valuable model for preliminary studies [1]. The analysis of the interactions between cells and materials at real time is helpful to provide information related to possible toxic effects of Fe and its degradation products formed after exposure to the biological media. RESULTS: Multidimensional microscopy (MDM) at real time was employed in this work to obtain images and videos of live non-stained macrophages in contact with bulk Fe. The acquired images were later analyzed using different algorithms. The MDM allows the study of the cell behavior in conditions where non-staining is required, avoiding a possible effect on cell metabolism.In order to study these interactions, murine macrophages (J774A.1) were exposed to rings of pure Feo (99.9% SpecPure) and their response was analyzed by MDM at real time. Images were taken every 15 minutes for 24 and 48 hours, and then the frames were mounted as a video film.Several algorithms were designed for the analysis of the sequences of images using free software FIJI and different parameters were obtained such as number of live/dead cells, displacement, average speed, number of divisions (splitting) and others morphological parameters. It could be noticed that the speed, displacement and cell division were altered with respect to control cells (in absence of Fe). As metal degrades, the macrophages capture and accumulate the degradation products as a dark deposit of insoluble material and cells die.CONCLUSIONS: The use of different algorithms enables to collect useful information from the images of non-stained live cells using MDM. Particularly, the application of these algorithms in the study of the effect of the degradation of Fe on macrophages, allowed to identify and select the most relevant parameters to describe the effects produced by the degradation of Fe on these cells.REFERENCES:1Fagali NS, Grillo CA, Puntarulo S, Fernández Lorenzo MA (2017). Is there any difference in the biological impact of soluble and insoluble degradation products of iron-containing biomaterials? Colloids and Surfaces B: Biointerfaces 160: 238?246ACKNOWLEDGMENTS: CONICET (PIP 2016-2018 GI 0601, P-UE 22920170100100CO), UNLP (11/I221), ANPCyT (PICT 2015-0232, PICT 2016-1424), MICINN (MAT2011-29152-C02-02)