Quantitative assessment of morphology and sub-cellular changes in macrophages and trophoblasts during inflammation

Abstract

In pregnancy during an inflammatory condition, macrophages present at the fetomaternal junction release an increased amount of nitric oxide (NO) and pro-inflammatory cytokines such as TNF-α and INF-γ, which can disturb the trophoblast functions and pregnancy outcome. Measurement of the cellular and sub-cellular morphological modifications associated with inflammatory responses are important in order to quantify the extent of trophoblast dysfunction for clinical implication. With this motivation, we investigated morphological, cellular and sub-cellular changes in externally inflamed RAW264.7 (macrophage) and HTR-8/SVneo (trophoblast) using structured illumination microscopy (SIM) and quantitative phase microscopy (QPM). We monitored the production of NO, changes in cell membrane and mitochondrial structure of macrophages and trophoblasts when exposed to different concentrations of proinflammatory agents (LPS and TNF-α). In vitro NO production by LPS-induced macrophages increased 22-fold as compared to controls, whereas no significant NO production was seen after the TNF-α challenge. Under similar conditions as with macrophages, trophoblasts did not produce NO following either LPS or the TNF-α challenge. Super-resolution SIM imaging showed changes in the morphology of mitochondria and the plasma membrane in macrophages following the LPS challenge and in trophoblasts following the TNF-α challenge. Label-free QPM showed a decrease in the optical thickness of the LPS-challenged macrophages while TNF-α having no effect. The vice-versa is observed for the trophoblasts. We further exploited machine learning approaches on a QPM dataset to detect and to classify the inflammation with an accuracy of 99.9% for LPS-challenged macrophages and 98.3% for TNF-α-challenged trophoblasts. We believe that the multi-modal advanced microscopy methodologies coupled with machine learning approach could be a potential way for early detection of inflammation.

Metformin Corrects Glucose Metabolism Reprogramming and NLRP3 Inflammasome‐Induced Pyroptosis via Inhibiting the TLR4/NF‐κB/PFKFB3 Signaling in Trophoblasts: Implication for a Potential Therapy of Preeclampsia

“…The research mentioned above collectively indicates the plausible metabolic switch to glycolysis in preeclamptic trophoblasts; however, whether it is associated with the TLR4/NF- κ B signaling has yet to be confirmed. In our studies, LPS-induced TLR4/NF- κ B activation destroyed mitochondrial structure and disrupted mitochondrial function in trophoblasts, which agrees with the latest research [ 64 ]. Meanwhile, TLR4/NF- κ B activation caused the metabolic reprogramming to the augmented glycolysis, characterized by the elevated secretion of lactate and the increased expression of PFKFB3, the pacemaker of glycolysis.…”

Oleuropein Attenuates Lipopolysaccharide-Induced Acute Kidney Injury In Vitro and In Vivo by Regulating Toll-Like Receptor 4 Dimerization

“…In LPS-induced macrophage, the overproduction of inflammatory factors such as nitric oxide (NO), nitrite, IL-6, IL-1β, TNF-α, and the overexpression of inflammation-related proteins such as iNOS and COX-2 are important manifestations of inflammation ( Singh et al, 2020 ; Zhang et al, 2020 ). Here, the effects of OP on LPS-stimulated J774A.1 cells were initially investigated.…”

Real-time study of spatio-temporal dynamics (4D) of physiological activities in alive biological specimens with different FOVs and resolutions simultaneously

“…The movement and energy parameters of mitochondria can be measured while cells interact with surrounding microbes. It will be interesting to have quantifiable data on how the position of microbes influences various organelles in the cells 33 – 35 . Another intriguing phenomenon is organelle transfer between cells like mitochondrial transfer, and it would be fascinating to quantify various parameters during this process.…”