Living systems respond to ambient pathophysiological changes by altering their phenotype, a phenomenon called 'phenotypic plasticity'. This program contains information about adaptive biological dynamism. Epithelial-mesenchymal transition (EMT) is one such process found to be crucial in development, wound healing, and cancer wherein the epithelial cells with restricted migratory potential develop motile functions by acquiring mesenchymal characteristics. In the present study, phase contrast microscopy images of EMT induced HaCaT cells were acquired at 24 h intervals for 96 h. The expression study of relevant pivotal molecules viz. F-actin, vimentin, fibronectin and N-cadherin was carried out to confirm the EMT process. Cells were intuitively categorized into five distinct morphological phenotypes. A population of 500 cells for each temporal point was selected to quantify their frequency of occurrence. The plastic interplay of cell phenotypes from the observations was described as a Markovian process. A model was formulated empirically using simple linear algebra, to depict the possible mechanisms of cellular transformation among the five phenotypes. This work employed qualitative, semi-quantitative and quantitative tools towards illustration and establishment of the EMT continuum. Thus, it provides a newer perspective to understand the embedded plasticity across the EMT spectrum.
Epithelial abnormality during the transformation of oral submucous fibrosis (OSF) into oral squamous cell carcinoma has been well studied and documented. However, the differential contribution of atrophy and hyperplasia for malignant potentiality of OSF is yet to be resolved. Existing diagnostic conjectures lack precise diagnostic attributes which may be effectively resolved by substantiation of specific molecular pathology signatures. Present study elucidates existence of cellular competitiveness in OSF conditions using computer-assisted neighbourhood analysis in quantitative immunohistochemistry (IHC) framework. The concept of field cancerization was contributory in finding correspondence among neighbouring cells of epithelial layers with reference to differential expression of cardinal cancer-related genes [c-Myc (oncogene), p53 (tumour suppressor), and HIF-1α (hypoxia regulator)] which are known to be important sensors in recognizing cellular competitive interface. Our analyses indicate that different states of OSF condition may be associated with different forms of competitiveness within epithelial neighbouring cells which might be responsible to shape the present and future of the pre-malignant condition. Analytical findings indicated association of atrophic epithelium with stress-driven competitive environment having low c-Myc, high-p53, and stable HIF-1α (the looser cells) which undergo apoptosis. Whereas, the cells with high c-Myc (winner cells) give rise to hyperplastic epithelium via possible mutation in p53. The epithelial dysplasia plausibly occurs due to clonal expansion of c-Myc and p53 positive supercompetitor cells. Present study proposes quantitative IHC along with neighbourhood analysis which might help us to dig deeper on to the interaction among epithelial cell population to provide a better understanding of field cancerization and malignant transformation of pre-malignancy.
We
report differential proliferation behavior of normal and fibrosis
associated human oral fibroblasts on micropillar honey embedded silk
fibroin substrates (HSF). Oral fibroblasts of different origins manifest
differences in proliferation rate, morphology, and the cytoskeletal
arrangement on HSF substrates with distinct topography (H, D, and
S), stiffness, and honey concentration. It is observed that the proliferation
rate is maximized for normal and inhibited for fibrosis associated
fibroblasts on a HSF substrate surface with moderate height of ∼8.5
μm and 2% honey concentration. Molecular expression analysis
reveals decrease in c-myc and p53 expression in later cells validating
the inhibition of their proliferation rate, which is further correlated
with the decreased Col I and Col III expression on this substrate.
A substrate with enhanced interspacing and intermediate mechanical
stiffness (0.57 ± 0.32 μN/nm) favors strong adhesion and
stable cell–matrix interaction for normal cells, while exhibiting
negative influence on fibrotic fibroblasts with poor adhesion and
spreading capability. Decrease in vimentin, fibronectin expression,
and cytoskeleton reorganization justify the poor stability of later
cells on the optimized substrate, thereby allowing selective modulation
of normal and fibrosis associated fibroblasts under the synergistic
influence of honey concentration, topography, and rigidity of HSF
substrates. The work highlights the possible therapeutic efficacy
of honey based micropatterned substrates as smart patches for fast
wound healing and in minimizing the chances of recurrence of precancer
post oral tumor resection surgeries.
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