Fibrosis Detection:a Combi-approach Of Optical Coherence Tomography, Histology, And Raman Spectroscopy In A Human Skin Scar Model
Rubinder Basson1, Martin Isabelle2, Mohamed Baguneid3, Ardeshir Bayat1.
1University of Manchester, Manchester, United Kingdom, 2Renishaw, Gloucestershire, United Kingdom, 3Al Ain Hospital, SEHA, United Arab Emirates.
BACKGROUND- After injury, normal cutaneous tissue is replaced by fibrous tissue known as remodelling. The principle component of extracellular matrix, collagen, is a key fibrosis marker. We propose it possible to identify the timing of secondary protein structural changes during remodelling using Raman spectroscopy (RS), validated by non-invasive measurements taken at the same time points using optical coherence tomography (OCT), corroborated by histology. METHODS- Samples from 20 human participants were utilised from a randomised, blinded trial comparing ‘treatment’ to ‘control’ topical. An excisional model using a 5mm punch created the scar; participants then received sequential 6mm biopsies of the scar. OCT measurements were taken bimonthly then RS performed on these samples; the results were correlated with H&E and Herovici stains. RESULTS- At week (W)16, RS demonstrated a shift in the amide I peak from 1667cm-1 to 1656cm-1 in the treatment group, representing a change in protein conformation from β-pleated sheets in elastin to α-helix collagen components. Previous Herovici staining showed a significant increase in the proportion of collagen I to collagen III at W16 (p=0.048, treatment group.) H&E demonstrated increased dermal thickness from scar formation at W2 significant at W16(p=0.0009), almost returning to normal skin thickness. OCT attenuation coefficient representing collagen deposition decreased in both groups from normal skin, then increased from W12 in all time points (p<0.002) and also significant between treatment and control groups; W4, and W16(p<0.05). CONCLUSIONS- We have demonstrated a combi-approach of non-invasive and histological techniques validating conformational changes in RS and how it can be applied to evaluate the effects of anti-scar therapeutics.
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