Potential of ATR-FTIR Spectroscopy and Molecular Parameters in Elucidating Immune Responses Associated with Burn...

Burn injuries are the fourth leading cause of trauma worldwide, with second-degree burns being highly prevalent. Early detection of immune response dynamics is crucial for effective burn management. This research aimed to develop a second-degree burn wound rat model and to investigate temporal immune responses using ATR-FTIR spectroscopy in combination with molecular and biochemical techniques over 28 days. Thermal burns were induced (0.8 cm diameter) in SD rats (N = 35) using a heated stainless-steel rod. ATR-FTIR spectrum of air-dried serum samples were acquired and integrated AUC associated with cellular (TNF-α, IFN-γ, IL-1β/6/10) and humoral (IgG/M/A) immunity biomarkers were evaluated along with PCA analysis. WB analysis of cellular immunity biomarkers was investigated. Histology revealed early infiltration of inflammatory cells followed by progressive re-epithelialization on days 7 and 14. ATR-FTIR analysis showed significant enhancement of IgA and IgM AUC's on day 3, indicating early immune homeostasis, while IFN-γ and IL-10 were found enhanced on days 3 and 21, suggesting biphasic immune modulation. Comparative analysis indicated partial agreement between ATR-FTIR and WB findings; however, the trends were not directly superimposable. SDS-PAGE identified day 14 as a critical stage for collagen remodelling. Haematological parameters showed increased levels of leukocytes and platelets indices after day 14, while serum calcium levels were found to be elevated during the proliferatory phase. These findings demonstrate that our developed model illustrated a broader understanding of burn wound healing dynamics. We believe that ATR-FTIR spectroscopy could serve as a complementary technology for monitoring immune response dynamics.