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Label-free objective quantification of collagen to assess burn wound healing

29 Mar | By Biophotonics.World
Label-free objective quantification of collagen to assess burn wound healing
Image source: Krishna Mahato

Burn injury is a common health problem in India with an estimate of 60-70 lakh cases reported every year. Depending on the sternness of the damage, 10 - 20% of these burn injuries require immediate medical attention and hospitalization to avoid mortality. During the hospital stay, these burn injuries need to be assessed periodically to monitor the progress of healing in the wounded tissue. Quick and timely assessment is indispensable for the aesthetic outcome of the healed tissue and improves the quality of life of the patient. 

Collagen, a major structural protein and vital component of the extracellular matrix widely accepted as a marker to monitor wound healing. Quantification of this marker protein is usually done by ‘gold standard' method through histopathological analysis on tissue excised from around the burnt area. Unfortunately, this method is an invasive and painful process that requires tissue biopsy and also necessitate a trained clinician to perform a subjective assessment of the stained and processed tissue sample. 

A team of researchers lead by Professor Krishna Kishore Mahato a physicist by training from School of Life Sciences, Manipal Academy of Higher Education (MAHE), Manipal, Karnataka, India has come up with a label-free spectroscopy method to quantify collagen in burn wound samples. By inspecting this native fluorescent marker in the unstained tissue biopsy samples, the progression of healing was monitored in real time. In collaboration with the clinicians in Kasturba Medical College, MAHE, Manipal, India, they used auto-fluorescence spectroscopy to examine 20 biopsy tissue samples collected from burn wound victims following clinical interventions. The fluorescence spectral findings from these samples were compared and correlated with the standard biochemical methods. The correlation of the label-free method was found to be 94%.

In the previous reports from the same group, the authors have been able to use laser-induced fluorescence to track collagen content and wound healing in vivo by directly implanting an optical fiber in the injured tissue.

The findings appeared in the peer-reviewed journal, Journal of Biophotonics, where in the authors report a quick, cost-effective and objective native-fluorescent spectral method of collagen quantification. "Besides the high correlation with the standard biochemical analyses, this method is found to be sensitive and consistent. Further, the laser-induced fluorescence method adds the benefits of being rapid, only taking 30 seconds to 1 minute per sample for recording (once set up) and does not require highly trained personnel. Any layman can do the experiment. It is not subjective; it is objective assessment" says K. K. Mahato. 

"The reported method is user-friendly and without the addition of any external dye molecule. This helps to maintain tissue architecture during the assessment of healing," said Vijendra Prabhu, first author of the study.

After successful testing on clinical samples, the team is ready for mass trial on burn wound patients. This method can also be applied to test collagen disorders in patients. The authors hope that this technique could eliminate the need for tissue biopsy in the future.

Author: Krishna Mahato

Area of application: Health, other applications
Methods and Techniques: Fluorescence spectroscopy

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