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Pilot study for detection of early changes in tissue associated with heterotopic ossification: moving toward clinical use of Raman spectroscopy
Authors:Mitchell Harris  Katherine Cilwa  Eric A Elster  Benjamin K Potter  Jonathan A Forsberg
Institution:1. Department of Surgery, Uniformed Services University of Health Science, Bethesda, MD, USA,;2. Regenerative Medicine Department, Naval Medical Research Center, Silver Spring, MD, USA,;3. Department of Surgery, Uniformed Services University of Health Science, Bethesda, MD, USA,;4. Department of Surgery, Walter Reed National Military Medical Center, Bethesda, MD, USA, and;5. Regenerative Medicine Department, Naval Medical Research Center, Silver Spring, MD, USA,;6. Department of Orthopedics, Walter Reed National Military Medical Center, Bethesda MD, USA
Abstract:Abstract

Over 60% of combat-wounded patients develop heterotopic ossification (HO). Nearly 33% of them require surgical excision for symptomatic lesions, a procedure that is both fraught with complications and can delay or regress functional rehabilitation. Relative medical contraindications limit widespread use of conventional means of primary prophylaxis, such as nonspecific nonsteroidal anti-inflammatory medications and radiotherapy. Better methods for risk stratification are needed to both mitigate the risk of current means of primary prophylaxis as well as to evaluate novel preventive strategies currently in development. We asked whether Raman spectral changes, measured ex vivo, could be associated with histologic evidence of the earliest signs of HO formation and substance P (SP) expression in tissue biopsies from the wounds of combat casualties. In this pilot study, we compared normal muscle tissue, injured muscle tissue, very early HO lesions (?<?16 d post-injury), early HO lesions (?>?16 d post-injury) and mature HO lesions. The Raman spectra of these tissues demonstrate clear differences in the Amide I and III spectral regions of HO lesions compared to normal tissue, denoted by changes in the Amide I band center (p?<?0.01) and the 1340/1270?cm?1 (p?<?0.05) band area and band height ratios. SP expression in the HO lesions appears to peak between 16 and 30 d post-injury, as determined by SP immunohistochemistry of corresponding tissue sections, potentially indicating optimal timing for administration of therapeutics. Raman spectroscopy may therefore prove a useful, non-invasive and early diagnostic modality to detect HO formation before it becomes evident either clinically or radiographically.
Keywords:Heterotopic ossification  muscle  Raman spectroscopy  substance P  war wounds
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