Current histological measurement techniques for interstitial
collagen, the basis of interstitial
fibrosis, are semi-quantitative at best and only provide a ratio of
collagen levels within tissues. The Genesis200 imaging system and supplemental image analysis software, FibroIndex from HistoIndex, is a novel, automated platform that uses second-harmonic generation (SHG) for imaging and characterization of interstitial
collagen deposition and additional characteristics, in the absence of any staining. However, its ability to quantify renal
fibrosis requires investigation. This study compared SHG imaging of renal
fibrosis in mice with unilateral ureteric obstruction (UUO), to that of Masson's trichrome staining (MTS) and immunohistochemistry (IHC) of
collagen I. Additionally, the platform generated data on
collagen morphology and distribution patterns. While all three methods determined that UUO-injured mice underwent significantly increased renal
fibrosis after 7 days, the HistoIndex platform additionally determined that UUO-injured mice had a significantly increased
collagen-to-tissue cross reticulation ratio (all P < .001 vs
sham group). Furthermore, in UUO-injured mice treated with the
relaxin family
peptide receptor-1 agonists,
relaxin (0.5 mg/kg/day) or B7-33 (0.25 mg/kg/day), or
angiotensin converting enzyme-inhibitor,
perindopril (1 mg/kg/day) over the 7-day period, only the HistoIndex platform determined that the drug-induced prevention of renal
fibrosis correlated with significantly reduced
collagen fiber thickness and
collagen-to-tissue cross reticulation ratio, but increased
collagen fiber counts.
Relaxin or B7-33 treatment also increased renal
matrix metalloproteinase-2 and reduced
tissue inhibitor of metalloproteinase-1 levels (all P < .01 vs UUO alone). This study demonstrated the diagnostic value of the HistoIndex platform over currently used staining techniques.