Abstract
Background: The confirmatory diagnosis of Osteogenesis Imperfecta (OI)
requires invasive, commonly bone biopsy, time consuming and destructive
methods. This paper proposes an alternative method using a combination
of two-photon excitation fluorescence (TPEF) and second-harmonic
generation (SHG) microscopies from easily obtained human skin biopsies.
We show that this method can distinguish subtypes of human OI.
Methodology/Principal Findings: Different aspects of collagen
microstructure of skin fresh biopsies and standard H&E-stained sections
of normal and OI patients (mild and severe forms) were distinguished by
TPEF and SHG images. Moreover, important differences between subtypes of
OI were identified using different methods of quantification such as
collagen density, ratio between collagen and elastic tissue, and
gray-level co-occurrence matrix (GLCM) image-pattern analysis. Collagen
density was lower in OI dermis, while the SHG/autofluorescence index of
the dermis was significantly higher in OI as compared to that of the
normal skin. We also showed that the energy value of GLCM texture
analysis is useful to discriminate mild from severe OI and from normal
skin.
Conclusions/Significance: This work demonstrated that nonlinear
microscopy techniques in combination with image-analysis approaches
represent a powerful tool to investigate the collagen organization in
skin dermis in patients with OI and has the potential to distinguish the
different types of OI. The procedure outlined in this paper requires a
skin biopsy, which is almost painless as compared to the bone biopsy
commonly used in conventional methods. The data presented here
complement existing clinical diagnostic techniques and can be used as a
diagnostic procedure to confirm the disease, evaluate its severity and
treatment efficacy.
Users
Please
log in to take part in the discussion (add own reviews or comments).