The severity of Osteogenesis imperfecta and type I collagen pattern in human skin as determined by nonlinear microscopy: proof of principle of a diagnostic method

Authors
Adur, Javier Fernando; D´Souza Li, Lilia; Pedroni, Marcus Vinícius; Steiner, Carlos E.; Pelagati, Vitor B.; de Thomaz, Andre A.; Carvalho, Hernandes F.; Cesar, Carlos L.
Publication Year
2013
Language
English
Format
article
Status
Published version
Description
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.
Fil: Adur, Javier Fernando. Universidad Nacional de Entre Ríos. Facultad de Ingeniería; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidade Estadual de Campinas; Brasil
Fil: D´Souza Li, Lilia. Universidade Estadual de Campinas; Brasil
Fil: Pedroni, Marcus Vinícius. Universidade Estadual de Campinas; Brasil
Fil: Steiner, Carlos E.. Universidade Estadual de Campinas; Brasil
Fil: Pelagati, Vitor B.. Universidade Estadual de Campinas; Brasil
Fil: de Thomaz, Andre A.. Universidade Estadual de Campinas; Brasil
Fil: Carvalho, Hernandes F.. Universidade Estadual de Campinas; Brasil
Fil: Cesar, Carlos L.. Universidade Estadual de Campinas; Brasil
Subject
OSTEOGENESIS IMPERFECTA
NONLINEAR MICROSCOPY
TEXTURE ANALYSIS
Otras Ciencias Físicas
Ciencias Físicas
CIENCIAS NATURALES Y EXACTAS
Access level
Open access
License
https://creativecommons.org/licenses/by/2.5/ar/
Repository
CONICET Digital (CONICET)
Institution
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identifier
oai:ri.conicet.gov.ar:11336/23785