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

Autor(es)

Adur, Javier ^{[1, 2]} ; DSouza-Li, Lilia ^[3] ; Pedroni, Marcus Vinicius ^[3] ; Steiner, Carlos E. ^[4] ; Pelegati, Vitor B. ^[5] ; de Thomaz, Andre A. ^[1] ; Carvalho, Hernandes F. ^{[6, 5]} ; Cesar, Carlos L. ^{[5, 1]}

Afiliação do(s) autor(es)

^[1] State Univ Campinas UNICAMP, Inst Phys Gleb Wataghin, Opt & Photon Res Ctr CEPOF, Biophoton Grp, Campinas, SP - Brazil

^[2] Natl Univ Entre Rios, Sch Bioengn, Microscopy Lab Appl Mol & Cellular Studies, Oro Verde, Entre Rios - Argentina

^[3] State Univ Campinas UNICAMP, CIPED, Pediat Endocrinol Lab, Campinas, SP - Brazil

^[4] State Univ Campinas UNICAMP, Fac Med Sci, Dept Med Genet, Campinas, SP - Brazil

^[5] INFABiC Natl Inst Sci & Technol Photon Appl Cell, Campinas, SP - Brazil

^[6] State Univ Campinas UNICAMP, Inst Biol, Dept Struct & Funct Biol, Campinas, SP - Brazil

• Tipo de documento: Artigo Científico
• Fonte: PLoS One; v. 8, n. 7 JUL 15 2013.
• Citações Web of Science: 10

Resumo

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. (AU)

• Processo FAPESP: 11/51591-3 - Javier Fernando Adur | Universidad Nacional de Entre Rios - Argentina
• Beneficiário: Carlos Lenz Cesar
• Modalidade de apoio: Auxílio à Pesquisa - Pesquisador Visitante - Internacional
• Processo FAPESP: 08/57906-3 - Instituto Nacional de Fotônica Aplicada à Biologia Celular - INFABIC
• Beneficiário: Hernandes Faustino de Carvalho
• Modalidade de apoio: Auxílio à Pesquisa - Temático