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Etude structurelle en Microscopie Confocale Raman du cartilage produit par ingénierie tissulaire

Abstract : Articular cartilage is connective tissue, non-innervated and non-vascularized, that covers the ends of the bones within the joints. The high stresses to which the cartilage is subjected often lead to the alteration of the articular matrix. Defects in articular cartilage are generally associated with pain and loss of joint mobility, and have an impact on quality of life, including physical, social and economic well-being. Due to the lack of spontaneous regeneration or healing of cartilage, the use of tissue engineering techniques appears to be a promising avenue to address the clinical needs of patients with osteoarthritis and temporomandibular joint disorders (TMD). To optimize cartilage tissue engineering, non-invasive, non-destructive, high-resolution (nanometric) techniques are needed to evaluate the quality and structural arrangement of cartilage manufactured in vitro. The objective of our study was to follow the maturation of artificial cartilage under mechanical loads with innovative microscopy methods, including confocal Raman microscopy, a non-contact, non-destructive analysis technique without any sample preparation. Using Raman spectra and hyperspectral images, we have been able to characterize the morphological differences of the chondrons according to the different zones of the cartilage matrix and to establish a representative mapping of the spatial distribution of the biochemical components of the said matrix over its entire depth. Calculation of the thickness of the pericellular matrix revealed increasing depth-dependent variations (superficial zone is = 2.7±0.8, intermediate zone = 3.6±1.1 and deep zone = 4.9±1.8, p<0.001). On the basis of Raman spectral data, it was possible to follow in real time the mechanisms underlying the development of cartilage tissue by tissue engineering, in particular the dedifferentiation of chondrocytes and mesenchymal stem cells (MSCs) differentiated into chondrocytes. The results were compared to immunohistological and polymerase chain reaction (RT-qPCR) assays. By comparing degraded cartilage versus normal cartilage, it was possible to characterize the subtle changes in the molecular structure of cartilage tissue preceding the morphological changes of the matrix in osteoarthritis. These results show both the ability and sensitivity of confocal Raman microscopy to characterize the structure of articular cartilage and provide valuable information that can be used as a basis for tissue-engineered cartilage tissue design and a diagnostic tool for early detection of subcellular changes related to osteoarthritis.
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https://tel.archives-ouvertes.fr/tel-03342131
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Submitted on : Monday, September 13, 2021 - 10:59:08 AM
Last modification on : Saturday, June 18, 2022 - 4:59:50 AM
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Jean N'Dre. Etude structurelle en Microscopie Confocale Raman du cartilage produit par ingénierie tissulaire. Médecine humaine et pathologie. Université Montpellier; Université Félix Houphouët-Boigny (Abidjan, Côte d'Ivoire), 2021. Français. ⟨NNT : 2021MONTT001⟩. ⟨tel-03342131⟩

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