The aim of this work was to develop an in vitro model to study mechanical compression effects on cartilage. A pressure-controlled compression device was used in this study. Cartilage explants obtained from human knee ...The aim of this work was to develop an in vitro model to study mechanical compression effects on cartilage. A pressure-controlled compression device was used in this study. Cartilage explants obtained from human knee were compressed at 1MPa/1Hz for 7 hours (30 min ON, 30 min OFF) under normoxia (5% CO2, 21% O2) or hypoxia (5% CO2, 5% O2). Cell viability was analyzed while nitric oxide (NO) and glycosaminoglycans (GAG) release was assayed in culture media. Mechanical stimulation increased NO production and GAG release by human cartilage explants under normoxia and hypoxia culture. In normoxia and hypoxia conditions, mechanical stimulation alters human OA cartilage metabolism. There is also, an increase in matrix degradation after compression, as shown by levels of GAG found in culture media. This study put in evidence the importance of mechanical compression in the progression of the osteoarthritis and present and in vitro model for mechanobiological and pharmacological studies.展开更多
In this work, we have considered a new multimodality imaging for macroscopy based on Second Harmonic Generation (SHG) method to monitor invasivelessly the matrix collagen. As the triple helicoidally structure of colla...In this work, we have considered a new multimodality imaging for macroscopy based on Second Harmonic Generation (SHG) method to monitor invasivelessly the matrix collagen. As the triple helicoidally structure of collagen molecules appearing as not centrosymetric, very organized and spatially oriented, collagen fibrils give rise to a very strong SHG signal and can be imaged without any exogenous dye. To integrate a multidimensional scale with a large field of view (non-sliced samples), we have adapted and validated an instrumental coupling between a two photon excitation laser and a macroscope to collect cartography of SHG signal. We introduced an index (F-SHG) based on decay time response measured by TCSPC for respectively Fluorescence (F) and Second Harmonic Generation (SHG) values. For various sample where protein collagen is the major component of extracellular matrix (vessel, skin, carotide vessel, rat femoral head cartilage, mouse tumor, human wharton’s jelly and rat tendon) or not (nacre), we compared the index distribution obtained with MacroSHG. In this work, we showed for the first time that multiscale large field imaging (Macroscopy) combined to Multimodality approaches (SHG-TCSPC) could be an innovative and non-invasive technique to detect and identify some biological interest molecules (collagen) in biomedical topics.展开更多
To overcome cartilage injury, strategies have been developed in the last few years based on tissue engineering to rebuild the defects. Cartilage engineering is principally based on three main biological factors: cells...To overcome cartilage injury, strategies have been developed in the last few years based on tissue engineering to rebuild the defects. Cartilage engineering is principally based on three main biological factors: cells (native cells (chondrocytes) or a more primitive ones as mesenchymal stem cells), scaffolds and functionalization factors (growth factors, mechanical stimulation and/or hypoxia). Cartilage tissue engineering strategies generally result in homogeneous tissue structures with little resemblance to native zonal organization of articular cartilage. The main objective of our work concerns the buildup of complex biomaterials aimed at reconstructing biological tissue with three dimensional cells construction for mimicking cartilage architecture. Our strategy is based on structures formation by simple and progressive spraying of mixed alginate hydrogel and human mesenchymal stem cells (hMSC). In this work, the comportment of cells and more precisely their chondrogenic differentiation potential is compared to a traditional making process: the mold. We report here that spraying method allowed to product a scaffold with hMSC that confer a favorable environment for neocartilage construction.展开更多
1 Introduction The cartilage i s a hydrated connective tissue in joint s that withstands and distributes mechanical forces.Chondrocytes utilize mechanical signals to maintain tissue homeostasis.They regulate their met...1 Introduction The cartilage i s a hydrated connective tissue in joint s that withstands and distributes mechanical forces.Chondrocytes utilize mechanical signals to maintain tissue homeostasis.They regulate their metabolic activity through complex biological and biophysical interactions with the extracellular matrix(ECM).Although some of the mechanisms of mechanotransduction are known today,there are certainly many others left unrevealed.Different topics of chondrocytes mechanobiology have led to the development of tissue engineering.It is the concept of substitute tissue developed in vitro,from bioresorbable or non2bioresorbable scaffolds and from cells harvested in a physiologic mechanical environment.展开更多
文摘The aim of this work was to develop an in vitro model to study mechanical compression effects on cartilage. A pressure-controlled compression device was used in this study. Cartilage explants obtained from human knee were compressed at 1MPa/1Hz for 7 hours (30 min ON, 30 min OFF) under normoxia (5% CO2, 21% O2) or hypoxia (5% CO2, 5% O2). Cell viability was analyzed while nitric oxide (NO) and glycosaminoglycans (GAG) release was assayed in culture media. Mechanical stimulation increased NO production and GAG release by human cartilage explants under normoxia and hypoxia culture. In normoxia and hypoxia conditions, mechanical stimulation alters human OA cartilage metabolism. There is also, an increase in matrix degradation after compression, as shown by levels of GAG found in culture media. This study put in evidence the importance of mechanical compression in the progression of the osteoarthritis and present and in vitro model for mechanobiological and pharmacological studies.
文摘In this work, we have considered a new multimodality imaging for macroscopy based on Second Harmonic Generation (SHG) method to monitor invasivelessly the matrix collagen. As the triple helicoidally structure of collagen molecules appearing as not centrosymetric, very organized and spatially oriented, collagen fibrils give rise to a very strong SHG signal and can be imaged without any exogenous dye. To integrate a multidimensional scale with a large field of view (non-sliced samples), we have adapted and validated an instrumental coupling between a two photon excitation laser and a macroscope to collect cartography of SHG signal. We introduced an index (F-SHG) based on decay time response measured by TCSPC for respectively Fluorescence (F) and Second Harmonic Generation (SHG) values. For various sample where protein collagen is the major component of extracellular matrix (vessel, skin, carotide vessel, rat femoral head cartilage, mouse tumor, human wharton’s jelly and rat tendon) or not (nacre), we compared the index distribution obtained with MacroSHG. In this work, we showed for the first time that multiscale large field imaging (Macroscopy) combined to Multimodality approaches (SHG-TCSPC) could be an innovative and non-invasive technique to detect and identify some biological interest molecules (collagen) in biomedical topics.
文摘To overcome cartilage injury, strategies have been developed in the last few years based on tissue engineering to rebuild the defects. Cartilage engineering is principally based on three main biological factors: cells (native cells (chondrocytes) or a more primitive ones as mesenchymal stem cells), scaffolds and functionalization factors (growth factors, mechanical stimulation and/or hypoxia). Cartilage tissue engineering strategies generally result in homogeneous tissue structures with little resemblance to native zonal organization of articular cartilage. The main objective of our work concerns the buildup of complex biomaterials aimed at reconstructing biological tissue with three dimensional cells construction for mimicking cartilage architecture. Our strategy is based on structures formation by simple and progressive spraying of mixed alginate hydrogel and human mesenchymal stem cells (hMSC). In this work, the comportment of cells and more precisely their chondrogenic differentiation potential is compared to a traditional making process: the mold. We report here that spraying method allowed to product a scaffold with hMSC that confer a favorable environment for neocartilage construction.
基金This work was partly supported by region de Lorraine(CPER)andFrench embassyin Beijing
文摘1 Introduction The cartilage i s a hydrated connective tissue in joint s that withstands and distributes mechanical forces.Chondrocytes utilize mechanical signals to maintain tissue homeostasis.They regulate their metabolic activity through complex biological and biophysical interactions with the extracellular matrix(ECM).Although some of the mechanisms of mechanotransduction are known today,there are certainly many others left unrevealed.Different topics of chondrocytes mechanobiology have led to the development of tissue engineering.It is the concept of substitute tissue developed in vitro,from bioresorbable or non2bioresorbable scaffolds and from cells harvested in a physiologic mechanical environment.
