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Jos Malda - One of the best experts on this subject based on the ideXlab platform.

  • Arthroscopic Determination of Cartilage Proteoglycan Content and Collagen Network Structure with Near-Infrared Spectroscopy
    Annals of Biomedical Engineering, 2019
    Co-Authors: Jaakko K. Sarin, Jos Malda, Jetze Visser, Virpi Tiitu, Harold Brommer, Isaac O. Afara, Olli Nykänen, Irina A. D. Mancini, P. René Weeren, Juha Töyräs
    Abstract:

    Conventional arthroscopic evaluation of articular Cartilage is subjective and insufficient for assessing early compositional and structural changes during the progression of post-traumatic osteoarthritis. Therefore, in this study, arthroscopic near-infrared (NIR) spectroscopy is introduced, for the first time, for in vivo evaluation of articular Cartilage thickness, proteoglycan (PG) content, and collagen orientation angle. NIR spectra were acquired in vivo and in vitro from Equine Cartilage adjacent to experimental Cartilage repair sites. As reference, digital densitometry and polarized light microscopy were used to evaluate superficial and full-thickness PG content and collagen orientation angle. To relate NIR spectra and Cartilage properties, ensemble neural networks, each with two different architectures, were trained and evaluated by using Spearman’s correlation analysis ( ρ ). The ensemble networks enabled accurate predictions for full-thickness reference properties (PG content: ρ _ in vitro, Val = 0.691, ρ _ in vivo = 0.676; collagen orientation angle: ρ _ in vitro, Val = 0.626, ρ _ in vivo = 0.574) from NIR spectral data. In addition, the networks enabled reliable prediction of PG content in superficial (25%) Cartilage ( ρ _ in vitro, Val = 0.650, ρ _ in vivo = 0.613) and Cartilage thickness ( ρ _ in vitro, Val = 0.797, ρ _ in vivo = 0.596). To conclude, NIR spectroscopy could enhance the detection of initial Cartilage degeneration and thus enable demarcation of the boundary between healthy and compromised Cartilage tissue during arthroscopic surgery.

  • Non-enzymatic cross-linking of collagen type II fibrils is tuned via osmolality switch.
    Journal of orthopaedic research : official publication of the Orthopaedic Research Society, 2018
    Co-Authors: Behdad Pouran, Jos Malda, Parisa R. Moshtagh, V. Arbabi, Jessica Snabel, Reinout Stoop, Jeffrey W. Ruberti, Amir A. Zadpoor, Harrie Weinans
    Abstract:

    An important aspect in Cartilage ageing is accumulation of advanced glycation end products (AGEs) after exposure to sugars. Advanced glycation results in cross-links formation between the collagen fibrils in articular Cartilage, hampering their flexibility and making Cartilage more brittle. In the current study, we investigate whether collagen cross-linking after exposure to sugars depends on the stretching condition of the collagen fibrils. Healthy Equine Cartilage specimens were exposed to l-threose sugar and placed in hypo-, iso-, or hyper-osmolal conditions that expanded or shrank the tissue and changed the 3D conformation of collagen fibrils. We applied micro-indentation tests, contrast enhanced micro-computed tomography, biochemical measurement of pentosidine cross-links, and Cartilage surface color analysis to assess the effects of advanced glycation cross-linking under these different conditions. Swelling of extracellular matrix due to hypo-osmolality made Cartilage less susceptible to advanced glycation, namely, the increase in effective Young's modulus was approximately 80% lower in hypo-osmolality compared to hyper-osmolality and pentosidine content per collagen was 47% lower. These results indicate that healthy levels of glycosaminoglycans not only keep Cartilage stiffness at appropriate levels by swelling and pre-stressed collagen fibrils, but also protect collagen fibrils from adverse effects of advanced glycation. These findings highlight the fact that collagen fibrils and therefore Cartilage can be protected from further advanced glycation ("ageing") by maintaining the joint environment at sufficiently low osmolality. Understanding of mechanochemistry of collagen fibrils provided here might evoke potential ageing prohibiting strategies against Cartilage deterioration. © 2018 The Authors. Journal of Orthopaedic Research Published by Wiley Periodicals, Inc. on behalf of Orthopaedic Research Society. J Orthop Res.

  • Crosslinkable Hydrogels Derived from Cartilage, Meniscus, and Tendon Tissue
    Tissue Engineering Part A, 2015
    Co-Authors: Jetze Visser, Peter A. Levett, Nikae C.r. Te Moller, Jeremy Besems, Mattie H.p. Van Rijen, Kristel W. M. Boere, Wouter J.a. Dhert, P. René Van Weeren, Janny C. De Grauw, Jos Malda
    Abstract:

    Decellularized tissues have proven to be versatile matrices for the engineering of tissues and organs. These matrices usually consist of collagens, matrix-specific proteins, and a set of largely undefined growth factors and signaling molecules. Although several decellularized tissues have found their way to clinical applications, their use in the engineering of Cartilage tissue has only been explored to a limited extent. We set out to generate hydrogels from several tissue-derived matrices, as hydrogels are the current preferred cell carriers for Cartilage repair. Equine Cartilage, meniscus, and tendon tissue was harvested, decellularized, enzymatically digested, and functionalized with methacrylamide groups. After photo-cross-linking, these tissue digests were mechanically characterized. Next, gelatin methacrylamide (GelMA) hydrogel was functionalized with these methacrylated tissue digests. Equine chondrocytes and mesenchymal stromal cells (MSCs) (both from three donors) were encapsulated and cultured in vitro up to 6 weeks. Gene expression (COL1A1, COL2A1, ACAN, MMP-3, MMP-13, and MMP-14), Cartilage-specific matrix formation, and hydrogel stiffness were analyzed after culture. The Cartilage, meniscus, and tendon digests were successfully photo-cross-linked into hydrogels. The addition of the tissue-derived matrices to GelMA affected chondrogenic differentiation of MSCs, although no consequent improvement was demonstrated. For chondrocytes, the tissue-derived matrix gels performed worse compared to GelMA alone. This work demonstrates for the first time that native tissues can be processed into crosslinkable hydrogels for the engineering of tissues. Moreover, the differentiation of encapsulated cells can be influenced in these stable, decellularized matrix hydrogels.

  • Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in Vitro redifferentiation
    2009
    Co-Authors: Wouter Schuurman, Mattie H.p. Van Rijen, Wouter J.a. Dhert, P. René Van Weeren, Debby Gawlitta, Travis J. Klein, Werner Ten Hoope, Jos Malda
    Abstract:

    Background: If chondrocytes from the superficial, middle, and deep zones of articular Cartilage could maintain or regain their characteristic properties during in vitro culture, it would be feasible to create constructs comprising these distinctive zones. ----- ----- Hypothesis: Zone-specific characteristics of zonal cell populations will disappear during 2-dimensional expansion but will reappear after 3-dimensional redifferentiation, independent of the culture technique used (alginate beads versus pellet culture).----- ----- Study Design: Controlled laboratory study.----- ----- Methods: Equine articular chondrocytes from the 3 zones were expanded in monolayer culture (8 donors) and subsequently redifferentiated in pellet and alginate bead cultures for up to 4 weeks. Glycosaminoglycans and DNA were quantified, along with immunohistochemical assessment of the expression of various zonal markers, including Cartilage oligomeric protein (marking cells from the deeper zones) and clusterin (specifically expressed by superficial chondrocytes).----- ----- Results: Cell yield varied between zones, but proliferation rates did not show significant differences. Expression of all evaluated zonal markers was lost during expansion. Compared to the alginate bead cultures, pellet cultures showed a higher amount of glycosaminoglycans produced per DNA after redifferentiation. In contrast to cells in pellet cultures, cells in alginate beads regained zonal differences, as evidenced by zone-specific reappearance of Cartilage oligomeric protein and clusterin, as well as significantly higher glycosaminoglycans production by cells from the deep zone compared to the superficial zone.----- ----- Conclusion: Chondrocytes isolated from the 3 zones of Equine Cartilage can restore their zone-specific matrix expression when cultured in alginate after in vitro expansion.

  • Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in vitro redifferentiation.
    The American journal of sports medicine, 2009
    Co-Authors: Wouter Schuurman, Mattie H.p. Van Rijen, Wouter J.a. Dhert, P. René Van Weeren, Debby Gawlitta, Travis J. Klein, Werner Ten Hoope, Jos Malda
    Abstract:

    If chondrocytes from the superficial, middle, and deep zones of articular Cartilage could maintain or regain their characteristic properties during in vitro culture, it would be feasible to create constructs comprising these distinctive zones. Zone-specific characteristics of zonal cell populations will disappear during 2-dimensional expansion but will reappear after 3-dimensional redifferentiation, independent of the culture technique used (alginate beads versus pellet culture). Controlled laboratory study. Equine articular chondrocytes from the 3 zones were expanded in monolayer culture (8 donors) and subsequently redifferentiated in pellet and alginate bead cultures for up to 4 weeks. Glycosaminoglycans and DNA were quantified, along with immunohistochemical assessment of the expression of various zonal markers, including Cartilage oligomeric protein (marking cells from the deeper zones) and clusterin (specifically expressed by superficial chondrocytes). Cell yield varied between zones, but proliferation rates did not show significant differences. Expression of all evaluated zonal markers was lost during expansion. Compared to the alginate bead cultures, pellet cultures showed a higher amount of glycosaminoglycans produced per DNA after redifferentiation. In contrast to cells in pellet cultures, cells in alginate beads regained zonal differences, as evidenced by zone-specific reappearance of Cartilage oligomeric protein and clusterin, as well as significantly higher glycosaminoglycans production by cells from the deep zone compared to the superficial zone. Chondrocytes isolated from the 3 zones of Equine Cartilage can restore their zone-specific matrix expression when cultured in alginate after in vitro expansion. Appreciation of the zonal differences can lead to important advances in Cartilage tissue engineering. Findings support the use of hydrogels such as alginate for engineering zonal Cartilage constructs.

P. René Van Weeren - One of the best experts on this subject based on the ideXlab platform.

  • Crosslinkable Hydrogels Derived from Cartilage, Meniscus, and Tendon Tissue
    Tissue Engineering Part A, 2015
    Co-Authors: Jetze Visser, Peter A. Levett, Nikae C.r. Te Moller, Jeremy Besems, Mattie H.p. Van Rijen, Kristel W. M. Boere, Wouter J.a. Dhert, P. René Van Weeren, Janny C. De Grauw, Jos Malda
    Abstract:

    Decellularized tissues have proven to be versatile matrices for the engineering of tissues and organs. These matrices usually consist of collagens, matrix-specific proteins, and a set of largely undefined growth factors and signaling molecules. Although several decellularized tissues have found their way to clinical applications, their use in the engineering of Cartilage tissue has only been explored to a limited extent. We set out to generate hydrogels from several tissue-derived matrices, as hydrogels are the current preferred cell carriers for Cartilage repair. Equine Cartilage, meniscus, and tendon tissue was harvested, decellularized, enzymatically digested, and functionalized with methacrylamide groups. After photo-cross-linking, these tissue digests were mechanically characterized. Next, gelatin methacrylamide (GelMA) hydrogel was functionalized with these methacrylated tissue digests. Equine chondrocytes and mesenchymal stromal cells (MSCs) (both from three donors) were encapsulated and cultured in vitro up to 6 weeks. Gene expression (COL1A1, COL2A1, ACAN, MMP-3, MMP-13, and MMP-14), Cartilage-specific matrix formation, and hydrogel stiffness were analyzed after culture. The Cartilage, meniscus, and tendon digests were successfully photo-cross-linked into hydrogels. The addition of the tissue-derived matrices to GelMA affected chondrogenic differentiation of MSCs, although no consequent improvement was demonstrated. For chondrocytes, the tissue-derived matrix gels performed worse compared to GelMA alone. This work demonstrates for the first time that native tissues can be processed into crosslinkable hydrogels for the engineering of tissues. Moreover, the differentiation of encapsulated cells can be influenced in these stable, decellularized matrix hydrogels.

  • Functional adaptation of Equine articular Cartilage: the formation of regional biochemical characteristics up to age one year.
    Equine veterinary journal, 2010
    Co-Authors: Pieter A.j. Brama, Johan M. Tekoppele, Ruud A. Bank, A. Barneveld, P. René Van Weeren
    Abstract:

    Biochemical heterogeneity of Cartilage within a joint is well known in mature individuals. It has recently been reported that heterogeneity for proteoglycan content and chondrocyte metabolism in sheep develops postnatally under the influence of loading. No data exist on the collagen network in general or on the specific situation in the horse. The objective of this study was to investigate the alterations in Equine articular Cartilage biochemistry that occur from birth up to age one year, testing the hypothesis that the molecular composition of Equine Cartilage matrix is uniform at birth and biochemical heterogeneity is formed postnatally. Water content, DNA content, glycosaminoglycan content (GAG) and biochemical characteristics of the collagen network (collagen content, hydroxylysine content and hydroxylysylpyridinoline [HP] crosslinks) were measured in immature articular Cartilage of neonatal (n = 16), 5-month-old foals (n = 16) and yearlings (n = 16) at 2 predefined differently loaded sites within the metacarpophalangeal joint. Statistical differences between sites were analysed by ANOVA (P

  • functional adaptation of Equine articular Cartilage the formation of regional biochemical characteristics up to age one year
    Equine Veterinary Journal, 2010
    Co-Authors: P A J Brama, Ruud A. Bank, Johan M. Tekoppele, A. Barneveld, P. René Van Weeren
    Abstract:

    Biochemical heterogeneity of Cartilage within a joint is well known in mature individuals. It has recently been reported that heterogeneity for proteoglycan content and chondrocyte metabolism in sheep develops postnatally under the influence of loading. No data exist on the collagen network in general or on the specific situation in the horse. The objective of this study was to investigate the alterations in Equine articular Cartilage biochemistry that occur from birth up to age one year, testing the hypothesis that the molecular composition of Equine Cartilage matrix is uniform at birth and biochemical heterogeneity is formed postnatally. Water content, DNA content, glycosaminoglycan content (GAG) and biochemical characteristics of the collagen network (collagen content, hydroxylysine content and hydroxylysylpyridinoline [HP] crosslinks) were measured in immature articular Cartilage of neonatal (n = 16), 5-month-old foals (n = 16) and yearlings (n = 16) at 2 predefined differently loaded sites within the metacarpophalangeal joint. Statistical differences between sites were analysed by ANOVA (P<0.01), and age correlation was tested by Pearson's product moment correlation analysis (P<0.01). In neonatal Cartilage no significant site differences were found for any of the measured biochemical parameters. This revealed that the horse has a biochemically uniform joint (i.e. the Cartilage) at birth. In the 5-month-old foals and yearlings, significant site differences, comparable to those in the mature horse, were found for DNA, GAG, collagen content and hydroxylysine content. This indicates that functional adaptation of articular Cartilage to weight bearing for these biochemical parameters takes place during the first months postpartum. Water content and HP crosslinks showed no difference between the 2 sites from neonatal horses, 5-month-old animals and yearlings. At both sites water, DNA and GAG decreased during maturation while collagen content, hydroxylysine content and HP crosslinks increased. We propose that a foal is born with a uniform biochemical composition of Cartilage in which the functional adaptation to weight bearing takes place early in life. This adaptation results in biochemical and therefore biomechanical heterogeneity and is thought to be essential to resist the different loading conditions to which articular Cartilage is subjected during later life. As collagen turnover is extremely low at mature age, an undisturbed functional adaptation of the collagen network of articular Cartilage at a young age may be of significant importance for future strength and resistance to injury.

  • Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in Vitro redifferentiation
    2009
    Co-Authors: Wouter Schuurman, Mattie H.p. Van Rijen, Wouter J.a. Dhert, P. René Van Weeren, Debby Gawlitta, Travis J. Klein, Werner Ten Hoope, Jos Malda
    Abstract:

    Background: If chondrocytes from the superficial, middle, and deep zones of articular Cartilage could maintain or regain their characteristic properties during in vitro culture, it would be feasible to create constructs comprising these distinctive zones. ----- ----- Hypothesis: Zone-specific characteristics of zonal cell populations will disappear during 2-dimensional expansion but will reappear after 3-dimensional redifferentiation, independent of the culture technique used (alginate beads versus pellet culture).----- ----- Study Design: Controlled laboratory study.----- ----- Methods: Equine articular chondrocytes from the 3 zones were expanded in monolayer culture (8 donors) and subsequently redifferentiated in pellet and alginate bead cultures for up to 4 weeks. Glycosaminoglycans and DNA were quantified, along with immunohistochemical assessment of the expression of various zonal markers, including Cartilage oligomeric protein (marking cells from the deeper zones) and clusterin (specifically expressed by superficial chondrocytes).----- ----- Results: Cell yield varied between zones, but proliferation rates did not show significant differences. Expression of all evaluated zonal markers was lost during expansion. Compared to the alginate bead cultures, pellet cultures showed a higher amount of glycosaminoglycans produced per DNA after redifferentiation. In contrast to cells in pellet cultures, cells in alginate beads regained zonal differences, as evidenced by zone-specific reappearance of Cartilage oligomeric protein and clusterin, as well as significantly higher glycosaminoglycans production by cells from the deep zone compared to the superficial zone.----- ----- Conclusion: Chondrocytes isolated from the 3 zones of Equine Cartilage can restore their zone-specific matrix expression when cultured in alginate after in vitro expansion.

  • Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in vitro redifferentiation.
    The American journal of sports medicine, 2009
    Co-Authors: Wouter Schuurman, Mattie H.p. Van Rijen, Wouter J.a. Dhert, P. René Van Weeren, Debby Gawlitta, Travis J. Klein, Werner Ten Hoope, Jos Malda
    Abstract:

    If chondrocytes from the superficial, middle, and deep zones of articular Cartilage could maintain or regain their characteristic properties during in vitro culture, it would be feasible to create constructs comprising these distinctive zones. Zone-specific characteristics of zonal cell populations will disappear during 2-dimensional expansion but will reappear after 3-dimensional redifferentiation, independent of the culture technique used (alginate beads versus pellet culture). Controlled laboratory study. Equine articular chondrocytes from the 3 zones were expanded in monolayer culture (8 donors) and subsequently redifferentiated in pellet and alginate bead cultures for up to 4 weeks. Glycosaminoglycans and DNA were quantified, along with immunohistochemical assessment of the expression of various zonal markers, including Cartilage oligomeric protein (marking cells from the deeper zones) and clusterin (specifically expressed by superficial chondrocytes). Cell yield varied between zones, but proliferation rates did not show significant differences. Expression of all evaluated zonal markers was lost during expansion. Compared to the alginate bead cultures, pellet cultures showed a higher amount of glycosaminoglycans produced per DNA after redifferentiation. In contrast to cells in pellet cultures, cells in alginate beads regained zonal differences, as evidenced by zone-specific reappearance of Cartilage oligomeric protein and clusterin, as well as significantly higher glycosaminoglycans production by cells from the deep zone compared to the superficial zone. Chondrocytes isolated from the 3 zones of Equine Cartilage can restore their zone-specific matrix expression when cultured in alginate after in vitro expansion. Appreciation of the zonal differences can lead to important advances in Cartilage tissue engineering. Findings support the use of hydrogels such as alginate for engineering zonal Cartilage constructs.

Nancy P. Camacho - One of the best experts on this subject based on the ideXlab platform.

  • Fourier transform infrared imaging spectroscopy investigations in the pathogenesis and repair of Cartilage.
    Biochimica et biophysica acta, 2006
    Co-Authors: Xu Yang, Mathias P.g. Bostrom, Nancy P. Camacho
    Abstract:

    Abstract Significant complications in the management of osteoarthritis (OA) are the inability to identify early Cartilage changes during the development of the disease, and the lack of techniques to evaluate the tissue response to therapeutic and tissue engineering interventions. In recent studies several spectroscopic parameters have been elucidated by Fourier transform infrared imaging spectroscopy (FT-IRIS) that enable evaluation of molecular and compositional changes in human Cartilage with progressively severe OA, and in repair Cartilage from animal models. FT-IRIS permits evaluation of early-stage matrix changes in the primary components of Cartilage, collagen and proteoglycan on histological sections at a spatial resolution of ∼6.25 μm. In osteoarthritic Cartilage, the collagen integrity, monitored by the ratio of peak areas at 1338 cm−1/Amide II, was found to correspond to the histological Mankin grade, the gold standard scale utilized to evaluate Cartilage degeneration. Apparent matrix degradation was observable in the deep zone of Cartilage even in the early stages of OA. FT-IRIS studies also found that within the territorial matrix of the Cartilage cells (chondrocytes), proteoglycan content increased with progression of Cartilage degeneration while the collagen content remained the same, but the collagen integrity decreased. Regenerative (repair) tissue from microfracture treatment of an Equine Cartilage defect showed significant changes in collagen distribution and loss in proteoglycan content compared to the adjacent normal Cartilage, with collagen fibrils demonstrating a random orientation in most of the repair tissue. These studies demonstrate that FT-IRIS is a powerful technique that can provide detailed ultrastructural information on heterogeneous tissues such as diseased Cartilage and thus has great potential as a diagnostic modality for Cartilage degradation and repair.

  • A novel method for determination of collagen orientation in Cartilage by Fourier transform infrared imaging spectroscopy (FT-IRIS)
    Osteoarthritis and cartilage, 2005
    Co-Authors: S.b. Doty, Nancy P. Camacho
    Abstract:

    Summary Objective The orientation of collagen molecules is an important determinant of their functionality in connective tissues. The objective of the current study is to establish a method to determine the alignment of collagen molecules in histological sections of Cartilage by polarized Fourier transform infrared imaging spectroscopy (FT-IRIS), a method based on molecular vibrations. Methods Polarized FT-IRIS data obtained from highly oriented tendon collagen were utilized to calibrate the derived spectral parameters. The ratio of the integrated areas of the collagen amide I/II absorbances was used as an indicator of collagen orientation. These data were then applied to FT-IRIS analysis of the orientation of collagen molecules in Equine articular Cartilage, in Equine repair Cartilage after microfracture treatment, and in human osteoarthritic Cartilage. Polarized light microscopy (PLM), the most frequently utilized technique to evaluate collagen fibril orientation in histological sections, was performed on picrosirius red-stained sections for comparison. Results and conclusion Thicknesses of each zone of normal Equine Cartilage (calculated based on differences in collagen orientation) were equivalent as determined by PLM and FT-IRIS. Comparable outcomes were obtained from the PLM and FT-IRIS analyses of repair and osteoarthritis tissues, whereby similar zonal variations in collagen orientation were apparent for the two methods. However, the PLM images of human osteoarthritic Cartilage showed less obvious zonal discrimination and orientation compared to the FT-IRIS images, possibly attributable to the FT-IRIS method detecting molecular orientation changes prior to their manifestation at the microscopic level.

Clemence Desjardin - One of the best experts on this subject based on the ideXlab platform.

  • next generation sequencing identifies Equine Cartilage and subchondral bone mirnas and suggests their involvement in osteochondrosis physiopathology
    BMC Genomics, 2014
    Co-Authors: Clemence Desjardin, Anne Vaiman, Xavier Mata, Rachel Legendre, Johann Laubier, Sean Kennedy, Denis Laloë, Eric Barrey, Claire Jacques
    Abstract:

    MicroRNAs (miRNAs) are an abundant class of small single-stranded non-coding RNA molecules ranging from 18 to 24 nucleotides. They negatively regulate gene expression at the post-transcriptional level and play key roles in many biological processes, including skeletal development and Cartilage maturation. In addition, miRNAs involvement in osteoarticular diseases has been proved and some of them were identified as suitable biomarkers for pathological conditions. Equine osteochondrosis (OC) is one of the most prevalent juvenile osteoarticular disorders in horses and represents a major concern for animal welfare and economic reasons. Its etiology and pathology remain controversial and biological pathways as well as molecular mechanisms involved in the physiopathology are still unclear. This study aims to investigate the potential role of miRNAs in Equine osteochondrosis (OC) physiopathology. Short-read NGS technology (SOLID™, Life Technologies) was used to establish a comprehensive repertoire of miRNA expressed in either Equine Cartilage or subchondral bone. Undamaged Cartilage and subchondral bone samples from healthy (healthy samples) and OC-affected (predisposed samples) 10-month Anglo-Arabian foals were analysed. Samples were also subjected or not to an experimental mechanical loading to evaluate the role of miRNAs in the regulation of mechano-transduction pathways. Predicted targets of annotated miRNAs were identified using miRmap. Epiphyseal Cartilage and subchondral bone miRNome were defined, including about 300 new miRNAs. Differentially expressed miRNAs were identified between bone and Cartilage from healthy and OC foals, as well as after an experimental mechanical loading. In Cartilage, functional annotation of their predicted targets suggests a role in the maintenance of Cartilage integrity through the control of cell cycle and differentiation, energy production and metabolism as well as extracellular matrix structure and dynamics. In bone, miRNA predicited targets were associated with osteoblasts and osteoclasts differentiation, though the regulation of energy production, vesicle transport and some growth factor signaling pathways. Taken together, our results suggest a role of miRNAs in Equine OC physiopathology and in the cellular response to biomechanical stress in Cartilage and bone. In silico target prediction and functional enrichment analysis provides new insight into OC molecular physiopathology.

  • Next-generation sequencing identifies Equine Cartilage and subchondral bone miRNAs and suggests their involvement in osteochondrosis physiopathology
    BMC Genomics, 2014
    Co-Authors: Clemence Desjardin, Anne Vaiman, Xavier Mata, Rachel Legendre, Johann Laubier, Sean Kennedy, Denis Laloë, Eric Barrey, Claire Jacques, Edmond Cribiu
    Abstract:

    Background: MicroRNAs (miRNAs) are an abundant class of small single-stranded non-coding RNA molecules ranging from 18 to 24 nucleotides. They negatively regulate gene expression at the post-transcriptional level and play key roles in many biological processes, including skeletal development and Cartilage maturation. In addition, miRNAs involvement in osteoarticular diseases has been proved and some of them were identified as suitable biomarkers for pathological conditions. Equine osteochondrosis (OC) is one of the most prevalent juvenile osteoarticular disorders in horses and represents a major concern for animal welfare and economic reasons. Its etiology and pathology remain controversial and biological pathways as well as molecular mechanisms involved in the physiopathology are still unclear. This study aims to investigate the potential role of miRNAs in Equine osteochondrosis (OC) physiopathology. Short-read NGS technology (SOLID (TM), Life Technologies) was used to establish a comprehensive repertoire of miRNA expressed in either Equine Cartilage or subchondral bone. Undamaged Cartilage and subchondral bone samples from healthy (healthy samples) and OC-affected (predisposed samples) 10-month Anglo-Arabian foals were analysed. Samples were also subjected or not to an experimental mechanical loading to evaluate the role of miRNAs in the regulation of mechano-transduction pathways. Predicted targets of annotated miRNAs were identified using miRmap. Results: Epiphyseal Cartilage and subchondral bone miRNome were defined, including about 300 new miRNAs. Differentially expressed miRNAs were identified between bone and Cartilage from healthy and OC foals, as well as after an experimental mechanical loading. In Cartilage, functional annotation of their predicted targets suggests a role in the maintenance of Cartilage integrity through the control of cell cycle and differentiation, energy production and metabolism as well as extracellular matrix structure and dynamics. In bone, miRNA predicited targets were associated with osteoblasts and osteoclasts differentiation, though the regulation of energy production, vesicle transport and some growth factor signaling pathways. Conclusion: Taken together, our results suggest a role of miRNAs in Equine OC physiopathology and in the cellular response to biomechanical stress in Cartilage and bone. In silico target prediction and functional enrichment analysis provides new insight into OC molecular physiopathology.

  • a method for proteomic analysis of Equine subchondral bone and epiphyseal Cartilage
    Proteomics, 2012
    Co-Authors: Clemence Desjardin, Thierry Balliau, Benoit Valot, Michel Zivy, Laurence Wimel, G Guerin, Edmond P Cribiu, Laurent Schibler
    Abstract:

    Proteomic analyses of Cartilage and, to a lesser extent, of bone have long been impaired because of technical challenges related to their structure and biochemical properties. We have developed a unified method based on phenol extraction, 2DE, silver staining, and subsequent LC-MS/MS. This method proved to be efficient to characterize the proteome of Equine Cartilage and bone samples collected in vivo. Since proteins from several cellular compartments could be recovered, our procedure is mainly suitable for in situ molecular physiology studies focused on the cellular content of chondrocytes, osteoblasts, and osteoclasts as well as that of the extracellular matrix, with the exception of proteoglycans. Our method alleviates some drawbacks of cell culture that can mask physiological differences, as well as reduced reproducibility due to fractionation. Proteomic comparative studies between Cartilage and bone samples from healthy and affected animals were thus achieved successfully. This achievement will contribute to increasing knowledge on the molecular mechanisms underlying the physiopathology of numerous osteoarticular diseases in horses and in humans.

Mattie H.p. Van Rijen - One of the best experts on this subject based on the ideXlab platform.

  • Crosslinkable Hydrogels Derived from Cartilage, Meniscus, and Tendon Tissue
    Tissue Engineering Part A, 2015
    Co-Authors: Jetze Visser, Peter A. Levett, Nikae C.r. Te Moller, Jeremy Besems, Mattie H.p. Van Rijen, Kristel W. M. Boere, Wouter J.a. Dhert, P. René Van Weeren, Janny C. De Grauw, Jos Malda
    Abstract:

    Decellularized tissues have proven to be versatile matrices for the engineering of tissues and organs. These matrices usually consist of collagens, matrix-specific proteins, and a set of largely undefined growth factors and signaling molecules. Although several decellularized tissues have found their way to clinical applications, their use in the engineering of Cartilage tissue has only been explored to a limited extent. We set out to generate hydrogels from several tissue-derived matrices, as hydrogels are the current preferred cell carriers for Cartilage repair. Equine Cartilage, meniscus, and tendon tissue was harvested, decellularized, enzymatically digested, and functionalized with methacrylamide groups. After photo-cross-linking, these tissue digests were mechanically characterized. Next, gelatin methacrylamide (GelMA) hydrogel was functionalized with these methacrylated tissue digests. Equine chondrocytes and mesenchymal stromal cells (MSCs) (both from three donors) were encapsulated and cultured in vitro up to 6 weeks. Gene expression (COL1A1, COL2A1, ACAN, MMP-3, MMP-13, and MMP-14), Cartilage-specific matrix formation, and hydrogel stiffness were analyzed after culture. The Cartilage, meniscus, and tendon digests were successfully photo-cross-linked into hydrogels. The addition of the tissue-derived matrices to GelMA affected chondrogenic differentiation of MSCs, although no consequent improvement was demonstrated. For chondrocytes, the tissue-derived matrix gels performed worse compared to GelMA alone. This work demonstrates for the first time that native tissues can be processed into crosslinkable hydrogels for the engineering of tissues. Moreover, the differentiation of encapsulated cells can be influenced in these stable, decellularized matrix hydrogels.

  • Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in Vitro redifferentiation
    2009
    Co-Authors: Wouter Schuurman, Mattie H.p. Van Rijen, Wouter J.a. Dhert, P. René Van Weeren, Debby Gawlitta, Travis J. Klein, Werner Ten Hoope, Jos Malda
    Abstract:

    Background: If chondrocytes from the superficial, middle, and deep zones of articular Cartilage could maintain or regain their characteristic properties during in vitro culture, it would be feasible to create constructs comprising these distinctive zones. ----- ----- Hypothesis: Zone-specific characteristics of zonal cell populations will disappear during 2-dimensional expansion but will reappear after 3-dimensional redifferentiation, independent of the culture technique used (alginate beads versus pellet culture).----- ----- Study Design: Controlled laboratory study.----- ----- Methods: Equine articular chondrocytes from the 3 zones were expanded in monolayer culture (8 donors) and subsequently redifferentiated in pellet and alginate bead cultures for up to 4 weeks. Glycosaminoglycans and DNA were quantified, along with immunohistochemical assessment of the expression of various zonal markers, including Cartilage oligomeric protein (marking cells from the deeper zones) and clusterin (specifically expressed by superficial chondrocytes).----- ----- Results: Cell yield varied between zones, but proliferation rates did not show significant differences. Expression of all evaluated zonal markers was lost during expansion. Compared to the alginate bead cultures, pellet cultures showed a higher amount of glycosaminoglycans produced per DNA after redifferentiation. In contrast to cells in pellet cultures, cells in alginate beads regained zonal differences, as evidenced by zone-specific reappearance of Cartilage oligomeric protein and clusterin, as well as significantly higher glycosaminoglycans production by cells from the deep zone compared to the superficial zone.----- ----- Conclusion: Chondrocytes isolated from the 3 zones of Equine Cartilage can restore their zone-specific matrix expression when cultured in alginate after in vitro expansion.

  • Zonal chondrocyte subpopulations reacquire zone-specific characteristics during in vitro redifferentiation.
    The American journal of sports medicine, 2009
    Co-Authors: Wouter Schuurman, Mattie H.p. Van Rijen, Wouter J.a. Dhert, P. René Van Weeren, Debby Gawlitta, Travis J. Klein, Werner Ten Hoope, Jos Malda
    Abstract:

    If chondrocytes from the superficial, middle, and deep zones of articular Cartilage could maintain or regain their characteristic properties during in vitro culture, it would be feasible to create constructs comprising these distinctive zones. Zone-specific characteristics of zonal cell populations will disappear during 2-dimensional expansion but will reappear after 3-dimensional redifferentiation, independent of the culture technique used (alginate beads versus pellet culture). Controlled laboratory study. Equine articular chondrocytes from the 3 zones were expanded in monolayer culture (8 donors) and subsequently redifferentiated in pellet and alginate bead cultures for up to 4 weeks. Glycosaminoglycans and DNA were quantified, along with immunohistochemical assessment of the expression of various zonal markers, including Cartilage oligomeric protein (marking cells from the deeper zones) and clusterin (specifically expressed by superficial chondrocytes). Cell yield varied between zones, but proliferation rates did not show significant differences. Expression of all evaluated zonal markers was lost during expansion. Compared to the alginate bead cultures, pellet cultures showed a higher amount of glycosaminoglycans produced per DNA after redifferentiation. In contrast to cells in pellet cultures, cells in alginate beads regained zonal differences, as evidenced by zone-specific reappearance of Cartilage oligomeric protein and clusterin, as well as significantly higher glycosaminoglycans production by cells from the deep zone compared to the superficial zone. Chondrocytes isolated from the 3 zones of Equine Cartilage can restore their zone-specific matrix expression when cultured in alginate after in vitro expansion. Appreciation of the zonal differences can lead to important advances in Cartilage tissue engineering. Findings support the use of hydrogels such as alginate for engineering zonal Cartilage constructs.