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See detailRhoGDIalpha-dependent balance between RhoA and RhoC is a key regulator of cancer cell tumorigenesis.
Ho, Thi Thanh Giang ULg; Stultiens, Audrey; Dubail, Johanne ULg et al

in Molecular Biology of the Cell (2011), 22(17), 3263-75

RhoGTPases are key signaling molecules regulating main cellular functions such as migration, proliferation, survival, and gene expression through interactions with various effectors. Within the RhoA ... [more ▼]

RhoGTPases are key signaling molecules regulating main cellular functions such as migration, proliferation, survival, and gene expression through interactions with various effectors. Within the RhoA-related subclass, RhoA and RhoC contribute to several steps of tumor growth, and the regulation of their expression affects cancer progression. Our aim is to investigate their respective contributions to the acquisition of an invasive phenotype by using models of reduced or forced expression. The silencing of RhoC, but not of RhoA, increased the expression of genes encoding tumor suppressors, such as nonsteroidal anti-inflammatory drug-activated gene 1 (NAG-1), and decreased migration and the anchorage-independent growth in vitro. In vivo, RhoC small interfering RNA (siRhoC) impaired tumor growth. Of interest, the simultaneous knockdown of RhoC and NAG-1 repressed most of the siRhoC-related effects, demonstrating the central role of NAG-1. In addition of being induced by RhoC silencing, NAG-1 was also largely up-regulated in cells overexpressing RhoA. The silencing of RhoGDP dissociation inhibitor alpha (RhoGDIalpha) and the overexpression of a RhoA mutant unable to bind RhoGDIalpha suggested that the effect of RhoC silencing is indirect and results from the up-regulation of the RhoA level through competition for RhoGDIalpha. This study demonstrates the dynamic balance inside the RhoGTPase network and illustrates its biological relevance in cancer progression. [less ▲]

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See detailDevelopment of a Chitosan Nanofibrillar Scaffold for Skin Repair and Regeneration.
Tchemtchoua Tateu, Victor ULg; Atanasova, G.; Aqil, Abdelhafid ULg et al

in Biomacromolecules (2011), 12

The final goal of the present study was the development of a 3-D chitosan dressing that would shorten the healing time of skin wounds by stimulating migration, invasion, and proliferation of the relevant ... [more ▼]

The final goal of the present study was the development of a 3-D chitosan dressing that would shorten the healing time of skin wounds by stimulating migration, invasion, and proliferation of the relevant cutaneous resident cells. Three-dimensional chitosan nanofibrillar scaffolds produced by electrospinning were compared with evaporated films and freeze-dried sponges for their biological properties. The nanofibrillar structure strongly improved cell adhesion and proliferation in vitro. When implanted in mice, the nanofibrillar scaffold was colonized by mesenchymal cells and blood vessels. Accumulation of collagen fibrils was also observed. In contrast, sponges induced a foreign body granuloma. When used as a dressing covering full-thickness skin wounds in mice, chitosan nanofibrils induced a faster regeneration of both the epidermis and dermis compartments. Altogether our data illustrate the critical importance of the nanofibrillar structure of chitosan devices for their full biocompatibility and demonstrate the significant beneficial effect of chitosan as a wound-healing biomaterial. [less ▲]

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See detailMicroRNA-21 Exhibits Antiangiogenic Function by Targeting RhoB Expression in Endothelial Cells.
Sabatel, Céline; Malvaux, Ludovic ULg; Bovy, Nicolas ULg et al

in PLoS ONE (2011), 6(2), 16979

BACKGROUND: MicroRNAs (miRNAs) are endogenously expressed small non-coding RNAs that regulate gene expression at post-transcriptional level. The recent discovery of the involvement of these RNAs in the ... [more ▼]

BACKGROUND: MicroRNAs (miRNAs) are endogenously expressed small non-coding RNAs that regulate gene expression at post-transcriptional level. The recent discovery of the involvement of these RNAs in the control of angiogenesis renders them very attractive in the development of new approaches for restoring the angiogenic balance. Whereas miRNA-21 has been demonstrated to be highly expressed in endothelial cells, the potential function of this miRNA in angiogenesis has never been investigated. METHODOLOGY/PRINCIPAL FINDINGS: We first observed in endothelial cells a negative regulation of miR-21 expression by serum and bFGF, two pro-angiogenic factors. Then using in vitro angiogenic assays, we observed that miR-21 acts as a negative modulator of angiogenesis. miR-21 overexpression reduced endothelial cell proliferation, migration and the ability of these cells to form tubes whereas miR-21 inhibition using a LNA-anti-miR led to opposite effects. Expression of miR-21 in endothelial cells also led to a reduction in the organization of actin into stress fibers, which may explain the decrease in cell migration. Further mechanistic studies showed that miR-21 targets RhoB, as revealed by a decrease in RhoB expression and activity in miR-21 overexpressing cells. RhoB silencing impairs endothelial cell migration and tubulogenesis, thus providing a possible mechanism for miR-21 to inhibit angiogenesis. Finally, the therapeutic potential of miR-21 as an angiogenesis inhibitor was demonstrated in vivo in a mouse model of choroidal neovascularization. CONCLUSIONS/SIGNIFICANCE: Our results identify miR-21 as a new angiogenesis inhibitor and suggest that inhibition of cell migration and tubulogenesis is mediated through repression of RhoB. [less ▲]

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See detailTendon lesion and platelet-rich plasma (PRP) injection: rat model
Kaux, Jean-François ULg; Drion, Pierre ULg; Renouf, Julien et al

in Annual Congress of the RBSPRM (2010, December 03)

Introduction: For a few years, the positive effect of platelets on the healing process of different tissues (skin, bones...) was demonstrated. In fact platelets contain lots of growth factors which can be ... [more ▼]

Introduction: For a few years, the positive effect of platelets on the healing process of different tissues (skin, bones...) was demonstrated. In fact platelets contain lots of growth factors which can be release locally and enhance the healing process. Thus the aim of our experiment was to ascertain by an original mechanical measure whether the use of PRP was of interest for accelerating the healing process of rats’ Achilles tendons after surgical induced lesion. Methods: Ninety rats’ Achilles tendons were sectioned. Forty-two rats beneficed of a PRP injection in situ. After 5, 15 and 30 days, 15 rats of both groups were euthanized after tendon sampling which were immediately submitted to a biomechanical tensile test until tendon rupture, using an original method of measurement (“cryo-jaw”). Histological and biochemical analyses were made as well as a quantification of collagen with an original procedure (quantification of the “greys” on histological cross-sections). Results: Tendons in the PRP group were more resistant to rupture than those in the control group. Histological findings showed in this group an increase of collagen proliferation and better collagen fibres reorganization. However, we did not find any biochemical difference neither in term of encoding gene expression for type III collagen, matrix metalloprotease 9 and tenomodulin. Conclusion: Our animal study demonstrated that an injection of PRP could accelerate the tendons healing process and improve its quality. [less ▲]

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See detailChitosan nanofiber membranes for tissue engineering - synthesis, characterization and properties
Toncheva, Natalia ULg; Aqil, Abdelhafid ULg; Croisier, Florence ULg et al

Poster (2010, November 29)

This poster was presented by Natalia Toncheva

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See detailIntrinsic modification of tendon structure after concentric or excentric training
Kaux, Jean-François ULg; Drion, Pierre ULg; Besançon, Benoît et al

Poster (2010, November 25)

Introduction: It is well known that eccentric training is a successful way of treating chronic tendinopathy. Although clinical results are very positive, beneficial morphological and histological effects ... [more ▼]

Introduction: It is well known that eccentric training is a successful way of treating chronic tendinopathy. Although clinical results are very positive, beneficial morphological and histological effects have not yet been elucidated. The aim of our experiment was to determine if there exist any intrinsic modifications in a tendon trained in concentric or eccentric modes, in a rat model, using an original method of measurement (cryo-jaws). Methods: 18 rats were divided into 3 groups: 6 for the control group, without physical restraint; 12 for a training of1 hour, 3 times a week, for 5 weeks, at a speed of 17m/min (1km/h), on a inclined treadmill: 6 rats running uphill at +15° for the concentric effort (group C) and 6 rats running downhill at -15° for the eccentric effort (group E). After this training period, the Achilles, patellar and tricipital tendons of both limbs were surgically removed in all 18 rats. Tendons taken from five rats of each group were subjected to a tensile test up to rupture using a “cryo” jaw. Tendons of the remaining rat of each group were subjected to a histological study. Results: The results showed significant changes in group E only: (1) an increase of the force required to rupture the patellar and tricipital tendons; (2) an improvement of the ratio between the force necessary to rupture the tricipital tendon and the body mass of the rats; (3) an increase of the surface area of the section of the tricipital tendon. No significant change was observed as far as constraint was concerned between groups. Histologically, we saw, in the group E, more peripheral blood vessels and a greater proportion of collagen. Conclusion: This study showed that the mechanical properties of tendon tissue are enhanced by eccentric training. Tendons become stronger, the amount of collagen increases and there is probably more interaction between collagen fibers (mechanotransduction). [less ▲]

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See detailTendon lesion and VEGF-111 injection
Kaux, Jean-François ULg; Drion, Pierre ULg; Libertiaux, Vincent ULg et al

Poster (2010, November 25)

Introduction: Tendon lesion is one of the most frequent pathology in sports and by physical workers. This pathology often becomes chronic. For this reason, it is of interest to develop new treatments ... [more ▼]

Introduction: Tendon lesion is one of the most frequent pathology in sports and by physical workers. This pathology often becomes chronic. For this reason, it is of interest to develop new treatments. Injection of platelet-rich plasma (PRP) seems to be a promising one by releasing growth factors (GF) locally. Among all the GF released by activated platelets, the vascular endothelial growth factor-A (VEGF-A) is known to induce positive effects on vascular function and angiogenesis, and could be implicated in the healing process of tendons. Recently, a novel VEGF-A isoform was identified, the VEGF-111, a biologically active and proteolysis-resistant VEGF-A isoform, also known to present beneficial effects on ischemic diseases. This prompted us to evaluate whether VEGF-111 would have a therapeutic interest within the framework of the tendon pathology. Methods: 60 Rats were divided into 2 groups: A: control (no injection), B: VEGF-111 treatment. A 5mm defect was surgically induced in rat Achilles tendon after resection of plantaris tendon. Rats received a local injection of VEGF-111 (100ng) in situ after the surgery and were placed in their cages without immobilization. After 5, 15 and 30 days, the traumatized Achilles tendons of 10 rats of both groups were removed and dissected during their healing process. Immediately after sampling, tendons were submitted to a biomechanical tensile test up to rupture, using a “Cryo-jaw”. Rats were then euthanized. Statistical analyses were made with an ANOVA. Values are significant when p-value is below 0.05. Results: Our results showed that the developed force necessary to induce tendon rupture during biomechanical tensile test was greater for tendons which had received an injection of 100ng of VEGF-111. These results were already noticed from day 5 onwards. The ratio between force and weight increased with time in both groups, but this ratio was greater for tendons which had been submitted to an injection of VEGF111. The surface area of the section of the tendons increased between 5 and 15 days followed by a stabilization. After 30 days, sections in both groups were similar. Thus, the constraint was similar after 5 and 15 days but was better for VEGF111 group after one month. Discussion - Conclusion: This experimentation has shown that a 100ng injection of VEGF-111 stimulated tendon healing process as suggested by the increased force needed to break tendons during its healing process and the increased of constraint in comparison with the control group. Other experimentations with different concentration of VEGF111 are now in process. Acknowledgement : This experimentation was partially financed by “Standard de Liège 2007” and “Lejeune-Lechien 2008” grants. [less ▲]

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See detailTendon lesion and platelet-rich plasma (PRP) injection
Kaux, Jean-François ULg; Drion, Pierre ULg; Renouf, Julien et al

Poster (2010, November 25)

Introduction: For a few years, the positive effects of platelets on the healing process of different tissues (skin, bones...) were demonstrated. In fact platelets contain lots of growth factors which can ... [more ▼]

Introduction: For a few years, the positive effects of platelets on the healing process of different tissues (skin, bones...) were demonstrated. In fact platelets contain lots of growth factors which can be release locally and enhance the healing process. Thus the aim of our experiment was to ascertain by an original mechanical measure whether the use of PRP was of interest for accelerating the healing process of rats’ Achilles tendons after surgical induced lesion. Methods: A 5mm defect was surgically induced in 90 rats’ Achilles tendon. Rats were divided into 2 groups of 45: (A) control (no treatment) and (B) PRP treatment. Rats of group B received a PRP injection in situ after the surgery. Afterwards, rats of both groups were placed in their cages without immobilization. After 5, 15 and 30 days, 10 traumatized Achilles tendons of each group were dissected and removed. Immediately after sampling, tendons were submitted to a biomechanical tensile test up to rupture, using a “Cryo-jaw”. After that, transcriptomic analyses were made on the tendon samples, to study the expression of type III collagen, matrix metalloproteases and tenomodulin. A hydroxyproline dosage was finally realised to quantify the collagen in the tendon during its healing process. Tendons of the 15 remaining rats of each group were subjected to a histological study, respectively at day 5, 15 and 30 (5 rats for each time). Results: We demonstrated that (1) the stress (F) during biomechanical tensile test up to tendon rupture was significantly greater for tendons which had been submitted to an injection of PRP compared to the control group; (2) the surface (S) area of the section of the tendon was greater in the PRP group during the 15 first days, but this section was similar after 30 days in the 2 groups; (3) the ration F/weight of the rat was significantly greater in the PRP group at each time; (4) constrain was similar after the 15 first days but was significantly greater in the PRP group after 30 days. Histological study showed that PRP could enhance cells proliferation, angiogenesis and collagen organisation. Our biochemical analyses did not explain beneficial effects of PRP. Indeed, there was no significant difference between the expressions of different studied genes. Conclusion: Our animal study demonstrated that an injection of PRP could accelerate the tendons healing process and improve its quality. [less ▲]

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See detailInfluence du mode de contraction sur le tendon : modèle animal
Kaux, Jean-François ULg; Drion, Pierre ULg; Besançon, Benoît et al

in 3ème Congrès Commun SFMS - SFTS (2010, October 01)

Introduction : Les tendinopathies sont fréquentes et touchent aussi bien les membres supérieurs que les membres inférieurs. La rééducation excentrique constitue une thérapeutique de choix dans le ... [more ▼]

Introduction : Les tendinopathies sont fréquentes et touchent aussi bien les membres supérieurs que les membres inférieurs. La rééducation excentrique constitue une thérapeutique de choix dans le traitement des tendinopathies. Malgré les résultats favorables en clinique, les effets morphologiques et biochimiques n’ont pas encore été élucidés. Matériel et méthodes : Dix-huit rats de souche Sprague-Dawley adultes ont été répartis en trois groupes : 6 témoins (groupe T) qui ne sont soumis à aucune contrainte physique ; 6 soumis à un effort concentrique (groupe C), course en montée ; 6 soumis à un effort excentrique (groupe E), course en descente. Les 12 rats des groupes C et E ont dû courir sur un tapis roulant incliné à +15° (groupe C) ou -15° (groupe E) à une vitesse de 17m/min (1km/h) pendant une heure à raison de trois séances par semaine pendant 5 semaines. Au terme de l’entraînement, les tendons achilléens, rotuliens et tricipitaux ont été prélevés bilatéralement. Les tendons de cinq rats de chaque groupe ont bénéficié d’une évaluation biomécanique (test de traction à l’aide de mors type « cryo-jaws »). Les tendons du sixième rat de chaque groupe ont permis l’étude histologique (coloration à l’hématoxyline-éosine et trichrome de Masson). Résultats : Le groupe E présente une augmentation de la tension de rupture des tendons rotuliens (29 ,5% ; p=0,047) et tricipitaux (72% ; p=0,018), une amélioration du rapport Force/Masse pour le tendon tricipital (54% ; p=0,043) ainsi qu’une augmentation de la section tendineuse tricipitale (74% ; p=0,008). Aucune variation pour aucun des tendons entre les groupes n’est relevée pour le paramètre contrainte (=Force/Section). Histologiquement, les tendons du groupe E se caractérisent par une plus grande quantité de vaisseaux sanguins périphériques ainsi qu’une plus grande proportion de collagène. Conclusion : Cette étude démontre que les propriétés mécaniques du tendon sont améliorées après un entraînement excentrique. Le tendon plus résistant augmente sa quantité de collagène et probablement les interactions entre les fibres de collagène. [less ▲]

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See detailInjection de concentrés plaquettaires et régénération tendineuse : modèle animal
Kaux, Jean-François ULg; Drion, Pierre ULg; Renouf, Julien et al

in 3ème Congrès Commun SFMS - SFTS (2010, October 01)

Introduction : La régénération tendineuse en traumatologie du sport demeure un processus actuellement difficile à gérer et de nouvelles voies thérapeutiques sont en cours d’exploration. La littérature ... [more ▼]

Introduction : La régénération tendineuse en traumatologie du sport demeure un processus actuellement difficile à gérer et de nouvelles voies thérapeutiques sont en cours d’exploration. La littérature récente fait état d’effets bénéfiques sur la régénération tendineuse de concentrés plaquettaires (platelet-rich plasma ou PRP), administrés in situ, dus au relargage de facteurs de croissance par activation des plaquettes et à leur activité stimulante au cours de la cicatrisation. Dès lors, nous avons souhaité tester l’effet bénéfique de ce traitement sur des rats préalablement lésés au niveau de leur tendon d’Achille. Matériel et Méthode : Une section unilatérale du tendon d’Achille a été réalisée chez 60 rats Sprague Dawley adultes. De ces 60 rats, 30 ont subi une cicatrisation naturelle (rats contrôles) et 30 rats ont bénéficié d’une injection in situ de PRP le jour de la lésion. Diverses études biomécaniques, biochimiques et histologiques ont été réalisées sur ces tendons d’Achille en cours de cicatrisation à respectivement J5, J15 et J30 après lésion. Dix rats supplémentaires ont servi de témoins sains (sans lésion tendineuse). L’étude biomécanique appréciait la résistance maximale des tendons à la traction à l’aide de mors type « cryo-jaws ». L’étude histologique évaluait l’évolution cellulaire pendant la phase de cicatrisation. L’analyse transcriptomique étudiait l’expression de gènes codant pour le collagène de type III, les métalloprotéases matricielles (MMP-9) et la ténomoduline (TNMD), ainsi qu’un dosage d’hydroxyproline permettant d’évaluer la quantité de collagène présente dans le tendon au cours de la cicatrisation. Résultats : L’étude biomécanique démontre la meilleure résistance des tendons traités avec du PRP par rapport aux tendons contrôles à J5 (+19%), J15 (+30%) et significativement à J30 (+43%). L’étude histologique suggère qu’une injection de PRP stimule la prolifération cellulaire, favorise l’organisation tissulaire, stimule l’angiogenèse et la réorganisation architectural du collagène. L’étude biochimique ne permet pas d’expliquer les effets bénéfiques puisqu’il n’y a pas de différence dans l’expression des gènes des différentes molécules matricielles (collagène de type III, MMP-9 et TNMD) ni dans la quantité d’hydroxyproline qui s’accroit au cours du temps de la cicatrisation de façon similaire dans les deux groupes. Conclusion : L’injection de PRP améliore et accélère la cicatrisation tendineuse et augmente la résistance aux contraintes mécaniques du tendon en cours de cicatrisation. [less ▲]

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