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IntroductionLes cicatrices chéloïdes (CC) sont des pathologies de la cicatrisation cutanée entraînant des gênes fonctionnelles et esthétiques souvent invalidantes. Elles sont liées à un excès de fabrication et une désorganisation du collagène lié en parti à un dérèglement du TGFβ. Les lasers thermiques permettent d'améliorer la cicatrisation par le biais d'une modification de la réponse inflammatoire. En effet, une élévation de la température entre 45 et 53°C entraine une hyperexpression des HSP 70, responsables d’une modification de l’expression du TGFβ. Le laser diode 810nm a déjà prouvé son efficacité mais il est contre indiqué chez les sujets de phototypes foncés qui sont les plus à risque de développer des CC. L'objectif de ce travail était d'évaluer l'effet d’un laser thermique sur les cicatrices chéloïdes chez l’animal et dans le cadre d’une étude clinique.Détermination des paramètres su laserDes études sur des explants de peau, puis sur des sujets volontaires sains ont permis de montrer que la longueur d’onde 1210 nm était la plus adaptée dans cette indication car elle est peu absorbée par la mélanine. Le choix des paramètres du laser ont également été déterminés afin d’obtenir une élévation contrôlée de la température cutanée. En théorie, un tir de laser diode 1210 nm, 5.1W/cm² pendant 10 secondes permet d’élever la température jusqu’à 53°C.Mise au point d’un modèle animal de CCIl n’existe pas de modèle animal de CC permettant d’étudier l’effet d’un laser. Nous avons donc mis au point un modèle pour le besoin de l’étude. Des fragments de CC humaines comprenant le derme et l’épiderme ont été greffés chez 40 souris nudes. Une évaluation clinique et histologique a permis de confirmer la bonne intégration du greffon et la persistance de son caractère chéloïde pendant 4 mois.Etude du laser diode 1210nm chez un modèle animal de CCUne étude du laser 1210 nm a été réalisée sur notre modèle animal. Il a été réalisé des tirs de laser directement sur les greffons et des évaluations cliniques et histologiques ont permis de montrer l’absence d’effet indésirable. La mesure de la température cutanée au moment du tir de laser était de 45°C en moyenne.Etude du laser diode 1210nm après excision intra chéloïdienne : étude cliniqueParallèlement à l’étude animale, il a été réalisé une étude pilote visant à évaluer la faisabilité et la sécurité d’un protocole de traitement des cicatrices chéloïdes utilisant le laser diode 1210nm. Il était réalisé une excision intra cicatricielle, puis la suture était irradiée par le laser 1210nm pendant environs 10 secondes. 20 patients ont été inclus dans l’étude. L’objectif de suivi était de 2 ans, mais l’étude est toujours en cours. Jusqu’à présent, il n’a été noté aucun effet délétère du laser. 8 patients ont bénéficié d’injections de corticoïdes en raison de récidive de la chéloïde à 6 mois.ConclusionCe travail à permis de mettre au point un modèle animal original de cicatrice chéloïde permettant pour la première fois l’étude des lasers in vivo. L’utilisation du laser 1210nm, 5.1W/cm2, pendant 10 secondes à entrainé une élévation de la température cutanée jusqu’à 45°C en moyenne sans aucun effet délétère sur la CC chez la souris. L’utilisation du laser thermique diode 1210 nm après une résection intra chéloïdienne n’a montré aucun signe de toxicité chez l’homme. La température cutanée était de 48°C en moyenne à la fin du tir. Une étude à plus grande échelle reste encore à réaliser afin de démontrer une efficacité de ce traitement prometteur. / Introduction Keloid scars (KS) are pathologies of skin healing causing often functional and aesthetic disturbances. They are linked to excess production and disorganization of collagen mostly due to TGF bêta; overproduction. The thermal lasers can improve healing through a modification of the inflammatory response. Indeed, a rise of temperature between 45 and 53 ° C leads to an overexpression of HSP 70, which causes a change in the expression of TGF bêta;. The 810nm laser diode has already proven to be effective but its wavelength cannot used on dark phototypes who are prone to developing KS.The objective of this study was to evaluate the effect of a thermal laser on keloids in animal and in the context of a clinical study.Determination of laser parameters Studies on skin explants, and on healthy volunteers have shown that 1210 nm was the most suitable wavelength for this indication because it is poorly absorbed by melanin. The laser parameters were also carefully determined in order to control the skin temperature elevation. For a 1210 nm laser diode, an irradiance of 5.1W/cm² for 10 seconds leads to a maximum temperature of 53 ° C.Development of a KS animal model. Since, there was no KS animal model of DC , an innovative one was developed for this specific study. Fragments of human KS including the dermis and epidermis were transplanted in 40 nude mice. A clinical and histological evaluation confirmed the successful integration of the graft. At 4 months, it was proven that the KS remained unchanged. Study of the 1210nm diode laser in an KS animal modelThe 1210 nm laser was evaluated on our KS animal model. Laser irradiation was performed on the grafts. Clinical and histological evaluations have shown no adverse reactions. A 45 ° C mean temperature was recorded during laser irradiation .Study of l the 1210nm diode laser after intrakeloid excision. Parallel to the animal study, a pilot study was performed to assess the feasibility and safety of a 1210nm diode laser irradiation. An intrakeloid excision was achieved; the suture was then irradiated for about 10 seconds. 20 patients were included in the study. The study is still ongoing. A 2-year follow up is scheduled. So far, it was noted that no harmful effects of the laser. 8 patients received injections of corticosteroids due to of keloid recurrence at six months.conclusionAn original KS animal model was developed for the first time to study of the in vivo effects of laser irradiation. It was demonstrated that a 1210nm diode laser (5.1W/cm², for 10 seconds) led to a 45°C skin temperature without any deleterious effect. The 1210 nm diode laser after intrakeloid excision showed no signs of toxicity in humans. In humans, the maximum temperature was 48 ° C +/- 3°C . A long term follow-up on a large series of patients is still necessary to confirm the effectiveness of this promising treatment.

Queimaduras severas causam trauma às vítimas, pela perda de líquido corpóreo, injúria no sistema vascular cutâneo e demora na cicatrização das lesões. A irradiação com laser em baixa intensidade vem sendo estudada como tratamento alternativo, por ser uma terapia não-invasiva e capaz de acelerar o processo de cicatrização. O objetivo deste estudo foi avaliar os efeitos do laser de baixa potência (λ = 660 nm) em queimaduras de pele por histomorfometria e fluxometria laser Doppler utilizando duas condições de irradiação. Trinta e seis animais com duas queimaduras criadas por vapor foram divididos em três grupos: no grupo dose fracionada (GF), as lesões foram irradiadas com dose de 1 J/cm2 nos dias 1, 3, 8 e 10; no grupo dose única (GU), as lesões foram irradiadas com dose de 4 J/cm2 no dia 1. O grupo controle (GC) não foi irradiado. O fluxo sanguíneo foi monitorado nos dias 3, 8, 10, 15 e 21 e três animais por grupo foram sacrificados nestes momentos. Os grupos irradiados mostraram um pico de vasos neoformados no dia 15, enquanto que o pico do GC foi no dia 21. O número de vasos no GC foi significantemente maior que GF e GU no dia 21. Com relação ao fluxo sangüíneo, os grupos irradiados mostraram um valor mais alto que o controle no dia 8, porém, sem diferenças significantes entre os grupos. Estes achados sugerem que o laser acelera a cicatrização de queimaduras, porém, sem diferenças significativas entre as doses estudadas. / Severe burn injuries cause extensive damage and are notoriously complicated by loss of body fluids, injury in the cutaneous vasculature and delayed wound healing. Low intensity laser therapy (LILT) has been studied as an alternative method since it is a non invasive treatment and is able to accelerate wound healing. The purpose of the present study was to evaluate the effects of LILT (λ = 660 nm) in burned skin with two different doses by histomorphometry and laser Doppler flowmetry. Thirdy six male adult Wistar rats with two burns created on the back using water vapor were divided into 3 groups. In the fractioned dose laser group (GF), the lesions were irradiated with 1 J/cm2 on days 1, 3, 8 and 10; in the single dose laser group (GU), the lesions were irradiated with 4 J/cm2 on day 1. On control group (GC) lesions were not irradiated. The blood flow was measured on days 1, 3, 8, 10, 15 and 21 and three animals per group were sacrificed in these moments. Irradiated groups showed a peak of new blood vessels formation on day 15 while for GC the peak was on day 21. At this moment, the number of vessels in GC was significantly higher than GF and GS. Regarding to blood flux, irradiated groups displayed a higher value than GC on day 8, though no significant differences were observed. These findings suggest that LILT may accelerate skin repair, however, no significant differences were observed between the studied doses.

La guérison des plaies cutanées appendiculaires chez le cheval, à la différence de celle des plaies corporelles, se complique régulièrement. Un retard de cicatrisation s’y observe et un tissu de granulation exubérant tend à s’y développer, le tout menant à une cicatrice pathologique hypertrophiée. La pathogénie exacte du tissu de granulation exubérant chez le cheval demeure inconnue à ce jour. Une hypoxie tissulaire pourrait favoriser son développement tout comme elle semble contribuer au développement de cicatrices cutanées pathologiques similaires observées chez l’Homme. L’objectif de cette étude était d’évaluer la perfusion vasculaire et la disponibilité locale en oxygène de plaies cutanées appendiculaires et corporelles en cours de cicatrisation normale et pathologique chez le cheval, à l’aide de la thermographie infrarouge et de la spectroscopie par réflectance dans le proche infrarouge. Six juments âgées de 3 à 4 ans ont été utilisées. Trois plaies cutanées ont été créées à l’aspect dorso-latéral du canon des membres thoraciques (plaies appendiculaires), et sur la paroi costale de l’un des hémithorax (plaies corporelles). Chez chaque jument, un canon a été aléatoirement bandé dans le but d’induire la formation de tissu de granulation exubérant dans les plaies s’y trouvant, tel que rapporté. La perfusion vasculaire et la disponibilité locale en oxygène ont été évaluées séquentiellement par thermographie infrarouge et spectroscopie par réflectance dans le proche infrarouge pour chaque plaie de chaque site (thorax; membre bandé; membre non bandé) au cours du processus de cicatrisation cutanée. Un modèle linéaire à doubles mesures répétées associé à une correction séquentielle de Bonferroni a révélé des différences significatives de perfusion vasculaire et de disponibilité locale en oxygène entre les plaies appendiculaires et corporelles. Ainsi la perfusion vasculaire et la disponibilité locale en oxygène étaient significativement plus élevées dans les plaies corporelles (P<0.05) et la perfusion vasculaire était significativement plus élevée dans les plaies appendiculaires non bandées que dans celles bandées (P<0.05). Nous avons récemment rapporté une plus grande occlusion de la micro-vascularisation au niveau des plaies appendiculaires chez le cheval. Nous rapportons maintenant que la perfusion vasculaire et la disponibilité locale en oxygène sont significativement inférieures dans les plaies appendiculaires, en particulier lorsqu’un tissu de granulation exubérant s’y développe. Compilés, ces résultats sous-tendent l’hypothèse que les plaies appendiculaires souffrent d’une altération de la perfusion vasculaire à l’origine possible d’une hypoxie tissulaire qui pourrait favoriser une cicatrisation cutanée anormale, telle la formation d’un tissu de granulation exubérant. / Cutaneous wound healing in horse limbs is often perturbed by the development of exuberant granulation tissue while body wounds tend to repair uneventfully. A delay in healing is usually observed in horse limb wounds, which tend to develop exuberant granulation tissue and excessive scarring. The events leading to exuberant granulation tissue formation in horses are not fully elucidated. Tissue hypoxia has been proposed as a major contributing factor as it appears to contribute to the development of similar abnormal skin scarring in human. The objective of this study was to investigate skin blood flow and tissue oxygen availability in normal / abnormal healing of full-thickness wounds created on the horse limb and body, using infrared thermography and near infrared reflectance spectroscopy. Six healthy, 3- to 4-year-old mares were used for the experiment. Three full-thickness cutaneous wounds were surgically created on the dorso-lateral surface of each metacarpal area (limb wounds) and on the lateral thoracic wall (body wounds). One randomly chosen distal forelimb was then left to heal by second intention without bandage, while the contralateral limb was bandaged postoperatively to induce the formation of exuberant granulation tissue and lead to excessive scarring. Thermal and spectroscopic data were collected from both anatomic sites (limb without bandage, bandaged limb, thoracic wall) at specific times following wounding. Mean changes in skin blood flow and tissue oxygen availability at specific wound sites over time were compared by use of repeated measures ANOVA with anatomical location and wound management as within-subject factors. A priori contrasts, submitted to Bonferroni sequential correction, were then used to compare pre-selected individual means. The statistical analysis revealed that there were significant differences in skin blood flow and tissue oxygen availability within wounds. Skin blood flow and tissue oxygen availability were significantly increased within body over limb wounds (P<0.05). Skin blood flow was significantly increased within unbandaged limb over bandaged limb wounds (P<0.05). We have recently reported a greater occlusion of microvessels in limb wounds in horses. We report here that skin blood flow and tissue oxygen availability are significantly inferior in limb wounds in horses, especially when exuberant granulation develops. Taken together, these results suggest that equine limb wounds may suffer from altered vascular perfusion and tissue hypoxia, which could favor abnormal skin healing and exuberant granulation tissue development.

Queimaduras severas causam trauma às vítimas, pela perda de líquido corpóreo, injúria no sistema vascular cutâneo e demora na cicatrização das lesões. A irradiação com laser em baixa intensidade vem sendo estudada como tratamento alternativo, por ser uma terapia não-invasiva e capaz de acelerar o processo de cicatrização. O objetivo deste estudo foi avaliar os efeitos do laser de baixa potência (&lambda; = 660 nm) em queimaduras de pele por histomorfometria e fluxometria laser Doppler utilizando duas condições de irradiação. Trinta e seis animais com duas queimaduras criadas por vapor foram divididos em três grupos: no grupo dose fracionada (GF), as lesões foram irradiadas com dose de 1 J/cm2 nos dias 1, 3, 8 e 10; no grupo dose única (GU), as lesões foram irradiadas com dose de 4 J/cm2 no dia 1. O grupo controle (GC) não foi irradiado. O fluxo sanguíneo foi monitorado nos dias 3, 8, 10, 15 e 21 e três animais por grupo foram sacrificados nestes momentos. Os grupos irradiados mostraram um pico de vasos neoformados no dia 15, enquanto que o pico do GC foi no dia 21. O número de vasos no GC foi significantemente maior que GF e GU no dia 21. Com relação ao fluxo sangüíneo, os grupos irradiados mostraram um valor mais alto que o controle no dia 8, porém, sem diferenças significantes entre os grupos. Estes achados sugerem que o laser acelera a cicatrização de queimaduras, porém, sem diferenças significativas entre as doses estudadas. / Severe burn injuries cause extensive damage and are notoriously complicated by loss of body fluids, injury in the cutaneous vasculature and delayed wound healing. Low intensity laser therapy (LILT) has been studied as an alternative method since it is a non invasive treatment and is able to accelerate wound healing. The purpose of the present study was to evaluate the effects of LILT (&lambda; = 660 nm) in burned skin with two different doses by histomorphometry and laser Doppler flowmetry. Thirdy six male adult Wistar rats with two burns created on the back using water vapor were divided into 3 groups. In the fractioned dose laser group (GF), the lesions were irradiated with 1 J/cm2 on days 1, 3, 8 and 10; in the single dose laser group (GU), the lesions were irradiated with 4 J/cm2 on day 1. On control group (GC) lesions were not irradiated. The blood flow was measured on days 1, 3, 8, 10, 15 and 21 and three animals per group were sacrificed in these moments. Irradiated groups showed a peak of new blood vessels formation on day 15 while for GC the peak was on day 21. At this moment, the number of vessels in GC was significantly higher than GF and GS. Regarding to blood flux, irradiated groups displayed a higher value than GC on day 8, though no significant differences were observed. These findings suggest that LILT may accelerate skin repair, however, no significant differences were observed between the studied doses.

La guérison des plaies cutanées appendiculaires chez le cheval, à la différence de celle des plaies corporelles, se complique régulièrement. Un retard de cicatrisation s’y observe et un tissu de granulation exubérant tend à s’y développer, le tout menant à une cicatrice pathologique hypertrophiée. La pathogénie exacte du tissu de granulation exubérant chez le cheval demeure inconnue à ce jour. Une hypoxie tissulaire pourrait favoriser son développement tout comme elle semble contribuer au développement de cicatrices cutanées pathologiques similaires observées chez l’Homme. L’objectif de cette étude était d’évaluer la perfusion vasculaire et la disponibilité locale en oxygène de plaies cutanées appendiculaires et corporelles en cours de cicatrisation normale et pathologique chez le cheval, à l’aide de la thermographie infrarouge et de la spectroscopie par réflectance dans le proche infrarouge. Six juments âgées de 3 à 4 ans ont été utilisées. Trois plaies cutanées ont été créées à l’aspect dorso-latéral du canon des membres thoraciques (plaies appendiculaires), et sur la paroi costale de l’un des hémithorax (plaies corporelles). Chez chaque jument, un canon a été aléatoirement bandé dans le but d’induire la formation de tissu de granulation exubérant dans les plaies s’y trouvant, tel que rapporté. La perfusion vasculaire et la disponibilité locale en oxygène ont été évaluées séquentiellement par thermographie infrarouge et spectroscopie par réflectance dans le proche infrarouge pour chaque plaie de chaque site (thorax; membre bandé; membre non bandé) au cours du processus de cicatrisation cutanée. Un modèle linéaire à doubles mesures répétées associé à une correction séquentielle de Bonferroni a révélé des différences significatives de perfusion vasculaire et de disponibilité locale en oxygène entre les plaies appendiculaires et corporelles. Ainsi la perfusion vasculaire et la disponibilité locale en oxygène étaient significativement plus élevées dans les plaies corporelles (P<0.05) et la perfusion vasculaire était significativement plus élevée dans les plaies appendiculaires non bandées que dans celles bandées (P<0.05). Nous avons récemment rapporté une plus grande occlusion de la micro-vascularisation au niveau des plaies appendiculaires chez le cheval. Nous rapportons maintenant que la perfusion vasculaire et la disponibilité locale en oxygène sont significativement inférieures dans les plaies appendiculaires, en particulier lorsqu’un tissu de granulation exubérant s’y développe. Compilés, ces résultats sous-tendent l’hypothèse que les plaies appendiculaires souffrent d’une altération de la perfusion vasculaire à l’origine possible d’une hypoxie tissulaire qui pourrait favoriser une cicatrisation cutanée anormale, telle la formation d’un tissu de granulation exubérant. / Cutaneous wound healing in horse limbs is often perturbed by the development of exuberant granulation tissue while body wounds tend to repair uneventfully. A delay in healing is usually observed in horse limb wounds, which tend to develop exuberant granulation tissue and excessive scarring. The events leading to exuberant granulation tissue formation in horses are not fully elucidated. Tissue hypoxia has been proposed as a major contributing factor as it appears to contribute to the development of similar abnormal skin scarring in human. The objective of this study was to investigate skin blood flow and tissue oxygen availability in normal / abnormal healing of full-thickness wounds created on the horse limb and body, using infrared thermography and near infrared reflectance spectroscopy. Six healthy, 3- to 4-year-old mares were used for the experiment. Three full-thickness cutaneous wounds were surgically created on the dorso-lateral surface of each metacarpal area (limb wounds) and on the lateral thoracic wall (body wounds). One randomly chosen distal forelimb was then left to heal by second intention without bandage, while the contralateral limb was bandaged postoperatively to induce the formation of exuberant granulation tissue and lead to excessive scarring. Thermal and spectroscopic data were collected from both anatomic sites (limb without bandage, bandaged limb, thoracic wall) at specific times following wounding. Mean changes in skin blood flow and tissue oxygen availability at specific wound sites over time were compared by use of repeated measures ANOVA with anatomical location and wound management as within-subject factors. A priori contrasts, submitted to Bonferroni sequential correction, were then used to compare pre-selected individual means. The statistical analysis revealed that there were significant differences in skin blood flow and tissue oxygen availability within wounds. Skin blood flow and tissue oxygen availability were significantly increased within body over limb wounds (P<0.05). Skin blood flow was significantly increased within unbandaged limb over bandaged limb wounds (P<0.05). We have recently reported a greater occlusion of microvessels in limb wounds in horses. We report here that skin blood flow and tissue oxygen availability are significantly inferior in limb wounds in horses, especially when exuberant granulation develops. Taken together, these results suggest that equine limb wounds may suffer from altered vascular perfusion and tissue hypoxia, which could favor abnormal skin healing and exuberant granulation tissue development.