For small diameter (<6 mm) blood vessel replacements, lack of collaterals and vascular disease preclude homografts; while synthetic analogs, ePTFE, expanded polytetrafluoroethylene, and PET, polyethyleneterephathalate, are prone to acute thrombosis and restenosis. It is postulated that the hierarchical assembly of cell populated matrices fabricated from protein analogs provides a new design strategy for generating a structurally viable tissue engineered vascular graft. To this end, synthetic elastin and collagen fiber analogs offer a novel strategy for creating tissue engineered vascular grafts with mechanical and biological properties that match or exceed those of native vessels. This work details techniques developed for the fabrication of prosthetic vascular grafts from a series of extracellular matrix analogs composed of nanofibrous collagen matrices and elastin-mimetic proteins, with and without cells, and subsequent evaluation of their biocompatibility and mechanical properties. The work details the fabrication and mechanical analysis of vascular grafts made from aforementioned protein analogs. Subesequent studies detail seeding and proliferation of rodent mesenchymal stem cells on protein-based composites to recapitulate the media of native vasculature. Finally detailing in vivo biocompatibility and stability of tissue engineered vascular grafts.

Cardiovascular disease is the main cause of death in the United States. Many of these conditions require the grafting or bypassing of compromised blood vessels. To this effect, biological vascular grafts (autografts and allografts) are the first line of action. However, when the patient lacks vasculature suitable for grafting use, several synthetic grafting options are available. The search for an inert biomaterial for vascular grafts has proven to be unsuccessful. This makes the interaction taking place on the blood-biomaterial interface critical for the success of the grafts. This thesis introduces a new bio-inspired approach to tackle the mechanical and biological challenges of vascular material design. The hypothesis of this research is that recombinant fibronectin protein can be stably incorporated onto elastin-mimetic polymers to increase endothelialization. Recombinant elastin, designed to recreate the mechanical properties of natural elastin as a candidate material for vascular graft fabrication, was used as a model surface. Recombinant fibronectin-functionalized elastin-mimetic polymer displayed significant improvement in cell adhesion. Quantification of surface bound recombinant fibronectin verified the concentration dependence of this cell adhesive behavior. Modified elastin-mimetic polymer also demonstrated an enhanced ability to support endothelial cell proliferation. Furthermore, the stability of recombinant fibronectin-modified polymers was assessed. These studies provide the foundation for fabricating elastin-mimetic vascular grafts with improved endothelialization and subsequent biological performance.

Biomimetic materials that recapitulate the complex mechanical and biochemical cues in load-bearing tissues are of significant interest in regenerative medicine and tissue engineering applications. Several investigators have endeavored to not only emulate the mechanical properties of the vasculature, but to also mimic the biologic responsiveness of the blood vessel in creating vascular substitutes. Previous studies in our lab generated the elastin-like protein polymer LysB10, which was designed with the capability of physical and chemical crosslinks, and was shown to display a range of elastomeric properties that more closely matched those of the native artery. While extensive validation of the mechanical properties of elastin-mimetic polymers has demonstrated their functionality in a number of tissue engineering applications, limited cell growth on the surfaces of the polymers has motivated further optimization for biological interaction. Recent biologically-inspired surface strategies have focused on functionalizing material surfaces with extracellular matrix molecules and bioactive motifs in order to encourage integrin-mediated cellular responses that trigger precise intracellular signaling processes, while limiting nonspecific biomaterial interactions. Consequently, this dissertation addresses three approaches to modulating cellular behavior on elastin-mimetic analogs with the goal of promoting vascular wall healing and tissue regeneration: genetic engineering of elastin-like protein polymers (ELPs) with cell-binding domains, biofunctionalization of elastin-like protein polymers via chemoselective ligation of bioactive ligands, and incorporation of matrix protein fibronectin for engineering of cell-seeded multilamellar collagen-reinforced elastin-like constructs. The synthesis of recombinant elastin-like protein polymers that integrate biologic functions of the extracellular matrix provides a novel design strategy for generating clinically durable vascular substitutes. Ultimately, the synthesis of model protein networks provides new insights into the relationship between molecular architecture, biomimetic ligand presentation, and associated cellular responses at the cell-material interface. Understanding how each of these design parameters affects cell response will contribute significantly to the rational engineering of bioactive materials. Potential applications for polymer blends with enhanced mechanical and biological properties include surface coatings on vascular grafts and stents, as well as composite materials for tissue engineered scaffolds and vascular substitutes.

The success of regenerative cardiac therapy requires reestablishing a capable blood supply via vasculature. The objective of this study was to develop an optimal scaffold formulation for de novo collateral vessel growth of aortic grafts using modified fibrin clots. This ex vivo vascular outgrowth model can be used to interrogate the complex cell or tissue interactions on the angiogenic front as vessels are formed. Based on formulation constraints, the methods used here may provide a clinically applicable option for guided collateral formation. Once understood, the methods and procedures can be tested and modified as necessary for in vivo, in situ regenerative therapy. Aortic segments from wild-type (C57BL/6J) and apolipoprotein-E deficient (ApoE) atherosclerosis-prone mice were cultured in a 3D environment created by various formulations of PEGylated fibrin. Aortic outgrowth was assessed and the optimal formulation was chosen to test the formation of de novo vascular circuits -- the first step necessary for collateral artery formation. The cultures were examined by conventional and confocal microscopy as well as by optical coherence tomography. Experiments testing the relationship between fibrin PEGylation and aortic vascular outgrowth showed that PEGylating fibrinogen prior to clot formation increased outgrowth over non-PEG control (n=6, p<.05) at lower fibrin concentrations. Lowering fibrin concentration to 10, 5, or 2.5mg/ml resulted in significantly higher outgrowth that was 1.92, 2.04, or 2.20 times that of 20mg/ml PEGylated fibrin gels. When multiple aortic segments are cultured in proximity, microvascular outgrowths visually anastamose suggesting that aorta-aorta conduits can be formed in fibrin based hydrogels. Anastomosing circuits appeared between wild-type aortic segments as well as between wild-type and atherosclerotic prone ApoE knockout segments. Fibrin gels, with or without PEGylation, form scaffolds suitable for regenerative vascular outgrowth ex vivo in normal and atherogenic environments. PEGylating fibrin prior to thrombin-initiated polymerization will allow the incorporation of growth factors or other bioactive components, making this a customizable therapy for guided collateral formation. Additionally, the incorporation of PEG itself does not limit and may actually increase the outgrowth from aortic segments in lower density gels. Finally, PEGylated fibrin gels offer an environment that will promote vascular extensions that visually anastamose, making this a viable model for ex vivo collateral formation. / text

The development of small diameter tissue engineered blood vessels (TEBVs) with low thrombogenicity, low immunogenicity, suitable mechanical properties, and a capacity to remodel to their environment could significantly advance the treatment of coronary and peripheral artery disease. Despite significant advances in the field of tissue engineering, autologous vessels are still primarily utilized as grafts during bypass surgeries. However, undamaged autologous tissue may not always be available due to disease or prior surgery. TEBVs lack long-term efficacy due to a variety of types of failures including aneurysmal dilations, thrombosis, and rupture; the mechanisms of these failures are not well understood. In vitro mechanical testing may help the understanding of these failure mechanisms. The typical mechanical tests lack standardized methodologies; thus, results vary widely. The overall goal of this study is to develop novel experimental and mathematical models to study the mechanical properties and failure mechanisms of TEBVs. Our results suggest that burst pressure tests, the current standard, are not sufficient to assess a TEBVs’ suitability as a coronary substitute; creep and/or cyclic loading tests are also required. Results from this model can help identify the most insightful experiments and quantities to be measured – ultimately reducing the overall number of experimental iterations. Improving the testing and characterization of TEBVs is critically important in decreasing the time necessary to validate the mechanical and functional responses of TEBVs over time, thus quickly moving TEBVs from the benchtop to the patient.

INTRODUCTION. La guérison rapide des sites donneurs des greffes cutanées favorise la survie des victimes de brûlures graves (>50 % de superficie brûlée). La mortalité élevée de ces patients est attribuable au fait que la superficie des brûlures excède celle de la peau saine. Des cultures épithéliales autologues (CEA) sont des feuillets de kératinocytes produits en culture à partir de la peau du patient. Cette étude a évalué l’effet des CEA sur l'épithélialisation des sites donneurs chez les grands brûlés. MÉTHODES. Tous les patients recevant des CEA ont été prospectivement inclus. Les plaies des sites donneurs ont été recouvertes de CEA, sauf pour une région contrôle randomisée de 7 x 7 cm. Des biopsies faites sur la greffe de peau ont permis de contrôler la profondeur des plaies sur les sites donneurs. Il y avait deux types de contrôles, avec gaze non adhérente trempée dans le milieu de culture ou dans le salin. L’épithélialisation était quantifiée globalement (% d’épithélialisation par photographie) et histologiquement (par biopsie au poinçon) à simple insu. La guérison des zones de contrôle et CEA était comparée par analyse de variance et par le test de Student. RÉSULTATS. Entre 2008 et 2009, 6 patients furent recrutés avec un total de 11 sites donneurs. Ces patients avaient en moyenne 43.5 ans, 56 % de superficie brûlée, 45% de brûlure pleine épaisseur, 66% avaient une brûlure d’inhalation, 75 jours de séjour. Il n’y a aucune corrélation entre le pourcentage d’épithélialisation et l’épaisseur du prélèvement des greffes (Pearson 0.19). Le score photographique est significativement influencé par le traitement (CEA vs Contrôle; p = 0,039) et par le jour postopératoire (p < 0,001). Le temps moyen pour atteindre un score photographique de guérison pour les zones contrôles fut de 10.2 jours contre 8.6 jours pour le CEA (p = 0,021). A l’évaluation histologique, les sites donneurs traités par le milieu de culture ont évolué aussi favorablement que ceux traités par des feuillets de CEA. CONCLUSION. L’utilisation de CEA sur les sites donneurs semble accélérer leur épithélialisation chez les victimes de brûlures graves. Cet effet est probablement le résultat d’une stimulation de la réépithélialisation innée de la plaie, plutôt que par une adhérence des feuillets de kératinocytes cultivés à la surface de la plaie. / RATIONALE: Prompt healing of split thickness skin graft donor sites is primordial to the survival of severely burned patients. Increased mortality of patients with >50 % TBSA is attributable to the limited availability of donor sites. This study evaluated the effect of Cultured Epithelial Autograft (CEA) application on skin graft donor site healing. METHODS: All burn patients receiving CEA were prospectively included. Donor site wounds were covered with CEA except a randomly designated 7x7 cm control region. Autograft biopsies were taken to document graft harvest thickness. One half of the controls were covered with non-adherent gauze soaked in the culture media and the other controls only received a non-adherent gauze dressing. Epithelialization was objectively evaluated by scoring blinded photographs with an analogue scale. Punch biopsies of the donor sites were evaluated histologically. Repeated measures ANOVA and T-test were used. RESULTS: Between 2008 and 2009, 6 patients were enrolled for a total of 11 donor sites. The patients averaged 43.5 years, 56 % TBSA, 45 % FT-TBSA, 66 % had inhalation injury and mean length of stay was 75 days. As expected, dermatome settings and autograft thickness measured by microscope did not correlate (Pearson 0.19). There was no correlation between the percentage of epithelialization of the punch biopsies of the donor sites and the thickness of the harvest. Photographic score was significantly influenced by its treatment CEA vs Control (p=0.039) and by postoperative day (p<0.001). Mean time to healing was 8.6 days for CEA compared to 10.2 days for controls (p=0.021). Infection was noted on only one donor site. On histologic analysis, the control sites dressed with gauze soaked in the culture media healed as nicely and promptly as the CEA sheet treated region. CONCLUSION: Use of CEA on donor sites appears to stimulate epithelialization. This effect is probably mediated by stimulation of local wound healing processes rather than by engraftment of keratinocytes from the CEA sheets.

2012 – 2013 metais Aleksandro Stulginskio Universiteto Pomologiniame sode, esančiame Kauno rajone, Ringaudų kaime, buvo vykdomi dviejų krypčių lauko bandymai: 1. Obelų vegetatyvinių poskiepių augimo dinamikos ir produktyvumo tyrimai; 2. Obelų vegetatyvinių poskiepių įtakos pirmamečių skiepų augimo dinamikai tyrimai. Obelų vegetatyvinių poskiepių augimo dinamikos ir produktyvumo tyrimai buvo atliekami poskiepių motininėje plantacijoje, siekiant įvertinti keleto M, MM ir B tipų poskiepių augimo dėsningumus vegetacijos metu, produktyvumą ir kokybinius rodiklius. Bandymas vykdytas 6 variantais, 3 pakartojimais. Pakartojimai išdėstyti sisteminiu būdu. Bandymas buvo išdėstytas tokia tvarka: 1 poskiepis – M.9 – 3 pakartojimai; 2 poskiepis – B.140 – 3 pakartojimai; 3 poskiepis – M.4 – 3 pakartojimai; 4 poskiepis – B.9 – 3 pakartojimai; 5 poskiepis – B.134 – 3 pakartojimai; 6 poskiepis – MM.106 – 3 pakartojimai. Obelų vegetatyvinių poskiepių įtakos pirmamečių skiepų augimo dinamikai tyrimai buvo atliekami ASU Pomologinio sodo medelyne. Šio bandymo tikslas buvo įvertinti, kaip skirtingų tipų poskiepiai įtakoja 'Auksio' ir 'Florina' obelaičių skiepų augimą bei kokybinius rodiklius. Bandymas vykdytas 6 variantais, 3 pakartojimais. Pakartojimai išdėstyti sisteminiu būdu. Bandymas išdėstytas tokia tvarka: 1 variantas 'Auksis'/B.396 – 3 pakartojimai; 'Florina'/B.396 – 3 pakartojimai. 2 variantas 'Auksis'/M.4 – 3 pakartojimai; 'Florina'/M.4 – 3... [toliau žr. visą tekstą] / The reaserch were carried out in periodo of 2012 – 2013 at the University of Aleksandras Stulginskis Pomology orchard in Ringaudai village, Kaunas destrict. The research were prosecuted in two directions: 1. Growth dynamics and productivity of apple trees vegetative rootstocks. 2. The influence of apple trees vegetative rootstocks to the firs year growth 'Auksis' and 'Florina' shoots. The aim of the first experiment was to investigate several M, MM and B groups rootstocks biometric parameters (height of shoots and diameter of stems), their productivity, quality index (leafs assimilation surface, leafs weight, count of leafs, count of roots, roots length). Treatment of the experiment: 1. Rootstock – M.9; 2. Rootstock – B.140; 3. Rootstock – M.4; 4. Rootstock – B.9; 5. Rootstock – B.134; 6. Rootstock – MM.106. The aim of the second experiment was to investigate biometric parameters (height of shoots, diameter of stems), quality index (leafs assimilation surface, weight of leafs) and quality of first year growth 'Auksis' and 'Florina' apple trees on different rootstocks. Treatmen of the experiment: 1. 'Auksis' – rootstock B.396; 'Florina' – rootstock B.396; 2. 'Auksis' – rootstock M.4;'Florina' – rootstock M.4; 3. 'Auksis' – rootstock MM.106;'Florina' – rootstock MM.106; 4. 'Auksis' – rootstock M.26;'Florina' – rootstock MM.106; 5. 'Auksis' – rootstock B.118;'Florina' – rootstock MM.106; 6. 'Auksis' – rootstock B.9;'Florina' – rootstock MM.106. It was determined that... [to full text]

Compliance mismatch is a significant hurdle to long-term patency in small-diameter arterial bypass grafts. Vascular tissue engineering has the potential to produce compliant, non-thrombogenic small-diameter grafts. However, current engineered grafts are relatively non-compliant, resulting in intimal hyperplasia and graft occlusion when subjected to arterial pressures. This research investigates the mechanical and biological properties of engineered constructs based on a biodegradable synthetic elastomer, poly(glycerol sebacate) (PGS). Several methods for fabricating porous PGS scaffolds in a tubular geometry were developed and compared. Adult baboon vascular cells were cultured in the scaffolds under various in vitro experimental conditions, including variations in initial cell seeding density, the type of scaffold used for culture, culture time, scaffold material, and hydrostatic pressure, and properties of the resultant constructs were compared. Scaffold fabrication using heat-shrinkable mandrels and glass tubes coated with hyaluronic acid significantly decreased tolerances of wall thickness and mechanical properties, improved handling, and decreased culture time required to reach luminal cellular confluence compared to scaffolds made with other fabrication techniques. Altering scaffold material from PGS to poly(lactide-co-glycolide) (PLGA), a benchmark biomaterial, did not affect scaffold yield, porosity, or luminal cellular confluence. Extracellular matrix (ECM) deposition increased with SMC-only culture time, and ECM deposition and remodeling during culture influenced construct compliance. Compared to PLGA scaffolds, PGS scaffolds promoted elastin crosslinking by SMCs and elastic tissue properties but attenuated collagen deposition. Hydrostatic pressure promoted ECM synthesis and deposition by SMCs and decreased construct compliance. Collagen and crosslinked elastin content in constructs correlated positively with construct burst pressure, and a negative correlation dependent on scaffold type was found between collagen content and construct compliance at low pressures. The systematic investigation of culture conditions in this research provides insights into the control of engineered blood vessel properties. The central hypothesis of this work, that grafts engineered from PGS scaffolds and adult vascular cells under biomimetic in vitro culture conditions can possess compliance comparable to autologous vessels, is true at pressures below 60 mmHg and demonstrates potential for PGS-based vascular tissue engineering. Overall, this work provides tools for engineering tubular soft tissues based on porous PGS scaffolds.

Σκοπός αυτής της πειραματικής μελέτης είναι ο έλεγχος βιολογικής συμπεριφοράς διαφόρων τύπων μοσχευμάτων που σήμερα χρησιμοποιούνται ευρέως ως υποκατάστατα οστοών. Υλικό–Μέθοδος: Στην παρούσα μελέτη χρησιμοποιήθηκαν 90 κουνέλια Νέας Ζηλανδίας, ηλικία 3.5 μηνών και βάρους 4(0.25)kg, τα οποία χωρίσθηκαν σε 6 ομάδες, η κάθε μία εκ των οποίων περιελάμβανε 15 κουνέλια. Υπό γενική αναισθησία, με ενδομυϊκή χορήγηση κεταμίνης 35mg/kg και ξυλαζίνης 5mg/kg, δημιουργήθηκε, με πλάγια χειρουργική προσπέλαση, μια οπή με φρέζα διαμέτρου 4.5mm και βάθους 8mm, στους μηριαίους κονδύλους των 2 οπισθίων άκρων του κάθε κουνελιού (σύνολο 180 οπές). Στις οπές αυτές τοποθετήθηκαν τα ακόλουθα μοσχεύματα: Ομάδα Ι-αυτομόσχευμα, Ομάδα ΙΙ-αλλομόσχευμα (Grafton®), Ομάδα ΙΙΙ-ξενομόσχευμα (Lubboc®), Ομάδα ΙV-συνθετικό υποκατάστατο οστικών μοσχευμάτων (Ceraform®), Ομάδα V- συνθετικό υποκατάστατο οστικών μοσχευμάτων (Οsteoset®), Ομάδα VI-χωρίς μόσχευμα. Μετά την τοποθέτηση των μοσχευμάτων, τα κουνέλια θυσιάστηκαν με ενδοφλέβια νατριούχο θειοπεντάλη 5ml (pentothal) 10%, σε 1, 3 και 6 μήνες όπου έγινε λήψη δειγμάτων (το κάτω τριτημόριο του μηριαίου) για ιστολογική μελέτη. Τα δείγματα αξιολογήθηκαν με μια ιστολογική κλίμακα βαθμολόγησης 15-point για να καθοριστεί η ποιότητα της πώρωσης, η παρουσία οστικού ελλείμματος, η νέοαγγειογένεση και η αντιδραστική παρουσία κυττάρων φλεγμονής, καθώς και ο βαθμός ενσωμάτωσης και ανακατασκευής του πώρου. Αποτελέσματα: Σύμφωνα με την ιστολογική κλίμακα το αυτομόσχευμα έδειξε τα καλύτερα αποτελέσματα σε όλες τις χρονικές στιγμές. Όλοι οι άλλοι τύποι μοσχεύματος έδειξαν σημαντικά κατώτερα αποτελέσματα σε σχέση με το αυτόλογο μόσχευμα (p≤0.05). Το Lubboc είχε σημαντικά καλύτερα αποτελέσματα σε σχέση με τα άλλα τρία μοσχεύματα (Grafton, Ceraform και Osteoset). Το Ceraform είχε τα κατώτερα αποτελέσματα σε όλες τις κατιγορίες Συμπεράσματα: Το αυτόλογο μόσχευμα παραμένει το πρότυπο αναφοράς “gold standard” των μοσχευμάτων, επιδεικνύοντας εξαιρετικές ικανότητες ενσωμάτωσης. Το βόειο ξενομόσχευμα (Lubboc®) συνέβαλλε στη σύνθεση του πεταλιώδους οστού πιο αποτελεσματικά από το αλλομόσχευμα (Grafton®). Τα υποκατάστατα οστών (Ceraform® και Οsteoset®) ήταν κατώτερα από τα αλλομοσχεύματα και τα ξενομοσχεύματα / Background: Different types of bone-graft substitutes have been developed and are on the market worldwide to eliminate the drawbacks of autogenous grafting. This experimental animal study was undertaken to evaluate the different histological properties of various bone graft substitutes utilized in this hospital. Material/Methods: Ninety New Zealand white rabbits were divided into six groups of 15 animals. Under general anesthesia, a 4.5 mm-wide hole was drilled into both the lateral femoral condyles of each rabbit, for a total of 180 condyles for analysis. The bone defects were filled with various grafts, these being 1) autograft, 2) DBM crunch allograft (Grafton(R)), 3) bovine cancellous bone xenograft (Lubboc(R)), 4) calcium phosphate hydroxyapatite substitute (Ceraform(R)), 5) calcium sulfate substitute (Osteoset(R)), and 6) no filling (control). The animals were sacrificed at 1, 3, and 6 months after implantation and tissue samples from the implanted areas were processed for histological evaluation. A histological grading scale was designed to determine the different histological parameters of bone healing. Results: The highest histological grades were achieved with the use of cancellous bone autograft. Bovine xenograft (Lubboc) was the second best in the histological scale grading. The other substitutes (Grafton, Ceraform, Osteoset) had similar scores but were inferior to both allograft and xenograft. Conclusions: Bovine xenograft showed better biological response than the other bone graft substitutes; however, more clinical studies are necessary to determine its overall effectiveness.

Nas últimas décadas tem aumentado o interesse pelos enxertos de pele humana radioesterilizadas, para aplicação principalmente em queimaduras extensas e profundas. Isto se deve ao fato destes enxertos apresentarem rápida aderência e menor potencial antigênico, em comparação com os demais tratamentos utilizados. A proposta deste estudo foi avaliar a histoarquitetura do enxerto de pele humana irradiada com doses de 25 kGy, 50 kGy e não irradiada, durante o processo de reparação tecidual, em camundongos Nude submetidos a enxertia de pele na região dorsal. Três grupos de animais receberam enxertos de pele humana irradiada (25 kGy e 50 kGy) e não irradiada e foram eutanasiados no 3º, 7º e 21º dia após a realização da cirurgia. Após os procedimentos histológicos de rotina, as amostras de tecido foram coradas com hematoxilina e eosina (HE) para a quantificação de queratinócitos, fibroblastos, células de defesa e vasos sanguíneos e a reação de imunofluorescência (IF) foi realizada para a determinação da expressão de colágeno do tipo I humano e do colágeno dos tipos I e III de camundongo. A quantificação, tanto das células quanto dos tipos de colágeno foi realizada por análise de imagem, utilizando o programa Image-Pro PLus 6.0. Os resultados histológicos demostraram que a pele humana irradiação, quando enxertada, influencia o aumento do número de células no local de cicatrização ao longo do tempo, principalmente na dose de 25 kGy, além de proporcionar uma melhor dispersão destas células. No 21º dia, os três grupos de animais com enxertia de pele humana tiveram parte do enxerto incorporado no processo de cicatrização. O grupo não irradiado apresentou maior incorporação do enxerto (43%), porém menor produção de colágeno do tipo III de camundongo (22%). Já os grupos com enxertia de pele irradiada apresentaram menor incorporação do enxerto (6 e 15%), mas com maior produção de colágeno do tipo III de camundongo (35% e 28%, para 25 kGy e 50 kGy, respectivamente). Com este estudo pôde-se concluir que o grupo irradiado a 25 kGy, apresenta maior proliferação celular e formação de vasos,além de melhor remodelamento da região de cicatrização. / Over the last few years it has increased the interest in the human skin grafts radio sterilized for application mainly in extensive and deep burns. Because these grafts quickly grip and present antigenic lower potential, compared with other treatments used. The purpose of this study was to evaluate the histoarchitecture of human skin grafts irradiated with doses 25 kGy, 50 kGy and non-irradiated during the pepair tissue process in nude mice submitted by skin grafting in the dorsal region. Three groups of animals received irradiated human skin grafts (25 kGy and 50 kGy) and non-irradiated and were euthanized on the 3rd, 7th and 21th day after the surgery. Indeed, routine histologic procedures, tissue samples were stained with hematoxylin and eosin (HE) for quantification of keratinocytes, fibroblasts, immune cells and blood vessels and immunofluorescence (IF) was performed to determine the expression human collagen type I and collagen type I and III mouse. Therefore, quantification of both the cells and the collagen types was performed by image analysis using Image-Pro Plus 6.0 software. Histologic results demonstrated at a dose of 25 kGy that human skin irradiation when grafted influences the increase in the number of cells in wound site over time and it provides better dispersion of these cells. In addition, on the 21st day, three groups of animals with human skin graft were embedded part of the graft in the healing process. On the other hand, the group not irradiated showed greater incorporation of the graft (43 %), but less production of collagen type III mouse (22 %). Since the groups irradiated skin graft showed lower graft incorporation (6 and 15%), but with greater production of collagen type III mice (35 % and 28 % to 25 kGy and 50 kGy, respectively). In conclusion, this study presented that the group irradiated to 25 kGy and it has a higher cell proliferation and vessel formation, and better remodeling of the healing area.