Two Systems were designed. The first permits tunable on-demand softening of a hydrogel network. The second permits reversible on demand ligand exchange within a hydrogel network. Both means were shown to be cytocompatible and their uses demonstrated in cell culture of mesenchymal stem cells and 3T3 fibroblast cells.

The extracellular matrix (ECM) governs many cellular processes through biochemical and mechanical cues. Particularly, the effect ECM mechanical properties on cells fate has been well established over the years. Many hydrogel systems have been used to mimic the dynamic stiffening processes occurring in ECM. However, changes in ECM stiffness does not fully recapitulate the mechanics of native ECM, as viscoelasticity is also a major factor contributing to ECM dynamic property. This thesis describes the design and characterization of an enzyme-crosslinked hydrogel system that is not only capable of being stiffened on demand, but also can be tuned to obtain viscoelasticity. The first objective of this thesis was to utilize horseradish peroxidase (HRP) to crosslink thiol-norbornene hydrogel and use mushroom tyrosinase (MT) to create secondary DOPA-dimer crosslinks that stiffened the hydrogel. The cytocompatibility of HRP-mediated thiol-norbornene gelation and the effect of stiffening on cell fate was evaluated. The second objective of this thesis represented the first step towards developing a hydrogel system whose viscoelasticity could be dynamically tuned. Thiol-norbornene hydrogel was designed to yield dynamically adaptable boronic ester bonds via partial enzymatic reaction. Thiol-norborne hydrogel was made to contain hydroxyl phenol as well as boronic acid residues within its network. MT, in this case was used to oxidize the hydroxy phenol moieties into DOPA, which then complexed with boronic acid, created dynamic bonds, introducing viscoelasticity to an initial elastic hydrogel.

Os extratos obtidos a partir da própolis vermelha brasileira (PVB) têm sido investigados devido às suas amplas atividades biológicas. Recentemente, em nosso grupo de pesquisa, demonstramos a viabilidade da incorporação de um extrato n-hexânico de PVB em nanoemulsões de uso tópico, bem como sua permeação/retenção em pele de orelha suína. No presente estudo, avaliamos as propriedades físico-químicas e reológicas de hidrogéis contendo essas nanoemulsões visando a obtenção de um produto semissólido adequado para aplicação tópica. Em uma primeira fase, foram preparadas nanoemulsões compostas de núcleo oleoso contendo extrato n–hexânico de PVB, miristato de isopropila, lecitina de ovo (NE) e DOTAP (NE/DT), e fase externa aquosa. O polímero gelificante hidroxietilcelulose foi incorporado às formulações após a sua obtenção por emulsificação espontânea (H-NE e H-NE/DT). As formulações apresentaram-se monodispersas com diâmetro médio na faixa de 200-300 nm, confirmado por microscopia eletrônica de transmissão. H-NE apresentou um potencial zeta negativo (-38mV), enquanto o mesmo parâmetro para H-NE/DT foi positivo (+36mV), devido à presença do lipídeo catiônico DOTAP na formulação. O teor de benzofenonas totais, determinado por cromatografia líquida de alta eficiência (CLAE), foi de cerca de 85 mg/g de extrato. Esses parâmetros mantiveram-se constantes durante 90 dias de armazenamento a 4C. As formulações H/NE e H-NE/DT apresentaram um comportamento não-Newtoniano pseudoplástico. Estudos de permeação/retenção das benzofenonas através da pele de orelha suína foram realizados utilizando células de difusão do tipo Franz. A maior retenção das benzofenonas na pele (18,11 μg/cm² após 8h) foi observada para a formulação H-NE/DT, demonstrando o efeito do lipídeo catiônico DOTAP nesse parâmetro. Em uma última etapa, investigou-se a capacidade das formulações de conferirem proteção à pele de orelha suína frente ao dano oxidativo gerado pela sua exposição à luz UVA/UVB. A proteção da pele de orelha suína foi evidenciada pelas técnicas de TBARS, carbonilação de proteínas e grupamentos tióis totais. Os resultados obtidos sugerem que os Hidrogéis contendo extrato de PVB apresentam propriedades físico-químicas e reológicas adequadas para serem utilizadas topicamente para a prevenção do dano oxidativo causado pela exposição à luz UVA/UVB. / Brazilian red propolis (BRP) extracts have been investigated due to their extensive biological activities. Recently, in our research group, we demonstrated the feasibility of incorporating an n-hexane extract of BRP into topical nanoemulsions, as well as the permeation/retention of these compounds in porcine ear skin. In the present study, we evaluated the physicochemical and rheological properties of hydrogels containing these nanoemulsions in order to obtain a semi-solid product suitable for topical application. In a first step, nanoemulsions composed of an oil nucleus containing BRP n-hexane extract, isopropyl myristate, egg lecithin (NE) and DOTAP (NE/DT), and an aqueous external phase were prepared. The hydroxyethylcellulose gelling polymer was incorporated into the formulations after being obtained by spontaneous emulsification (H-NE and H-NE/DT). The formulations were monodisperse exhibiting a mean diameter in the 200-300 nm range, confirmed by transmission electron microscopy. H-NE presented a negative zeta potential (-38mV), while H-NE/DT showed a positive value (+ 36mV) due to the presence of the cationic lipid DOTAP in the formulation. The total benzophenone content, determined by high performance liquid chromatography (HPLC), was about 85 mg/g extract. These parameters were maintained constant for 90 days of storage at 4°C. The formulations H/NE and H-NE/DT presented a non-Newtonian pseudoplastic behavior. Permeation/retention studies of benzophenones through porcine ear skin were performed using Franz type diffusion cells. The highest retention of benzophenones in the skin (18.11 μg/cm² after 8h) was observed for the H-NE/DT formulation, demonstrating the effect of the DOTAP cationic lipid on this parameter. In a last step, the ability of the formulations to confer protection of porcine ear skin against oxidative damage, generated by its exposure to UVA/UVB light, was investigated. The protection was evidenced by the TBARS, carbonylation of proteins, and total thiol groups techniques. The results obtained suggest that the hydrogels containing BRP have adequate physicochemical and rheological properties to be used topically for the prevention of oxidative damage caused by exposure to UVA/UVB light.

A papaína é uma enzima proteolítica empregada no debridamento e cicatrização de feridas. Contudo, problemas de estabilidade na forma farmacêutica, bem como reações alérgicas reportadas por pacientes submetidos à tratamentos com a enzima, culminaram na restrição aos produtos contendo papaína para uso tópico por órgãos regulatórios internacionais. Este trabalho objetivou desenvolver hidrogel nanoestruturado contendo complexo de papaína e ciclodextrina visando obter forma farmacêutica estável e eficaz como curativo dérmico, com redução da resposta imunológica. A síntese do hidrogel foi realizada combinando fenômenos de cristalização e/ou reticulação e esterilização simultânea induzida por radiação gama, de modo a promover nanoestruturação adequada da membrana para veiculação da papaína nativa e do complexo. O complexo e o produto final tiveram suas propriedades biológicas e físico-químicas avaliadas. O hidrogel a base de PVA contendo complexo de papaína-ciclodextrina apresentou características adequadas para aplicação como curativo, além de apresentar indícios de redução na resposta imunológica e melhora na citocompatibilidade quando comparado à papaína nativa, isso devido ao encapsulamento molecular com a ciclodextrina e à alta retenção do complexo por parte da matriz. Por outro lado, a irradiação, não alterou o perfil citotóxico da enzima, mas acarretou leve diminuição em seu potencial imunogênico. O hidrogel se mostrou promissor para uso como curativo e demonstrou potencial redução nas reações adversas desencadeadas pelo uso da papaína. / Papain is a proteolytic enzyme applied for wound healing and debridement. However, stability issues as well as allergenic reactions reported by patients submitted to papain pharmaceutics led to restriction of papain containing products for topical use by international regulatory agencies. This work aimed the development of nanostructured hydrogel containing papain and cyclodextrin complex or papain in order to obtain stable and suitable pharmaceutical form as a wound dressing with reduced allergenic properties. The hydrogel synthesis was performed by combining freezing cycles and the simultaneous crosslinking and sterilization process promoted by gamma irradiation to achieve a nanostructured hydrogel for the loading of the papain and cyclodextrin complex. The biological and physical-chemical properties of the complex and the final product were assayed. The PVA based hydrogel containing cyclodextrin-papain complex presented desirable characteristics for wound dressing purposes. In addition, an in vitro shift in the immunological response and an increase in the cytocompatibility if compared to native papain were observed as a function of the molecular encapsulation with cyclodextrin. The process of irradiation was not capable of altering papain cytotoxicity, but conferred a slight decrease in the immunogenic properties. In conclusion, the developed hydrogel was promising as a novel papain containing dressing with potential reduced adverse reactions.

Bromelina é um nome coletivo para enzimas proteolíticas encontradas no talo, fruto e folhas do abacaxi (Ananas comosus Merr). A bromelina possui propriedades anti-inflamatórias, de debridamento, entre outras. Para a produção da bromelina deve-se, preferencialmente, usar resíduos do abacaxi, visto que os produtos do fruto têm aplicação comercial. Este trabalho teve como objetivo a extração de bromelina a partir de cascas de abacaxi através de sistema de duas fases aquosas (SDFA), e sua aplicação em hidrogel polimérico. Foram realizados estudos de estabilidade da bromelina comercial, em que se observou maior estabilidade em pH 5,0 com menor perda da atividade relativa em todas as temperaturas estudadas (20, 30, 40 e 50°C). O estudo da extração da bromelina em SDFA formado por polietileno glicol (PEG) e ácido poliacrílico (PAA) (com auxílio da análise de variância de parâmetros como rendimento, fator de purificação e coeficiente de partição) proporcionou rendimento de 335% e fator de purificação de 25,8. Os hidrogéis poliméricos à base de PEG estudados apresentaram-se flexíveis, com pouca elasticidade e taxa de absorção superior a 1000%. Hidrogel carreado de bromelina pelo método de turgescência proporcionou a maior liberação da enzima, assim como a maior atividade (80% da bromelina liberada em 24 h e 278 ± 89 U/mL). / Bromelain is a collective name for the proteolytic enzymes found in the stem, fruit and leaves of pineapple (Ananas comosus Merr.). Bromelain possesses anti-inflammatory properties, debridement, among others. For bromelain production one should preferably use the waste materials, whereas pineapple fruit products have commercial application. This study aimed to extract bromelain from pineapple peels using aqueous two-phase system (ATPS), and its application in polymeric hydrogels. Stability studies of commercial bromelain were performed, which found greater stability at pH 5.0 with minor loss of relative activity at all temperatures studied. The study of bromelain extraction in ATPS composed by polyethylene glycol (PEG) and poly acrylic acid (PAA) (with assistance of variance analysis of parameters such as yield, purification factor and partition coefficient) showed yield 335% and purification factor of 25.8. The PEG-based hydrogels studied presented flexibility, low elasticity and swelling ratio higher than 1000%. Hydrogel containing bromelain, loading by embedding (solvent sorption) method, yielded the highest enzyme release, as well as the highest activity (80% bromelain released over 24 h and 278 ± 89 U / mL).

Oropharyngeal candidiasis is a localised infection in the oropharynx region caused by Candida species, predominately C. albicans. It is commonly spread among immunocompromised patients and aggravated by hyposalivation or xerostomia. Current treatment is by systemic antifungals, which might be accompanied by gastrointestinal tract disorders, headache, allergic reactions and drug interactions or Candida becoming resistant to them. In the present work, the anti-candida activity of chlorhexidine diacetate (CHD) was tested as the drug of choice, it has no systemic side effects and microorganisms do not develop resistance against it. Thymol and farnesol were also tested individually and in combination with CHD to investigate a synergistic effect against Candida planktonic cells. The effects of CHD and thymol were investigated against C. albicans biofilm after two hours exposure by testing the metabolic stress, vacuolar activity and protein content. The results of the anti-Candida activity of CHD and thymol based on the minimum inhibitory concentration (MIC) and the minimum biocidal concentration (MBC) were 2.5 and 5 μg/ml for the former and 125 and 250 μg/ml for the later. Farnesol did not show an MIC and MBC at the investigated concentrations, however, it increased the MIC and MBC of CHD to 5 and 40 μg/ml and of thymol to 250 and >250 μg/ml, respectively. The antibiofilm activity of CHD and thymol was concentration dependent and CHD was more potent than thymol. A concentration of 20 μg/ml and 2 hours treatment of Candida biofilm grown for 24 hours showed an 85% decrease in oxidative stress, 78% and 60% loss of vacuolar activity and protein content, respectively. The combination of both drugs showed a limited increase in the activity. The cytotoxic effects of CHD and thymol were tested on human embryo kidney epithelial cell line (HEK 293); the metabolic stress, lysosomal activity and protein content were tested. The cytotoxic effects were also concentration dependent and the combination have increased the cytotoxicity. A concentration of 20 μg/ml and 2 hours treatment showed a 40% decrease in oxidative stress and neither the lysosomal activity nor protein content of HEK 293 cells was affected by the treatment Finally, a mucoadhesive hydrogel buccal tablets for the controlled release of CHD were designed and prepared to increase the residence time of an effective concentration of CHD in the oral cavity for two hours. They were prepared using Poloxamer 407 (P407), hydroxypropyl methylcellulose (HPMC) and either sorbitol, mannitol or xylitol at different ratios. The tablets were investigated for their physical properties, ex vivo mucoadhesion, the rate of hydration, gelling efficiency using image analysis, differential scanning calorimetry (DSC), Fourier transforms infrared spectroscopy (FTIR), X-ray diffractometry (XRD) and in vitro dissolution using Apparatus I and a novel method based on controlled flow rate to mimic salivary drug delivery in the oral cavity. Based on the antibiofilm activity and the cytotoxic effect of CHD a concentration of 20 μg/mL was chosen to be released from the tablets to maintain both efficacy and safety. Accordingly, to maintain this concentration the final formulations were prepared with a 2.5 mg dose of CHD. Tablets analysis showed no chemical interaction with the excipient based on DSC, FTIR and XRD. Furthermore, a novel dissolution method was developed based on a constant flow rate of the dissolution media to mimic oral salivary flow. By comparing CHD release using App I and the flow rate method it was shown that hydrogel-forming tablets successfully controlled the release of CHD regardless of the volume of the dissolution media with approximately 90% release and an average release concentration of 19 μg/ml and 1 ml/min flow rate. This making it a potential candidate for future application for treatment of candidiasis in all types of patients.

Hidrogéis são macromoléculas tridimensionais formadas por polímeros hidrofílicos, os quais são entrecruzados para manter sua estrutura. Quando expostos a água apresentam grande absorção, porém sem sofrer a dissolução. Nesta tese foram preparados hidrogéis para serem aplicados como materiais de liberação controlada de fertilizantes e condicionadores de solo. Os hidrogéis foram sintetizados através de reações de entrecruzamento de goma gelana (GGHA e GGLA) com (i) quitosana (CH), (ii) Jeffamina (JEF), (iii) L-lisina (LYS), (iv) gelatina (GEL) e alginato (ALG) com quitosana (CH). A formação de redes poliméricas foi confirmada por espectroscopia FT-IR e análises térmicas, também evidenciando a estabilidade térmica dessas amostras acima de 200°C. As morfologias dos hidrogéis liofilizados foram observadas por MEV, evidenciando as estruturas abertas e porosas. Os resultados de hidratação dos hidrogéis mostraram altos valores de absorção de água em relação ao seu peso seco, sendo 218 vezes para GGHA:CH, 164 vezes para GGHA:LYS, 145 vezes para GGHA:JEF, 113 vezes para GGHA:GEL e 80 vezes para ALG:CH. Amostras de GGLA apresentaram baixos valores de absorção de água. Os testes de perda de água dos hidrogéis ALG:CH apresentam tempo 59% maior e os hidrogéis de GGHA:CH 100% maior e, aproximadamente 60% maior para os hidrogéis GGHA:LYS, GGHA:JEF e GGHA:GEL quando comparados com a evaporação de água pura. A liberação completa do fertilizante comercial monopotássio de fosfato (MKP) ocorreu em 8 h de imersão dos hidrogéis em água e foi estimada em aproximadamente 400 mg por um grama de polímero, independente do hidrogel utilizado, enquanto os ensaios de liberação controlada do fertilizante nitrogênio, fósforo e potássio (NPK) revelaram liberação de aproximadamente 300 mg de fertilizante por um grama de hidrogel, também independente do hidrogel utilizado. Os resultados obtidos demonstraram a possível utilização de hidrogéis entrecruzados a base de macromoléculas naturais em sistemas de controle de umidade do solo e liberação controlada de fertilizantes. / New hydrogels based on natural macromolecules such as gellan gum, chitosan, alginate, gelatin and lysine were synthetized and characterized by spectroscopic, thermal and microscopic analysis. The samples were also investigated in terms of water absorption and fertilizers release. The samples were obtained by crosslinking reactions of gellan gum (GGHA and GGLA) with (i) chitosan (CH), (ii) Jeffamine (JEF), (iii) L-lysine (LYS) and (iv) gelatin (GEL). The samples of alginate (ALG) and chitosan (CH) were also obtained and characterized. The formation of polymer networks was confirmed by FT-IR spectroscopy and thermal analysis, which revealed the thermal stability of the samples above 200°C. The morphology of lyophilized hydrogels was observed by SEM presenting open and porous structure. Moreover, the results of swelling showed high water absorption values in relation to the samples dry weight. These values were of 218 times for GGHA:CH, 164 times for GGHA:LYS, 145 times for GGHA:JEF, 113 times for GGHA:GEL and 80 times for ALG:CH. GGLA samples showed lower values of water absorption when compared with other samples. The water retention time of hydrogel ALG:CH hydrogels was 59% higher than pure water. For the sample GGHA:CH the water retention time was 100% higher and about 60% higher for the hydrogels GGHA:LYS, GGHA:JEF and GGHA:GEL. The samples were charged with commercial fertilizers such as phosphate monopotassium (MKP) and phosphate monopotassium (MKP), and they were tested as controlled fertilizer release materials for agriculture. It was observed for all samples that a complete release of MKP occurred after 8 h of its immersion in water and was estimated in about 400 mg per one gram of polymer. The controlled release of NPK was in about 300 mg per one gram of hydrogel, independent of the hydrogel composition. In conclusion, all the obtained results revealed that the natural macromolecules-based hydrogels are very promising alternatives for humidity control system and controlled fertilizers release in agriculture.

O açaizeiro é uma palmeira com potencial comercial valorizado principalmente pela polpa do fruto que possui propriedades químicas benéficas à saúde humana. Os produtores necessitam corresponder ao incremento de demanda e a produção de mudas deve contemplar essa tendência de mercado. O conhecimento a respeito do manejo hídrico de mudas da espécie ainda pode ser ampliado e a utilização de polímero hidroabsorvente (hidrogel) na produção de mudas da espécie surge como uma opção para redução de custos em irrigação associada a uma boa qualidade das plantas. O objetivo deste estudo é gerar informações que correlacionem a influência de doses de hidrogel à produção de mudas de açaizeiro em diferentes regimes hídricos. O experimento foi conduzido no Departamento de Engenharia de Biossistemas da Escola Superior de Agricultura \"Luiz de Queiroz\". O delineamento utilizado foi o de blocos ao acaso sob os seguintes tratamentos, quatro doses de hidrogel (0; 1,0; 2,0; 3,0 g L-1) e três níveis de depleção (0-5; 25; 50% da capacidade de retenção de água do substrato com o polímero) com quatro repetições. O experimento foi conduzido por 9 meses e ao final foram realizadas avaliações biométricas e análises destrutivas para obtenção de valores de massa seca. Para embasamento estatístico dos dados aplicou-se o teste F, em 1 e 5% de probabilidade, análise de regressão polinomial e o teste de Tukey em 5% de probabilidade. O hidrogel aumentou a capacidade de retenção hídrica do substrato e os resultados mostraram que a presença do polímero acrescentou qualidade à muda de forma crescente em função das doses; todavia, doses de 3g L-1 podem acarretar problemas com drenagem devido à perda de eficiência de absorção do polímero. Os níveis de depleção (25 e 50%) afetaram substancialmente, de forma negativa, o desempenho da muda quando comparados ao tratamento de menor depleção (0-5%), demonstrando que a espécie é bastante sensível a regimes com menor disponibilidade hídrica. Não foi observado efeito de interação entre as doses do polímero e os níveis de depleção para a grande maioria das variáveis analisadas, com exceção apenas da área foliar específica (AFE) e da irrigação aplicada (IA). O Kc médio da cultura, para cultivo protegido, apresentou tendência crescente ao longo do experimento. Conclui-se que a melhor dose do polímero é de 2 g L-1 e que o nível de depleção de 0-5% apresentou o melhor desempenho. O Kc determinado mostra tendência de acordo com a fase de desenvolvimento da espécie, variando de 0,76 até 1,41. / The assai palm is a plant with commercial potential mainly valued by the pulp of the fruit which has chemical properties beneficial to human health. Producers need to match the demand and the production increase of seedlings should address this market trend. The knowledge of the water management of species of plants can be further extended and the use of hydrogel polymer in seedling production of the specie appears as cost reduction option in irrigation associated with good quality plants. The aim of this study is to generate information to correlate the influence of hydrogel doses to the production of assai seedlings in different water regimes. The experiment was conducted in the Department of Biosystems Engineering of the Luiz de Queiroz College of Agriculture. The design was a randomized blocks under the following treatments, four hydrogel doses (0, 1.0, 2.0, 3.0 g L-1) and three depletion levels (0-5; 25; 50% the water retention capacity of the substrate with the polymer) with four replications. The experiment was conducted for 9 months and in the end were held biometric evaluations and destructive analysis to obtain dry matter values. For statistical method of data applied the F test at 1 and 5% probability, polynomial regression analysis and the Tukey test at 5% probability. The hydrogel increased water retention capacity of the substrate and the results showed that the presence of polymer added quality changes as function of increasing doses, however, 3g L-1 doses may lead to problems with drainage because of loss of efficiency of polymer absorption. Depletion levels (25 and 50%) affected substantially negatively the performance changes when compared to untreated lower depletion (0-5%), indicating that the specie is very sensitive to lower water availability. There was no effect of interaction between the polymer doses and depletion levels for most of the variables analyzed, except only the specific leaf area (SLA) and the applied irrigation (AI). The average Kc culture for protected cultivation, presented a growing trend throughout the experiment. We conclude that the best dose of the polymer is 2 g L-1 and the depletion level of 0-5% showed the best performance. Kc was determined and shows tendency according to the phase of development, ranging from 0.76 to 1.41.

"Recently, hydrogel have found to be promising biomaterials since their porous structure and hydrophilicity enables them to absorb a large amount of water. In this study the role of water on the mechanical properties of hydrogel are studied using ab-initio molecular dynamics (MD) and coarse-grained simulations. Condensed-Phased Optimized Molecular Potential (COMPASS) and MARTINI force fields are used in the all-atom atomistic models and coarse-grained simulations, respectively. The crosslinking process is modeled using a novel approach by cyclic NPT and NVT simulations starting from a high temperature, cooling down to a lower temperature to model the curing process. Radial distribution functions for different water contents (20%, 40%, 60% and 80%) have shown the crosslinks atoms are more hydrophilic than the other atoms. Diffusion coefficients are quantified in different water contents and the effect of crosslinking density on the water diffusion is studied. Elasticity parameters are computed by constant strain energy minimization in mechanical deformation simulations. It is shown that an increase in the water content results in a decrease in the elastic. Finally, continuum hyper elastic model of contact lens is studied for three different loading scenarios using Finite Element Model. "

O β-cariofileno é um composto presente no óleo de diversas espécies vegetais, como as copaibeiras, e sua atividade frente a cicatrização cutânea ainda não está elucidada. O objetivo deste estudo foi produzir um hidrogel contendo β-cariofileno nanoemulsionado, e avaliar os perfis de liberação/permeação cutânea in vitro e a atividade cicatrizante in vivo. Para a nanoemulsão empregou-se um homogeneizador a alta pressão, e para o hidrogel, dispersão direta com Natrosol®. As formulações foram caracterizadas de acordo com o tamanho de gotícula, PDI, potencial zeta e teor (CG/MS). Nos estudos de liberação/permeação cutânea utilizou-se células tipo Franz. No estudo da atividade cicatrizante, empregou-se o modelo de feridas dorsais em ratos Wistar, avaliando o tamanho das lesões, marcadores de atividades anti-inflamatória, antioxidante e análise histológica, durante 12 dias. Após caracterizadas, as formulações desenvolvidas apresentaram-se estáveis durante 60 dias. No estudo de permeação cutânea, observou-se que o β-cariofileno puro não foi capaz de atravessar o estrato córneo da pele. Não foram observadas diferenças significativas entre as formulações na quantidade de β-cariofileno retido na derme, sugerindo haver uma situação de saturação. O estudo de liberação indicou maior cedência do β-cariofileno a partir da forma nanoemulsionada do que do hidrogel derivado. No estudo da atividade cicatrizante, o fechamento inicial da ferida foi alcançado em menos tempo no grupo tratado com o hidrogel contendo β-cariofileno nanoemulsionado (GHNβ), indicando um efeito anti-inflamatório nesta formulação. Já a análise histológica indicou que o GHNβ apresentou resultados semelhantes ao do grupo controle positivo (Dersani®), mostrando-se eficaz para a reparação tecidual cutânea. / β-caryophyllene is a compound present in the oil of many plant species, such as copaibeiras, and the healing activity is still being studied. The objectives of this study was to produce a hydrogel containing β-caryophyllene nanoemulsified and to evaluate the in vitro release and permeation profile, and the in vivo healing activity, yet unexplored. For the nanoemulsion, a high pressure homogenizer was used, and for the hydrogel, direct dispersion with Natrosol®. The formulations were characterized according to droplet size, PDI, zeta potential and content (GC/MS). In the release/permeation studies, Franz type cells were used. The study of the healing activity, the dorsal wound model was used in Wistar rats, evaluating lesion size, markers of anti-inflammatory and antioxidant activities, and histological analysis, for 12 days. After characterization, the formulations developed were stable for 60 days. In skin permeation study, was observed that the pure β-caryophyllene was not able to cross the stratum corneum. No significant differences were observed between the formulations in the amount of β-caryophyllene retained in the dermis, suggesting a saturation condition. The release study indicated higher yield of β-caryophyllene from the nanoemulsion form than from the derived hydrogel. In the study of healing activity, the initial closure of the wound was achieved in less time in the group treated with the hydrogel containing β-caryophyllene nanoemulsion (GHNβ), indicating an anti-inflammatory effect in this formulation. The histological analysis indicated that GHNβ presented similar results that of the positive control group (Dersani®), proving to be effective for cutaneous tissue repair.