A presente dissertação encontra-se subdividida em três capítulos que abordam cada uma das etapas da unidade curricular “Estágio” do Mestrado Integrado em Ciências Farmacêuticas. O primeiro capítulo aborda a componente de investigação, desenvolvida no Centro de Investigação de Ciências da Saúde (CICS), da Universidade da Beira Interior. Este focase na verificação da atividade regenerativa de amostras de mel com extrato de própolis em fibroblastos humanos da derme, seguido de uma análise de um possível sinergismo entre as propriedades bioativas de diferentes misturas de mel com extrato de própolis. Ao longo do capítulo, as características fitoquímicas e biológicas das amostras são determinadas e descritas e os resultados e conclusões apresentados. Neste sentido, os compostos fenólicos foram determinados através do método clássico de Folin-Ciocalteu. A determinação dos flavonoides foi efetuada por um método colorimétrico que usa o cloreto de alumínio. Procedeu-se de seguida à determinação da atividade antioxidante por dois métodos com princípios e mecanismos distintos - o método do DPPH baseado na sequestração de radicais livres e o sistema ßcaroteno/ácido linoleico que se baseia na atividade anti-peroxidação lipídica. Recorreram-se a dois métodos para avaliar a atividade anti-inflamatória e a viabilidade celular - um método baseado na desnaturação proteica e o método MTT, respectivamente. Por fim para avaliar a capacidade cicatrizante foi utilizado um ensaio de cicatrização - scratch wound healing assay. O segundo capítulo descreve o estágio em Farmácia Comunitária realizado na Farmácia Holon Covilhã entre 16 de setembro de 2019 e 29 de novembro de 2019, sob orientação da Dra. Patrícia Amaral. Neste capítulo são abordadas as atividades realizadas que contribuíram para a minha experiência durante o estágio, assim como ilustradas as competências e funções dos farmacêuticos. Por último o terceiro capítulo refere-se ao estágio em Farmácia Hospitalar realizado no Centro Hospitalar Universitário Cova da Beira, no período de 4 de dezembro de 2019 a 24 de janeiro de 2020 sob orientação da Dra. Olímpia Fonseca. Neste capítulo são descritas as competências adquiridas assim como o funcionamento dos serviços farmacêuticos.

The objective of this study was to investigate the effects of bevacizumab, an inhibitor of the vascular endothelial growth factor activity, after intralesional administration in eyelid scars of rabbits. Identical bilateral linear wounds were created with a #15 scalpel blade in the periocular region of each rabbit. The wounds healed by second intention, and 1 month postoperatively, apparently similar scar formation was confirmed bilaterally on inspection. On postoperative day 30 (between the scar maturation phase and the beginning of the remodeling phase), all scars on the right side were administered with a subcutaneous injection of bevacizumab (2.5 mg/0.1 mL) in the middle third of the total scar extension. The left side (contralateral) was not treated, and it served as a control. One month later, the eyelid scars and general health of the rabbits were evaluated. After checking for intact and morphologically similar wounds through simple inspection, the animals were euthanized. Subsequently, the periocular scar material was collected by removing its entire length with a scalpel blade and sent for histological evaluation of the quantity of new vessels, collagen subtypes, and scar elevation index. This study included nine 5-month-old New Zealand rabbits weighing 4 to 4.5 kg. The mean scar elevation index score was 1.35 on the treated side and 1.80 on the untreated side (p = 0.001). Further, neovascularization was lesser (88,9% vs 22,2%) (p = 0.006) in the eyelids treated with bevacizumab compared to controls. In this animal model, the administration of bevacizumab was associated with a reduced rate of scar elevation and reduced neovascularization.

Aloe vera gel is a very popularly plant used in burns treatment. This study aimed to evaluate the healing activity of Aloe vera L. gel provided by a cosmetics company in 3rd degree burns caused experimentally in rats. This study used 45 male Wistar rats, adults who were divided into 3 groups with 05 subgroups, each subgroup with n = 03. Group 1, treated with 1% silver sulfadiazine; group 2, treated with aqueous solution containing 1% preservative from the Aloe vera gel and group 3, treated with 98% Aloe vera gel. The animals were anesthetized, undergo shaving and cleaning the cervical dorsal region with saline solution and burn injury 3rd degree with 4cm² using boiling water to about 96 C in contact with the animal's skin for 14 seconds. The animals received an occlusive dressing on all experimental days. The wounds were evaluated on the 3rd, 7th, 14th, 21st and 30th days after the burn and the animals were euthanized for morphological, macroscopic and microscopic analysis. The evaluation of macroscopic results, whose parameters were crust, hyperemia, granulation tissue and fibrosis were no statistically significant differences between groups 1, 2 and 3 when compared over time. Morphometry, which analyzed the wounds contraction degrees were also no statistically significant differences between groups 1, 2 and 3 in the evaluated periods. The evaluation of microscopic results found some statistically significant results in 3th day: hyperemia and fibroblasts were higher in group 1 when compared to group 2; on the 30th day , angiogenesis was higher in groups 2 and 3 , when compared to group 1. Other parameters: necrosis, hyperemia, hemorrhage, fibrin and collagen, showed no statistically significant differences when comparing the groups over time. The 98% Aloe vera gel of the present study did not show the expected efficacy in relation to the anti-inflammatory action and healing among the three groups compared the experimental days.

INTRODUCTION: Vascular ulcers are a major public health problem, affecting around 1% of the general population, with a higher prevalence among the aged. Treatment is time consuming and costly, they can cause physical, social, economic and emotional problems. They are considered complex and difficult to heal, have a high number of relapses, and most of them are critically colonized or infected. Service to people with vascular ulcers is challenging to both health professionals and the public administration by virtue of its complexity, including the absence of public policies for this population, structuring services, access to care, professional qualification, and appropriate resources. GENERAL OBJECTIVE: To analyze the outcomes of healthcare for people with vascular ulcers treated in the examination rooms of the primary care system from the perspective of the evolution of healing over a period of six months. METHODOLOGY: Analytic longitudinal study with nested cross-sectional studies, in the examination rooms of the public health service of the city of Aparecida de Goiania, Goias, Brazil, from August 2012 to October 2013. Population consists of users receiving treatment by nurses, and professionals who provided nursing care in the study setting. Data collection occurred through structured nonparticipant observation, interview, physical examination, and use of Pressure Ulcer Scale for Healing (PUSH), planigraphy of ulcers, and photographic record. Absolute frequencies and percentages were used for data collection. The study complies with guidelines for research ethics, being approved under protocol 085/2012. RESULTS: The three rooms designed to care for people with vascular ulcers in the studied scenario showed non-washable walls, windows not screened, without adequate hand hygiene resources, without proper procedures for cleaning and disinfection of the environment, and lack of essential resources and inconsistent supply thereof. As for users, there was gender parity, prevalence among 60 years or older, and social and economic conditions in the lower middle class and below. The majority had lesions for more than one year, which were usually painful. Microbiological examination showed predominantly Gram-negative rods, including Pseudomonas aeruginosa. Among professionals, most were females, aged between 18 and 36 years and "reasonable experience" in treating people with vascular ulcers. CONCLUSION: Conditions were inadequate for serving people with vascular ulcers and contribute to care not compliant with current recommendations, which creates a discontinuity of care and diminished capacity to resolve these issues. Therefore, it is necessary to structure the service with respect to people with chronic ulcers, and maintain supplies and appropriate resources for care.

The economic relevance of equine breeding in Brazil indicates the importance of the development of new economically viable treatments to promote tissue healing. Skin grafts and Platelet Rich Plasma (PRP) are available alternatives for improving the wound healing process in these animals. This study aimed to establish a simple method for autologous PRP production, as well as to evaluate its action on cutaneous wounds containing skin autografts in horses. Eight healthy horses were used, ranging in age from 3 to 18 years and mean live weight of 350 kg. For PRP obtention, a double centrifugation protocol was used. Two 6 x 6 cm wounds were performed on each side of the gluteal region. After the wound was filled by granulation tissue, punch autografts were obtained, with fragments harvested from the neck. Left-sided wounds were treated with PRP (GT) and right-sided wounds were not treated (GC). The protocol used to obtain PRP resulted in platelet concentration, on average, three times higher than that of whole blood. In the evaluation of PRP action in the healing of autografts, macroscopic and microscopic variables were considered. There was no significant difference between the groups (p> 0.05) for the variables inflammatory infiltrate, young fibroblasts, collagenization, epidermal thickness, macroscopic and microscopic integration of the autografts and retraction of the wound edges. However, neovascularization was significantly greater (p = 0.0191) in the PRP group, on the 14 th day after insertion of the autografts. It is concluded that the double centrifugation method results in PRP with adequate platelet concentration for therapeutic use. As for the action of PRP, this favors the process of skin repair with autografts in horses, since it increases the neovascularization in the initial phase of wound healing. Moreover, PRP seems to positively influence the integration of skin autografts and retraction of the wound edges, which is of fundamental importance for the reduction of hospitalization time and consequently of the therapeutic costs.

Analyze the conditions of care to people with vascular ulcers and the evolution of venous ulcers and associated factors among users of the public healthcare system in an interior city. The research integrated a cross- sectional design with a cohort study, developed from July 2011 to October 2013, in an interior Brazilian Midwest city. It involved people with vascular ulcers, the wound dressing units where this population was cared for and nursing professionals who worked there. Data were collected using interviews, clinical examinations, the Pressure Ulcer Scale Healing (PUSH), planigraphy, photographic records of the lesions, samples collected from the lesions for microbiological analysis, and a script for observation of the units. The greatest ulcer was elected as the main lesion. Criteria were established to detect ulcers considered to be infected and to classify the risk for cross- contamination in the units. A questionnaire was used to approach the nursing professionals. Associations were analyzed using a marginal homogeneity test, McNemar and the chi-square test, with p<0.05. The research proposal was approved by the Research Ethics Committee of the Federal University of Goiás (protocol 026/2011). Initially,31 people with vascular ulcers undergoing treatment at the 13 wound dressing units of the studied city were included. All units presented conditions classified as highly susceptible to cross- contamination. Of the 31 assessed ulcers, 29 (93.5%) presented evidence of infection. Samples could be collected from 25 lesions, and a positive result in culture was found for 23 (92.0%), with presence of one to three microorganisms. Seven (20.0%) occurrences were found for oxacillin- resistant staphylococcus, four (11.4%) forampicillin-resistant enterococci, and two (5.7%) for beta-lactam-resistant pseudomonasaeruginosa. Of the 26 participants with venous ulcers, 22 were included in the cohort and presented a mean age of 57.9 years, ulcers of median duration of 24 months (SD±32.8), covering an area up to 12 cm 2 (63.5%). An 18.2% healing rate was found in the follow-up from 6 to 15 months, with improvement in the condition of the ulcers, as evidenced by decreased total scores in the PUSH (p=0.041). Aggravation in healing conditions was associated with clinical signs of infection in the lesions (p=0.019). Regarding self-care, there was a prevalence of reports of care with feeding and rest, which did not agree with the recommendations in literature, and the use of medications to control systemic diseases. Among the 53 nursing professionals who worked in the units, nursing technicians (56.6%) prevailed. Of the 411 activities referred in the care of people with vascular ulcers, 392 were mapped in theNursing Intervention Classification, with 53.3% corresponding to the “wound/skin control” class, demonstrating that care is focused on the topical treatment of the lesion. In a context of limitations in the care of users, the evolution of the healing of ulcers in follow-up was found favorable, however, the healing rate was significantly low. This context points to the need to reorganize public policies for the care of people with vascular ulcers, as well as the model of management of care to this population.

The relevance of cutaneous wounds in the veterinary routine indicates the importance of the expansion of economically viable therapeutic techniques aimed at its treatment. The application of ozonized solutions, especially ozonated water and oil, are presented as techniques that may contribute in the tissue repair process. This study aimed to evaluate and compare the effects of ozonated water and oil on the tissue repair of experimentally induced wounds in rats. Forty-eight female Wistar albino rats weighing between 250 and 300 grams were used. Single 9 mm in diameter wounds were performed on the dorsum of the animals, between the scapulae. The wounds were treated daily in the different experimental groups, with 12 mice treated with ozonated water (GA), 12 mice with ozonated oil (GO), 12 mice with 0,2% allantoin cream (GAL) and 12 rats treated with 0,9% sodium chloride (GCL). The wounds were submitted to macroscopic morphometry and microscopic evaluation with HE, picrosirius red and immunostaining for vascular endothelial growth factor (VEGF). At the macroscopic evaluation, the GA group animals presented a higher percentage of retraction than the rats of the other groups at days three, eight and 15 after induction, and the GA retraction was statistically larger than that of the rats of the GO and GAL groups on days three and eight (p<0,05). Regarding the type of extracellular matrix (ECM) in the wounds, there was no statistical difference between the studied group. However, picrosirius red staining showed that the GA and GO groups showed higher total collagen deposition at 15 and 22 days of treatment, respectively. Moreover, at these times of evaluation, these groups presented a higher percentage of type I collagen fibers compared to other groups (p <0.05). Moreover, the neovascularization variable was higher in the GO rats wound on all evaluated days, being statistically higher at three and eight days compared to the GCL group, at 15 days compared to the GCL and GAL. At 22 days, the GA group had a significantly smaller number of vessels compared to the other groups, indicating that this group went into scar remodeling more quickly. These findings were corroborated by VEGF immunohistochemistry marking, which was higher in the GO group at all times, which generated significantly greater immunostaining in the animal's wounds at 8 days compared to the GCL group, at 15 days compared to the group GA, and at 22 days for the GA and GCL groups. It is concluded that ozone water and oil at the tested concentrations contribute to the tissue repair process in experimentally induced skin wounds in rats. Ozonized water enhances wound retraction as well as promotes better epithelial organization and accelerates the onset of healing in the maturation and remodeling phase, taking into account the greater deposition of type I collagen at 15 days and less vascularization at 22 days; while ozonized oil at 418,48 mEq/kg of peroxides promotes greater neovascularization during tissue repair, as well as greater deposition of type I collagen from the third week of treatment.

Leishmaniasis is a neglected disease, caused by protozoa of the genus Leishmania, which affects organs (visceral leishmaniasis), skin and mucous (American tegumentary leishmaniasis, ATL). The treatment of ATL is effective, however causes several collateral effects, and the absence treatment or incomplete treatment is associated with relapse. Alternative treatments should, therefore, be explored, and low potency lasers indicate the possibility of a noninvasive therapy without collateral effects and that can act together with conventional therapy. LLLT (Low-Level Laser Therapy) was applied on experimental mice BALB/c lesions, caused by Leishmania (Leishmania) amazonensis (MAB-6) in order to evaluate the healing time. The cytokines profile expression was evaluated by real-time PCR and ELISA, and Immunohistochemistry was performed to quantify M2 macrophages using MP-23 antibody. In the lesions treated with laser, it was observed a lower healing time when compared with the group of animals with untreated injuries. Moreover, it was observed an increased expression of regulatory and anti-inflammatory cytokines in the inflammatory infiltrate. We evaluated the effect of laser on macrophages RAW264.7, differentiated or not for the M1 or M2 profile, and infected in vitro. Also, we evaluated the phagocytic activity by quantification of the infected macrophages and the number of amastigotes per infected macrophage at 4 and 24 hours after infection; and nitric oxide (NO) production. The laser treatment increased the NO production and phagocytic activity of macrophages in different profiles, including those macrophages that were not differentiated before the infection. The 660nm laser stimulates macrophages and accelerates the healing of experimental injuries caused by L. amazonensis. Depending on the stage of the infection that the treatment is initiated, the laser may affect the inflammatory phase for the elimination of the parasite or stimulate the proliferative phase regulators and anti-inflammatory cytokines.

Studies with medicinal plants as alternative treatment for animals and people have been developing, because of the low cost income and due to the emergence of multi-resistant bacteria to synthetic antibiotics. Among the great popular use of plants is the Coité or Crescentia cujete. It is a Bignoniaceae, also known as gourd or cuieira. It is a native specie of tropical America and can be easily found in Brazil. It is used to combat high blood pressure, diabetes, urinary problems, coughs and anemia, among others. There are reports of popular Coité extract use for the treatment of wounds in humans and animals, but no scientific proof of this use is currently available. The study aimed to evaluate the healing process of skin wounds in rats treated with Coité (Crescentia cujete), and its bactericidal power. The second chapter, it tested the therapeutic potential of Coité in skin wounds in rats. To this end, we used 18 rats, female, 45 days old and 115 grams, on average. The animals were divided into three groups according to the proposed type of treatment (Coité alcoholic extract of 25% commercial neomycin healing ointment base to 5% and negative control saline - 0.9% NaCl). To carry out the wounded was instituted general anesthesia and skin excision in the dorsal cervical region with punch 0.6 cm. The animals of all groups were treated daily, every 12 hours until complete healing of the tissue. The results showed that the Coité extract contributed to the healing process evolved satisfactorily. In the third chapter, in addition to the healing potential, it evaluated the antibacterial power plant. The test animals included the assessment of the wounds the same way as described in chapter 2, with the difference that these lesions were contaminated with polymicrobial solution of fresh feces of rats own immediately after surgical skin removal. It was observed that the aqueous solution containing the ethanol extract of Coité to 25% helped the scar healing process with less complications and early centripetal contraction of the lesions when compared with the ointment neomycin and brine. To demonstrate the antibacterial effect of Coité, a study was conducted in vitro on Escherichia coli strains, Staphylococcus aureus, Streptococcus spp. and Enterobacter spp. by immunodiffusion in agar, with the construction of a hole for placing the liquid to be tested. The aqueous solution of Coité extract showed inhibitory activity (inhibition zone formation) in all the bacteria used. The protocol used is an effective alternative for treatment of skin lesions.

The use of Hevea brasiliensis latex in the wound treatment have been widely studied as a biomembrane bandage. Nowadays it is comercially available as Regederm®, a gel-ointment indicated in the use of cutaneous wounds treatment. Also the Hevea brasiliensis látex has been used in the wound healing because of its angiogenic and anti-inflammatory ativities. The presente study aimed the comparison of the healing effects of H. speciosa biomembrane and Regederm®. 70 Wistar rats divided in G1 – control group, treated with physiologic solution, G2 – treated with Regederm®, G3 – treated with H. speciosa biomembrane, G4 – mixed treatment with Regederm® and physiologic solution. The euthanasia occurred at 3, 7, 14 and 21 days after the induction (DAI) of the incisional injury. Morphometric, microscopic and macroscopic analysis were performed evaluating the general pathologic processes. It was possible to observe that the H. speciosa biomembrane decreased necrosis and inflammation during the inflammatory and proliferative phases of the healing process. The angiogenesis observed was similar to the one found in the Regederm® treated group. During the maturation phase of the healing process the Regederm® continued to stimulate the angiogenesis which was not observed in the H. speciosa biomembrane treated group. There was an intense stimulus of fibroblastos with the Regederm® treated group and the mixed treatment group in the proliferative and maturation phases of the healing process while in the H. speciosa biomembrane treated group there was an inhibition of these cells. It was possible to observe that in the Regederm® treated group there was a continuous inflammation of the tissue during the maturation phase of the healing process. Therefore it is possible to conclude that Regederm® contributed to the healing process in its inflammatory and proliferative phases but tended to perpetuate the inflammation process. While the H. speciosa biomembrane decreased necrosis and inflammation during the initial phases of the healing process, stimulated angiogenesis and interrupted these stimuly during the maturation phase of the healing process. The mixed treatment group decreased the delayed angiogenic effects of Regederm® however the inflammation was sustained leading to a delay in the healing process with less wound contraction.