RESUMEN
Objective: The aim of this research was to evaluate the efficacy of a topical formulation containing chitosan-chamomile microparticles in cutaneous healing in rats. Method: Male Wistar rats (n=57) were randomly distributed into three groups: treatment; vehicle; and control. Evaluations were performed on days 2, 7 and 14 after the surgical procedure using skin lesion photography, and histological and biochemical analyses. Results: The results showed that there was no difference in the healing index and in the histological analysis of the inflammatory infiltrate among groups. Fibrogenesis was more significant in the group treated with the test formulation at day 7, and angiogenesis was greater in the vehicle and chamomile groups at day 2. The quantification of hydroxyproline showed a higher amount of collagen in the group treated with chamomile, mainly at day 14, although the histological quantification of collagen showed no difference between the groups. Conclusion: From the results of this study, it can be concluded that the formulation, although it had no effect on the healing time, improved the quality of the cicatricial tissue formed with a greater quantity of fibroblasts and collagen.
Asunto(s)
Quitosano , Ratas , Masculino , Animales , Ratas Wistar , Quitosano/farmacología , Manzanilla , Cicatrización de Heridas , Colágeno/farmacología , PielRESUMEN
Wound healing studies are intricate, mainly because of the multifaceted nature of the wound environment and the complexity of the healing process, which integrates a variety of cells and repair phases, including inflammation, proliferation, reepithelialization and remodelling. There are a variety of possible preclinical models, such as in mice, rabbits and pigs, which can be used to mimic acute or impaired for example, diabetic and nutrition-related wounds. These can be induced by many different techniques, with excision or incision being the most common. After determining a suitable model for a study, investigators need to select appropriate and reproducible methods that will allow the monitoring of the wound progression over time. The assessment can be performed by non-invasive protocols such as wound tracing, photographic documentation (including image analysis), biophysical techniques and/or by invasive protocols that will require wound biopsies. In this article, we provide an overview of some of the most often needed and used: (a) preclinical/animal models including incisional, excisional, burn and impaired wounds; (b) methods to evaluate the healing progression such as wound healing rate, wound analysis by image, biophysical assessment, histopathological, immunological and biochemical assays. The aim is to help researchers during the design and execution of their wound healing studies.
Asunto(s)
Fibroblastos/patología , Queratinocitos/patología , Piel/patología , Cicatrización de Heridas , Animales , Diferenciación Celular , Movimiento Celular , Proliferación Celular , Células Cultivadas , Modelos Animales de Enfermedad , Fibroblastos/metabolismo , Humanos , Queratinocitos/metabolismo , Transducción de Señal , Piel/lesiones , Piel/metabolismoRESUMEN
Natural rubber latex (NRL) is a natural polymer which has arisen large interest in the biomedical field, mostly, due to its ability to facilitate angiogenesis and therefore, tissue repair. Moxifloxacin (MXF) is a broad-spectrum antibiotic orally administrated. Considering the biological properties of the NRL and its ability to deliver a wide range of compounds, the present study aimed to develop a novel device for infected chronic wound treatment. MXF-loaded NRL was obtained by a casting method. The results demonstrated that the incorporation of MXF in NRL did not promote any molecular interaction, preserving the integrity of the compounds. The mechanical properties of the biomaterial did not show any significant change, indicating enough elasticity for dermal application. The microbiological assays confirmed the ability of the polymer to deliver the drug without influencing its pharmacological properties. Moreover, it has expressed activity against major bacterial strains presented in wound infections. Finally, the biomaterial shown biocompatibility from the in vitro study. Thus, the present work has shown that MXF-loaded NRL membrane is a promising biomaterial to infected wound treatment.
Asunto(s)
Vendajes , Sistemas de Liberación de Medicamentos/instrumentación , Moxifloxacino/farmacología , Polímeros/química , Infección de Heridas/terapia , Animales , Antibacterianos/farmacología , Bacterias/efectos de los fármacos , Línea Celular , Escherichia coli/efectos de los fármacos , Fibroblastos/microbiología , Humanos , Queratinocitos/microbiología , Látex/química , Ratones , Pruebas de Sensibilidad Microbiana , Pseudomonas aeruginosa/efectos de los fármacos , Goma/química , Cicatrización de HeridasRESUMEN
The use of mesenchymal stem cells (MSCs) in tissue engineering has been extensively investigated. The greater the proliferation of this cellular group, the greater the regenerative and healing capacity of the tissue to which they belong. In this context, photobiomodulation (PBM) is an efficient technique in proliferation of distinct cell types. However, its parameters and mode of action are still unclear and require further investigation. This study aimed to evaluate the PBM action with different energies in MSCs of adipose tissue (hASCs). We used hASCs, seeded in 24-well plates, with 3 × 104 cells per well, in culture media. We used a total of four experimental groups, one with hASCs and simulated PBM and three other groups, which received PBM irradiation at 24, 48, and 72 h, with a 660-nm laser and power of 40 mW and energy of 0.56, 1.96, and 5.04 J. We performed analyses of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidefor) and trypan blue to evaluate cell proliferation and viability, 1 h after PBM irradiation. Software Graph PadPrism 7.0 was used. Intergroup comparisons were performed with ANOVA two-way and we used the Tukey post hoc test. Mitochondrial activity evaluated by MTT revealed the statistical difference in the first 24 h for group with more high energy when compared to control group; and in the 72 h for two irradiated groups when compared to the control group. The trypan blue test showed significant differences at the end of the experiment for two irradiated groups LG1 (4.52 × 104 ± 0.2) and LG2 (4.85 × 104 ± 0.8), when compared to the control group (1.87 × 104 ± 0.7). Both tests failed to be statistically different at the end of the experiment for groups LG1 and LG2 and observed a reduction in cellular mitochondrial growth and activity for group LG3. We conclude that PBM with energy close to 0.56 and 1.96 J promote proliferation of hASCs, and higher energy, such as 5.04 J, can be harmful.
Asunto(s)
Tejido Adiposo/citología , Terapia por Luz de Baja Intensidad , Células Madre Mesenquimatosas/citología , Células Madre Mesenquimatosas/efectos de la radiación , Proliferación Celular/efectos de la radiación , Supervivencia Celular/efectos de la radiación , Células Cultivadas , Humanos , Rayos Láser , Mitocondrias/metabolismo , Mitocondrias/efectos de la radiaciónRESUMEN
OBJECTIVES: Show that hidden endemic leprosy exists in a municipality of inner São Paulo state (Brazil) with active surveillance actions based on clinical and immunological evaluations. METHODS: The study sample was composed by people randomly selected by a dermatologist during medical care in the public emergency department and by active surveillance carried out during two days at a mobile clinic. All subjects received a dermato-neurological examination and blood sampling to determine anti-PGL-I antibody titers by enzyme-linked immunosorbent assay (ELISA). RESULTS: From July to December 2015, 24 new cases of leprosy were diagnosed; all were classified as multibacillary (MB) leprosy, one with severe Lucio's phenomenon. Seventeen (75%) were found with grade-1 or 2 disability at the moment of diagnosis. Anti-PGL-I titer was positive in 31/133 (23.3%) individuals, only 6/24 (25%) were positive in newly diagnosed leprosy cases. CONCLUSIONS: During the last ten years before this study, the average new case detection rate (NCDR) in this town was 2.62/100,000 population. After our work, the NCDR was raised to 42.8/100,000. These results indicate a very high number of hidden leprosy cases in this supposedly low endemic area of Brazil.
Asunto(s)
Anticuerpos Antibacterianos/sangre , Lepra/epidemiología , Mycobacterium leprae/inmunología , Adolescente , Adulto , Anciano , Brasil/epidemiología , Enfermedades Endémicas , Ensayo de Inmunoadsorción Enzimática , Femenino , Humanos , Lepra/diagnóstico , Masculino , Tamizaje Masivo , Persona de Mediana Edad , Adulto JovenRESUMEN
CONTEXT: The quantification of total collagen is of major importance in a wide range of research areas, including the study of cutaneous wound healing and new drugs trials. OBJECTIVE: The total collagen content in skin biopsies was compared by biochemical hydroxyproline assay and by two computer-aided histomorphometric analyses of histological sections. MATERIALS AND METHODS: Two methods were used to evaluate collagen formation: the hydroxyproline assay, as the gold standard and histomorphometric image analysis of the filled areas by corresponding stained collagen fibres, using picrosirius and Gomori's trichrome staining. The image analyses were determined by digital densitometry recognition using computer-aided ImageJ software. One-way ANOVA, simple linear regression and ANCOVA were applied for the statistical analysis and correlation. RESULTS: In a simple linear regression analysis carried out on the 14th day period after the induction of skin injury, three techniques, picrosirius red (F = 33.57, p = 0.00), Gomori's trichrome (F = 81.61, p = 0.00) and hydroxyproline content (F = 16.85, p = 0.00) were able to detect collagen production. After scale adjustment, there were no significant differences among either the slopes (F = 1.17, p = 0.32) or the intercepts (F = 0.69, p = 0.51) of the estimated regression lines. It seems that a highly significant correlation exists between the histomorphometrical analysis and hydroxyproline assay. DISCUSSION AND CONCLUSION: The morphometric analysis proved to be adequate and can be used as a simple, rapid, low-cost technology for evaluating total collagen in cutaneous wound specimens, compared with the gold standard hydroxyproline assay.
Asunto(s)
Colágeno/análisis , Piel/lesiones , Cicatrización de Heridas/fisiología , Animales , Hidroxiprolina/análisis , Procesamiento de Imagen Asistido por Computador , Masculino , Ratas , Ratas Wistar , Piel/químicaRESUMEN
This study aimed to compare the phototherapy effects on wound healing in rats submitted to normal and high-fat diets. Thirty-six rats received normal lipidic diet (NL) and 36 high lipidic (HL) diet for 45 days. The nutritional status was measured by body mass, blood glucose, total cholesterol, and triglycerides levels. Four experimental groups were performed according light (L) therapy applied "on" or "off" (660 nm, 100 mW, 70 J/cm(2), 2 J) on 1.5-mm-punched dorsum skin wounds as NLL+, NLL-, HLL+, and HLL-. The wound healing rate (WHR) and oxidative stress markers were analyzed on 2nd, 7th, and 14th days. Despite no difference among body mass, the HL rats presented higher blood glucose, total cholesterol, and triglycerides levels than NL rats. Respectively, on the 2nd and 14th days, the HLL+ group presented the highest WHRs (0.38 ± 0.16/0.97 ± 0.02) among all groups, while the HLL- (-0.002 ± 0.12/0.81 ± 12.1) the lowest WHRs. Hydroxyproline level was lower in HLL- (6.41 ± 1.09 µg/mg) than HLL+ (7.71 ± 0.61 µg/mg) and also NLL+ (9.33 ± 0.84 µg/mg). HLL+ presented oxidative stress markers similar to normal control group (NLL-) during follow up and highest antioxidant defense on 7th day. The results showed phototherapy accelerated the cutaneous wound healing by modulating oxidative stress in rats with metabolic disorders under a high-fat diet.
Asunto(s)
Láseres de Semiconductores/uso terapéutico , Terapia por Luz de Baja Intensidad , Repitelización/efectos de la radiación , Animales , Colágeno/biosíntesis , Dieta Alta en Grasa/efectos adversos , Glutatión/sangre , Masculino , Enfermedades Metabólicas/sangre , Enfermedades Metabólicas/etiología , Enfermedades Metabólicas/fisiopatología , Estrés Oxidativo , Ratas WistarRESUMEN
The in vitro cultivation of M. leprae has not been possible since it was described as causing leprosy, and the limitation of animal models for clinical aspects makes studies on leprosy and bacteria-human host interaction a challenge. Our aim was to standardize the ex vivo skin model (hOSEC) to maintenance and study of M. leprae as an alternative animal model. Bacillary suspensions were inoculated into human skin explants and sustained in DMEM medium for 60 days. Explants were evaluated by RT-PCR-16SrRNA and cytokine gene expression. The viability and infectivity of bacilli recovered from explants (D28 and D60) were evaluated using the Shepard's model. All explants were RT-PCR-16SrRNA positive. The viability and infectivity of recovered bacilli from explants, analyzed after 5 months of inoculation in mice, showed an average positivity of 31%, with the highest positivity in the D28 groups (80%). Furthermore, our work showed different patterns in cytokine gene expression (TGF-ß, IL-10, IL-8, and TNF-α) in the presence of alive or dead bacilli. Although changes can be made to improve future experiments, our results have demonstrated that it is possible to use the hOSEC to maintain M. leprae for 60 days, interacting with the host system, an important step in the development of experimental models for studies on the biology of the bacillus, its interactions, and drug susceptibility.
RESUMEN
Liquid crystalline nanoparticles (LCNs) are an attractive drugs topical delivery system due to the great internal ordering, wide interfacial area and structural similarities with the skin. In this work, LCNs were designed to encapsulate triptolide (TP) and to complex on its surface small interfering RNAs (siRNA) targeting TNF-α and IL-6, aiming at topical co-delivery and regulating multi-targets in psoriasis. These multifunctional LCNs showed appropriate physicochemical properties for topical application, such as a mean size of 150 nm, low polydispersion, TP encapsulation greater than 90% and efficient complexation with siRNA. The internal reverse hexagonal mesostructure of LCNs was confirmed by SAXS while their morphology was assessed by cryo-TEM. In vitro permeation studies revealed an increase of more than 20-fold in the distribution of TP through the porcine epidermis/dermis was achieved after the application of LCN-TP or LCN TP in hydrogel. In cell culture, LCNs showed good compatibility and rapid internalization, which was attributed to macropinocytosis and caveolin-mediated endocytosis. Anti-inflammatory potential of multifunctional LCNs was assessed by reducing of TNF-α, IL-6, IL-1ß and TGF-ß1 levels in LPS-stimulated macrophages. These results support the hypothesis that the co-delivery of TP and siRNAs by LCNs may be a new strategy for psoriasis topical therapy.
Asunto(s)
Nanopartículas , Psoriasis , Porcinos , Animales , ARN Interferente Pequeño , Factor de Necrosis Tumoral alfa , Interleucina-6 , Dispersión del Ángulo Pequeño , Difracción de Rayos X , Psoriasis/tratamiento farmacológico , Nanopartículas/químicaRESUMEN
Vegetable oils offer excellent biological properties, but their high lipophilicity limits their bioavailability. This work aimed to develop nanoemulsions based on sunflower and rosehip oils and to evaluate their wound-healing activity. The influence of phospholipids of plant origin on nanoemulsions' characteristics was investigated. A nanoemulsion prepared with a mixture of phospholipids and synthetic emulsifiers (Nano-1) was compared with another prepared only with phospholipids (Nano-2). The healing activity was evaluated in wounds induced in human organotypic skin explant culture (hOSEC) based on histological and immunohistochemical analysis. The hOSEC wound model was validated, showing that high nanoparticle concentration in the wound bed interferes with cell mobility and the ability to respond to the treatment. Nanoemulsions were 130 to 370 nm, with a concentration of 1013 particles/mL, and a low potential to induce inflammatory processes. Nano-2 was three times larger than Nano-1 but less cytotoxic and could target the oils to the epidermis. Nano-1 permeated intact skin to the dermis and showed a more prominent healing effect than Nano-2 in the hOSEC wound model. Changes in the lipid nanoemulsion stabilizers impacted the cutaneous and cellular penetration of the oils, cytotoxicity, and healing kinetics, resulting in versatile delivery systems.
RESUMEN
Introduction: Hansen's disease (HD) is the most common cause of treatable peripheral neuropathy in the world that may or may not involve skin manifestations, and physical examination based on simplified neurologic evaluation is a subjective and inaccurate procedure. High-resolution ultrasound (HRUS) can be used to evaluate peripheral nerves and is a validated technique of good reproducibility, permitting a detailed and precise examination. Objectives: We proposed to establish objective criteria for absolute values of the measurement of the CSA of peripheral nerves and their indices of the ΔCSA and ΔTpT in the diagnosis of Hansen's disease neuropathy as compared with healthy voluntaries. Materials and methods: In municipalities from different regions of Brazil, we randomly selected 234 volunteer Brazilian patients diagnosed with leprosy to be submitted to peripheral nerve echography and compared with 49 healthy Brazilian volunteers. Results: Hansen Disease assessed by high resolution ultrasound is a primarily neural disease that leads to multiple hypertrophic mononeuropathy characterized by CSA values exceeding normal limits (Med CT = 10.2 mm2; UT = 9.8 mm2; UPT = 9.3 mm2; CFFH = 18.3 mm2; T = 9.6 mm2), and the pattern of asymmetry (ΔCSA>2.5 mm2 with RR 13) and focality (ΔTPT > 2.5 mm2 with RR 6.4) of this thickening has higher sensitivity (76,1%) and specificity (87,8 %) for its early diagnosis that laboratory tests. Analyzing each subject, the percentage of thickened nerves detected among the total number of nerves assessed was higher among patients with HD than among healthy individuals (p < 0.0001). Individuals with two or more thickened nerves were at 24.1 times higher relative risk (95% CI: 6.74-88.98) of HD.
RESUMEN
Introduction: Hansen's disease (HD) primarily infects peripheral nerves, with patients without HD being free of peripheral nerve damage. Household contacts (HHCs) of patients with HD are at a 5-10 times higher risk of HD than the general population. Neural thickening is one of the three cardinal signs that define a case of HD according to WHO guidelines, exclusively considering palpation examination that is subjective and may not detect the condition in the earliest cases even when performed by well-trained professionals. High-resolution ultrasound (HRUS) can evaluate most peripheral nerves, a validated technique with good reproducibility allowing detailed and accurate examination. Objective: This study aimed to use the peripheral nerve HRUS test according to the HD protocol as a diagnostic method for neuropathy comparing HHCs with healthy volunteers (HVs) and patients with HD. Methods: In municipalities from 14 different areas of Brazil we selected at random 83 HHC of MB-patients to be submitted to peripheral nerve ultrasound and compared to 49 HVs and 176 HD-patients. Results: Household contacts assessed by HRUS showed higher median and mean absolute peripheral nerve cross-sectional area (CSA) values and greater asymmetries (ΔCSA) compared to HVs at the same points. Median and mean absolute peripheral nerve CSA values were higher in patients with HD compared to HCCs at almost all points, while ΔCSA values were equal at all points. Mean ± SD focality (ΔTpT) values for HHCs and patients with HD, respectively, were 2.7 ± 2.2/2.6 ± 2.2 for the median nerve, 2.9 ± 2.7/3.3 ± 2.9 for the common fibular nerve (p > 0.05), and 1.3 ± 1.3/2.2 ± 3.9 for the ulnar nerve (p < 0.0001). Discussion: Considering HRUS findings for HHCs, asymmetric multiple mononeuropathy signs (thickening or asymmetry) in at least 20% of the nerves evaluated could already indicates evidence of HD neuropathy. Thus, if more nerve points are assessed in HHCs (14 instead of 10), the contacts become more like patients with HD according to nerve thickening determined by HRUS, which should be a cutting-edge tool for an early diagnosis of leprosy cases.
RESUMEN
Purpose: Nanoparticles are resources of advanced nanotechnology being present in several products. Titanium dioxide nanoparticles are among the five most widely used NP currently expanding their benefits from the oil industry to the areas of diagnostic medicine due to their properties and small size. However, its impact on human health is still controversial in the literature. We aimed to evaluate the cytotoxicity of a new titanium NP functionalized with sodium carboxylic ligand (COOH-Na+) in human keratinocytes (HaCaT) and human fibroblasts (HDFn). Methods: The physical-chemical characterization was performed by the transmission electron microscopy (TEM), dynamic light scattering (DLS) and zeta potential techniques, respectively. MTT and LDH assays were used to assess cytotoxicity and cell membrane damage respectively, ELISA to identify the inflammatory profile and, reactive oxygen species assay and cytometry to detect reactive oxygen species and their relationship with apoptosis/necrosis mechanisms. Results: The results demonstrated a decrease in cell viability at the highest concentrations tested for both cell lines, but no change in LDH release was detected for the HaCaT. The cell membrane damage was found only at 100.0 µg/mL for the HDFn. It was demonstrated that cytotoxicity in the highest concentrations evaluated for both cell lines for the 72 h period. The HDFn showed damage to the cell membrane at a concentration of 100 µg/mL followed by a significant increase in reactive oxygen species production. No inflammatory profile was detected. The HaCaT showed apoptosis when exposed to the highest concentration evaluated and HDFn showed both apoptosis and necrosis for the same concentration. Conclusion: Thus, it is possible to conclude that the cytotoxicity mechanism differs according to the cell type evaluated, with HDFn being the most sensitive line in this case, and this mechanism can be defined in a dose and time dependent manner, since the highest concentrations also triggered death cell.
Asunto(s)
Nanopartículas del Metal , Nanopartículas , Apoptosis , Supervivencia Celular , Humanos , Nanopartículas del Metal/química , Nanopartículas del Metal/toxicidad , Nanopartículas/química , Nanopartículas/toxicidad , Necrosis/inducido químicamente , Estrés Oxidativo , Especies Reactivas de Oxígeno/metabolismo , Titanio/química , Titanio/toxicidadRESUMEN
Acute injuries, such as surgical and traumatic, heal normally in an organized and rapid manner. Studies point to the healing activity of hyaluronic acid in all phases of healing. The aim was to evaluate the effectiveness of hyaluronic acid in skin abrasions on the dorsum of rats to compare to usual products on the market. Seventy-two Wistar rats were subjected to excoriation of approximately 2.0 cm2 on the back by dermabrasion. According to the treatment, 3 groups were established: saline, chlorhexidine digluconate and 0.2% hyaluronic acid for 14 days. Animals were photographed on the 2nd, 7th, 10th and 14th postinjury days, and the index of healing of the abrasions was calculated. Biochemically, myeloperoxidase measurements of skin biopsies in addition to histological studies to assess the crust and epidermal layers were performed. The group treated with hyaluronic acid showed better re-epithelialization from the other groups (p < 0.05) on the 7th and 10th days. For the thickness of the crust, the hyaluronic acid group presented thinner crust than other groups on the 10th and 14th days (p < 0.05), but in the epidermis, no difference was observed between the groups studied. All groups showed an increase in myeloperoxidase enzyme on the 2nd day, but a decreasing on the 7th day. On the 10th day, there was a difference in the hyaluronic acid group compared to the other groups (p < 0.05). The application of 0.2% hyaluronic acid significantly accelerated the re-epithelialization of skin abrasions compared to saline and chlorhexidine digluconate.
RESUMEN
BACKGROUND: This study evaluates implementation strategies for leprosy diagnosis based on responses to a Leprosy Suspicion Questionnaire (LSQ), and analyzes immunoepidemiological aspects and follow-up of individuals living in a presumptively nonendemic area in Brazil. METHODOLOGY/PRINCIPAL FINDINGS: Quasi-experimental study based on LSQ throughout Jardinópolis town by community health agents, theoretical-practical trainings for primary care teams, dermatoneurological examination, anti-PGL-I serology, RLEP-PCR, and spatial epidemiology. A Leprosy Group (LG, n = 64) and Non-Leprosy Group (NLG, n = 415) were established. Overall, 3,241 LSQs were distributed; 1,054 (32.5%) LSQ were positive for signs/symptoms (LSQ+). Among LSQ+ respondents, Q2-Tingling (pricking)? (11.8%); Q4-Spots on the skin? (11.7%); Q7-Pain in the nerves? (11.6%); Q1-Numbness in your hands and/or feet? (10.7%) and Q8-Swelling of hands and feet? (8.5%) were most frequently reported symptoms. We evaluated 479 (14.8%) individuals and diagnosed 64 new cases, a general new case detection rate (NCDR) of 13.4%; 60 were among 300 LSQ+ (NCDR-20%), while 4 were among 179 LSQ negative (NCDR-2.23%). In LG, Q7(65%), Q2(60%), Q1(45%), Q4(40%) and Q8(25%) were most frequent. All 2x2 crossings of these 5 questions showed a relative risk for leprosy ranging from 3 to 5.8 compared with NLG. All patients were multibacillary and presented hypochromatic macules with loss of sensation. LG anti-PGL-I titers were higher than NLG, while 8.9% were positive for RLEP-PCR. The leprosy cases and anti-PGL-I spatial mappings demonstrated the disease spread across the town. CONCLUSIONS/SIGNIFICANCE: Implementation actions, primarily LSQ administration focused on neurological symptoms, indicate hidden endemic leprosy in a nonendemic Brazilian state.
Asunto(s)
Agentes Comunitarios de Salud/educación , Lepra/diagnóstico , Lepra/epidemiología , Mycobacterium leprae/inmunología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Brasil/epidemiología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Mycobacterium leprae/genética , Mycobacterium leprae/aislamiento & purificación , Prevalencia , Encuestas y CuestionariosRESUMEN
Alternative models to replace animals in experimental studies remain a challenge in testing the effectiveness of dermatologic and cosmetic drugs. We proposed a model of human organotypic skin explant culture (hOSEC) to assess the profile of cutaneous drug skin distribution, adopting dacarbazine as a model, and respective new methodologies for dermatokinetic analysis. The viability tests were evaluated in primary keratinocytes and fibroblasts, and skin by MTT and TTC assays, respectively. Then, dacarbazine was applied to the culture medium, and the hOSEC method was applied to verify the dynamics of skin distribution of dacarbazine and determine its dermatokinetic profile. The results of cell and tissue viability showed that both were considered viable. The dermatokinetic results indicated that dacarbazine can be absorbed through the skin, reaching a concentration of 36.36 µg/mL (18,18%) of the initial dose (200 µg/mL) after 12 h in culture. Histological data showed that the skin maintained its structure throughout the tested time that the hOSEC method was applied. No apoptotic cells were observed in the epidermal and dermal layers. No visible changes in the dermo-epidermal junction and no inflammatory processes with the recruitment of defense cells were observed. Hence, these findings suggest that the hOSEC concept as an alternative ex vivo model for assessing the dynamics of skin distribution of drugs, such as dacarbazine, and determining their respective dermatokinetic profiles.
Asunto(s)
Dacarbazina , Preparaciones Farmacéuticas , Animales , Fibroblastos , Humanos , Queratinocitos , PielRESUMEN
Skin wounds have been a public health concern of high frequency, in addition to requiring intensive and expensive care. The natural rubber latex (NRL) from Hancornia speciosa Gomes has been used to treat many problems in traditional medicine and also present healing properties, antifungal and anti-inflammatory activity and antinociceptive effects. The purpose of this study was to characterize the new biomembrane from the NRL of H. speciosa (HS) by Fourier transform infrared (FTIR) and mechanical strength test and to investigate its biological properties by the cytotoxicity assay and in vivo healing activity. The results showed that the HS biomembrane exhibited characteristic bands of the main component cis-1,4-polyisoprene. Besides, its Young modulus was close to human skin with adhesive-compatible mechanical characteristics. The cytotoxicity assays revealed that the HS biomembrane was not toxic to fibroblast cells neither using agar diffusion test nor MTT assay. Furthermore, the HS biomembrane stimulated the inflammatory cells and the angiogenesis, increased significantly the collagenesis and improved the quality of heal until the remodeling phase induced by implants in mice. Thus, this biomembrane has proven to be a safe and biocompatible biomaterial with healing potential, becoming an effective and low-cost alternative for the treatment of skin wounds.
Asunto(s)
Apocynaceae/química , Látex , Ensayo de Materiales , Membranas Artificiales , Cicatrización de Heridas/efectos de los fármacos , Células 3T3 , Animales , Látex/química , Látex/farmacología , Masculino , RatonesRESUMEN
BACKGROUND: Regarding the leprosy transmission through the upper airways, overcrowded locations such as prisons can become a risk to get sick. Like the leprosy hidden endemic demonstrated in male prison population, being interesting to assess the leprosy scene also among confined women. METHODS: A prospective descriptive study conducted at Female Penitentiary, Brazil. Leprosy Suspicion Questionnaire (LSQ) were applied to the participants, and submitted to specialized dermatoneurological exam, peripheral nerve ultrasonography, and anti-PGL-I serology. FINDINGS: 404 female inmates were evaluated, 14 new cases were diagnosed (LG-leprosy group), a new case detection rate (NCDR) of 3.4%, 13 multibacillary, while another 390 constituted the Non-Leprosy group (NLG). Leprosy cases were followed up during multidrug therapy with clinical improvement. The confinement time median was 31 months in LG, similar to NLG, less than the time of leprosy incubation. Regarding LSQ, the neurological symptoms reached the highest x2 values as Q1-numbness (5.6), Q3-anesthetizes areas in the skin (7.5), Q5-Stinging sensation (5.8), and Q7-pain in the nerves (34.7), while Q4-spots on the skin was 4.94. When more than one question were marked in the LSQ means a 12.8-fold higher to have the disease than a subject who marked only one or none. The high 34% rate of anti-PGL-I seropositivity in the penitentiary, higher levels in LG than NLG. Three additional leprosy cases each were diagnosed on the second (n = 66) and third (n = 14) reevaluations 18 and 36 months after the initial one. Semmes-Weinstein monofilaments demonstrated lower limbs (32.2%) more affected than the upper limbs (25%) with improvement during the follow-up. INTERPRETATION: The NCDR in this population showed an hidden endemic of leprosy as well as the efficacy of a search action on the part of a specialized team with the aid of the LSQ and anti-PGL-I serology as an auxiliary tracking tools.
Asunto(s)
Lepra/diagnóstico , Prisiones/estadística & datos numéricos , Adulto , Brasil/epidemiología , Femenino , Estudios de Seguimiento , Humanos , Leprostáticos/efectos adversos , Leprostáticos/uso terapéutico , Lepra/tratamiento farmacológico , Lepra/epidemiología , Lepra/microbiología , Persona de Mediana Edad , Mycobacterium leprae/efectos de los fármacos , Mycobacterium leprae/fisiología , Estudios Prospectivos , Adulto JovenRESUMEN
OBJECTIVE To present a methodology for apportioning Union resources to the federative units (FU - 26 states and one federal district) within the Brazilian Unified Health System (SUS) based on health needs measured by demographic, socioeconomic, epidemiological and geographical dimensions. METHODS The apportionment methodology proposal prioritizes the health needs axis, based on Law 141/2012. We adopted a proxy of needs that measures relative inequalities between, socioeconomic, geographic demographic and epidemiological conditions of the populations of the Brazilian Federative Units (FU) for 2015. We first used an adjustment so that the populations of the 27 FU are corrected by their relative needs regarding age and gender. To calculate the health needs axis, the multivariate techniques factorial analysis and principal components were used, and, based on such correction, we applied the health needs index. Subsequently, this index was applied to simulate the resources that should be transferred by the Ministry of Health to states in 2015. RESULTS As we made the methodological choice of transferring a single per capita amount to all states, so the proposal required population correction. Thus, in the analysis of health needs, the FUs that had their population corrected by a factor higher than the national average because of their greater relative need, were the states of: Maranhão, Piauí, Alagoas, Paraíba, Ceará, Pará, Bahia, Acre, Pernambuco, Rio Grande do Norte, Sergipe, Amazonas, Tocantins and Roraima. For the simulation aggregating all the financing blocks, without reducing the resources already distributed to the remaining states in 2015, indicated the additional need of R$ 4.6 billion. CONCLUSIONS The proposal addresses the absence of studies presenting quantitative simulations of federal resources distribution within the scope of SUS to the FUs, based on the apportionment criteria defined by Law 141/2012, in order to contribute to the reduction health inequalities and mitigate the effects of the economic crisis.
Asunto(s)
Recursos en Salud , Brasil , Geografía , Factores SocioeconómicosRESUMEN
BACKGROUND: Dermabrasion is related with mechanical and surgical traumas on the skin; usually topical antiseptics and/or saline have been used for healing. Natural products for wound healing can also be used for abrasions, such as latex from Hevea brasiliensis. OBJECTIVE: This study aimed to evaluate the in vitro viability and migratory/proliferative effects of latex serum from H. brasiliensis and to compare with a commercially available standard antiseptic solution and saline in experimental dermabrasion on rats. METHODS: For in vitro evaluation, MTT and scratch assays were used. In vivo testing was performed in 72 rats submitted to dermabrasion, treated with saline, antiseptic, or latex serum. This study evaluated re-epithelialization, neutrophilic infiltration, and the quantification of crust and epidermis. RESULTS: Latex showed viability at 1% and 0.1% concentrations and migratory/proliferative activity at 0.01% concentrations. The re-epithelialization was highest in latex group on 7th day. The latex group displayed lower thickness of crusts and greater extent of epidermal layers. The latex and antiseptic groups showed increases of myeloperoxidase levels on the 2nd day and showed important reductions from the 7th day. STUDY LIMITATIONS: Acute superficial wound model in rats and non-use of gel-cream (medium) without latex. CONCLUSION: In conclusion, non-toxic latex stimulated migration/proliferation of keratinocytes in vitro and significantly accelerated wound healing in animal excoriation models compared to chlorhexidine or saline.