RESUMO
Bufadienolides are digitalis-like aglycones mainly found in skin secretions of toads. Among their biological properties, the mechanisms of antiproliferative action on tumor cells remain unclear for many compounds, including against leukemia cells. Herein, it was evaluated the mechanisms involved in the antiproliferative and genotoxic actions of hellebrigenin on tumor cell lines and in silico capacity to inhibit the human topoisomerase IIa enzyme. Firstly, its cytotoxic action was investigated by colorimetric assays in human tumor and peripheral blood mononuclear cells (PBMC). Next, biochemical and morphological studies were detailed by light microscopy (trypan blue dye exclusion), immunocytochemistry (BrdU uptake), flow cytometry and DNA/chromosomal damages (Cometa and aberrations). Finally, computational modelling was used to search for topoisomerase inhibition. Hellebrigenin reduced proliferation, BrdU incorporation, viability, and membrane integrity of HL-60 leukemia cells. Additionally, it increased G2/M arrest, internucleosomal DNA fragmentation, mitochondrial depolarization, and phosphatidylserine externalization in a concentration-dependent manner. In contrast to doxorubicin, hellebrigenin did not cause DNA strand breaks in HL-60 cell line and lymphocytes, and it interacts with ATPase domain residues of human topoisomerase IIa, generating a complex of hydrophobic and van der Waals interactions and hydrogen bonds. So, hellebrigenin presented potent anti-leukemic activity at concentrations as low as 0.06 µM, a value comparable to the clinical anticancer agent doxorubicin, and caused biochemical changes suggestive of apoptosis without genotoxic/clastogenic-related action, but it probably triggers catalytic inhibition of topoisomerase II. These findings also emphasize toad steroid toxins as promising lead antineoplasic compounds with relatively low cytotoxic action on human normal cells.
Assuntos
Antineoplásicos , Bufanolídeos , Leucemia , Humanos , Leucócitos Mononucleares , Bromodesoxiuridina/farmacologia , Dano ao DNA , Antineoplásicos/farmacologia , Bufanolídeos/química , Células HL-60 , Apoptose , DNA/farmacologia , Doxorrubicina/farmacologiaRESUMO
OBJETIVO: Descrever a trajetória de desenvolvimento dos curativos biológicos oriundos de pele de tilápia (Oreochromis niloticus) em glicerol e liofilizada para uso em cirurgias externas e, posteriormente, uma matriz proteica acelular (scaffold) para uso interno. RESULTADOS: A pele de tilapia no glicerol e liofilizada foi aplicada com sucesso em mais de 550 pacientes queimados. A pele de tilápia liofilizada obteve sucesso no tratamento de 53 mulheres em vaginoplastias, em 160 pacientes na redesignação sexual e na preparação do leito da ferida na autoenxertia em 15 portadores da Síndrome de Apert. O scaffold está sendo empregado na oftalmologia na medicina veterinária na reconstrução de córnea em 420 cães, nas duroplastias na neurocirurgia nos testes em animais, e em estudos para uso cirúrgico em 10 especialidades médicas. CONCLUSÕES: O curativo de pele de tilápia supera desafios do tratamento de queimados do Brasil. É barato, biossustentável, efetivo e reduz a dor do paciente, propiciando melhores resultados com potencial redução de custos, contribuindo para a qualidade de vida dos pacientes. O sucesso da pesquisa confirma a pele de tilápia como um novo biomaterial de grande potencial em medicina regenerativa.
OBJECTIVE: To describe the development trajectory of biological dressings made from tilapia (Oreochromis niloticus) skin in glycerol and freeze-dried for use in external surgeries and, subsequently, an acellular protein matrix (scaffold) for internal use. RESULTS: Tilapia skin in glycerol and freeze-dried was successfully applied to more than 550 burn patients. Freeze-dried tilapia skin was successful in the treatment of 53 women undergoing vaginoplasty, in 160 patients in sexual reassignment and in preparing the wound bed in self-grafting in 15 patients with Apert Syndrome. The scaffold is being used in ophthalmology, veterinary medicine in corneal reconstruction in 420 dogs, in duraplasty in neurosurgery in animal tests, and in studies for surgical use in 10 medical specialties. CONCLUSIONS: The tilapia skin dressing overcomes challenges in treating burns in Brazil. It is cheap, biosustainable, effective and reduces patient pain, providing better results with potential cost reduction, contributing to patients' quality of life. The success of the research confirms tilapia skin as a new biomaterial with great potential in regenerative medicine.
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OBJETIVO: Produzir um scaffold baseado em matriz extracelular (SMEC) biocompatível, atóxico e estéril, para tratamento de queimaduras e feridas. Explorou-se a utilização da pele de tilápia como alternativa, ressaltando suas propriedades semelhantes à pele humana e sua aplicação bem-sucedida em diferentes áreas médicas. MÉTODO: Descreve o processo de preparação dos SMEC de pele de tilápia, incluindo etapas de desengorduramento, descontaminação, descelularização e irradiação por raios gama a 25kGy para esterilização. São realizados testes laboratoriais para avaliar a toxicidade celular in vitro pelo método do MTT, análises histológicas com coloração de hematoxilina-eosina, análises microbiológicas e de quantificação de DNA. RESULTADOS: Destacam que os SMEC produzidos foram descelularizados de maneira eficaz, preservando a matriz extracelular e mostrando-se não citotóxicos. Além disso, a análise microbiológica evidenciou a esterilidade dos materiais após a irradiação, comprovando sua adequação para aplicação clínica. A quantificação de DNA revelou a efetividade da descelularização, reduzindo significativamente o conteúdo de DNA original do tecido. CONCLUSÕES: Foi possível o desenvolvimento de uma matriz oriunda da pele de tilápia, sendo ela não citotóxica, estéril, portadora de morfologia adequada para aplicação clínica e acelular. Assim, contribuindo para inovação da ciência brasileira.
OBJECTIVE: To produce a biocompatible, non-toxic, and sterile scaffold based on extracellular matrix (ECM) for the treatment of burns and wounds. The utilization of tilapia skin was explored as an alternative, highlighting its similar properties to human skin and its successful application in different medical areas. METHODS: The process of preparing tilapia skin-derived ECM scaffolds is described, including steps of degreasing, decontamination, decellularization, and gamma ray irradiation at 25kGy for sterilization. Laboratory tests were conducted to assess in vitro cellular toxicity using the MTT method, histological analyses with hematoxylin-eosin staining, microbiological analyses, and DNA quantification. RESULTS: It is emphasized that the produced ECM scaffolds were effectively decellularized, preserving the extracellular matrix and demonstrating non-cytotoxicity. Furthermore, microbiological analysis evidenced the sterility of the materials after irradiation, confirming their suitability for clinical application. DNA quantification revealed the effectiveness of decellularization, significantly reducing the original DNA content of the tissue. CONCLUSIONS: The development of a tilapia skin-derived matrix was achieved, which is non-cytotoxic, sterile, possesses suitable morphology for clinical application, and is acellular. Thus, contributing to the innovation of Brazilian science.
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BACKGROUND: Penile cancer is one of the most aggressive male tumors. Although it is preventable, the main etiologic causes are lifestyle behaviors and viral infection, such as human papillomavirus (HPV). Long-term epigenetic changes due to environmental factors change cell fate and promote carcinogenesis, being an important marker of prognosis. We evaluated epidemiological aspects of penile squamous cell carcinoma (SCC) and the prevalence of HPV infection using high-risk HPV (hrHPV) and p16INK4A expression of 224 participants. Global DNA methylation was evaluated through 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC). RESULTS: The incidence of HPV was 53.2% for hrHPV and 22.32% for p16INK4a. hrHPV was not related to systemic or lymph node metastasis and locoregional recurrence, nor influenced the survival rate. P16INK4a seems to be a protective factor for death, which does not affect metastasis or tumor recurrence. Lymph node and systemic metastases and locoregional recurrence increase the risk of death. An increased 5mC mark was observed in penile SCC regardless of HPV infection. However, there is a reduction of the 5hmC mark for p16INK4a + (P = 0.024). Increased 5mC/5hmC ratio (> 1) was observed in 94.2% of penile SCC, irrespective of HPV infection. Despite the increase in 5mC, it seems not to affect the survival rate (HR = 1.06; 95% CI 0.33-3.38). CONCLUSIONS: P16INK4a seems to be a good prognosis marker for penile SCC and the increase in 5mC, an epigenetic mark of genomic stability, may support tumor progression leading to poor prognosis.
Assuntos
Alphapapillomavirus , Carcinoma de Células Escamosas , Infecções por Papillomavirus , Neoplasias Penianas , Masculino , Humanos , Neoplasias Penianas/genética , Neoplasias Penianas/epidemiologia , Neoplasias Penianas/patologia , Infecções por Papillomavirus/complicações , Infecções por Papillomavirus/genética , Infecções por Papillomavirus/epidemiologia , Inibidor p16 de Quinase Dependente de Ciclina/genética , Prognóstico , 5-Metilcitosina , Metilação de DNA , Recidiva Local de Neoplasia/genética , Papillomaviridae/genética , Carcinoma de Células Escamosas/metabolismo , Alphapapillomavirus/genética , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Epigênese Genética , DNA ViralRESUMO
Maternal separation (MS) is a risk factor for major depressive disorder. Both cancer and depression seem to share a common biological link. Here, we evaluated the progression of melanoma and the underlying mechanisms related to this progression, namely cell proliferation and apoptosis, in adult female mice exposed to MS. Female C57BL/6 mice were exposed to MS for 60 min/day during the first 2 postnatal weeks (here called MS mice) or left undisturbed (here called non-MS mice). Melanoma cells were inoculated subcutaneously into the axillary region of adult animals, and tumor progression was evaluated for 25 days. Adult MS mice presented depressive-like behavior and working memory deficits. MS accelerated murine melanoma growth by mechanisms related to decreased apoptosis and increased cell proliferation rate, such as increased expression of IL-6 and mTOR. MS stimulated eukaryotic elongation factor 2 expression and increased the number of circulating monocytes and DNA damage in peripheral blood leukocytes, an effect associated with oxidative DNA damage. In conclusion, MS accelerated the progression of murine melanoma by mechanisms related to tumor proliferation and apoptosis, revealing a relationship between adverse childhood experiences and cancer progression, particularly melanoma.
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Avaliação do Impacto na Saúde , Imunidade , Privação Materna , Melanoma/imunologia , Melanoma/patologia , Animais , Apoptose , Comportamento Animal , Biomarcadores , Proliferação de Células , Dano ao DNA , Modelos Animais de Doenças , Progressão da Doença , Suscetibilidade a Doenças , Feminino , Contagem de Leucócitos , Melanoma/metabolismo , Melanoma Experimental , Camundongos , Neuroimunomodulação , Fatores Sexuais , Estresse FisiológicoRESUMO
Background: Wounds are disruptions of the normal continuity of anatomic structures, generally due to local trauma. Theyare extremely prevalent in animals, especially horses, and a common reason for seeking veterinary attention. Their management aims to restore the function and integrity of the affected area in the shortest possible time and cost, while providingsatisfactory cosmetic results. This task becomes challenging when working with horses, considering the contact betweenwounds and contaminated environment is common. Thus, the present study aims to report the case of a traumatic equinewound treated with Nile Tilapia Fish Skin (NTFS).Case: A male 27-year-old horse previously castrated, with no defined breed (NDB), and weighing 400 kg presented a 6.0 x5.5 cm superficial wound in the distal left anterior limb (LAL) due to skin laceration. The animal belonged to the cavalry ofthe Military Police of Ceará, a public institution in Fortaleza, Brazil. Although in excellent general health, with no previouscomorbidities or restriction of movement, the animal was removed from its role in equine-assisted therapy (EAT) until completewound healing, aiming adequate evaluation of the novel biomaterial via lower influence of external factors. After informedconsent from the owner was obtained, Nile Tilapia Fish Skin was applied to the lesion. The Ethics Committee on the use ofanimals of the Drug Research and Development Center of the Federal University of Ceará, Fortaleza, Brazil, approved the studyprotocol. Compliance with regulations on the ethical treatment of animals was performed. Nile Tilapia Fish Skin applicationfollowed a protocol similar to that established in human clinical studies. Initially, the horse was submitted to wound cleaningwith tap water and 2% chlorhexidine gluconate, with no requirement of pre-treatment surgical...(AU)
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Animais , Transplante de Pele/veterinária , Cavalos/cirurgia , Ferimentos e Lesões/cirurgia , Ferimentos e Lesões/veterinária , Ciclídeos , Técnicas de Fechamento de Ferimentos/veterináriaRESUMO
Background: Wounds are disruptions of the normal continuity of anatomic structures, generally due to local trauma. Theyare extremely prevalent in animals, especially horses, and a common reason for seeking veterinary attention. Their management aims to restore the function and integrity of the affected area in the shortest possible time and cost, while providingsatisfactory cosmetic results. This task becomes challenging when working with horses, considering the contact betweenwounds and contaminated environment is common. Thus, the present study aims to report the case of a traumatic equinewound treated with Nile Tilapia Fish Skin (NTFS).Case: A male 27-year-old horse previously castrated, with no defined breed (NDB), and weighing 400 kg presented a 6.0 x5.5 cm superficial wound in the distal left anterior limb (LAL) due to skin laceration. The animal belonged to the cavalry ofthe Military Police of Ceará, a public institution in Fortaleza, Brazil. Although in excellent general health, with no previouscomorbidities or restriction of movement, the animal was removed from its role in equine-assisted therapy (EAT) until completewound healing, aiming adequate evaluation of the novel biomaterial via lower influence of external factors. After informedconsent from the owner was obtained, Nile Tilapia Fish Skin was applied to the lesion. The Ethics Committee on the use ofanimals of the Drug Research and Development Center of the Federal University of Ceará, Fortaleza, Brazil, approved the studyprotocol. Compliance with regulations on the ethical treatment of animals was performed. Nile Tilapia Fish Skin applicationfollowed a protocol similar to that established in human clinical studies. Initially, the horse was submitted to wound cleaningwith tap water and 2% chlorhexidine gluconate, with no requirement of pre-treatment surgical...
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Animais , Cavalos/cirurgia , Ferimentos e Lesões/cirurgia , Ferimentos e Lesões/veterinária , Transplante de Pele/veterinária , Ciclídeos , Técnicas de Fechamento de Ferimentos/veterináriaRESUMO
This study aims to evaluate the efficacy of Nile tilapia skin as a xenograft for the treatment of partial-thickness burn wounds in children. This is an open-label, monocentric, randomized phase II pilot study conducted in Fortaleza, Brazil. The study population consisted of 30 children between the ages of 2 and 12 years with superficial "partial-thickness" burns admitted less than 72 hours from the thermal injury. In the test group, the tilapia skin was applied. In the control group, a thin layer of silver sulfadiazine cream 1% was applied. Tilapia skin showed good adherence to the wound bed, reducing the number of dressing changes required, the amount of anesthetics used, and providing benefits for the patients and also for healthcare professionals, by reducing the overall work load. The number of days to complete burn wound healing, the total amount of analgesics required throughout the treatment, burn improvement on the day of dressing removal, and pain throughout the treatment were similar to the conventional treatment with silver sulfadiazine. Thus, tilapia skin can be considered an effective and low-cost extra resource in the therapeutic arsenal of pediatric superficial partial thickness burns.
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Queimaduras/cirurgia , Transplante de Pele/métodos , Tilápia , Animais , Brasil , Criança , Pré-Escolar , Feminino , Xenoenxertos , Humanos , Lactente , Masculino , Projetos Piloto , Sulfadiazina de Prata/uso terapêutico , CicatrizaçãoRESUMO
Mayer-Rokitansky-Küster-Hauser syndrome is the second most common cause of primary amenorrhea, trailing only to gonadal dysgenesis. Neovaginoplasty is an appropriate treatment option for patients who have failed dilation therapy. Several biomaterials have been used in this procedure, including peritoneum, amnion, skin grafts, and myocutaneous flaps. Nile Tilapia Fish Skin has noninfectious microbiota, morphologic structure comparable to human skin, and high in vivo bioresorption. In addition, it showed good outcomes when used as a xenograft for burn treatment. Thus, we suggest it as a new biologic graft for vaginal agenesis management. In this descriptive study, neovaginoplasty using Nile Tilapia Fish Skin offered 3 patients an anatomic and functional neovagina via a simple method with potential long-term effectiveness. When postsurgical dilation was performed correctly, a vaginal length greater than 6 cm was maintained at 180 days follow-up. Histologic and immunohistochemical analyses revealed the presence of stratified squamous epithelium with high expression of cytokeratins and fibroblast growth factor, matching the characteristics of normal adult vaginal tissue. We believe that further studies will show Nile Tilapia Fish Skin to be a relevant option in the therapeutic arsenal of Mayer-Rokitansky-Küster-Hauser syndrome.
Assuntos
Transtornos 46, XX do Desenvolvimento Sexual/cirurgia , Ciclídeos , Anormalidades Congênitas/cirurgia , Ductos Paramesonéfricos/anormalidades , Procedimentos de Cirurgia Plástica/métodos , Transplante de Pele/métodos , Vagina/anormalidades , Administração Intravaginal , Adolescente , Adulto , Animais , Produtos Biológicos/uso terapêutico , Brasil , Dilatação/métodos , Feminino , Humanos , Ductos Paramesonéfricos/cirurgia , Procedimentos de Cirurgia Plástica/efeitos adversos , Transplante de Pele/efeitos adversos , Retalhos Cirúrgicos , Transplante Heterólogo/efeitos adversos , Transplante Heterólogo/métodos , Transplante Heterotópico/efeitos adversos , Transplante Heterotópico/métodos , Resultado do Tratamento , Vagina/cirurgia , Adulto JovemRESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Extracts, essential oils and molecules from Casearia sylvestris have popularly shown pharmacological actions against chronic diseases, as anxiety, inflammation, cancer and dyslipidemia. In the context of antitumoral therapy, we investigated in vitro, ex vivo and in vivo toxicological changes induced by a Fraction with Casearins (FC) and its component Casearin X isolated from C. sylvestris on animal and vegetal cells, and upon invertebrates and mammals. MATERIAL AND METHODS: Cytotoxicity was carried out using normal lines and absorbance and flow cytometry techniques, Artemia salina nauplii, Danio rerio embryos and meristematic cells from Allium cepa roots. Acute and 30 days-mice analysis were done by behavioral, hematological and histological investigations and DNA/chromosomal damages detected by alkaline Cometa and micronucleus assays. RESULTS: FC was cytotoxic against lung and fibroblasts cells and caused DNA breaks, loss of integrity and mitochondrial depolarization on ex vivo human leukocytes. It revealed 24 h-LC50 values of 48.8 and 36.7⯵g/mL on A. salina nauplii and D. rerio embryos, reduced mitotic index of A. cepa roots, leading to cell cycle arrest at metaphase and anaphase and micronuclei. FC showed i.p. and oral LD50 values of 80.9 and 267.1â¯mg/kg body weight. Subacute i.p. injections induced loss of weight, swelling of hepatocytes and tubules, tubular and glomerular hemorrhage, microvesicular steatosis, lung inflammatory infiltration, augment of GPT, decrease of albumin, alkaline phosphatase, glucose, erythrocytes, and lymphocytes, and neutrophilia (pâ¯>â¯0.05). FC-treated animals at 10â¯mg/kg/day i.p. caused micronuclei in bone marrow and DNA strand breaks in peripheral leukocytes. CONCLUSIONS: This research postulated suggestive side effects after use of FC-related drugs, demonstrating FC as antiproliferative and genotoxic on mammal and meristematic cells, including human leukocytes, teratogenicity upon zebrafish embryos, myelosuppression, clastogenicity, and morphological and biochemical markers indicating liver as main target for FC-induced systemic toxicity.