RESUMEN
A method for simultaneous and quantitative determination of Cr, Mn, Fe, Co, Cu, Zn, Se and Cd elements in the subcellular fractions of nuclei, mitochondria, lysosome, microsome and cytosol of wound granulation tissue of severe burn patients by octopole reaction system (ORS) inductively coupled plasma mass spectrometry (ICP-MS) was established. Using differential centrifugation, the sample is separated into different subcellular fractions. The subcellular fraction was digested by HNO3 + H2O2 with microwave digestion followed by dilution with ultrapure water then the above 8 trace elements in the solution were analyzed directly by ICP-MS. In the presented method, using ORS eliminates the polyatomic interferences caused by the matrixes. Rh as internal standard element was used to compensate matrix effect and signal drift. The detection limits of the 8 elements are in the range of 0.72-33.05 ng x L(-1), and the RSD is less than 8.4%. The results showed that the levels of some elements in subcellular fractions of wound granulation tissues were significantly different from those of normal skin tissues. ORS-ICP-MS is a useful tool for simultaneous determination of multi-elements in wound granulation tissue of severe burn patients, and could be widely used in other biological samples analysis.
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Quemaduras/patología , Tejido de Granulación/química , Oligoelementos/análisis , Humanos , Espectrometría de Masas , Espectrofotometría AtómicaRESUMEN
Vascular lesions are commonly encountered in routine pathologic practice and often pose diagnostic challenges owing to their morphologic diversity. Although WT-1 expression was reported in some vascular tumors, little is known about its staining patterns in a spectrum of vascular lesions from various locations. We examined WT-1 immunostain in 95 cases of vascular lesions including angiosarcomas (AS, 19 cases), hemangioendotheliomas (HE, 5), Kaposi's sarcomas (KS, 4), cavernous hemangiomas (CVH, 12), capillary hemangiomas (CPH, 7), pyogenic granulomas (PG, 4), lymphangiomas (LA, 4), hemangiopericytomas (HP, 5), glomus tumors (GT, 8), vascular malformation (VM, 13) and granulation tissue (GRT, 14). Strong WT-1 cytoplasmic stain was invariably observed in all cases of malignant and borderline vascular tumors including AS (19/19), KS (4/4) and HE (5/5). WT-1 was also consistently expressed in CPH (7/7), PG (4/4), and GRT (14/14), while it became weaker in VM (10/13) and often negative in CVH (2/12) and LA (0/4). WT1 stain was not demonstrated in HP (0/5) and rarely in GT (2/8). We conclude that consistent and diffuse WT-1 cytoplasmic stain in AS, HE and KS can be useful in distinguishing these tumors from poorly differentiated tumors with mimicking features. On the other hand, reliable WT-1 stain in CPH, PG and GRT may help in differential diagnosis with non-endothelial vascular tumors such as GT and HP. Recognizing the WT-1 cytoplasmic stain in a broad spectrum of benign and neoplastic tissues is critical in formulating appropriate immunohistochemical panels and avoiding misinterpretation of results.
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Biomarcadores de Tumor/análisis , Citoplasma/química , Tejido de Granulación/química , Granuloma Piogénico/metabolismo , Inmunohistoquímica , Linfangioma/química , Neoplasias de Tejido Vascular/química , Neovascularización Patológica , Malformaciones Vasculares/metabolismo , Proteínas WT1/análisis , Adolescente , Adulto , Anciano , Biopsia , Niño , Preescolar , Diagnóstico Diferencial , Femenino , Tejido de Granulación/patología , Granuloma Piogénico/patología , Humanos , Linfangioma/patología , Masculino , Persona de Mediana Edad , Neoplasias de Tejido Vascular/patología , Valor Predictivo de las Pruebas , Malformaciones Vasculares/patología , Adulto JovenRESUMEN
Accurate and rapid assessment of the healing status of a wound in a simple and noninvasive manner would enable clinicians to diagnose wounds in real time and promptly adjust treatments to hasten the resolution of nonhealing wounds. Histologic and biochemical characterization of biopsied wound tissue, which is currently the only reliable method for wound assessment, is invasive, complex to interpret, and slow. Here we demonstrate the use of Raman microspectroscopy coupled with multivariate spectral analysis as a simple, noninvasive method to biochemically characterize healing wounds in mice and to accurately identify different phases of healing of wounds at different time-points. Raman spectra were collected from "splinted" full thickness dermal wounds in mice at 4 time-points (0, 1, 5, and 7 days) corresponding to different phases of wound healing, as verified by histopathology. Spectra were deconvolved using multivariate factor analysis (MFA) into 3 "factor score spectra" (that act as spectral signatures for different stages of healing) that were successfully correlated with spectra of prominent pure wound bed constituents (i.e., collagen, lipids, fibrin, fibronectin, etc.) using non-negative least squares (NNLS) fitting. We show that the factor loadings (weights) of spectra that belonged to wounds at different time-points provide a quantitative measure of wound healing progress in terms of key parameters such as inflammation and granulation. Wounds at similar stages of healing were characterized by clusters of loading values and slowly healing wounds among them were successfully identified as "outliers". Overall, our results demonstrate that Raman spectroscopy can be used as a noninvasive technique to provide insight into the status of normally healing and slow-to-heal wounds and that it may find use as a complementary tool for real-time, in situ biochemical characterization in wound healing studies and clinical diagnosis.
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Espectrometría Raman/métodos , Cicatrización de Heridas/fisiología , Heridas y Lesiones/clasificación , Heridas y Lesiones/patología , Animales , Biopsia , Análisis Factorial , Tejido de Granulación/química , Inflamación/metabolismo , Análisis de los Mínimos Cuadrados , Ratones , Ratones Endogámicos BALB C , Análisis Multivariante , Piel/químicaRESUMEN
INTRODUCTION: Apical periodontitis is an inflammatory disease of the periradicular tissues caused by the host's immune response to infection of the root canal system. MicroRNAs (miRNAs) have been shown to play an important role in the regulation of inflammation and the immune response; however, their role in the pathogenesis of endodontic periapical disease has not been explored. The purpose of this study was to examine the differential expression of miRNAs in diseased periapical tissues as compared with healthy controls. METHODS: We first compared miRNA profiles in diseased periapical tissues collected from patients undergoing endodontic surgery with those of healthy pulps by using microarray analyses. The target genes of the differentially expressed miRNAs were identified by using miRWalk and PubMed. Selected miRNAs linked to inflammation and the immune response were then confirmed in a separate cohort of diseased and healthy tissues by using quantitative reverse transcription-polymerase chain reaction. Healthy pulps and periodontal ligaments were used as controls. Data were normalized to the level of SNORD 44, which served as an endogenous control. RESULTS: Of the 381 miRNAs identified by using microarray, 24 miRNAs were down-regulated in diseased periapical tissues compared with controls (n = 13) (P < .003). The down-regulation of 7 miRNAs was confirmed from 9 selected miRNAs by using quantitative real-time polymerase chain reaction (n = 19) (P < .05). Target genes of these miRNAs include key mediators in the immune and inflammatory response such as interleukin-6, matrix metalloproteinase-9, and transforming growth factor-ß. CONCLUSIONS: These findings offer new insight into the pathogenesis of endodontic disease and have the potential to impact the development of new methods for prevention, diagnosis, and treatment of apical periodontitis.
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Enfermedades de la Pulpa Dental/genética , MicroARNs/análisis , Periodontitis Periapical/genética , Adolescente , Adulto , Anciano , Apicectomía/métodos , Biología Computacional , Pulpa Dental/química , Enfermedades de la Pulpa Dental/inmunología , Regulación hacia Abajo , Femenino , Tejido de Granulación/química , Humanos , Interleucina-6/genética , Masculino , Metaloproteinasa 9 de la Matriz/genética , Análisis por Micromatrices , Persona de Mediana Edad , Periodontitis Periapical/inmunología , Ligamento Periodontal/química , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factor de Crecimiento Transformador beta/genética , Adulto JovenRESUMEN
Carica papaya L. (Linn) (Caricaceae) is traditionally used to treat various skin disorders, including wounds. It is widely used in developing countries as an effective and readily available treatment for various wounds, particularly burns. This study evaluated the wound-healing and antimicrobial activity of C. papaya seed extract. Ethanol extract of C. papaya seed (50 mg/kg/day) was evaluated for its wound-healing activity in Sprague-Dawley rats using excision wound model. Animals were randomly divided into four groups of six each (group 1 served as control, group 2 treated with papaya seed extract, group 3 treated with a standard drug mupirocin and papaya seed extract (1:1 ratio) and group 4 treated with a mupirocin ointment. Rate of wound contraction and hydroxyproline content were determined to assess the wound-healing activity of the seed extract. The group 2 animals showed a significant decrease in wound area of 89% over 13 days when compared with groups 1 (82%), 3 (86%) and 4 (84%) respectively. The hydroxyproline content was significantly higher with the granulation tissue obtained from group 2 animals which were treated with C. papaya seed extract. Histological analysis of granulation tissue of the group 2 animals showed the deposition of well-organized collagen. The extract exhibited antimicrobial activity against Salmonella choleraesuis and Staphylococcus aureus. Our results suggest that C. papaya promotes significant wound healing in rats and further evaluation for this activity in humans is suggested.
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Carica/química , Extractos Vegetales/farmacología , Cicatrización de Heridas/efectos de los fármacos , Animales , Colágeno/análisis , Etanol , Tejido de Granulación/química , Hidroxiprolina/análisis , Masculino , Ratas , Ratas Sprague-Dawley , Salmonella/efectos de los fármacos , Semillas/química , Staphylococcus aureus/efectos de los fármacosRESUMEN
The objective of our study is to investigate the wound healing activity of the bark extract of Carapa guianensis in rats using three different wound models. The animals were randomly divided into two groups of six each in all the models. Test group animals were treated topically with the bark extract (200 mg/kg of body weight), and the controls were treated with petroleum jelly in the excision wound model. In the incision and dead space wound models the test group animals were treated with the extract of C. guianensis (200 mg/kg/day) orally by mixing in drinking water, and the control group animals were maintained with plain drinking water. Healing was assessed by the rate of wound contraction, period of epithelialization, skin breaking strength, granulation tissue weight, and hydoxyproline content. Antimicrobial activities of the extract against the microorganisms were also assessed. On day 15 extract-treated animals exhibited 99% reduction in the wound area compared to controls (93%). The extract-treated wounds were found to epithelialize faster than controls (P < .02). The skin breaking strength was significantly higher in extract-treated animals compared to controls (P < .001). The weight and hydroxyproline content of granulation tissue were significantly increased compared to controls. These observations support the use of C. guianensis is in the management of wound healing.
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Meliaceae/química , Fitoterapia , Corteza de la Planta/química , Extractos Vegetales/uso terapéutico , Cicatrización de Heridas , Administración Oral , Administración Tópica , Animales , Tejido de Granulación/química , Tejido de Granulación/patología , Hidroxiprolina/análisis , Masculino , Extractos Vegetales/administración & dosificación , Extractos Vegetales/química , Ratas , Ratas Sprague-Dawley , Cicatrización de Heridas/fisiología , Heridas y Lesiones/tratamiento farmacológico , Heridas y Lesiones/patología , Heridas y Lesiones/fisiopatologíaRESUMEN
BACKGROUND: Frog skin has been sequentially and scientifically evaluated by our group for its wound healing efficiency. Owing to the complex structure of skin, attempts were being made to analyse the role of individual constituents in different phases of healing. Our earlier papers have shown the significance of frog skin not only in wound healing but also enhancing the proliferating activity of the epidermal and dermal cells which are instrumental for normal healing process. We also have identified for the first time novel antimicrobial peptides from the skin of Rana tigerina and thereby reduce the complications involved in the sepsis. PURPOSE OF THE STUDY AND RESULTS: The current study envisages the role of frog skin lipids in the inflammatory phase of wound healing. The lipid moiety of the frog skin dominated by phospholipids exhibited a dose dependent acceleration of healing irrespective of the mode of application. The efficiency of the extract is attributed partially to the anti-inflammatory activity as observed by the histochemical and immunostimulatory together with plethysmographic studies. CONCLUSIONS: Thus, frog skin for the first time has been demonstrated to possess lipid components with pharmaceutical and therapeutic potential. The identification and characterization of such natural healing molecules and evaluating their mechanism of action would therefore provide basis for understanding the cues of Nature and hence can be used for application in medicine.
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Antiinflamatorios no Esteroideos/uso terapéutico , Lípidos/uso terapéutico , Materia Medica , Ranidae , Piel/química , Piel/efectos de los fármacos , Extractos de Tejidos/uso terapéutico , Cicatrización de Heridas/efectos de los fármacos , Administración Cutánea , Animales , Antiinflamatorios no Esteroideos/administración & dosificación , Antiinflamatorios no Esteroideos/análisis , Antiinflamatorios no Esteroideos/inmunología , Relación Dosis-Respuesta a Droga , Descubrimiento de Drogas , Edema/inducido químicamente , Edema/tratamiento farmacológico , Femenino , Tejido de Granulación/química , Tejido de Granulación/efectos de los fármacos , Hipersensibilidad/tratamiento farmacológico , Hipersensibilidad/inmunología , Inmunidad Humoral/efectos de los fármacos , India , Inyecciones Intraperitoneales , Lípidos/administración & dosificación , Lípidos/análisis , Lípidos/inmunología , Medicina Tradicional , Ratas , Ratas Wistar , Piel/lesiones , Extractos de Tejidos/administración & dosificación , Extractos de Tejidos/química , Extractos de Tejidos/inmunologíaRESUMEN
We prepared full thickness skin defects in rats fed on a protein-free diet as a hypoproteinaemia model, then switched the animals to a diet containing a normal protein level 1, 6 or 12 days after wounding (inflammatory, granulation and rearrangement phases of the wound healing process) to examine whether improvement in the low-protein state promotes subsequent wound healing. The interval until wound healing in rats fed on a normal protein diet was significantly shorter, whereas that in rats continuously fed on a protein-free diet was significantly longer than those of other groups. Early correction tended to accelerate wound healing. Although wound contraction in groups receiving a protein-corrected or protein-free diet remained similar until 15 days after wounding, thereafter the duration of the rearrangement phase was significantly longer in the protein-free group than in the other groups. The collagen level per unit of granulation tissue area during wound healing was significantly lower in the protein-free group than in the other groups. These findings indicate that protein correction at any time after wounding accelerates wound healing, although early correction is more effective, and reduces the duration of the rearrangement phase more than those of the inflammatory and granulation phases because of the deposit of collagen.
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Dieta con Restricción de Proteínas/efectos adversos , Proteínas en la Dieta/uso terapéutico , Deficiencia de Proteína/dietoterapia , Cicatrización de Heridas/fisiología , Heridas y Lesiones/dietoterapia , Animales , Proteínas Sanguíneas/metabolismo , Peso Corporal , Caseínas/uso terapéutico , Colágeno/análisis , Dieta con Restricción de Proteínas/métodos , Modelos Animales de Enfermedad , Ingestión de Energía , Tejido de Granulación/química , Masculino , Evaluación Nutricional , Deficiencia de Proteína/sangre , Deficiencia de Proteína/complicaciones , Deficiencia de Proteína/diagnóstico , Ratas , Ratas Wistar , Albúmina Sérica/metabolismo , Estadísticas no Paramétricas , Factores de Tiempo , Heridas y Lesiones/complicaciones , Heridas y Lesiones/patologíaRESUMEN
BACKGROUND: The receptor activator for nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG) are the key factors controlling the osteoclast and osteoblast action in the bone. PURPOSE: The study objective was to investigate the expression level of RANKL and OPG in cholesteatoma and granulation tissue, and to assess the relationship between their expression levels and osteolysis. MATERIAL AND METHODS: Patients with chronic otitis media with cholesteatoma (n = 28) and without cholesteatoma (n = 24) treated surgically at the Department of Otolaryngology of the Medical University of Gdansk were included in the study. RANKL and OPG expressions were analyzed by immunohistochemistry and Western blot. RESULTS: RANKL and OPG were expressed in all cholesteatoma and granulation tissues. RANKL expression was mainly observed in cholesteatoma subepithelial stroma, whereas OPG-positive cells originated from the epithelium. The number of OPG-positive cells in the normal skin was significantly higher than in cholesteatoma tissues. The RANKL protein level in cholesteatoma tissues was 1.8- and 1.5-fold higher than in the auditory canal skin and granulation tissues, respectively. The number of RANKL-positive cells in cholesteatoma tissues was significantly higher than in the normal skin. No substantial differences were found in average OPG protein levels between cholesteatoma tissues and the normal auditory canal skin. The ratio of RANKL/OPG was significantly higher in cholesteatoma tissues (2.93 ± 0.79) than in the skin samples (1.36 ± 0.34). CONCLUSIONS: Altered ratio of RANKL/OPG protein level in cholesteatoma tissues suggests that these proteins might be somehow involved in the pathogenesis of cholesteatoma. However, to resolve this issue a study on a larger group of patients should be conducted.
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Osteoprotegerina/análisis , Otitis Media/metabolismo , Ligando RANK/análisis , Adulto , Anciano , Anciano de 80 o más Años , Western Blotting , Colesteatoma/complicaciones , Colesteatoma/metabolismo , Enfermedad Crónica , Conducto Auditivo Externo/química , Epitelio/química , Femenino , Tejido de Granulación/química , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Piel/químicaRESUMEN
The wound healing potential of the aqueous, alcoholic extracts, and the butanolic fraction of the alcoholic extract obtained from the bark of Schrebera swietenioides were evaluated in the dexamethasone suppressed wound healing model. The work was conducted on rodents using incision, excision, and dead space wound models. The extracts of S swietenioides enhanced the breaking strength of incision wounds significantly (P < .05). Faster epithelization and contraction of excision wounds were observed in the treated groups (P < .05). Dead space wound model demonstrated an increase in breaking strength of granulation tissue and weight of dried granulation tissue after treatment with the extracts.The extracts attenuated the effect of dexamethasone on healing.The total RNA isolated from the granulation tissues of the extract-treated animals was significantly higher than in both dexamethasone and normal groups, (P < 0.05). It was observed that the DNA was intact in all the groups. These findings suggest that dexamethasone suppresses wound healing, possibly through an inappropriate transcription rather than causing DNA damage.The S swietenioides extracts have the capacity to reverse this effect.
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Dexametasona/uso terapéutico , Oleaceae , Fitoterapia/métodos , Extractos Vegetales/uso terapéutico , Cicatrización de Heridas/efectos de los fármacos , Heridas y Lesiones/tratamiento farmacológico , Administración Oral , Animales , ADN/análisis , Dexametasona/administración & dosificación , Modelos Animales de Enfermedad , Glucocorticoides/administración & dosificación , Glucocorticoides/uso terapéutico , Tejido de Granulación/química , Tejido de Granulación/patología , Inyecciones Intramusculares , Masculino , Ratones , Extractos Vegetales/administración & dosificación , Ratas , Ratas Wistar , Resultado del Tratamiento , Cicatrización de Heridas/genética , Heridas y Lesiones/genética , Heridas y Lesiones/patologíaRESUMEN
OBJECT: In this paper the authors' goal was to identify histological and immunohistochemical differences between cervical disc hemrniation and spondylosis. METHODS: A total of 500 cervical intervertebral discs were excised from 364 patients: 198 patients with disc herniation and 166 patients with spondylosis. We examined en bloc samples of endplate-ligament-disc complexes. Types of herniation and graded degrees of disc degeneration on MR images were examined histologically and immunohistochemically. RESULTS: The herniated discs showed granulation tissue, newly developed blood vessels, and massive infiltration of CD68-positive macrophages, which surrounded the herniated tissue mainly in the ruptured outer layer of the anulus fibrosus. The vascular invasion was most significant in uncontained (extruded)-type herniated discs. Chondrocytes positive for matrix metalloproteinase (MMP)-3, tumor necrosis factor (TNF)-alpha, basic fibroblast growth factor (bFGF), and vascular endothelial growth factor (VEGF) were abundant in both herniated and spondylotic discs. Free nerve fibers, positive for nerve growth factor (NGF), neurofilament 68, growth-associated protein (GAP)-43, and substance P, were strongly apparent in and around the outer layer of uncontained (extruded)-type herniated discs, with enhanced expression of NGF. The authors observed that herniated discs showed more advanced degeneration in the outer layer of the anulus fibrosus around the granulation tissue than spondylotic discs. On the other hand, spondylotic discs showed more advanced degeneration in the cartilaginous endplate and inner layer of the anulus fibrosus than herniated discs. Spondylotic discs also had thicker bony endplates and expressed TNFalpha and MMP-3 more diffusely than herniated discs, especially in the inner layer of the anulus fibrosus. CONCLUSIONS: The authors' results indicate that herniated and spondylotic intervertebral discs undergo different degenerative processes. It is likely that TNFa, MMP-3, bFGF, and VEGF expression is upregulated via the herniated mass in the herniated intervertebral discs, but by nutritional impairment in the spondylotic discs. Macrophage accumulation around newly formed blood vessels in the herniated disc tissues seemed to be regulated by MMP-3 and TNFalpha expression, and both herniated and spondylotic discs exhibited marked neoangiogenesis associated with increased bFGF and VEGF expression. Nerve fibers were associated with NGF overexpression in the outer layer of the anulus fibrosus as well as in endothelial cells of the small blood vessels.
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Vértebras Cervicales , Desplazamiento del Disco Intervertebral/patología , Osteofitosis Vertebral/patología , Condrocitos/química , Condrocitos/patología , Factor 2 de Crecimiento de Fibroblastos/análisis , Proteína GAP-43/análisis , Tejido de Granulación/química , Humanos , Immunoblotting , Inmunohistoquímica , Disco Intervertebral/patología , Imagen por Resonancia Magnética , Metaloproteinasa 3 de la Matriz/análisis , Fibras Nerviosas/química , Factor de Crecimiento Nervioso/análisis , Proteínas de Neurofilamentos/análisis , Sustancia P/análisis , Factor de Necrosis Tumoral alfa/análisis , Factor A de Crecimiento Endotelial Vascular/análisisRESUMEN
The aim of the study was to observe the effects of dibutyrylchitin (DBC) on the repair processes and to explain the mechanisms of its action in comparison with other dressing materials made of butyrylchitin (BC), regenerated chitin (RC), and chitosan. The results showed that DBC implanted subcutaneously to the rats increased weight of the granulation tissue. Increased cell number isolated from the wound and cultured on the DBC films was also revealed. The DBC was proved to reduce also the necrotic cells number in the culture. DBC elevates the glycosaminoglycans (GAG) level in the granulation tissue. The total collagen content in the wound was not influenced by all applied dressing materials. However, a low level of the poorly polymerized soluble collagen in the wounds treated with DBC and BC indicated better polymerization of the remaining part of that protein. Both DBC and chitosan increased the weight of granulation tissue. However, chitosan contrary to DBC lowered GAG content and increased water capacity in the wound. The study documents the beneficial influence of DBC on the repair, which could be explained by the modification of the extracellular matrix and cells number. The best effects were observed after application of DBC with [eta] DBC-1 = 1.75 dL/g.
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Vendajes , Quitina/análogos & derivados , Tejido de Granulación/efectos de los fármacos , Cicatrización de Heridas/efectos de los fármacos , Animales , Biopolímeros/química , Biopolímeros/farmacología , Celulosa/química , Celulosa/farmacología , Quitina/química , Quitina/farmacología , Quitosano/química , Quitosano/farmacología , Colágeno/análisis , Glicosaminoglicanos/análisis , Tejido de Granulación/química , Masculino , Ratas , Ratas WistarRESUMEN
The experimental data indicate that melatonin actively influences time-causes of changes of lipid content in rats tissue during the inflammation process. Its effect depends on a dose, modes of administration (intraperitonial, hypodermic or local) and duration of treatment. A single dose intraperitonial administration of melatonin (4 mg/kg) did not influence lipid content in the granular-fibrose tissue, while repeated injections of this hormone limited the increase in contents of lipids and phospholipids at the 5th and 8th days of regeneration. Long-term subcutaneous injections of melatonin caused distinct changes of lipids: at the dose of 0.3 mg/kg it prevented, and at the dose of 4 mg/kg it promoted the increase of lipid content in the granular-fibrose tissue. Local application of a melatonin solution (1.5 mg/ml) at early periods of regeneration caused insignificant changes of total lipids and total phospholipids in the granular-fibrose tissue. However, the higher concentration (15 mg/ml) of melatonin caused the decrease of total lipids due to reduced content of cholesterol and triglycerides and the increase of total phospholipids and some of their fractions.
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Tejido de Granulación/efectos de los fármacos , Lípidos/análisis , Melatonina/farmacología , Administración Cutánea , Administración Tópica , Animales , Relación Dosis-Respuesta a Droga , Tejido de Granulación/química , Técnicas In Vitro , Inyecciones Intraperitoneales , Masculino , Melatonina/administración & dosificación , Ratas , Ratas Wistar , Regeneración , Cicatrización de HeridasRESUMEN
The present investigation was undertaken to determine the efficacy of topical administration of flavone of sea buckthorn (Hippophae rhamnoides L.) on cutaneous wound healing in rats. Four full-thickness excision wounds were created on the back of rat and 1.0% w/v flavone prepared in propylene glycol was applied topically. Control animals received the vehicle alone in an identical manner. The healing of the wound was assessed by the rate of wound contraction, period of epithelialization, hydroxyproline, hexosamine, antioxidants estimation and histopathology of the granulation tissue. The sea buckthorn flavone promoted the wound healing activity as indicated by improved rate of wound contraction, decreased time taken for epithelialization (16.3 days versus 24.8 days in controls) and significant increase in hydroxyproline (26.0%) and hexosamine (30.0%) content. These findings were also confirmed by histopathological examinations. In addition, it was observed that sea buckthorn flavone possesses potent antioxidant properties as evidenced by significant increase in reduced glutathione (55.0%), vitamin C (70.0%) and catalase (20.0%) activities in wound granulation tissue. The flavone treatment also resulted in significant decrease in lipid peroxide levels (39.0%). The results suggest that the sea buckthorn flavone promotes wound healing activity.
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Dermis/patología , Flavonas/farmacología , Hippophae/química , Cicatrización de Heridas/efectos de los fármacos , Animales , Antioxidantes/análisis , Flavonoides/análisis , Tejido de Granulación/química , Tejido de Granulación/ultraestructura , Hexosaminas/análisis , Hidroxiprolina/análisis , Fenoles/análisis , Polifenoles , Ratas , Ratas Sprague-Dawley , Piel , Cicatrización de Heridas/fisiologíaRESUMEN
The aim of this study is to determine whether and to which extent pentosidine (PEN) is influenced in the case of advanced osteoarthritis (OA) in urine and in some tissues. Reverse phase HPLC method for PEN determination with fluorescence monitoring was utilized. Urinary pentosidine (U-PEN) and pyridinoline (U-PD) were determined in the group (N=67, age 67.1+/-11.7 years) in patients with OA before and after surgery (knee or hip replacement), and in healthy age-matched controls (N=18, age 67.0+/-6.0 years). Cartilage, synovial membrane and granulation tissue were also treated. In OA group significantly higher (P<0.001) U-PEN concentrations before (7.5+/-5.1 nmol/mmol creat.) and after (4.9+/-2.6 nmol/mmol creat.) surgery were found. U-PD was not significantly different between the groups (62.5+/-33.0 and 55.0+/-29.51 nmol/mmol creat., respectively), but differed significantly (P<0.0002 and P<0.01) from healthy subjects (40.2+/-16.5 nmol/mmol creat.). In the newly formed granulation tissue, PEN concentrations are significantly lower than in cartilage and in synovial tissues.
Asunto(s)
Arginina/análogos & derivados , Líquidos Corporales/química , Cromatografía Líquida de Alta Presión/métodos , Reactivos de Enlaces Cruzados/análisis , Tejido de Granulación/química , Lisina/análogos & derivados , Osteoartritis/metabolismo , Líquido Sinovial/química , Anciano , Aminoácidos/análisis , Aminoácidos/orina , Arginina/análisis , Biomarcadores/análisis , Biomarcadores/química , Biomarcadores/metabolismo , Biomarcadores/orina , Estudios de Casos y Controles , Femenino , Humanos , Lisina/análisis , Masculino , Persona de Mediana Edad , Osteoartritis/patología , Osteoartritis/cirugíaRESUMEN
Hippophae rhamnoides L. (family Elaeagnaceae), commonly known as seabuckthorn, is a wild shrub growing at high altitude (1200-4500 meters) in adverse climatic conditions. The aim of the present study was to evaluate healing potential of seabuckthorn leaves in a preclinical study on rats using a cutaneous excision-punch wound model. Four full-thickness excision-type wounds of 8.0 mm diameter were created on the dorsal surface of rats under aseptic conditions. The aqueous lyophilized extract of seabuckthorn leaves, at doses of 0.5%, 1.0%, and 1.5% w/v prepared in propylene glycol, were applied topically twice daily for 7 days. Control animals received the vehicle alone in an identical manner. Wound granulation tissues were excised on eighth day postwounding, and the hydroxyproline, hexosamine, total protein content, and antioxidant levels were determined. Wound surface area was also measured on the eighth day before wound excision to determine wound contraction. Topical application of 1.0% seabuckthorn leaf extract statistically significantly augmented the healing process, as evidenced by increases in the content of hydroxyproline and protein as well as the reduction in wound area when compared with similar effects in response to treatment using povidone-iodine ointment (standard care). The reduced glutathione, vitamin C, superoxide dismutase, catalase, and glutathione peroxidase activities showed significant increases in seabuckthorn leaf extract-treated wounds as compared to controls. The lipid peroxide levels were significantly decreased in leaf extract-treated wounds. The results suggest that aqueous leaf extract of seabuckthorn promotes wound healing, which may be due to increased antioxidant levels in the granulation tissue.
Asunto(s)
Hippophae , Extractos Vegetales/farmacología , Hojas de la Planta , Piel/lesiones , Cicatrización de Heridas/efectos de los fármacos , Animales , Antioxidantes/análisis , Tejido de Granulación/química , Hidroxiprolina/análisis , Masculino , Malondialdehído/análisis , Ratas , Ratas Sprague-DawleyRESUMEN
Wound healing occurs as a fundamental response to tissue injury. Several natural products have been shown to accelerate the healing process. The present investigation was undertaken to determine the efficacy of topical administration of an alcoholic bark extract of Butea monosperma (B. monosperma) on cutaneous wound healing in rats. Full-thickness excision wounds were made on the back of rat and B. monosperma extract was administered topically. The granulation tissue formed on days 4, 8, 12 and 16 (post-wound) was used to estimate total collagen, hexosamine, protein, DNA and uronic acid. The extract increased cellular proliferation and collagen synthesis at the wound site, as evidenced by increase in DNA, total protein and total collagen content of granulation tissues. The extract treated wounds were found to heal much faster as indicated by improved rates of epithelialization and wound contraction, also confirmed by histopathological examinations. Also, the tensile strength of drug-treated wounds was increased significantly. In addition, we show that B. monosperma possesses antioxidant properties, by its ability to reduce lipid peroxidation. The results clearly substantiate the beneficial effects of the topical application of B. monosperma in the acceleration of wound healing.
Asunto(s)
Butea , Extractos Vegetales/farmacología , Piel/lesiones , Cicatrización de Heridas/efectos de los fármacos , Administración Tópica , Animales , Colágeno/efectos de los fármacos , ADN/efectos de los fármacos , Evaluación Preclínica de Medicamentos , Tejido de Granulación/química , Tejido de Granulación/metabolismo , Hexosaminas/metabolismo , Masculino , Corteza de la Planta/química , Plantas Medicinales/química , Proteínas/efectos de los fármacos , Ratas , Ratas Wistar , Piel/metabolismo , Piel/patología , Resistencia a la Tracción , Ácidos Urónicos/metabolismoRESUMEN
The subcellular localization of cerium and lanthanum in the intestinal mucosa was studied after oral administration of cerium chloride or lanthanum chloride or lanthanum chloride followed 30 minutes after of cerium chloride to young adults Wistar rats. Two methods of observation and microanalysis were used. The transmission electron microscopy revealed the presence of dense electron granulations in the lysosmes of the duodenum enterocyte, when these elements were administrated simultaneously. The ion mass microanalysis permits to detect the presence of La and Ce as bright points outlining the intestinal villi. These points correspond to the lysosomes containing the granulations previously described. These granulations are formed by the cerium and the lanthanum associated to the phosphor and forming probably insoluble salts of Ce/La phosphate.
Asunto(s)
Cerio/administración & dosificación , Duodeno/efectos de los fármacos , Mucosa Intestinal/efectos de los fármacos , Lantano/administración & dosificación , Administración Oral , Animales , Cerio/análisis , Cerio/farmacocinética , Evaluación Preclínica de Medicamentos , Duodeno/química , Duodeno/ultraestructura , Microanálisis por Sonda Electrónica , Enterocitos/química , Enterocitos/efectos de los fármacos , Enterocitos/ultraestructura , Tejido de Granulación/química , Tejido de Granulación/efectos de los fármacos , Tejido de Granulación/ultraestructura , Absorción Intestinal , Mucosa Intestinal/química , Mucosa Intestinal/ultraestructura , Lantano/análisis , Lantano/farmacocinética , Lisosomas/química , Lisosomas/efectos de los fármacos , Lisosomas/ultraestructura , Microscopía Electrónica de Transmisión de Rastreo , Ratas , Ratas Wistar , Soluciones , Factores de TiempoRESUMEN
BACKGROUND: We investigated the effect of preoperative rectal irrigation with short-chain fatty acids on irradiated colonic anastomosis in rats. METHODS: Sixty male Wistar rats were divided into four groups. Group I (control group, n = 15) underwent left colon resection and primary anastomosis. Group II (Short-chain fatty acids pretreatment group, n = 15) had short-chain fatty acids rectal irrigation for five days preoperatively. Group III (preoperative radiotherapy group, n = 15) underwent irradiation to the whole pelvis eight and four days before the operation, for a total dose of 20 Gy. Group IV (preoperative radiotherapy group + short-chain fatty acids pretreatment group, n = 15) had rectal irrigation with short-chain fatty acids for five days after the second irradiation. Within each group, animals were anesthetized to assess the clinical, mechanical, histologic, and biochemical parameters of anastomotic healing on either the third or seventh postoperative days. RESULTS: The mean bursting pressure was significantly low in Group III on Day 3 and was significantly high in Group IV on Day 7 (P = 0.001, P = 0.021). The burst occurred at the anastomoses in all animals tested on the third postoperative day, and outside of the anastomoses in all animals tested on the seventh postoperative day. The histologic parameters of anastomotic healing, such as epithelial regeneration and formation of granulation tissue, were significantly improved by use of preoperative rectal irrigation with short-chain fatty acids on Day 7. The amount of total and salt-soluble collagen concentrations significantly increased in Group IV compared with the control group on Day 3 (P = 0.008, P = 0.004). CONCLUSION: Some mechanical and histologic aspects of colonic anastomotic healing can be adversely affected by preoperative radiotherapy, but rectal irrigation with short-chain fatty acids may improve anastomotic healing.
Asunto(s)
Neoplasias del Colon/terapia , Modelos Animales de Enfermedad , Ácidos Grasos Volátiles/administración & dosificación , Terapia Neoadyuvante/efectos adversos , Cuidados Preoperatorios/métodos , Radioterapia Adyuvante/efectos adversos , Recto , Irrigación Terapéutica/métodos , Cicatrización de Heridas , Anastomosis Quirúrgica/efectos adversos , Animales , Colectomía/efectos adversos , Colágeno/análisis , Colágeno/química , Colágeno/efectos de los fármacos , Colágeno/efectos de la radiación , Evaluación Preclínica de Medicamentos , Enema/métodos , Ácidos Grasos Volátiles/farmacología , Tejido de Granulación/química , Tejido de Granulación/efectos de los fármacos , Tejido de Granulación/efectos de la radiación , Masculino , Ratas , Ratas Wistar , Dehiscencia de la Herida Operatoria/etiología , Infección de la Herida Quirúrgica/etiología , Resistencia a la Tracción , Factores de Tiempo , Pérdida de Peso , Cicatrización de Heridas/efectos de los fármacos , Cicatrización de Heridas/efectos de la radiaciónRESUMEN
In this study, we investigated the role of nerve growth factor (NGF)-incorporated collagen on wound healing in rats. Full-thickness excision wounds were made on the back of female rats weighing about 150-160 g. Topical application of NGF-incorporated collagen, at a concentration of 1 microg/1.2 mg collagen/cm(2), once a day, for 10 days resulted in complete healing of wounds on the 15th day. The concentrations of collagen, hexosamine and uronic acid in the granulation tissue were determined. The NGF-incorporated collagen-treated rats required shorter duration for the healing with an increased rate of wound contraction. Histological and electron microscopical evaluations were also performed, which reveal the activation of fibroblasts and endoplasmic reticulum and therefore increased level of collagen synthesis due to NGF application. These results clearly indicate that the topical application of NGF-incorporated collagen enhanced the rate of healing of excision wounds.