Hemoperitoneum in a cow with lumbar muscle hemangiosarcoma

Background: Hemangiosarcoma is a malignant neoplasm of endothelial cells with an infiltrative growth pattern. Hemangiosarcomas are frequently reported in canines and rare in felines, sheep, goats, swine, horses and cattle. Few cases of hemangiosarcoma were reported in cattle. In the present report, we describe the clinicopathological findings of a bovine muscle hemangiosarcoma. Case: A 6-year-old, Girolando cow from the Dairy Cattle Sector of the Federal Rural University of Rio de Janeiro (UFRRJ), Seropédica, presented sternal decubitus. Clinical signs were markedly pale mucous membranes, moderate dehydration, respiratory distress, and increased heart rate. The hematological examination revealed intense regenerative anemia. Due to the worsening of the clinical condition, the cow was submitted to euthanasia. The necropsy and collection of various fragments of organs were performed, which were sent to the "Setor de Anatomia Patológica" (SAP-UFRRJ). Tissues were fixed in 10 % buffered formalin, routinely processed for histology and stained with Hematoxylin and Eosin (HE). The external mucous membranes were markedly pale. Multifocal areas of 1.5 x 1.0 cm, irregular and dark red were observed dissecting the quadratus lumborum muscle (hemangiosarcoma) fibers. These neoplasms were associated with an extensive cruoric clot adhered to the muscle fibers. The extensive, red, friable mass measured approximately 76 x 55 x 20 cm on the serous surfaces of the organs of the peritoneal cavity (hemoperitoneum). The spleen was moderately reduced. The bone marrow was markedly pale. Histologically, it was observed that there was an extensive proliferation of endothelial cells in the quadratus lumbar muscle mass dissecting the epimysium and perimysium. Endothelial cells had moderate pleomorphism, organized in vascular channels and forming multifocally solid areas with a significant amount of eosinophilic fibrillar material (fibrin). Sections of muscle neoplasm were subjected to immunohistochemistry with anti-von Willebrand factor primary antibody, which showed a multifocal moderate cytoplasmic immunolabeling of neoplastic endothelial cells. Discussion: There are few reports of striated muscle hemangiosarcoma in cattle. Muscular hemangiosarcomas were reported in a 4-month-old calf in the left cervical trapezius muscle and a 6-year-old Holstein cow with left pelvic limb mass lateral and distal to the knee. Some reports presented hemangiosarcoma in the iliopsoas muscle, left cervical trapezius muscle, pelvic limb muscles and right cervical muscle of the bovine. In the presented report, hemoperitoneum occurred as a result of hemorrhages from muscle hemangiosarcoma. Other studies have demonstrated cavity hemorrhages in joint, pelvic, pleural and cranial cavities associated with hemangiosarcoma. Hemangiosarcoma with regenerative anemia must be distinguished from other diseases that cause anemia. The main differential diagnoses of bovine with anemia are vena cava syndrome, coumarin derivatives poisoning, acute poisoning by Pteridium spp., tick fever, anaplasmosis, babesiosis and trypanosomiasis. Hemangiosarcoma should be differentiated from other lesions as hemangioma, vascular tumor of lymphatic endothelium and perivascular wall tumors. Cases with poorly differentiated morphology should be submitted for immunohistochemistry. In the present hemangiosarcoma case, we have used the von Willebrand factor for immunohistochemistry diagnosis. Expression of angiogenic growth factors such as CD31, vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and angiopoietin-1 (Ang-1) have also been used in the diagnosis of vascular proliferation lesions. Hemangiosarcoma in cattle should be included mainly in the differential diagnosis of diseases that cause acute anemia in cattle.

Cordycepin prevents the esophageal stricture formation in the alkali-burn rat model by exerting anti-inflammatory and antifibrotic effects

Purpose To investigate the efficacy of cordycepin, an adenosine analogue, on prevention of esophageal damage and stricture formation due to esophageal caustic burns in rat model comparing with prednisolone. Methods Caustic esophageal burn was introduced by 37.5% of NaOH to distal esophagus. Thirty-two Wistar albino rats were divided in four groups: sham rats undergone laparotomy, treated with 0.9% NaCl; control rats injured with NaOH without cordycepin treatment; cordycepin group injured with NaOH, treated with 20 mg/kg cordycepin; prednisolone group injured with NaOH, treated with 1 mg/kg prednisolone for 28 days. Efficacy was assessed by histopathological and immunohistochemical analysis of esophageal tissues. Results Cordycepin treatment significantly decreased inflammation, granulation tissue and fibrous tissue formation and prevented formation of esophageal strictures shown by histopathological damage score and stenosis indexes compared to control group (p 0.01). These effects are relatively more substantial than prednisolone, probably based on attenuation of elevation of proinflammatory cytokines hypoxia-inducible factor 1-alpha (HIF-1?), tumor necrosis factor alpha (TNF-?), proliferative and fibrotic factor fibroblast growth factor 2 (FGF2) and angiogenic factor vascular endothelial growth factor A (VEGFA) (p 0.05). Conclusions The findings suggest that cordycepin has a complex multifactorial healing process in alkali-burned tissue, more successful than prednisolone in preventing the formation of esophageal strictures and may be used as a therapeutic agent in the acute phase of esophageal alkali-burn.(AU)

Effect of Supplementary Methionine on Feather Growth and Related Indicators of Pigeon Squabs

We assessed the effect of health sand dietary supplementation with methionine (Met) on White King pigeons. Paired pigeons (n = 180) were fed one of five diets; group T1 received no added Met, while T2, T3, T4 and T5 received 30, 60, 90 and 120 g of supplemental DL-Met/kg, respectively. Each treatment was replicated three times with 24 pairs in each replicate. The results showed that supplementary Met had a minor effect on the length of the fourth primary wing feather in 28-day-old squabs (p>0.05), but the length of 14-day-old squabs in T2 was significantly longer (p=0.010). Dietary Met had a minor effect on Wnt-7a and fibroblast growth factor receptors-2 (FGFR-2) mRNA levels in 28-day-old squabs (p>0.05). The IGF-1 concentration in plasma was highest in T4 and lowest in T2 (p=0.012), but there was no difference between T1, T2 and T5 (p>0.05). In the chest muscle, the expression of IGF-1 in T3 and T4 was higher than in T1 (p=0.172 and 0.015, respectively). In the leg muscle, IGF-1 mRNA level was higher in T4 and T3, and lower in T2 (p>0.05). The results indicate that the optimal Met supplement for increasing fourth primary wing feather length was 30 g/kg Met in health sand, and the feathers were the longest in 14-day-old squabs. Adding 90 g/kg Met to health sand can improve the concentration of IGF-1, which is important for growth performance of pigeon squabs.(AU)

Effect of Supplementary Methionine on Feather Growth and Related Indicators of Pigeon Squabs

We assessed the effect of health sand dietary supplementation with methionine (Met) on White King pigeons. Paired pigeons (n = 180) were fed one of five diets; group T1 received no added Met, while T2, T3, T4 and T5 received 30, 60, 90 and 120 g of supplemental DL-Met/kg, respectively. Each treatment was replicated three times with 24 pairs in each replicate. The results showed that supplementary Met had a minor effect on the length of the fourth primary wing feather in 28-day-old squabs (p>0.05), but the length of 14-day-old squabs in T2 was significantly longer (p=0.010). Dietary Met had a minor effect on Wnt-7a and fibroblast growth factor receptors-2 (FGFR-2) mRNA levels in 28-day-old squabs (p>0.05). The IGF-1 concentration in plasma was highest in T4 and lowest in T2 (p=0.012), but there was no difference between T1, T2 and T5 (p>0.05). In the chest muscle, the expression of IGF-1 in T3 and T4 was higher than in T1 (p=0.172 and 0.015, respectively). In the leg muscle, IGF-1 mRNA level was higher in T4 and T3, and lower in T2 (p>0.05). The results indicate that the optimal Met supplement for increasing fourth primary wing feather length was 30 g/kg Met in health sand, and the feathers were the longest in 14-day-old squabs. Adding 90 g/kg Met to health sand can improve the concentration of IGF-1, which is important for growth performance of pigeon squabs.

The interaction of Wnt signaling members with growth factors in cultured granulosa cells

Wnt family members have recently been distinguished in the adult ovary with potential roles in ovarian function. Though particular growth factors interact with Wnt signaling members in extraovarian cell types, it is unclear whether this interaction is applicable in the granulosa cells. Therefore, the current study aimed to determine the effect of insulin-like growth factor-1 (IGF-I), epidermal growth factor (EGF) and basic fibroblast growth factor (FGF-β) on Wnt ligands WNT2 and WNT4 and Wnt receptor Frizzled-4 (FZD4) protein levels in cultured mouse granulosa cells. Granulosa cells were isolated from antral follicles of adult Balb/C mice and cultured for 24 hours in the presence of 100 ng/mL of IGF-I, or EGF or FGF-β. WNT2, WNT4 and FZD4 protein levels were evaluated through western blotting after the culture process. IGF-I treated granulosa cells had significantly the highest level of WNT2 and WNT4 as well as FZD4 when compared to FGF-β and EGF groups. FGF-β group had a significantly higher level of WNT2, WNT4 and FZD4 expression when compared to EGF group. FZD4 expression was at the highest level in the IGF-I group and this difference was statistically significant for all groups including uncultured cells and vehicle group. In addition, FGF-β was shown to positively affect the adhesion of granulosa cells. This study demonstrates that IGF-I, FGF-β and EGF have differential effects on the expressions of WNT2, WNT4, and FZD4 in cultured mouse granulosa cells, suggesting that particular growth factors related to ovarian function might conduct their roles in the ovary through Wnt signaling.

The interaction of Wnt signaling members with growth factors in cultured granulosa cells

Wnt family members have recently been distinguished in the adult ovary with potential roles in ovarian function. Though particular growth factors interact with Wnt signaling members in extraovarian cell types, it is unclear whether this interaction is applicable in the granulosa cells. Therefore, the current study aimed to determine the effect of insulin-like growth factor-1 (IGF-I), epidermal growth factor (EGF) and basic fibroblast growth factor (FGF-β) on Wnt ligands WNT2 and WNT4 and Wnt receptor Frizzled-4 (FZD4) protein levels in cultured mouse granulosa cells. Granulosa cells were isolated from antral follicles of adult Balb/C mice and cultured for 24 hours in the presence of 100 ng/mL of IGF-I, or EGF or FGF-β. WNT2, WNT4 and FZD4 protein levels were evaluated through western blotting after the culture process. IGF-I treated granulosa cells had significantly the highest level of WNT2 and WNT4 as well as FZD4 when compared to FGF-β and EGF groups. FGF-β group had a significantly higher level of WNT2, WNT4 and FZD4 expression when compared to EGF group. FZD4 expression was at the highest level in the IGF-I group and this difference was statistically significant for all groups including uncultured cells and vehicle group. In addition, FGF-β was shown to positively affect the adhesion of granulosa cells. This study demonstrates that IGF-I, FGF-β and EGF have differential effects on the expressions of WNT2, WNT4, and FZD4 in cultured mouse granulosa cells, suggesting that particular growth factors related to ovarian function might conduct their roles in the ovary through Wnt signaling.(AU)

Suppresses of Astragalus Polysaccharide on E. coli-Induced Injured Intestinal Microvascular through TLR4-NF-B Signal Pathways in Chickens

To investigate the hypothesis that APS can attenuate E. coli-induced microvascular dysfunction in chicken intestine, 60 0-day old male chickens were divided into three groups with 5 replications of 4 chicks. Chicken in the APS group were treated with 15 mg APS daily while the others were given 0 mg APS for 6 days. Then all 7-day old chicken were injected intraperitoneally by E. coli, except for the chicken in the control group. After 4 h, all chickens intestinal samples were collected to detect gene expressions involved in inflammatory factors and adhesion molecules. Results showed that APS attenuated the signs of edema and hemorrhage in 7-day old chicken intestinal mucosa which were induced by E. coli injection. Consistently, APS significantly reduced the increasing mRNA levels of inflammatory factors such as Tumor necrosis factor-a (TNF-a), interleukin (IL) -1 and inducible nitric oxide synthase (iNOS) (p 0.05), the same results were observed in vascular adhesion molecules such as E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Moreover, we observed that APS supplementation in water suppressed significantly (p 0.05) the decline of reparative factors such as epithelial growth factor (EGF) and basic fibroblast growth factor (bFGF) in E. coli injected group. Furthermore, supplementation with APS substantially blocked (p 0.05) the increase in Toll-like receptor-4 (TLR4) and Nuclear factor (NF)-B mRNA abundance (p=0.087) induced by E. coli infection. This study indicated that microvascular injured chicken intestine induced by E. coli would be attenuated with feeding APS, and the mechanism of repairing were probably mediated through TLR4-NF-B signal pathways.(AU)

Suppresses of Astragalus Polysaccharide on E. coli-Induced Injured Intestinal Microvascular through TLR4-NF-B Signal Pathways in Chickens

To investigate the hypothesis that APS can attenuate E. coli-induced microvascular dysfunction in chicken intestine, 60 0-day old male chickens were divided into three groups with 5 replications of 4 chicks. Chicken in the APS group were treated with 15 mg APS daily while the others were given 0 mg APS for 6 days. Then all 7-day old chicken were injected intraperitoneally by E. coli, except for the chicken in the control group. After 4 h, all chickens intestinal samples were collected to detect gene expressions involved in inflammatory factors and adhesion molecules. Results showed that APS attenuated the signs of edema and hemorrhage in 7-day old chicken intestinal mucosa which were induced by E. coli injection. Consistently, APS significantly reduced the increasing mRNA levels of inflammatory factors such as Tumor necrosis factor-a (TNF-a), interleukin (IL) -1 and inducible nitric oxide synthase (iNOS) (p 0.05), the same results were observed in vascular adhesion molecules such as E-selectin, intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). Moreover, we observed that APS supplementation in water suppressed significantly (p 0.05) the decline of reparative factors such as epithelial growth factor (EGF) and basic fibroblast growth factor (bFGF) in E. coli injected group. Furthermore, supplementation with APS substantially blocked (p 0.05) the increase in Toll-like receptor-4 (TLR4) and Nuclear factor (NF)-B mRNA abundance (p=0.087) induced by E. coli infection. This study indicated that microvascular injured chicken intestine induced by E. coli would be attenuated with feeding APS, and the mechanism of repairing were probably mediated through TLR4-NF-B signal pathways.

Combination of heterologous fibrin sealant and bioengineered human embryonic stem cells to improve regeneration following autogenous sciatic nerve grafting repair

Background Peripheral nerve injury is a worldwide clinical problem, and the preferred surgical method for treating it is the end-to-end neurorrhaphy. When it is not possible due to a large nerve gap, autologous nerve grafting is used. However, these surgical techniques result in nerve regeneration at highly variable degrees. It is thus very important to seek complementary techniques to improve motor and sensory recovery. One promising approach could be cell therapy. Transplantation therapy with human embryonic stem cells (hESCs) is appealing because these cells are pluripotent and can differentiate into specialized cell types and have self-renewal ability. Therefore, the main objective of this study was to find conditions under which functional recovery is improved after sciatic nerve neurorrhaphy. We assumed that hESC, either alone or in combination with heterologous fibrin sealant scaffold, could be used to support regeneration in a mouse model of sciatic nerve injury and repair via autografting with end-to-end neurorrhaphy. Methods Five millimeters of the sciatic nerve of C57BL/6 J mice were transected off and rotated 180 degrees to simulate an injury, and then stumps were sutured. Next, we applied heterologous fibrin sealant and/or human embryonic stem cells genetically altered to overexpress fibroblast growth factor 2 (FGF2) at the site of the injury. The study was designed to include six experimental groups comprising neurorrhaphy (N), neurorrhaphy + heterologous fibrin sealant (N + F), neurorrhaphy + heterologous fibrin sealant + doxycycline (N + F + D), neurorrhaphy + heterologous fibrin sealant + wild-type hESC (N + F + W), neurorrhaphy + heterologous fibrin sealant + hESC off (N + F +T), and neurorrhaphy + heterologous fibrin sealant + hESC on via doxycycline (N + F + D + T). We evaluated the recovery rate using Catwalk and von Frey functional recovery tests, as well as immunohistochemistry analysis. Results The experiments indicated...(AU)

Combination of heterologous fibrin sealant and bioengineered human embryonic stem cells to improve regeneration following autogenous sciatic nerve grafting repair

Background Peripheral nerve injury is a worldwide clinical problem, and the preferred surgical method for treating it is the end-to-end neurorrhaphy. When it is not possible due to a large nerve gap, autologous nerve grafting is used. However, these surgical techniques result in nerve regeneration at highly variable degrees. It is thus very important to seek complementary techniques to improve motor and sensory recovery. One promising approach could be cell therapy. Transplantation therapy with human embryonic stem cells (hESCs) is appealing because these cells are pluripotent and can differentiate into specialized cell types and have self-renewal ability. Therefore, the main objective of this study was to find conditions under which functional recovery is improved after sciatic nerve neurorrhaphy. We assumed that hESC, either alone or in combination with heterologous fibrin sealant scaffold, could be used to support regeneration in a mouse model of sciatic nerve injury and repair via autografting with end-to-end neurorrhaphy. Methods Five millimeters of the sciatic nerve of C57BL/6 J mice were transected off and rotated 180 degrees to simulate an injury, and then stumps were sutured. Next, we applied heterologous fibrin sealant and/or human embryonic stem cells genetically altered to overexpress fibroblast growth factor 2 (FGF2) at the site of the injury. The study was designed to include six experimental groups comprising neurorrhaphy (N), neurorrhaphy + heterologous fibrin sealant (N + F), neurorrhaphy + heterologous fibrin sealant + doxycycline (N + F + D), neurorrhaphy + heterologous fibrin sealant + wild-type hESC (N + F + W), neurorrhaphy + heterologous fibrin sealant + hESC off (N + F +T), and neurorrhaphy + heterologous fibrin sealant + hESC on via doxycycline (N + F + D + T). We evaluated the recovery rate using Catwalk and von Frey functional recovery tests, as well as immunohistochemistry analysis. Results The experiments indicated...

Analysis of BMSCs-intervened viscoelasticity of sciatic nerve in rats with chronic alcoholic intoxication

To investigate the impact of bone mesenchymal stem cells (BMSCs) intervention on the viscoelasticity of sciatic nerve in rats with chronic alcohol intoxication (CAI). Methods: The CAI rat models were prepared, divided into model groups, and treated with either BMSCs or basic fibroblast growth factor (bFGF). Then the rats underwent electrophysiological test and the serum levels of malondialdehyde (MDA), superoxide dismutase (SOD), and metallothionein (MT) were measured. Histological observation, stress relaxation test, and creep test were performed for the sciatic nerve of the CAI model in each group. Results: The MDA level of group BMSC was significantly lower (p 0.05) than that of groups MOD (the CIA model) and bFGF. The SOD and MT levels were higher in group BMSC than in groups MOD and bFGF (p 0.05). The motor nerve conduction velocity and amplitude were higher in group BMSC than in groups MOD and bFGF (p 0.05). The amounts of 7200s stress reduction and 7200 s strain increase of the sciatic nerve in group BMSC were greater than those in groups bFGF and MOD (p 0.05). Conclusion: Bone mesenchymal stem cells can improve the metabolism of free radicals, restore the tissue morphology and viscoelasticity of the chronic alcohol intoxication animal model, and positively affect the repairing of the injured sciatic nerve.(AU)

Pravastatin induces cell cycle arrest and decreased production of VEGF and bFGF in multiple myeloma cell line / Pravastatina induz parada no ciclo celular e diminuição na produção de VEGF e bFGF em linhagem de Mieloma Multiplo

Abstract Multiple myeloma (MM) is a B cell bone marrow neoplasia characterized by inflammation with an intense secretion of growth factors that promote tumor growth, cell survival, migration and invasion. The aim of this study was to evaluate the effects of pravastatin, a drug used to reduce cholesterol, in a MM cell line.Cell cycle and viability were determinate by Trypan Blue and Propidium Iodide. IL6, VEGF, bFGF and TGF were quantified by ELISA and qRT-PCR including here de HMG CoA reductase. It was observed reduction of cell viability, increase of cells in G0/G1 phase of the cell cycle and reducing the factors VEGF and bFGF without influence on 3-Methyl-Glutaryl Coenzyme A reductase expression.The results demonstrated that pravastatin induces cell cycle arrest in G0/G1 and decreased production of growth factors in Multiple Myeloma cell line.(AU)

Resumo O Mieloma Múltiplo é uma neoplasia de linfócitos B da medula óssea, caracterizada por inflamação com uma intensa secreção de fatores de crescimento que promovem o aumento do volume do tumor, sobrevivência celular, migração e invasão. O objetivo deste estudo foi avaliar os efeitos da pravastatina, uma droga usada para reduzir o colesterol, em um linhagem de MM. O ciclo celular e viabilidade foram determinadas por Trypan Blue e iodeto de propídio. IL6, VEGF, bFGF e TGF foram quantificadas por ELISA e qRT-PCR, incluindo aqui de HMG CoA redutase. Observou-se a redução da viabilidade das células, aumento de células na fase G0/G1 do ciclo celular e redução no VEGF e bFGF, sem influência na expressão da enzima 3-Metil-Glutaril Coenzima A redutase. Os resultados demonstraram que a pravastatina induz parada no ciclo celular em G0/G1 e diminuição da produção de fatores de crescimento em várias linhas de células de Mieloma.(AU)

Pravastatin induces cell cycle arrest and decreased production of VEGF and bFGF in multiple myeloma cell line

Abstract Multiple myeloma (MM) is a B cell bone marrow neoplasia characterized by inflammation with an intense secretion of growth factors that promote tumor growth, cell survival, migration and invasion. The aim of this study was to evaluate the effects of pravastatin, a drug used to reduce cholesterol, in a MM cell line.Cell cycle and viability were determinate by Trypan Blue and Propidium Iodide. IL6, VEGF, bFGF and TGF were quantified by ELISA and qRT-PCR including here de HMG CoA reductase. It was observed reduction of cell viability, increase of cells in G0/G1 phase of the cell cycle and reducing the factors VEGF and bFGF without influence on 3-Methyl-Glutaryl Coenzyme A reductase expression.The results demonstrated that pravastatin induces cell cycle arrest in G0/G1 and decreased production of growth factors in Multiple Myeloma cell line.

Resumo O Mieloma Múltiplo é uma neoplasia de linfócitos B da medula óssea, caracterizada por inflamação com uma intensa secreção de fatores de crescimento que promovem o aumento do volume do tumor, sobrevivência celular, migração e invasão. O objetivo deste estudo foi avaliar os efeitos da pravastatina, uma droga usada para reduzir o colesterol, em um linhagem de MM. O ciclo celular e viabilidade foram determinadas por Trypan Blue e iodeto de propídio. IL6, VEGF, bFGF e TGF foram quantificadas por ELISA e qRT-PCR, incluindo aqui de HMG CoA redutase. Observou-se a redução da viabilidade das células, aumento de células na fase G0/G1 do ciclo celular e redução no VEGF e bFGF, sem influência na expressão da enzima 3-Metil-Glutaril Coenzima A redutase. Os resultados demonstraram que a pravastatina induz parada no ciclo celular em G0/G1 e diminuição da produção de fatores de crescimento em várias linhas de células de Mieloma.

Fator de crescimento fibroblástico 18 (FGF18) modula a expressão de CTGF em CCOs e embriões bovinos produzidos in vitro

Em 1995 a via Hippo foi inicialmente descoberta na Drosofila melanogaster, e desde então diversos pesquisadores vêm tentando entender melhor essa via. A via Hippo está relacionada a processos de proliferação celular, tanto em células foliculares ovarianas como no desenvolvimento embrionário. Os efetores dessa via, YAP e TAZ atuam como coativadores de transcrição de genes de proliferação e sobrevivência celular como CTGF e CYR61. Porém o fator de crescimento de fibroblasto também induz a expressão de CTGF. E segundo estudos recentes foi descoberto que o fator de crescimento fibroblastico 18 (FGF18) está presente na tuba uterina durante o desenvolvimento embrionário inicial. Isso nos levou a pensar que o FGF18 teria alguma função durante o desenvolvimento embrionário. Portanto, o objetivo deste estudo foi determinar se o FGF18 modula a via Hippo através da expressão de genes alvos de proliferação celular (CTGF e CYR61) durante a maturação oocitária e desenvolvimento embrionário inicial. Para isso três experimentos foram realizados. No primeiro experimento complexos cumulus oocito (CCOs) de bovinos foram maturados in vitro durante 6, 12 e 24 horas na presença de concentrações graduais de FGF18(0, 10 e 100ng/mL). No segundo experimento os CCOs foram maturados 3, 6 e 9 horas na presença de 0 ou 100ng/mL de FGF18. No experimento três CCOs foram colocados para maturação e foram divididos em três grupos. Um grupo considerado grupo controle no qual CCOs foram maturados, realizada a fertilização in vitro (FIV) e depois cultivados in vitro (CIV) sem a adição de FGF18. Um grpo no qual 100nf/mL de FGF18 foram adicionados durante as 24 horas da maturação in vitro e epós foi realizado a fertilização e o cultivo em meios sem adição de FGF18. E um terceiro grupos os CCOs foram cultivados (CIV) por sete dias na presença de 100ng/mL de FGF18. Neste grupo a MIV e FIV foram realizadas na ausência de FGF18. Os genes avaliados nos três experimentos foram os genes de proliferação celular, CTGF e CYR61. O experimento um que teve o objetivo de avaliar a adição de concentrações graduais durante a maturação houve um aumento (p<0,05) da expressão gênica para CTGF no grupo tratado com 100ng/mL as 12horas. No experimento dois em que o objetivo era avaliar a concentração de 100ng/mL houve uma diminuição (p<0,05) do grupo tratado em relação ao grupo controle as três horas. E no experimento três em que o objetivo era avaliar a adição de FGF18 durante o desenvolvimento embrionário, houve diferença estatística (p<0,05) do grupo tratado com FGF18 durante a CIV. Desde forma concluímos que FG18 modula a expressão de CTGF em período críticos da maturação nuclear do oócito, da expansão do cumulus e do desenvolvimento embrionário inicial

In 1995, the Hippo pathway was initially discovered in Drosophila melanogaster, and since then several researchers have been trying to better understand this pathway. The Hippo pathway is involved in cell proliferation processes, both in ovarian follicular cells in the early embryonic development. The effectors of this pathway YAP e TAZ act as transcription coactivators of cell proliferation and survival genes such as CTGF and CYR61. However, the fibroblast growth factor also induces CTGF expression. However, recent studies by the group found that fibroblast growth factor 18 (FGF18) is present in the fallopian tube during early embryonic development. This led us to think that FGF18 would have some role during embryonic development. Therefore, the aim of the following study was to determine whether FGF18 modulates the Hippo pathway through the expression of target genes for cell proliferation (CTGF and CYR61) during oocyte maturation and early embryonic development. To acquire the results of the objective, three experiments were carried out, with in vitro maturation (IVM) of cumulus oocyte complexes (COCs) in the first and second experiments, with the addition of FGF18 during IVM, and the structures were collected at different times. In the first experiment, gradual concentrations of FGF18 (0, 10 and 100ng / mL) were added and there was a control group without the addition of FGF18, and 6, 12 and 24 hours of maturation were collected. In the second experiment, a concentration of 100ng / mL of FGF18 was added during IVM, there was a control group and the structures were collected at 0, 3, 6 and 9 hours of maturation. In the experiment, three COCs were put to maturation and were divided into three groups. A group considered a control group in which COCs were matured, with in vitro fertilization (IVF) and then in vitro culture (IVC) without the addition of FGF18. A group in which, during maturation, FGF18 was added only during IVM and afterwards fertilization and culture were carried out in media without the addition of FGF18. In a third group, COCs were put to mature, fertilized and, when put to culture, FGF18 was added in the medium. The genes evaluated in the three experiments were the cell proliferation genes, CTGF and CYR61. In experiment one, which aimed to evaluate the addition of gradual concentrations during maturation, there was an increase (p <0.05) in CTGF gene expression in the group treated with 100ng / mL at 12 hours. In experiment two, in which the objective was to assess the concentration of 100ng / mL, there was a decrease (p <0.05) in the treated group compared to the control group at three hours. And in experiment three in which the objective was to evaluate the addition of FGF18 during embryonic development, there was a statistical difference (p <0.05) in the group treated with FGF18 during IVC. Therefore, we conclude that FGF18 modulates CTGF expression in critical periods of oocyte nuclear maturation, cumulus expansion and early embryonic development in cattle.

Efeito dos fatores peptídicos de crescimento recombinantes (PDGF-BB e/ou VEGF165) no secretoma de células-tronco mesenquimais e em modelo animal de cicatrização de pele

A Medicina Regenerativa visa alcançar tratamentos que acelerem as diferentes fases do processo de cicatrização em humanos e animais e a manipulação da composição dos fatores de crescimento (FC) pode melhorar ou modificar o processo de reparo e remodelação dos tecidos lesados, bem como proporcionar cicatrizes mais estéticas e funcionais. O presente estudo objetivou avaliar o efeito dos fatores peptídicos de crescimento recombinantes humanos derivado de plaquetas (PDGF- BB) e/ou de crescimento endotelial e/ou vascular (VEGF165) em células-tronco mesenquimais do tecido adiposo (canino) e de medula óssea (equino) e seu secretoma, e em modelo de cicatrização de pele em ratos nude Rowett. Os FC foram obtidos a partir de meios purificados por cromatografia de afinidade a heparina e caracterizados por ELISA e Western blot, e foram utilizados em tampão como veículo ou em hidrogel de alginato 2%. As células foram obtidas, caracterizadas, cultivadas e tratadas com FC e o meio condicionado foi utilizado para isolamento de exossomos. O modelo animal de cicatrização de feridas foi estabelecido em ratos nude Rowett, onde foram aplicados 3g/mL de FC sozinhos ou combinados em feridas no dorso dos animais, e após 7 dias a evolução da cicatrização foi avaliada macroscópica e histopatologicamente. O presente estudo demonstrou pela primeira vez o potencial da suplementação de rhPDGF-BB e/ou rhVEGF165 na cicatrização de feridas cutâneas em sete dias, estimulando angiogênese, reação de fibroblastos e reepitelização. O hidrogel de alginato 2% mostrou ser a melhor abordagem para ser usada como veículo para aplicações de FC in vivo em relação ao tampão veículo, mas não em ensaios in vitro. O tratamento com ambos os fatores combinados melhorou o perfil terapêutico de células-tronco mesenquimais de tecido adiposos estimulando a migração celular e a secreção de exossomos. O estudo demonstrou pela primeira vez que o tratamento com rhVEGF165 estimulou proliferação celular, o perfil angiogênico e a secreção de exossomos por células-tronco mesenquimais de medula óssea equina. Juntos os dados embasam futuras investigações sobre efeito da suplementação com FC in vitro, no conteúdo celular e molecular do secretoma de células-tronco mesenquimais, e in vivo em um modelo seguro e reprodutível, abrindo uma nova perspectiva para abordagem terapêutica livre de células para cicatrização de feridas. Palavras-chave: Cicatrização de feridas. PDGF-BB. VEGF. Células-tronco mesenquimais. Exossomos.

Regenerative Medicine aims to achieve treatments that accelerate the different stages of the healing process in both humans and animals, and the manipulation of the growth factor (GF) composition can improve or modify the repair and remodeling process of injured tissues, as well as providing more aesthetic and functional scars. The present study aimed to evaluate the effect of recombinant human platelet- derived growth factor (PDGF-BB) and/or endothelial vascular growth factor (VEGF165) on adipose (canine) and bone marrow (equine) mesenchymal stem cells and their secretome, as well as in a model of skin healing in nude rats. GF were obtained from purified media by heparin affinity chromatography and characterized by ELISA and Western blot, and were used in vehicle buffer or in 2% alginate hydrogel. Mesenchymal stem cells were obtained, characterized, cultured and treated with GF and exosomes were isolated from conditioned media. The animal model of wound healing was established in Rowett nude rats, where 3g/mL of GF were applied alone or combined in wounds on the animals' backs, and after 7 days the healing evolution was evaluated macroscopically and histopathologically. The present study demonstrated for the first time the potential of rhPDGF-BB and/or rhVEGF165 supplementation in the healing of skin wounds in seven days, stimulating angiogenesis, fibroblast reaction and re-epithelialization, but a long-term evaluation is necessary. The 2% alginate hydrogel proved to be the best approach to be used as a vehicle for GF applications in vivo in relation to the vehicle buffer, but not in vitro assays. The treatment with both factors combined improved the therapeutic profile of adipose mesenchymal stem cells by stimulating cell migration and exosome secretion. The study demonstrated for the first time that treatment with rhVEGF165 stimulated cell proliferation, angiogenic profile and secretion of exosomes by equine bone marrow mesenchymal stem cells. Together, the data support future investigations into the effect of GF supplementation in vitro, on cellular and molecular content of the mesenchymal stem cell secretome and in vivo, in a safe and reproducible model, opening a new perspective for a cell-free therapeutic approach for wound healing.

Repair of surgical wounds in rats using a 10 por cento unripe Musa sapientum peel gel

To investigate the efficacy of a 10% gel of unripe banana (Musa sapientum) peel in treating surgical wounds in rats. A longitudinal, prospective, randomized triple-blind study was conducted with 60 Wistar rats (Rattus norvegicus albinus) weighing approximately 400g. The animals were randomly divided into: control group (treated with gel containing no active ingredient) and study group (treated with 10% gel of unripe banana peel). The gel was applied every three days to a 4x4-cm surgical wound created on the back of each animal (day 0) in both groups. Tissue samples were collected for histological analysis on days 14, 21 and 28. On day 14, more extensive vascular proliferation (p=0.023), presence of mononuclear cells (p=0.000), fibroblast proliferation (p=0.012), re-epithelialization (p=0.000), and decreased presence of polymorphonuclear cells (p=0.010) were observed in the study group than in controls. No significant between-group difference in the presence of polymorphonuclear cells was found on day 21. Fibroblast proliferation was significantly greater (p=0.006) in the study group than in the control group on day 28. The 10% gel of unripe banana peel showed anti-inflammatory activity and stimulated wound healing in rat skin when compared with a gel containing no active ingredient.(AU)

Comparative histological study on wound healing on rat's skin treated with Mitomycin C or Clobetasol propionate

To compare histologically the action of Mitomycin C and that of Clobetasol propionate for surgical wound healing in rats. A circular skin fragment was surgically removed from 57 Wistar rats. Twenty-two animals were treated with Mitomycin C with topical medication in a single dose, 22 with Clobetasol propionate with a cream medication once a day for 15 days and 13 did not receive any medication. The animals were euthanized 30 and 60 days, and the scars subjected to histological examination. The histological analysis on the samples did not show statistically significant differences regarding the quantities of fibroblasts, fibrocytes and vascular proliferation in the three groups, in the evaluations after 30 and 60 days. In the treated groups with Mitomycin C and Clobetasol there was a decrease in collagen concentration over the 30-day period and an increase in collagen concentration over the 60-day period, in comparison with the control group. The actions of Mitomycin C and Clobetasol were equivalent and not interfere in fibroplasias and in angiogenesis. Both drugs initially cause a decrease in collagen over a 30-day period and an increase over a 60-day period, demonstrating a delay in the wound healing.(AU)

Platelet rich plasma to faster navel healing in lambs / Plasma rico em plaquetas para acelerar a cicatrização umbilical em cordeiros

Onfaloflebitis is common in lambs born in poor hygienic conditions and occurs where inadequate treatment of umbilical healing is placed. Males are more affected lambs probably because of contact with urine that slows healing umbilical and removes some medications or topical antibiotics applied facilitating the bacteria entrance, mainly Fusobacterium necrophorum affecting joints, lungs, kidneys, liver, leading to animal death. Platelets are blood cells that interact with several receptors and modulate platelet function. It promotes hemostasis through four mechanisms: adhesion, release of platelet secretion, aggregation and pro-coagulation action. Complex network of growth factors are involved in tissue homeostasis such as transforming growth factor (TGF)-b, platelet-derived growth factor (PDGF), insulin-like growth factor (IGF), fibroblast growth factor (FGF)-b and hepatocyte growth factor (HGF). These factors and other bioactive molecules are contained in platelet alpha granules, and the use of intra-articular injections of platelet concentrate has been proposed as a minimally invasive solution to promote cartilage, osteo and muscular healing. Its use in regenerative medicine represents a simple, low-cost, and minimally invasive approach that might fasten healing processes. Regenerative treatment with autologous products, including the PRP (Platelet Rich Plasma) have been widel

O artigo não apresenta resumo em português.

Reconstruction of abdominal wall defects using small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor

To construct a new biomaterial-small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor, and to evaluate the new biomaterials for the reconstruction of abdominal wall defects. Thirty six Sprague-Dawley rats were used in the animal experiments and randomly divided into three groups. The new biomaterial was constructed by combining small intestinal submucosa with gelatin hydrogel for basic fibroblast growth factor release. Abdominal wall defects were created in rats, and repaired using the new biomaterials (group B), compared with small intestinal submucosa (group S) and ULTRAPROTM mesh (group P). Six rats in each group were sacrificed at three and eight weeks postoperatively to examine the gross effects, inflammatory responses, collagen deposition and neovascularization. After implantation, mild adhesion was caused in groups B and S. Group B promoted more neovascularization than group S at three weeks after implantation, and induced significantly more amount of collagen deposition and better collagen organization than groups S and P at eight weeks after implantation. Small intestinal submucosa coated with gelatin hydrogel incorporating basic fibroblast growth factor could promote better regeneration and remodeling of host tissues for the reconstruction of abdominal wall defects.(AU)

TGF- B1 on induced osteogenic differentiation of human dermal fibroblast

PURPOSE: To evaluate the role of transforming growth factor beta 1 (TGF-β1) on the induced osteogenic differentiation of human dermal fibroblasts. METHODS: We performed four groups with cultured dermal fibroblasts according to the culture medium: CONTROL (DMEM culture medium); TGF-β1 (DMEM culture medium with 10 ng/ml of TGF-β1); OSTEOG (DMEM culture medium with 0.5 µg/ml of ascorbic acid, 10 mmol/l of β-glycerophosphate and 10 nmol/L of dexamethasone); and OSTEOG/TGF-β1 (osteogenic medium with 10 ng/ml of TGF-β1). Alkaline phosphatase (ALP) activity and the amount of osteocalcin (OC) in the supernatant, as well as the capability to form calcium phosphate deposits, were analysed for 28 days RESULTS: There were significant differences (p<0.05) between CONTROL and TGF-β1 groups in comparison with OSTEOG and OSTEOG/TGF-β1 groups in the ALP activity and OC amount. Although, both osteogenic groups had the same behavior with regard the expression curve during the experimental time, the OSTEOG/TGF-β1 group achieved significantly higher ALP and OC levels and showed no significant difference in the levels of mineralized deposits and in comparison with the levels found in the OSTEOG group. CONCLUSION: The addition of transforming growth factor beta 1 to the osteogenic culture medium increased the activity of alkaline phosphatase and the amount of osteocalcin, but TGF-β1 did not alter the presence of mineralized calcium phosphate deposits. .(AU)