RESUMO
Nanoplastics (NPs) affect fertility. We evaluated the effects of NPs treatment on luteal and endothelial cells. We examined crucial markers of growth and redox status. NPs treatment did not induce changes in ATP levels in luteal cells, while it increased (p< 0.05) their proliferation. In endothelial cells, no change in proliferation was detected, while an increase (p<0.05) in ATP levels was observed. The increase of reactive oxygen species, superoxide anion (p<0.05) and nitric oxide (p<0.001) was detected in both cell types, which also showed changes in superoxide dismutase enzyme activity as well as an increase of non-enzymatic antioxidant power (p<0.05). A decrease (p<0.05) in progesterone production as well as an increase of vascular endothelial growth factor A levels were detected (p<0.05). In addition, a dose-dependent accumulation of NPs in endothelial cells was shown, that likely occurred through adhesion and internalization. Results underline potential risk of NPs for corpus luteum functionality.
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BACKGROUND: This study aimed to investigate variations of the oxidative status in cats affected by urethral obstruction (UO) under Feline Idiopathic Cystitis (FIC) and Bacterial Cystitis (BC), in comparison with a group of healthy subjects. In both groups, the levels of several markers (either direct or indirect) indicative of the oxidative attack and of the antioxidant response were analyzed on plasma and urine samples. In particular, the plasma samples were evaluated for nitric oxide (NO), hydroperoxides derived by reactive oxygen activity (d-ROMs test), superoxide anion (O2-), glutathione peroxidase activity (GPx), superoxide dismutase activity (SOD), and ferric reducing antioxidant power (FRAP test); while on urine the levels of NO, d-ROMs, FRAP, SOD, malondialdehyde (MDA) and 8-hydroxydeoxyguanosine (8-OHdG) were measured. Urine of UO patients was also subjected to urine-culture test. RESULTS: The analytical data on plasma showed that UO, independently of the FIC or BC etiology, induced the insurgence of oxidative stress conditions at the systemic level. In the urine of the UO patients, except for SOD that increased, the markers of redox status were markedly decreased due probably their compromised filtration, thus suggesting involvement of renal function (assessed also by the high levels of plasma creatinine and proteinuria) with no oxidative damage of the lower urinary tract. Moreover, the adoption of a novel oxidative stress index' (OSI) allowed to establish, by means of a numerical value, the different degrees of oxidative stress conditions for single UO patients, both in terms of oxidative attack and antioxidant response. CONCLUSIONS: Feline urethral obstruction, induced by Idiopathic Cystitis and Bacterial Cystitis, causes oxidative stress conditions at the systemic level that do not interest the lower urinary tract. Despite to the high variability of the profiles of oxidative stress indexes both in healthy and UO patients, the determination of OSI made possible the evaluation of their single degrees of oxidative stress. Possibly the results of this investigation can be compared with those of correspondent pathologies both in humans and in other animal species.
Assuntos
Biomarcadores , Doenças do Gato , Estresse Oxidativo , Obstrução Uretral , Animais , Gatos , Biomarcadores/urina , Biomarcadores/sangue , Obstrução Uretral/veterinária , Obstrução Uretral/urina , Obstrução Uretral/sangue , Doenças do Gato/urina , Doenças do Gato/sangue , Masculino , Feminino , Cistite/veterinária , Cistite/urina , Cistite/sangue , Cistite/microbiologia , 8-Hidroxi-2'-Desoxiguanosina/urina , 8-Hidroxi-2'-Desoxiguanosina/sangue , Superóxido Dismutase/sangueRESUMO
Mesenchymal Stromal Cells (MSCs)-based therapies are rapidly gaining interest in veterinary medicine. Cellular therapy represents a new challenge for practitioners and requires precise coordination between the cell processing laboratory and the veterinary clinic. Cryopreservation is the best method to provide fast, in-time, and long-distance delivery of cells for therapeutic applications. However, potentially toxic cryoprotectants and xenobiotic products make the direct administration of cells impracticable for patients. Alternatively, the cells may be resuspended in a ready-to-use vehicle and shipped to the veterinary clinic. In this study, two nutrient-poor vehicles (physiologic saline and ringer lactate solutions) and two nutrient-rich vehicles (the releasate derived from autologous Platelet Poor Plasma and Platelet Rich Plasma) were tested on adipose tissue-derived canine MSCs (AD-MSCs). AD-MSCs stored for 2, 4, or 24 h in the different media were compared regarding mortality, metabolic activity, and replicative capacity. Furthermore, antioxidant activity and the pattern of expression of genes related to AD-MSCs function were performed following 24 h of storage. The results showed that all the different vehicles preserve cell vitality and replication following short-term storage. In long-term storage, the vehicle and cell density affect cell vitality, proliferation, and gene expression (CCL-2, CXCR-4, and TSG-6). Nutrient-rich vehicles seem better suited to preserve cell functionalities in this contest.
Assuntos
Tecido Adiposo , Células-Tronco Mesenquimais , Humanos , Animais , Cães , Diferenciação Celular , Terapia Baseada em Transplante de Células e Tecidos , Crioprotetores/farmacologia , Células-Tronco Mesenquimais/metabolismo , Proliferação de Células , Células CultivadasRESUMO
Melatonin is a hormone mainly produced by the pineal gland in the absence of light stimuli. The light, in fact, hits the retina, which sends a signal to the suprachiasmatic nucleus, which inhibits the synthesis of the hormone by the epiphysis. Mostly by interacting with MT1/MT2 membrane receptors, melatonin performs various physiological actions, among which are its regulation of the sleep-wake cycle and its control of the immune system. One of its best known functions is its non-enzymatic antioxidant action, which is independent from binding with receptors and occurs by electron donation. The hormone is also an indicator of the photoperiod in seasonally reproducing mammals, which are divided into long-day and short-day breeders according to the time of year in which they are sexually active and fertile. It is known that melatonin acts at the hypothalamic-pituitary-gonadal axis level in many species. In particular, it inhibits the hypothalamic release of GnRH, with a consequent alteration of FSH and LH levels. The present paper mainly aims to review the ovarian effect of melatonin.
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Plastic is an important environmental issue and a more critical aspect concerns plastic fragments, mainly in term of nanoplastics (NPs). We demonstrated that NPs interfere with reproductive and adipose stromal cells. Since several research underlined an increased cardiovascular risk due to NPs, present study was undertaken to investigate their effect on aortic endothelial cells (AOC). We explored the specificity of their interaction with endothelial cells, quantifying their load in treated cells. Then, NPs effect was assessed on cell growth, generation of free radicals and antioxidant defence. Our data demonstrate that NPs colocalize with AOC. We found a significant (p < 0.01) increase both in metabolic activity and Vascular Endothelial Growth Factor (VEGF) production (p < 0.01). Redox status appeared to be disrupted (p < 0.05) by NPs. Taken together, the normal function of cultured AOC appeared negatively affected by AOC. Since NPs have been detected in blood, our present data appear of particular interest.
Assuntos
Células Endoteliais , Fator A de Crescimento do Endotélio Vascular , Fator A de Crescimento do Endotélio Vascular/metabolismo , Microplásticos , Estresse Oxidativo , AortaRESUMO
PFOA is mainly employed in products with water and oil repellent properties. Due to its persistence, bioaccumulation and critical effects on health, its use has been restricted in several countries. This research was intended to explore PFOA action on the main functions of swine ovarian granulosa cells, a valuable model for translational medicine. Moreover, since we previously demonstrated a disruptive effect on free radical generation we sought to explore PFOA effects on the main antioxidant enzymes. PFOA inhibited cell proliferation (p < 0.001), assessed by BrdU uptake. Steroidogenesis was disrupted: PFOA also stimulated 17ß-estradiol production (p < 0.05), increased progesterone production (p < 0.05) at the lowest dose while it displayed an inhibitory effect at higher concentrations (p < 0.05). SOD (p < 0.001), catalase (p < 0.05) and peroxidase (p < 0.01) activities were stimulated. Therefore, our study supports a disruptive effect of PFOA in cultured swine granulosa cells.
Assuntos
Ovário , Progesterona , Feminino , Animais , Suínos , Antioxidantes/farmacologia , Células da Granulosa , Estradiol/farmacologia , Células CultivadasRESUMO
Irisin is a hormone able to reproduce some of the positive effects of physical activity and diet. Recently, we demonstrated the presence of Irisin at the ovarian level as a potential physiological regulator of follicular function. Adipose tissue is crucial for reproductive function through its metabolic activity and the production of adipokines. At present, the exact nature of adipocyte precursors is still under debate, but an important role has been assigned to the population of adipose tissue mesenchymal stromal cells (ASCs) of perivascular origin. It should be noted that, when appropriately stimulated, ASCs can differentiate into preadipocytes and, subsequently, adipocytes. Therefore, this present study was undertaken to explore the potential effect of Irisin on ASCs, known for their high differentiative potential. Since Irisin expression in ASCs was confirmed by PCR, we tested its potential effects on the main functional activities of these cells, including proliferation (BrdU uptake); metabolic activity (ATP production); redox status, evaluated as the generation of free molecules such as superoxide anion and nitric oxide; and scavenger activities, assessed as both enzymatic (superoxide dismutase) and non-enzymatic antioxidant power. Moreover, we tested the effect of Irisin on ASCs adipogenic differentiation. BrdU uptake was significantly (p < 0.001) inhibited by Irisin, while ATP production was significantly (p < 0.05) increased. Both superoxide anion and nitric oxide generation were significantly increased (p < 0.001) by Irisin, while scavenger activity was significantly reduced (p < 0.05). Irisin was found to significantly (p < 0.05) inhibit ASCs adipogenic differentiation. Taken together, the present results suggest a potential local role of Irisin in the regulation of adipose tissue function.
Assuntos
Fibronectinas , Superóxidos , Animais , Suínos , Fibronectinas/metabolismo , Superóxidos/metabolismo , Óxido Nítrico/metabolismo , Bromodesoxiuridina/metabolismo , Tecido Adiposo/metabolismo , Células Estromais , Trifosfato de Adenosina/metabolismoRESUMO
Triclosan is a chlorinated biphenolic with a broad spectrum of antiseptic activities used in cosmetics and hygiene products. Continuous exposure can lead to absorption and bioaccumulation of this substance with harmful health effects. In fact, previous studies have shown that Triclosan acts as an endocrine-disrupting chemical on reproductive organs, with consequent negative effects on reproductive physiology. Therefore, to assess potential adverse impacts on fertility, we tested Triclosan on swine granulosa cells, a model of endocrine reproductive cells. We examined its effects on the main features of granulosa cell functions such as cell growth (BrdU incorporation and ATP production) and steroidogenesis (17-ß estradiol and progesterone secretion). Moreover, since oxidant−antioxidant balance plays a pivotal role in follicular function, redox status markers (superoxide, hydrogen peroxide and nitric oxide production, enzymatic and non-enzymatic scavenging activity) were studied. Our results show that Triclosan significantly inhibits cell growth (p < 0.001), steroidogenesis (p < 0.001), superoxide and nitric oxide production (p < 0.001), while it increases (p < 0.05) enzymatic defense systems. Collectively, these data suggest a disruption of the main granulosa cell functions, i.e., proliferation and hormone production, as well as an imbalance in redox status. On these bases, we can speculate that Triclosan would impair granulosa cell functions, thus exerting negative effects on reproductive function. Further studies are needed to explore lower Triclosan concentrations and to unravel its mechanisms of action at gene level.
RESUMO
Perfluorooctanoic acid (PFOA) is employed in the production and processing of several plastic materials, mainly during the production of waterproof fabrics or nonstick cookware. PFOA is identified as a substance of very high concern, as it is classified as a persistent, bioaccumulative, and toxic (PBT) substance because of its persistence in the environment and its potential accumulation in organisms. Thus, safe levels of exposure cannot be established, and PFOA emissions should be minimized. PFOA has recently been linked to several health concerns in humans. In particular, a disruptive effect on redox status homeostasis has been documented, with a potential impairment of normal reproductive function that requires adequate oxidative balance. Therefore, the aim of the present study was to evaluate the effects of PFOA (2, 20, and 200 ng/mL) on ovarian granulosa cells, a model of reproductive cells. The obtained results reveal that PFOA stimulated cell viability (p < 0.05). Regarding the effects on free radical production, O2−, NO, and H2O2 were significantly inhibited (p < 0.05), while the nonenzymatic antioxidant power was not significantly modified. Collectively, the present results deserve attention since free radical molecules play a crucial role in ovarian follicle development leading to a successful ovulation.
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In addition to the well-known central modulatory role of orexins, we recently demonstrated a peripheral involvement in swine granulosa cells for orexin A and in adipose tissue for orexin B (OXB). The aim of present research was to verify immunolocalization of OXB and its potential role in modulating the main features of swine granulosa cells. In particular, we explored the effects on granulosa cell proliferation (through the incorporation of bromodeoxyuridine), cell metabolic activity (as indirect evaluation by the assessment of ATP), steroidogenic activity (by immunoenzymatic examination) and redox status (evaluating the production of superoxide anion by means of the WST test, production of nitric oxide through the use of the Griess test and the non-enzymatic reducing power by FRAP test). Our data point out that OXB does not modify granulosa cell growth, steroidogenesis and superoxide anion generation. On the contrary, the peptide stimulates (p < 0.05) nitric oxide output and non-enzymatic reducing power. Since new vessel growth is crucial for ovarian follicle development, a further aim of this study was to explore the expression of prepro-orexin and the effects of OXB on swine aortic endothelial cells. We found that the peptide is ineffective in modulating cell growth, while it inhibits redox status parameters. In addition, we demonstrated a stimulatory effect on angiogenesis evaluated in fibrin gel angiogenesis assay. Taken together, OXB appears to be potentially involved in the modulation of redox status in granulosa and endothelial cells and we could argue an involvement of the peptide in the follicular angiogenic events.
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Alzheimer's disease (AD) is the most common cause of dementia in humans and, currently, a valid treatment is lacking. Our goal is to demonstrate the importance and benefits of the relationship with companion animals (considered as co-therapists), intended as a means of facilitating social relations and promoting evident wellbeing in AD patients. The study involved 30 randomly chosen patients with Alzheimer's disease (group T) and three dogs. The group participated in a total of 24 animal-assisted interventions (AAIs) sessions over a span of 12 weeks, using the Mini-Mental State Examination (MMSE), Wellness and Cognitive Ability Questionnaire (Brief Assessment Cognition or BAC), and Alzheimer's Disease Assessment Scale (ADAS) as assessment tests. A second group (group C), consisting of 10 people with AD, was enrolled as control group and underwent the same assessment tests but did not benefit from the presence of the dogs. Tests were carried out at time T0 (before starting sessions), T1 (end of sessions), and T2 (two months after last session). People belonging to group T achieved an overall improvement in their perceived state of wellbeing, even on a cognitive and mnemonic plane. However, two months after the end of the sessions, the test results in people suffering from AD decreased towards the baseline (T0). The study shows how such progress can be achieved through activities based on the relationship with an animal, as long as the animal is a steady presence in the life of the patient receiving the intervention. Dogs involved in other dog-assisted therapies have been found suitable also for assisting patients with AD.
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Single-board computers (SBCs) and microcontroller boards (MCBs) are extensively used nowadays as prototyping platforms to accomplish innovative tasks. Very recently, implementations of these devices for diagnostics applications are rapidly gaining ground for research and educational purposes. Among the available solutions, Raspberry Pi represents one of the most used SBCs. In the present work, two setups based on Raspberry Pi and its CMOS-based camera (a 3D-printed device and an adaptation of a commercial product named We-Lab) were investigated as diagnostic instruments. Different camera elaboration processes were investigated, showing how direct access to the 10-bit raw data acquired from the sensor before downstream imaging processes could be beneficial for photometric applications. The developed solution was successfully applied to the evaluation of the oxidative stress using two commercial kits (d-ROM Fast; PAT). We suggest the analysis of raw data applied to SBC and MCB platforms in order to improve results.
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In the present study, the Eurasian magpie (Pica pica), was evaluated as a possible bioindicator of environmental pollution by heavy metals (HMs). Levels of Ni, Pb, Cd, and Hg in feathers of 64 magpies (31 males and 33 females) were measured by ICP-MS technique. Plasmatic biomarkers of oxidative stress (OS) were also assessed. The birds were captured in the province of Parma (Italy), in different capture sites within 1 km from urban area (UZ), and farther than 5 km from urban area (RZ). Median HM levels were 0.68 mg/kg (0.18-2.27), 2.80 mg/kg (0.41-17.7), Assuntos
Plumas
, Metais Pesados
, Animais
, Monitoramento Ambiental
, Poluição Ambiental/análise
, Plumas/química
, Feminino
, Itália
, Masculino
, Metais Pesados/análise
, Pica
RESUMO
A physiological equilibrium exists between pro- and antioxidant factors. When the oxidant factors exceed the capacity of their removal or inactivation, oxidative stress (OS) occurs. The OS levels were assayed in plasma obtained from 2 bird species. Blood samples were collected from 20 healthy domestic chicken hens, 10 living in an intensive farming environment and 10 free-range, and from 18 healthy Eurasian magpies (Pica pica; 7 females and 11 males, with an estimated age of >1 year of age). For OS biomarker assessment, the determinable reactive oxygen metabolites (d-ROMs) were measured, and the plasmatic antioxidant test (PAT) was performed; the OS index (OSI) was then calculated (d-ROMs/PAT × 1000) as a parameter of overall oxidative stress. Moreover, lipid peroxidation was assessed by measuring plasmatic malondialdehyde (MDA) levels. A hematological evaluation was also performed on each bird with a hemocytometer, on which a blood sample was placed to obtain both a total and differential white blood cell (WBC) count. In hens, OSI and MDA levels were significantly higher (P = .04, and P = .004) in subjects from intensive farming (14.7 ± 7.1 and 27.2 ± 10.4 nmol/mL) than in those bred in rural conditions (5.6 ± 10.3 and 8.2 ± 13.3 nmol/mL). In magpies, a positive correlation between the total WBC count and OS was found, and both d-ROMs and OSI were significantly higher (P = .03) in subjects with a total WBC count greater than the median value (20.4 × 103 cells/µL) with respect to those with a total WBC count less than the median value. The results generated from this study indicate that higher OS levels occurred in hens bred in an intensive indoor farm environment compared with outdoor free-range conditions. Possibly the higher OS levels could be related to the higher stocking density and dust levels found in the indoor facility. Additionally, the correlation between OS biomarker levels in magpies and total WBC count suggests that OS level is influenced by immune response, in agreement with previous studies. Collectively, present data seem to be promising for the application of OS measurement in avian medicine for health and animal welfare monitoring.
Assuntos
Galinhas , Pica , Animais , Antioxidantes , Feminino , Masculino , Malondialdeído , Estresse OxidativoRESUMO
Triclosan is a chlorinated phenolic, used in many personal and home care products for its powerful antimicrobial effect. Several studies have shown triclosan toxicity and the American Food and Drug Administration (FDA) in 2016 has limited its use. It has been recently included in endocrine-disrupting chemicals (EDCs), a list of chemicals known for their ability to interfere with hormonal signaling with particular critical effects on reproduction both in animals and humans. In order to deepen the knowledge in this specific field, the present study was undertaken to explore the effect of different concentrations of triclosan (1, 10, and 50 µM) on cultured luteal cells, isolated from swine ovaries, evaluating effects on growth Bromodeoxyuridine (BrDU) incorporation and Adenosine TriPhosphate (ATP) production, steroidogenesis (progesterone secretion) and redox status (superoxide and nitric oxide production, enzymatic and non-enzymatic scavenging activity). A biphasic effect was exerted by triclosan on P4 production. In fact, the highest concentration inhibited, while the others stimulated P4 production (p < 0.05). Triclosan significantly inhibited cell proliferation, metabolic activity, and enzymatic scavenger activity (p < 0.05). On the contrary, nitric oxide production was significantly increased by triclosan (p < 0.01), while superoxide anion generation and non-enzymatic scavenging activity were unaffected.
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Based on our previous study in follicles, the first aim of this work was to evaluate the effect of melatonin in the swine corpus luteum (CL). Luteal cells were exposed to 10 and 20pg mL-1 melatonin. We evaluated the effect on proliferation (bromo-deoxy-uridine uptake), steroidogenesis (progesterone) and redox status by means of Griess test (nitric oxide production), WST-1 test (superoxide anion generation) and FRAP test (non-enzymatic antioxidant power). The results showed a significant increase in antioxidant power, as well as a reduction in the other parameters analysed. These data and the expression of MT2 observed in luteal cells allow us to hypothesise a physiological role of melatonin in the regulation of CL functionality. The reproductive function is dependent on energy reserves stored in adipose tissue. Therefore, we sought to verify the effect of melatonin on adipose stromal cells (ASCs). MT2 receptor expression was detected in ASCs and the presence of gene markers (PPARγ and leptin) before and after adipogenic differentiation was verified. The differentiation was significantly inhibited by melatonin, as well as cell viability. In conclusion, present results suggest that melatonin exerts a potential inhibitory action on luteal function and adipogenesis, possibly mediated by MT2.
Assuntos
Tecido Adiposo/efeitos dos fármacos , Corpo Lúteo/efeitos dos fármacos , Melatonina/farmacologia , Células Estromais/efeitos dos fármacos , Adipogenia/efeitos dos fármacos , Tecido Adiposo/citologia , Tecido Adiposo/metabolismo , Animais , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Corpo Lúteo/citologia , Corpo Lúteo/metabolismo , Feminino , Leptina/metabolismo , Óxido Nítrico/metabolismo , Oxirredução , PPAR gama/metabolismo , Progesterona/biossíntese , Receptor MT2 de Melatonina/agonistas , Receptor MT2 de Melatonina/genética , Receptor MT2 de Melatonina/metabolismo , Células Estromais/metabolismo , Sus scrofaRESUMO
The World Health Organization defined leishmaniasis as one of the priority attention diseases. Aiming to clarify some aspects of its pathogenetic mechanisms, our study focused on the assessment of redox status in dogs, the main reservoir for Leishmania infantum. Forty-five dogs from an endemic area in southern Italy were divided into four different groups (from mild disease with negative to low positive antibody levels to very severe disease with medium to high positive antibody levels) according to the LeishVet group guidelines. Their plasma and/or sera were tested for reactive oxygen species (ROS), namely the superoxide anion (O2-), reactive nitrogen species (RNS), such as nitric oxide (NO) and hydroperoxides (ROOH), as well as activity of the detoxifying enzyme superoxide dismutase (SOD), and total nonenzymatic antioxidant capacity, as determined by the ferric reducing-antioxidant power (FRAP) assay. O2- generation was significantly (p < 0.05) reduced in leishmaniasis-affected dogs independently of the clinical stage, while NO production was stimulated (p < 0.05) only in II and III stage patients. No difference could be found for the levels of hydroperoxides and SOD activity between healthy and pathological subjects. FRAP values were lower in affected dogs but only in stage II. Taken together, although we demonstrated that several redox status parameters are altered in the plasma of dog affected by leishmaniasis, the oxidative stress changes that are observed in this disease, are possibly mainly due to cellular blood components i.e., neutrophils responsible for the elimination of the parasite. Further studies are required to assess the clinical values of the collected data.
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Mesenchymal stem cells (MSCs) have been recently introduced in veterinary medicine as a potential therapeutic tool for several pathologies. The large-scale in vitro expansion needed to ensure the preparation of a suitable number of MSCs for clinical application usually requires the use of xenogeneic supplements like the fetal bovine serum (FBS). The substitution of FBS with species-specific supplements would improve the safety of implanted cells, reducing the risk of undesired immune responses following cell therapy. We have evaluated the effectiveness of canine adipose tissue-derived stromal vascular fraction (SVF) and MSCs (ADMSCs) expansion in the presence of canine blood-derived supplements. Cells were cultured on traditional plastic surface and inside a 3D environment derived from the jellification of different blood-derived products, i.e., platelet-poor plasma (PPP), platelet-rich plasma (PRP), or platelet lysate (PL). PPP, PRP, and PL can contribute to canine ADMSCs in vitro expansion. Both allogeneic and autologous PPP and PL can replace FBS for ADMSCs culture on a plastic surface, exhibiting either a similar (PPP) or a more effective (PL) stimulus to cell replication. Furthermore, the 3D environment based on homospecific blood-derived products polymerization provides a strong stimulus to ADMSCs replication, producing a higher number of cells in comparison to the plastic surface environment. Allogeneic or autologous blood products behave similarly. The work suggests that canine ADMSCs can be expanded in the absence of xenogeneic supplements, thus increasing the safety of cellular preparations. Furthermore, the 3D fibrin-based matrices could represent a simple, readily available environments for effective in vitro expansion of ADMSCs using allogeneic or autologous blood-products.
Assuntos
Tecido Adiposo/metabolismo , Meios de Cultura/metabolismo , Fibrina/metabolismo , Células-Tronco Mesenquimais/metabolismo , Plásticos/metabolismo , Xenobióticos/farmacologia , Tecido Adiposo/efeitos dos fármacos , Animais , Plaquetas/efeitos dos fármacos , Plaquetas/metabolismo , Técnicas de Cultura de Células/métodos , Cães , Células-Tronco Mesenquimais/efeitos dos fármacos , Plasma Rico em Plaquetas/efeitos dos fármacos , Plasma Rico em Plaquetas/metabolismo , Soro/metabolismoRESUMO
Recent studies have demonstrated the surprising ability of reproductive endocrine cells to express receptors of innate immunity useful to sense danger in order to avoid disruption of tissue homeostasis. Present research demonstrates the presence of pattern recognition receptors, i.e. toll like receptors (TLR) TLR2, TLR4 and TLR 5 and NOD like receptors (NLR) NOD1 and NOD2 in swine granulosa cells from ovarian follicles> 5â¯mm. Therefore, our second goal was to expose granulosa cells to different concentrations (1000, 100 and 10â¯ng/ml) of lipopolysaccharide (LPS) and N-Palmitoyl-S-[2,3-bis(palmitoyloxy)-(2RS)- propyl]-[R]-cysteinyl-[S]-seryl-[S]-lysyl-[S]-lysyl-[S]-lysyl-[S]- lysine (Pam3CSK4), two substances associated with pathogen molecular patterns. Their potential effects on the main functional parameters were monitored: proliferation (through the incorporation of Bromo-deoxy-Uridine), cell viability (by testing the metabolization of MTT salt), steroidogenic activity (by immunoenzymatic examination) and redox status (evaluating the production of superoxide anion by means of the WST test, production of nitric oxide through the use of the Griess test, and the non-enzymatic reducing power, by FRAP test). The data collected show a significant inhibition (pâ¯<â¯0.01) of cell proliferation after treatment with both LPS and Pam3CSK4, while cell viability has not been modified. As for steroidogenesis, treatment with both LPS and Pam3CSK4 significantly inhibited (pâ¯<â¯0.05) the production of 17ß-estradiol and progesterone. LPS and Pam3CSK4 stimulated (pâ¯<â¯0.05) the production of superoxide anion and nitric oxide, while inhibited (pâ¯<â¯0.05) the antioxidant power. In conclusion, the study shows that the functionality of granulosa cells is compromised by the exposure to molecular profiles associated with pathogens; the knowledge gathered could lay the theoretical basis for the definition of therapeutic treatments related to diseases that can affect normal reproductive processes.
Assuntos
Células da Granulosa/fisiologia , Proteínas NLR/fisiologia , Suínos , Receptores Toll-Like/fisiologia , Animais , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Lipopolissacarídeos/toxicidade , Óxido Nítrico , Oligopeptídeos , Esteroides/metabolismo , Superóxidos , Receptor 2 Toll-Like/agonistas , Receptor Toll-Like 9/agonistasRESUMO
The extract of the seeds from Indian celery, Apium greaveolens (CSE), tested in experimental animals (rodents), and in humans affected by chronic osteoarthritic diseases, exhibits anti-inflammatory effects that can be compared, to some degree, to those of non-steroid anti-inflammatory drugs (NSAID). In view of a potential use of CSE in the equine species, it was tested on horses affected by chronic articular pathologies. The trial was performed on 20 horses divided into three different groups, orally treated with 0 (controls), 7.0 or 30 g of CSE BID. Basic orthopedic examinations were conducted, vital signs were observed, and blood samples collected. Improvement was observed at the highest dosage tested (30 g of CSE BID), as reflected in the score values of three clinical parameters, (i) amplitude and (ii) sensitivity to passive flexion and (iii) flexion test. Since the improvement of these parameters can be correlated with a lower perception of the pain, the present data suggest that the CSE treatment can have an analgesic effect in horses affected by chronic osteoarthritic diseases.