Pluronic F127 composite hydrogel for the repair of contraction suppressed full-thickness skin wounds in a rabbit model.

Hydrogels are commonly used as carriers for cell delivery due to their similarities to the extracellular matrix. A contraction-suppressed full-thickness wound model was used to evaluate the therapeutic potential of Pluronic F127 (PF127) hydrogel loaded with adipose-derived stromal vascular fraction (AdSVF), mesenchymal stem cells (AdMSC), and conditioned media (AdMSC-CM) for the repair of wounds in a rabbit model. The experimental study was conducted on forty-eight healthy adult New Zealand white rabbits randomly divided into eight groups with six animals each and treated with AdSVF, AdMSC, and AdMSC-CM as an injectable or topical preparation. The healing potential of different adipose-derived cell-based and cell-free therapeutics was evaluated based on percentage wound healing, period of epithelialization, epidermal thickness, scar evaluation, histopathology analysis, histochemical evaluation, immunohistochemistry (collagen type I), and hydroxyproline assay by comparing with the positive and negative control. Collagen density analysis using different staining methods, immunohistochemistry, and hydroxyproline assay consistently showed that delivering AdMSC and AdMSC-CM in PF127 hydrogel enhanced epithelialization, collagen production, and organization, contributing to improved tissue strength and quality. Even though allogeneic AdSVF was found to promote wound healing in rabbits, it has a lower potential than AdMSC and AdMSC-CM. The wound healing potential of AdMSC and AdMSC-CM was enhanced when loaded in PF127 hydrogel and applied topically. Even though wounds treated with AdMSC outperformed AdMSC-CM, a significant difference in the healing quality was not observed in most instances, indicating almost similar therapeutic potential. The findings indicate that the wound healing potential of AdMSC and AdMSC-CM was enhanced when loaded in PF127 hydrogel and applied topically. These treatments promoted collagen production, tissue organization, and epidermal regeneration, ultimately improving overall healing outcomes.

Chitosan-TPP encapsulated quercetin nanoparticles: amplified protection mechanisms unveiled against Ethion-induced developmental toxicity through comprehensive in-vivo and in-silico elucidation.

Aim: The study investigated Ethion-induced developmental toxicity in Wistar albino rats and the potential ameliorative effects of quercetin and nano-quercetin co-administration. Further, In-silico docking of Ethion and quercetin with MCL-1 was conducted. Methodology: Quercetin nanoparticles were synthesized by ionic-gelation method. The encapsulated quercetin nanoparticles were characterized for Zeta size, UV-Vis spectroscopy, encapsulation efficiency, and TEM studies. Male rats were administered Ethion (high/low dose), quercetin, and nano-quercetin alone or in combination for 60 days. Female rats were introduced for mating on the 61st day, and pregnant females were observed for 20 gestational days. On GD 20, rats were sacrificed and evaluated for body/organ weight, reproductive indices, fetal morphology, skeletal, and visceral deformities.In silico binding energies of ethion and quercetin with MCL-1 were determined. Results: Nanoparticle size was 363.2 ± 1.23 nm on day 0 and 385.63 ± 1.53 nm on day 60, with PDI of 0.247 and charge of 22.9 mV. Absorbance maxima were at 374 nm, with encapsulation efficacy of 85.16 ± 0.33%. EHD male crossed females showed decreased body/organ weights, reduced fertility, hematoma, cleft palate, tail curling, and absence of extremity. Nano-quercetin co-administration normalized parameters comparable to controls. Both Ethion and quercetin interacted with MCL-1, with quercetin exhibiting stronger binding energy. Conclusion: Nano-quercetin demonstrated stronger antioxidant properties than quercetin, counteracting ethion-induced maternal/fetal abnormalities.

Fipronil-induced genotoxicity and DNA damage in vivo: Protective effect of vitamin E.

Fipronil, an insecticide of the phenylpyrazole class has been classified as a carcinogen by United States Environmental Protection Agency, yet very limited information is available about its genotoxic effects. Adult male and female animals were gavaged with various doses of fipronil (2.5, 12.5, and 25 mg/kg body weight (bw)) to evaluate micronucleus test (mice), chromosome aberration (CA), and comet assay (rats), respectively. Cyclophosphamide (40 mg/kg bw; intraperitoneal) was used as positive control. Another group of animals were pretreated with vitamin E orally (400 mg/kg bw) for 5 days prior to administration of fipronil (12.5 mg/kg). Fipronil exposure in both male and female mice caused significant increase in the frequency of micronuclei (MN) in polychromatic erythrocytes. Similarly, structural CAs in bone marrow cells and DNA damage in the lymphocytes was found to be significantly higher in the male and female rats exposed to fipronil as compared to their respective controls. The average degree of protection (male and female animals combined together) shown by pretreatment of vitamin E against fipronil-induced genotoxicity was 63.28%: CAs; 47.91%: MN formation; and 74.70%: DNA damage. Findings of this study demonstrate genotoxic nature of fipronil regardless of gender effect and documents protective role of vitamin E.

Fipronil induced oxidative stress involves alterations in SOD1 and catalase gene expression in male mice liver: Protection by vitamins E and C.

In the present investigation, hepatic oxidative stress induced by fipronil was evaluated in male mice. We also investigated whether pretreatment with antioxidant vitamins E and C could protect mice against these effects. Several studies conducted in cell lines have shown fipronil as a potent oxidant; however, no information is available regarding its oxidative stress inducing potential in an animal model. Out of 8 mice groups, fipronil was administered to three groups at low, medium, and high dose based on its oral LD50 (2.5, 5, and 10 mg/kg). All three doses of fipronil caused a significant increase in the serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) level with concomitant increase in the absolute and relative weight of liver. High dose of fipronil caused significant down-regulation in the hepatic mRNA expression of superoxide dismutase 1 (SOD1) and catalase (0.412 ± 0.01 and 0.376 ± 0.05-fold, respectively) as well as an increase in the lipid peroxidation (LPO). Also, decrease in the activity of antioxidant enzymes; SOD, catalase, and glutathione-S-transferase (GST) and the content of nonantioxidant enzymes; glutathione and total thiol were recorded. Histopathological examination of liver revealed dose dependant changes such as severe fatty degeneration and vacuolation leading to hepatocellular necrosis. Prior administration of vitamin E or vitamin C against fipronil high dose caused decrease in lipid peroxidation and increased activity of antioxidant enzymes. Severe reduction observed in functional activities of antioxidant enzymes was aptly substantiated by down-regulation seen in their relative mRNA expression. Thus results of the present study imply that liver is an important target organ for fipronil and similar to in vitro reports, it induces oxidative stress in the mice liver, which in turn could be responsible for its hepatotoxic nature. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1147-1158, 2016.

Firpronil induced spermotoxicity is associated with oxidative stress, DNA damage and apoptosis in male rats.

The present study is the first to investigate and characterize the fipronil (FPN) induced spermotoxicity in male rats. Male rats were orally given FPN (2.5, 5.0 and 10 mg/kg/day) for 4 weeks. Epididymal sperms were collected and remaining testis was processed for histopathological evaluation. FPN treatment significantly reduced sperm density, motility, viability and per cent intact acrosome along with concomitant increase in spermatozoa abnormalities. Exposure of FPN caused excessive ROS generation, lipid peroxidation and alteration in mitochondrial membrane potential leading to apoptosis of spermatozoa in dose dependent manner. Higher FPN doses (5 and 10 mg/kg) markedly reduced the DNA integrity of spermatozoa. These data suggest that FPN causes male reproductive toxicity through oxidative stress induced DNA damage and apoptosis of spermatozoa.

Fipronil induced oxidative stress in kidney and brain of mice: protective effect of vitamin E and vitamin C.

Fipronil is a relatively new insecticide of the phenpyrazole group. Fipronil-induced effects on antioxidant system and oxidative stress biomarkers are yet to be studied in vivo. The present study was undertaken to evaluate fipronil-induced alterations in the blood biochemical markers and tissue antioxidant enzymes after oral exposure in mice and to explore possible protective effect of pre-treatment of antioxidant vitamins against these alterations. Mice were divided into eight groups containing control, test and amelioration groups. Mice in the test groups were exposed to different doses of fipronil, i.e., 2.5, 5 and 10 mg/kg bw, respectively for 28 days. Mice in the amelioration groups were treated with vitamin E or vitamin C (each at 100 mg/kg) 2 h prior to high dose (10 mg/kg) of fipronil. Fipronil exposure at three doses caused significant increase in the blood biochemical markers, lipid peroxidation and prominent histopathological alterations; while level of antioxidant enzymes was severely decreased both in kidney and brain tissues. Prior administration of vitamin E or vitamin C in the fipronil exposed mice led to decrease in lipid peroxidation and significant increase in activities of antioxidants, viz., glutathione, total thiol, superoxide dismutase and catalase. Vitamin E and vitamin C administration in fipronil exposed mice also improved histological architecture of the kidney and brain when compared with fipronil alone treated groups. Thus, results of the present study demonstrated that in vivo fipronil exposure induces oxidative stress and pre-treatment with vitamin E or C can protect mice against this oxidative insult.

Toxicopathological alterations induced by high dose dietary T-2 mycotoxin and its residue detection in Wistar rats.

T-2 toxin is one of the most potent cytotoxic and food-borne mycotoxins. Most experimental studies on the T-2 toxin have been performed at extremely low doses (ppb level). However, several field reports of contaminated feed have shown concentration of T-2 toxin to be as high as ≥20 ppm. Therefore, the impact of high dose T-2 toxin (20 ppm) after subacute exposure was investigated in an experimental setup with respect to growth performance, oxidative stress, and detailed pathomorphology in young male Wistar rats. Furthermore, to see the effect of such a high dose on the accumulation of T-2 toxin, its residues in various organs were quantified by high-performance thin-layer chromatography (HPTLC). Apart from obvious clinical toxicosis, rats in the toxin-fed group showed significant hemato-biochemical alterations and increased levels of biological markers of oxidative stress with concomitant decrease in levels of serum and tissue catalase and superoxide dismutase. These alterations were strongly supported by histopathological changes, such as hyperkeratosis and hyperplasia of the squamous gastric mucosa, oxidative damage to hepatocytes, atrophy of the thymus and spleen, and overall decrease in the spermatogenic activity of testes. An economical, simple, reliable, and quick method for the detection and quantification of T-2 toxin residues by HPTLC is also reported here. No residual T-2 toxin was detected in any of the organs tested, suggesting that T-2 toxin does not accumulate in tissues even at such a high exposure level.

Studies on apoptotic changes in combined toxicity of citrinin and endosulfan in pregnant wistar rats and their fetuses.

BACKGROUND: Citrinin (mycotoxin) and endosulfan (pesticide) both environmental contaminants easily enter the food chain and are caoomon causes of various toxicities. MATERIALS AND METHODS: In the present investigation, citrinin (CIT) (10 mg/kg feed) and endosulfan (1 mg/kg body weight) were administered orally alone and in combination to pregnant Wistar rats from gestational day 6 to 20 to study their effect to cause apoptosis in the pregnant Wistar rats and their fetuses. Apoptosis was assessed in dams by agarose gel electrophoresis, flow cytometry and electron microscopy, while in the fetuses it was assessed by flow cytometry only. RESULT: Citrinin and endosulfan in the combination group caused apoptosis in an additive manner as there was increased number of apoptotic cells as compared to the individual toxin and control groups. The fetuses also showed increased number of apoptotic cells in the combination groups, which also indicated that both the toxins crossed the placental barrier. CONCLUSION: So it was concluded that apoptosis played a significant role in the pathogenesis of endosulfan and citrinin toxicity.

Curcumin protects against cypermethrin-induced genotoxicity in rats.

Cypermethrin is a synthetic pyrethroid insecticide used worldwide in agriculture, home pest control, protection of foodstuff and disease vector control. The aim of the present study was to investigate the protective effect of curcumin on cypermethrin-induced genotoxicity in rats. Administration of cypermethrin (25mg/kg, p.o.) for 28 days resulted in significant increase in the frequency of micronuclei formation in bone marrow cells and DNA damage in blood cells. Curcumin (100mg/kg, p.o.) administration caused significant reduction in micronuclei formation and, marked reduction in DNA damage. The present study revealed that presence of curcumin could diminish cypermethrin-induced genotoxicity in rats.

Comparative plasma pharmacokinetics of meloxicam in sheep and goats following intravenous administration.

Meloxicam, a novel cyclooxygenase-2 selective nonsteroidal anti-inflammatory drug (NSAID), has been used extensively in humans and recently in some domestic animal species. Although it is an attractive NSAID for use in small ruminants, meloxicam pharmacokinetics have not been investigated in sheep and goats and this information is essential for rational therapeutic use of the drug in these species. In this investigation, comparative pharmacokinetic properties of meloxicam were studied in sheep and goats after a single intravenous dose of 0.5 mg kg(-1) body mass. Blood samples were collected via jugular venepuncture into heparinised tubes at predetermined times after drug administration. Plasma concentrations of meloxicam were determined by reversed-phase high performance liquid chromatography. The plasma concentrations of meloxicam were detectable in sheep and goats up to 72 and 48 h, respectively. The plasma concentration versus time data of meloxicam in both sheep and goats were adequately described by a two-compartment open model. The values obtained for sheep and goats for distribution half-life, volume of distribution at steady state and volume of the central compartment were almost similar in sheep and goats. The elimination half-life (t(1/2beta)), area under the plasma concentration-time curve (AUC), mean residence time (MRT) and total systemic clearance (Cl(B)) in sheep were significantly different from those of goats. The mean+/-S.E. values of t(1/2beta), MRT, AUC and Cl(B) in sheep were 10.85+/-1.21 h, 15.13+/-1.67 h, 31.88+/-2.97 microg h mL(-1) and 0.016+/-0.002 L h(-1) kg(-1), respectively whereas the respective values in goats were 6.73+/-0.58 h, 9.37+/-0.83 h, 19.23+/-2.23 microg h mL(-1) and 0.03+/-0.01 L h(-1) kg(-1). The results indicate that elimination kinetics of meloxicam differ significantly between sheep and goats and the elimination of the drug tends to be faster in goats compared to sheep.

Teratogenic effects in rabbits of simultaneous exposure to ochratoxin A and aflatoxin B1 with special reference to microscopic effects.

To elucidate the teratogenic effects, ochratoxin A (OTA) and aflatoxin B1 (AFB1) were dissolved in corn oil and administered in combination to New Zealand White rabbits during 6-18 days of gestation orally with the dose levels of OTA+AFB1, 0.05+0.05 and 0.1+0.1mg/kg body weight. To assess pathomorphological features of the anomalies, the fetal serial sections were histologically examined. There was no mortality in any of the treated groups. Body weights and body weight gains of dams in the combined treatment groups were comparable with those of controls and individual treatments. The mean crown to rump lengths in both the combination dose groups and mean fetal weights in high dose combination group were significantly decreased. In the high dose combination, there was increase in the percent of implants resorbed and significant increase in the incidence of visceral anomalies. The combination treatment resulted in various gross, skeletal and visceral anomalies such as wrist drop, scoliosis, bent metacarpals, rudimentary ribs, cardiac defects and microphthalmia. There was a dose-related increase in the percent of litters showing the histopathological changes in the fetal tissues. The incidence of histopathological changes in the tissue sections prepared from fetal liver, kidneys, brain, heart and eyes was found increased in the high dose combination group. The comparative evaluation of the results of combination versus individual treatments revealed that certain anomalies observed in the individual treatment of OTA such as knuckling of fetlock, rudimentary tail or agenesis of tail, wavy ribs, hydrocephalus and agenesis of kidney and AFB1 as enlarged eye sockets and enlarged liver were absent in the combination treatment. However, some new manifestations such as cardiac defects and scoliosis were seen. The results of the present study indicated that in combination, OTA and AFB1 have antagonistic interaction. The presence of subtle lesions histologically due to an interference with normal development suggested that microscopic examination of the fetal tissues could provide additional, useful information to a developmental toxicity study.

Effects of aflatoxin B1 on embryo fetal development in rabbits.

Aflatoxin B1 (AFB1), is a food borne mycotoxin produced by fungal species of the genera Aspergillus. To elucidate the teratogenic effects, AFB1 was dissolved in corn oil and given orally to New Zealand White rabbits during 6-18 days of gestation with the dose levels of 0.025, 0.05 and 0.1 mg/kg body weight. To assess pathomorphological features of the anomalies induced by AFB1, the fetal serial sections were histologically examined. There was no maternal mortality in any group. There was non-significant decrease in percent of live fetuses and increase in the percent resorptions and post-implantation losses at 0.1 mg/kg dose group as compared with those of controls. The mean crown to rump lengths of 0.05 and 0.1 mg/kg dose groups were significantly reduced than that of the control. The mean fetal weights were significantly reduced in 0.1 mg/kg dose group than that of other treated groups. The gross anomalies observed included wrist drop and enlarged eye socket whereas, skeletal anomalies were agenesis of caudal vertebrae, incomplete ossification of skull bones and bent metacarpals. The visceral anomalies of microphthalmia and cardiac defects were seen at 0.1 mg/kg dose group. The characteristic histological findings of fetal tissues were distortion of normal hepatic cord pattern and reduced megakaryocytes in liver, fusion of auriculo-ventricular valves, mild degenerative changes in myocardial fibers, microphthalmic eyes and lenticular degeneration. The results of this study indicated that AFB1 was found to be teratogenic in rabbits when given by oral route during gestation days 6-18 and the dose of 0.1 mg/kg could be considered as the minimum oral teratogenic dose. The histological examination of the fetal tissues indicated its importance in identifying the visceral anomalies which were otherwise not visible.

Influence of hypothyroid state on 45Ca(2+) influx and sensitivity of rat uterus to nifedipine and diltiazem.

The influence of methimazole-induced hypothyroidism on spontaneous rhythmic contractions and Ca2+ channel function of rat uterus was examined. Hypothyroidism significantly reduced the amplitude and frequency of spontaneous rhythmic contractions. Nifedipine (10(-12)-10(-6) M) and diltiazem (10(-9)-10(-4) M) caused concentration-related inhibition of the myogenic responses of the oestrogenised rat uterus obtained from both eu- and hypothyroid rats. However, nifedipine was less potent (IC(50); 5.4 x 10(-9) M; n=6) in hypothyroid rat uterus as compared to euthyroid controls (IC(50): 8.13 x 10(-12) M; n=9) to inhibit the rhythmic contractions. Similarly, diltiazem was less potent (IC(50): 4.57 x 10(-6) M; n=9) to inhibit the uterine spontaneous contractions in hypothyroid than in euthyroid rat uterus (IC(50): 6.4 x 10(-8) M; n=6). A similar decrease in the sensitivity to nifedipine and diltiazem for reversal of K+ (100 mM)-induced tonic contraction was observed in uterus obtained from hypothyroid rats compared to the controls. Both nifedipine and diltiazem were less potent for causing concentration-related inhibition of K+-stimulated 45Ca2+ influx in uterine strips taken from the hypothyroid rats. Thus, the IC(50) values of nifedipine (1.83 x 10(-8) M; n=12) and diltiazem (1.8 x 10(-6) M; n=9) were significantly greater in tissues obtained from hypothyroid rats compared to the controls (IC(50) of nifedipine, 1.15 x 10(-11) M; n=12, diltiazem, 8.1 x 10(-8) M; n=8). Nifedipine-sensitive influx of 45Ca2+ - stimulated either by K+ (100 mM) or Bay k8644 (1,4-dihydro-2,6-dimethyl-5-nitro-4-[2'-(trifluromethyl)phenyl]-3-pyridine carboxylic acid methyl ester) (10(-8) M) was significantly less in uterine strips from hypothyroid rats compared to the controls. The results of the present study suggest that the inhibition of uterine rhythmic contractions may be attributable to a reduction in rat myometrial Ca2+ channel function in the hypothyroid state.

Anti-inflammatory activity of Dalbergia sissoo leaves.

The possible anti-inflammatory activity of the 90% ethanolic extract of Dalbergia sissoo leaves (DSELE) was studied in different models of inflammation in rats after oral administration at doses of 100, 300 and 1000 mg/kg. DSELE significantly inhibited carrageenin, kaolin and nystatin-induced paw oedema, as well as the weight of granuloma induced by a cotton pellet. It also inhibited dye leakage in acetic acid-induced vascular permeability test in mice. DSELE was devoid of ulcerogenic effect on the gastric mucosa of rats in acute and chronic tests. In acute toxicity studies, it was found to be safe up to 10.125 g/kg, p.o. in the rat. It was concluded that the D. sissoo leaf extract possessed significant anti-inflammatory activity (in acute, sub-acute and chronic models of inflammation) without any side effect on gastric mucosa.

Modulation of vascular KATP channels in hypothyroidism.

The role of vascular KATP channels in hypothyroidism-induced decrease in myogenic activity of rat portal vein was examined by using pharmacologically relevant concentrations of K+ channel ligands. As compared to controls, a significant decrease in the myogenic tone and noradrenaline (10(-9)-10(-5) M)-induced contractions was observed in portal veins from hypothyroid rats. In both euthyroid and hypothyroid states, pinacidil (10(-9)-10(-5) M) and cromakalim (10(-9)-10(-5) M) caused concentration-related inhibition of the myogenic tone (frequency and amplitude). However, hypothyroidism caused a leftward shift in the concentration-response curves of the K+ channel openers with a corresponding decrease in their IC50 values both in the absence and presence of the KATP channel blocker, glibenclamide (10(-7) M). Further, concentration-dependent increase in the frequency of myogenic tone by glibenclamide (10(-8)-3 x 10(-6) M) was greater in tissues from hypothyroid rats (EC50 = 2.07 x 10(-7) M; 95% CL, 1.06-4.05 x 10(-7) M) in comparison to controls (EC50 = 8.07 x 10(-7) M; 95% CL, 0.53-1.22 x 10(-6) M). These results suggest that a decrease in the myogenic tone of rat portal vein may possibly be related to an enhanced opening of the KATP channels in hypothyroidism.