scholarly journals 159 Laparoscopic insemination method in sheep allows the use of an animal protein-free and inexpensive freezing medium

2020 ◽  
Vol 32 (2) ◽  
pp. 206
Author(s):  
L. Gavin-Plagne ◽  
L. Boyer ◽  
A. Baudot ◽  
M. Guedes Teixeira ◽  
G. Louis ◽  
...  
Keyword(s):  
Pregnancy Rate ◽  
Propidium Iodide ◽  
Random Effect ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Positive Control ◽  
Negative Control ◽  
Motion Parameter ◽  
Control Medium ◽  
Cooling Rates

Animal-derived products are widely used in sperm cryopreservation for their cryoprotective properties. These components, however, must be replaced because of sanitary risks. STEMALPHA.CRYO3 (Ref. 5617, Stem Alpha), called CRYO3, is a chemically defined preservation medium currently used for freezing human tissue and adult stem cells. The aim of this study was to evaluate the effects of a CRYO3-based medium and of two cooling rates on invitro parameters and invivo fertility of ram sperm. Six rams (Blanche du Massif Central) were subjected to sperm collection four times using an artificial vagina. Sperm were split and frozen in three media: an egg yolk and milk-based medium (positive control), a CRYO3-based medium (tested medium), and a medium without additives (negative control). The two cooling rates were related to the distance between the straws and the surface of liquid nitrogen during the freezing process (5 and 20cm). Sperm membrane integrity (propidium iodide/SYBR-14), acrosome integrity (fluorescein isothiocyanate-peanut agglutinin/propidium iodide; FITC-PNA/PI), and mitochondrial membrane potential (JC-1) were assessed using flow cytometry, whereas functional membrane integrity was assessed using a hypo-osmotic swelling test and motion characteristics were evaluated using computer-assisted sperm analysis. Pregnancy rate, parturition rate, and prolificacy were evaluated after performing laparoscopic inseminations (n=75 ewes). Moreover, we characterised the freezing media thermodynamically using a differential scanning calorimeter. Statistical analyses were performed using R software. Invitro parameters were assessed using a mixed model including the time and the medium as fixed effects and the ram as a random effect. Pregnancy and parturition rates, following a binomial distribution, and prolificacy, assumed to follow a Poisson distribution, were analysed using generalised linear models, including the medium as a fixed effect and the ram as a random effect. Differences with P<0.05 were considered statistically significant. The cooling rates had no significant effect except on the wobble motion parameter. The positive control medium showed significantly higher results than the CRYO3-based medium and the negative control medium for all invitro parameters except for straightness motion parameter. Conversely, field trials showed no significant difference between the media for pregnancy rate (71, 64, and 74%), parturition rate (68, 61, and 74%) and prolificacy (2.0, 2.1, and 1.7), for the positive control, CRYO3-based medium, and the negative control, respectively. This study showed that the product, CRYO3, cannot replace egg yolk and milk in freezing extenders. Moreover, we showed that laparoscopic inseminations allowed a 74% parturition rate due to an easy and inexpensive medium comprising only a Tris buffer and glycerol. Although it could not be used on a large scale, this medium remains an option for international transport or long-term storage of genetic diversity.

130 EFFECTIVENESS OF BUTYLATED HYDROXYTOLUENE AS EGG YOLK SUBSTITUTE FOR CRYOPRESERVATION OF BOAR SPERMATOZOA

2007 ◽  
Vol 19 (1) ◽  
pp. 182 ◽  
Author(s):  
L. Rodriguez-Vilar ◽  
M. Hernandez ◽  
C. Lopez-Sanchez ◽  
J. M. Vazquez ◽  
E. A. Martinez ◽  
...  
Keyword(s):  
Protective Effect ◽  
Mixed Model ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Positive Control ◽  
Negative Control ◽  
Butylated Hydroxytoluene ◽  
Main Effects ◽  
Boar Spermatozoa

Butylated hydroxytoluene (BHT) has proven to be efficient as a supplement for cryopreservation boar spermatozoa (Roca et al. 2004 J. Androl. 25, 397–405). Moreover, it has been successfully used as an egg yolk substitute to cryopreserve goat spermatozoa (Khalifa and El-Saidy 2006 Anim. Reprod. Sci. 93, 303–315). The objective of this study was to evaluate the effectiveness of BHT as an egg yolk substitute for freezing boar spermatozoa. Nine sperm-rich ejaculate fractions were collected from 3 boars (3 ejaculates per boar) using the gloved-hand method. After centrifugation (2400g for 3 min), the sperm pellet of each ejaculate was split into 5 aliquots. The aliquots were diluted (to a final concentration of 1 � 109 sperm/mL) in a Tris-citric-glucose extender with 3% glycerol and supplemented with 20% egg yolk (positive control, PC aliquot) or BHT at the final concentrations of 0 (negative control, NC aliquot), 0.2, 0.4, and 0.8 mM. Diluted semen samples were dispensed into 0.5-mL straws, and frozen in a programmable cell freezer at 20�C min. Thawing was carried out in a water bath at 70�C for 8 s. Post-thaw sperm survival was assessed according to total sperm motility (TSM, %) using a CASA system (SCA�; Microptic, Barcelona, Spain), and plasma membrane integrity (PMI, %) and acrosome membrane integrity (AMI, %) using a flow cytometric procedure (SYBR-14/propidioum iodide/FITC-phycoerythrin), at 30 and 150 min post-thawing in diluted Beltsville thawing solution with spermatozoa held in a waterbath at 37�C (3 straws per ejaculate). Data were analyzed using a ANOVA mixed model including the main effects of aliquot, boar, post-thaw assessment time, and their interactions, with ejaculate and straw as random effects. All main effects had significant influence (P ≤ 0.01) in all post-thaw sperm assessments. However, no interactions (P ≥ 0.05) among main effects were shown. Data were combined for the 2 post-thaw assessment times. The best (P ≤ 0.05) post-thaw sperm quality (mean � SEM) was achieved in PC aliquots (47.11 � 3.10, 58.98 � 2.78, and 51.35 � 3.42 for TSM, PMI, and AMI, respectively). In NC aliquots, the percentage of TSM, PMI, and AMI were always below 1% (P ≤ 0.05). BHT has a beneficial (P ≤ 0.05) effect on post-thaw sperm assessments, and no differences (P ≥ 0.05) among concentrations were shown. The mean post-thaw sperm quality in the BHT aliquots was 8.50 � 0.80, 20.29 � 0.53, and 16.03 � 0.55 for TSM, PMI, and AMI, respectively. On the basis of these data, we can conclude that BHT has a protective effect for boar spermatozoa during the cryopreservation process. However, BHT alone is insufficient to replace the protective effect of egg yolk. This work was supported by CICYT (AGF2005-00706), Madrid, Spain.

scholarly journals 117 Functionality of a novel consensus bacterial 6-phytase variant in enhancing phosphorus digestibility in gestating and lactating sows

2020 ◽  
Vol 98 (Supplement_4) ◽  
pp. 109-109
Author(s):  
Deepak Velayudhan ◽  
Xandra Benthem de Grave ◽  
Katie Waller ◽  
Leon Marchal ◽  
Yueming Dersjant-Li
Keyword(s):  
Soybean Meal ◽  
Random Effect ◽  
Positive Control ◽  
Negative Control ◽  
Combined Effect ◽  
Restricted Feeding ◽  
Sunflower Meal ◽  
Tukey Test ◽  
Lactating Sows ◽  
Ad Libitum

Abstract Two experiments were conducted to determine the efficacy of phytase on apparent total tract digestibility (ATTD) of phosphorus (P) in gestating and lactating sows. In Exp. 1, a total of 40 sows (parity 2–6) at d 78 of gestation were randomly assigned, based on parity, over 4 experimental diets. In Exp. 2, 40 sows (parity 2.0–6.0) at d 4 of lactation were randomly assigned based on parity, over 4 experimental diets. Both studies had a positive control (PC) diet with adequate levels of calcium (Ca) and digestible P (6.5 and 2.5 g/kg and 7.0 and 3.2 g/kg for gestation and lactation diets, respectively), a negative control (NC) diet with low Ca and digestible P levels (5.0 and 1.0 g/kg and 5.0 and 1.5 g/kg for gestation and lactation diets, respectively), and NC supplemented with a novel consensus bacterial 6-phytase variant (PhyG) or a commercial Buttiauxella sp. phytase (PhyB), both at 500 FTU/kg. Diets were corn, soybean meal and sunflower meal based (phytate P of 3.0 g/kg) with restricted feeding during gestation and ad libitum during lactation. In both studies, fecal samples were collected after an adaptation of 14 d on 4 successive days to determine ATTD of P. Data were analyzed using ANOVA, treatment mean comparison using Tukey test using JMP 14. In addition, data from the Exp. 1 and 2 were pooled to further investigate their combined effect, using trial as random effect. In both studies, supplementation of both phytases improved (P < 0.05) ATTD of P vs NC. With data combined from two trials, sows fed PhyG showed greater (P < 0.05) ATTD of P when compared those fed PhyB. In conclusion, PhyG at 500 FTU/kg showed a greater ATTD of P in sows when compared to the PhyB at 500 FTU/kg, when data combined from two trials.

20 Effect of Adding Bioactive Peptide in Combination of Pharmaceutical Zinc Oxide or Organic Acids on Growth Performance, Hematology Profile, and Nutrient Digestibility in Nursery Pigs

2021 ◽  
Vol 99 (Supplement_1) ◽  
pp. 22-23
Author(s):  
Richard A Mudarra ◽  
Tsung Cheng Cheng Tsai ◽  
Kristopher Bottoms ◽  
Thomas S Shieh ◽  
Casey Bradly ◽  
...  
Keyword(s):  
Growth Performance ◽  
Phase 1 ◽  
Random Effect ◽  
Positive Control ◽  
Negative Control ◽  
Nutrient Digestibility ◽  
Bioactive Peptide ◽  
Nursery Pigs ◽  
Cell Counts ◽  
High Zinc

Abstract To evaluate the effect of bioactive peptide (P) in combination with high level of zinc (HZ) or acidifiers on growth performance, complete blood cell counts (CBC) and nutrient digestibility in nursery pigs, a total of 288 weaned pigs (PIC1050xDNA600) were stratified by initial BW within gender and allotted to 1of 7 treatments. Treatments for phase 1&2 were: 1) nutrient adequate positive control with HZ (PC), 2) nutrient deficient negative control with HZ (NC, -0.13% SID Lysine by reducing fish meal), 3) NC+0.25% peptide (0.25PZ), 4) NC+0.5% peptide (0.5PZ), 5) NC+0.25% peptide with standard zinc (0.25P), 6) NC+0.5% peptide with standard zinc (0.5P), 7) as 5 + 0.1% sodium butyrate and 0.5% benzoic acid (PSB). All pigs were fed a common low Zn diet (197 ppm) during phase 3. The whole blood was obtained from a close-to-average pen-BW pig repeatedly at weaning, and at the end of phase 2 and 3 to determine CBC. Titanium dioxide was used as an indigestible marker to determine nutrient digestibility. Data were analyzed using the Mixed procedures of SAS as a RCBD with treatment as fixed effect, and BW block as random effect. In overall phase 1&2, pigs fed PSB had similar ADG and BW when compared to pigs fed 0.25PZ and both were greater than NC pigs (Table 1). With the same inclusion rate of peptide, pigs fed a high zinc diet had greater BW and ADG than pigs fed a standard zinc diet. PSB pigs had the greatest G:F ratio and nitrogen digestibility among treatments. Increasing peptide in high zinc diets gradually decreased Neutrophil-to-lymphocyte ratio. This study indicates that the improvement in growth performance from pigs fed peptide is pharmaceutical zinc dependent and acidifiers can be an alternative to replace ZnO without affecting growth performance.

76 THE EFFECT OF GLYCEROL CONCENTRATIONS IN EGG-YOLK CITRATE EXTENDER ON THE QUALITY OF CRYOPRESERVED NGUNI BULL SEMEN

2013 ◽  
Vol 25 (1) ◽  
pp. 185
Author(s):  
M. M. Seshoka ◽  
M. L. Mphaphathi ◽  
F. V. Ramukhithi ◽  
T. R. Netshirovha ◽  
C. Hlungwani ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Rural Areas ◽  
Egg Yolk ◽  
Positive Control ◽  
Negative Control ◽  
Glycerol Concentration ◽  
Bull Semen ◽  
Frozen Semen ◽  
Thawing Process ◽  
Motility Rate

There are bull shortages in South African poor rural areas. Artificial-insemination technology could play a significant role on breeding emerging farmer’s cattle. The objective of this study was to compare glycerol concentrations (0, 4, 8, or 12%) during freezing of Nguni bull semen to conduct AI in different villages. Semen was collected by electro ejaculator from 2 Nguni bulls of known and proven fertility. Collected semen samples were kept in a thermo flask (37°C) and transported to the laboratories within 10 min after collections. Semen samples were pooled and evaluated by Sperm Class Analyser® and allocated randomly per treatment. Semen was then diluted (1 : 2 v:v) with egg-yolk citrate extender supplemented with either 0% (negative control), 4, 8, and 12% of glycerol concentration or AndroMed® (positive control). Semen samples were equilibrated for 4 h at 5°C. After equilibration period, samples were loaded into 0.5-mL straws and placed horizontally into the controlled rate (–5, –8, –10, –12, –15, –25, –35°C min–1) from 5°C until target temperature of –80°C is reached. The frozen semen straws were stored in a liquid nitrogen tank (–196°C) until thawing. Treatment means were separated using Fisher’s protected t-test least, and data are presented as mean ± SD. There was a significant differences (P < 0.05) between raw total sperm motility (83.3 ± 19.3) and frozen–thawed sperm with either 0% (0.0 ± 0.0), 4% (30.2 ± 13.4), 8% (47.9 ± 12.5), or 12% (61.5 ± 4.7) of glycerol and on AndroMed® (27.7 ± 17.8) group. Regardless of the glycerol concentrations used, the freezing-thawing process reduced (P < 0.05) the Nguni total sperm motility rate compared to uncryopreserved sperm (83.3 ± 19.3). In conclusion, egg-yolk citrate extender supplemented with 12% glycerol yielded a better (P < 0.05) total sperm motility rate (61.5 ± 4.7) as compared with the 0% (0.0 ± 0.0), 4% (30.2 ± 13.4), 8% (47.9 ± 12.5), and AndroMed® (27.7 ± 17.8) group. Further studies are required to test other levels of glycerol concentrations (>12%) on freezing Nguni semen and conducting AI.

scholarly journals Yumurtacı Tavuk Yemlerine Biberiye (Rosmarinus officinalis L.) Uçucu Yağ İlavesinin Performans, Yumurta Kalite Kriterleri ve Yumurta Sarısı Lipit Oksidasyonu Üzerine Etkisi

2016 ◽  
Vol 4 (2) ◽  
pp. 113 ◽  
Author(s):  
Tülay Çimrin ◽  
Murat Demirel
Keyword(s):  
Vitamin E ◽  
Essential Oil ◽  
Storage Time ◽  
Egg Quality ◽  
Egg Yolk ◽  
Positive Control ◽  
Negative Control ◽  
Feed Conversion ◽  
Treatment Groups ◽  
Rosemary Essential Oil

This experiment was designed to investigate the effects of rosemary essential oils in various doses on hen performance, egg quality, lipid peroxidation malondialdehyde (MDA) level in fresh and stored egg yolk. The experiment was carried out in random parcel experimental design. In the experiment, 6 groups were formed and each group was divided into 5 replicates. In total 240 Bovans hite laying hens, each replicate included 8 hens, 32 weeks old, were used. Treatment groups were control (Negative Control), 500 mg//kg antibiotic (Positive Control 1), 200 mg/kg vitamin E (Positive Control 2), 100 mg/kg rosemary essential oil, 200 mg/kg rosemary essential oil, 300 mg/kg rosemary essential oil. There was no significant difference in live weight, egg production, egg weight, damaged egg ratio, mortality and egg quality but feed comsumption, feed conversion ratio (FCR) and egg yolk MDA values were affected significantly in between control and treatment groups. At all treatment groups feed comsumption decreased significantly and other groups improved in feed conversion ratio except 500 mg/kg antibiotic and 100 mg/kg rosemary doses groups compared to negative control significantly. 100 mg/kg rosemary essential oil addition during the storage time for 42 and 56 days and 200 mg/kg vitamin E addition for all storage time to ration decreased egg yolk MDA values significantly. As a result, laying birds fed 200 mg/kg vitamin E, and 100 mg/kg of rosemary essential oil additives can use to prevent lipid oxidation.

scholarly journals Body Weight, Cholesterol Changes and Sub-Chronic Toxicity of Mice Treated with An Emulsion Product Rich in b-Carotene

2020 ◽  
Vol 4 (1) ◽  
pp. 19
Author(s):  
Bohari Bohari ◽  
Fenny Dian Lestari ◽  
Anton Rahmadi
Keyword(s):  
Chronic Toxicity ◽  
Herbal Medicines ◽  
Egg Yolk ◽  
Positive Control ◽  
Visual Observation ◽  
Negative Control ◽  
Additional Treatment ◽  
Average Weight ◽  
Β Carotene ◽  
Standard Ration

Background: One of the conditions for releasing standardized herbal medicines is the presence of pre-clinical testing that can be conducted with mice. Emulsion products with the main composition of pumpkin, red palm oil, and dragon fruit have high levels of β-carotene. The purpose of this study was to observe changes in weight, cholesterol, and sub-chronic toxicity from mice treated with emulsion products.Materials and Methods: Mice observed consisted of 6 groups: (1) untreated control (given standard ration); (2) negative control (given standard ration and 2.8% (v/b) egg yolk); (3) positive control (given standard ration and 0.4% (v/b) β-carotene); (4) standard ration, 0.4% (v/b) β-carotene and 2.8% (v/b) egg yolk; (5) given a standard ration and 2% (v/b) emulsion products; and (6) given a standard ration, 2% (v/b) emulsion products and 2.8% (v/b) egg yolk. Observations of sub-chronic toxicity were done by measuring creatinine levels, kidney weight, and visual observation of kidney swelling.Results: The use of β-carotene-rich emulsions does not cause a decrease in the average weight of mice. In general, β-carotene and emulsion can overcome cholesterol increases on the 7th day, but not for the 15th day as a result of standard food intake. The increase in creatinine levels only occurs in mice treated with egg yolk control. The additional treatment of β-carotene and emulsion successfully prevents the increase in creatinine levels.Conclusion: Emulsion products did not cause changes in weight, were able to reduce cholesterol levels in a limited manner and were not toxic to mice.Keywords: cholesterol changes, sub-chronic toxicity, b-carotene, emulsion

scholarly journals Cytotoxicity evaluation of four endodontic sealers

2008 ◽  
Vol 19 (3) ◽  
pp. 228-231 ◽  
Author(s):  
Paulo Tadeu da Silva ◽  
Fernanda Geraldes Pappen ◽  
Erick Miranda Souza ◽  
João Eduardo Dias ◽  
Idomeo Bonetti Filho ◽  
...  
Keyword(s):  
Peritoneal Macrophages ◽  
Positive Control ◽  
Negative Control ◽  
Phenol Red ◽  
Control Medium ◽  
Griess Reaction ◽  
Phosphate Buffered Saline ◽  
Mouse Peritoneal Macrophages ◽  
Endodontic Sealers

This study evaluated in vitro the cytotoxicity of four root canal sealers (Topseal, EndoRez, TubliSeal and Kerr Pulp Canal Sealer E.W.T.) and their effects on reactive oxygen/nitrogen intermediate induction by mouse peritoneal macrophages. Thioglycollate-induced cells were obtained from Swiss mice by peritoneal lavage with 5 mL 10 mM phosphate-buffered saline, washed twice and resuspended (106 cells/mL) in appropriate medium for each test. Cytotoxicity was determined by the presence of hydrogen peroxide (H2O2) and nitric oxide (NO) by the peroxidase-dependent oxidation of phenol red and Griess reaction, respectively. Sealer suspensions were obtained in two different concentrations from each material: 18 mg/mL and 9 mg/mL, established according to compatibility parameters following MTT assay. Comparing the sealers, H2O2 release at concentrations of 9 mg/mL and 18 mg/mL was similar: Topseal > positive control (medium + cells + 5 mg/mL zimozan solution) > EndoRez > TubliSeal > Kerr Pulp E.W.T. > negative control (medium + cells). NO release at concentration of 9 mg/mL was: positive control (medium + cells + 10 µg/mL LPS solution) > Topseal > Kerr Pulp E.W.T. > TubliSeal = EndoRez > negative control (medium + cells); at concentration of 18 mg/mL was: positive control > Topseal > Kerr Pulp E.W.T > TubliSeal > EndoRez > negative control. Based on the results, it may be concluded that Topseal presented the highest cytotoxicity among the tested sealers, releasing higher concentrations of NO and H2O2 in macrophage culture.

scholarly journals Quality of Sexed Sperm of Bali Bull in Regional Artificial Insemination Center of Riau Province

2019 ◽  
Vol 43 (2) ◽  
Author(s):  
Yendraliza Yendraliza ◽  
Anwar Efendi Harap ◽  
July Handoko ◽  
Muhammad Rodiallah
Keyword(s):  
Plasma Membrane ◽  
Pregnancy Rate ◽  
Artificial Insemination ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Female Offspring ◽  
Plasma Membrane Integrity ◽  
Frozen Semen ◽  
Calving Rate

This study aimed to evaluate the quality of frozen semen of Bali bull resulted from sexing procedure on calf or offspring production with desired sex. The tested sperm of Bali bull were collected from Bali bull raised at Regional Artificial Insemination Center of Riau Province (BIBD Riau). The study was carried out in 2 stages. The first stage was X and Y chromosome separation by albumin method. The extender used in the sexing procedure is trice citrate fructose and egg yolk. The second stage was mainly testing the sexed sperm collected in 60 Bali cow in Langkat Village, Bengkalis Regency. To determine the quality of post thawing frozen semen collected from the sexing procedure, the study evaluated motility, viability, mortality, abnormality and plasma membrane integrity of the spermatozoa. The pregnancy rate, calving rate, and birth accuracy of inseminated sexed sperm to offspring’ sex were also evaluated. The evaluation resulted in motility (66.3-75.3%), viability (70-78.5%), plasma membrane integrity (60-65.8%), abnormality (6.05-8.05%), mortality (20.05-30.05%), and pregnancy rate (83.33-90%). The calving rate on this study was 100% with the birth accuracy of 81.8% for male offspring and 40% for female offspring. As conclusion, the sexed sperm evaluated on this study have fairly good fertility.

scholarly journals PSII-3 Effect of molasses-based block supplementation on cattle fed endophyte-infected Tall Fescue (Festuca arundinacea) seed

2019 ◽  
Vol 97 (Supplement_2) ◽  
pp. 227-228
Author(s):  
Luis Felipe Barbosa Braga. Feitoza ◽  
Hans Christian Muller ◽  
Tom Williams ◽  
James Drouillard
Keyword(s):  
Tall Fescue ◽  
Festuca Arundinacea ◽  
Fixed Effects ◽  
Treatment Time ◽  
Random Effect ◽  
Positive Control ◽  
Negative Control ◽  
Time Interaction ◽  
Doppler Ultrasound Imaging ◽  
Protein Block

Abstract Ergot-alkaloid toxicosis induces persistent vasoconstriction in cattle, interfering with efficient thermoregulation. Our objective was to evaluate impact of cooked molasses-based block supplements (MBS) on performance and physiological measurements in cattle fed ergot-containing Tall Fescue seed (TFS). Crossbred steers (n = 95; 287 ± 6.4 kg) were blocked by body weight, randomly assigned to individual indoor feeding pens and treatments consisting of: Negative Control (NC; prairie hay only); Positive Control (PC; hay plus TFS); and molasses block treatments fed hay and TFS with ad libitum access to a 38% protein block (CB); 38% protein block containing 0.3% crystalline menthol (MB); or a 38% protein block containing a proprietary blend of mannan oligosaccharide and capsaicin (AB). The TFS was mixed with molasses (9:1), and amount fed was increased gradually at 3-d intervals from 45 to 520 g/animal daily by day 42. Temperature differential between ocular conjunctiva and ear tip (ΔT) was determined weekly using thermographic imaging. Coccigeal vein diameter (CVD) was measured with Doppler ultrasound imaging (DU) on days 63 and 84. Data were analyzed as mixed models with block as a random effect and treatment, time, and treatment by time interaction as fixed effects. Block intakes did not differ among MBS treatments (P > 0.05). Cattle fed MBS had greater ADG, DMI, gain:feed (P < 0.01) compared to NC and PC. For the second half of the study MBS treatments had lower ΔT (P < 0.05). Supplementing cooked molasses-based block supplements low-moisture molasses block can improve performance and potentially enhance peripheral blood irrigation in cattle consuming ergot-infested feeds.

Multiparametric In Vitro Evaluation of Cytocompatibility of 1% Strontium-Containing Nanostructured Hydroxyapatite

2014 ◽  
Vol 631 ◽  
pp. 345-350
Author(s):  
Daiana Reis ◽  
Daniela Silva ◽  
Juliana Côrtes ◽  
Letícia Hummel ◽  
Elena Mavropoulos ◽  
...  
Keyword(s):  
Biological Effects ◽  
Membrane Integrity ◽  
Bone Substitutes ◽  
Biological Properties ◽  
Positive Control ◽  
Negative Control ◽  
In Vitro Tests ◽  
Xtt Assay ◽  
Experimental Control

Hydroxiapatite (HA), one of the most widely employed bioceramic bone substitutes, when applied on its nanostructured form (nHA) may contribute to achieve a crystalline structure which is closer to the size and morphology of biological apatite. Furthermore, HA might also be doped with several different cations with biological effects including Sr2+. Therefore, a biomaterial based on nanostructured HA containing 1% Strontium (nSrHA) could present interesting biological properties, as strontium is described as a modulator of both osteoblast and osteoclast activities, presenting an important regulatory role on bone resorption. However, such modifications may also affect the biocompatibility of this material, which should be accessed initially by in vitro methods. Therefore, the present work aimed to evaluate the in vitro biocompatibility of 1% nSrHA discs with human primary osteoblasts through a multiparametric assay which assesses simultaneously metabolic activity (XTT assay), membrane integrity (NR test) and cell density (CVDE). Extracts of nSrHA, latex fragments (positive control), polystyrene beads (negative control) and nHA (for comparison) were prepared and exposed to 104 cells for 24h at 37°C/5% CO2on test plates, according to ISO 10993-5:2009, on quintuplicates. Cells exposed to unconditioned media were used as experimental control. After exposure, cells were tested for viability with a commercial multiparametric kit (In Cytotox, Xenometrix, Germany). The positive and negative controls presented the expected results, validating the assay. Both nHA and SrnHA were considered biocompatible, since the presented a cell viability after exposure statistically similar to the experimental control. In conclusion, the synthesized nSrHA discs are cytocompatible and, consequently, adequate for further in vitro tests on cell adhesion, proliferation and differentiation.

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