32 Vitrification of in vitro-matured bovine oocytes in triacetate cellulose hollow fibres

2019 ◽  
Vol 31 (1) ◽  
pp. 142
Author(s):  
E. V. Kornienko ◽  
A. B. Romanova ◽  
M. A. Ikonopistseva ◽  
G. P. Malenko
Keyword(s):  
Ethylene Glycol ◽  
Fertilization Rate ◽  
Control Group ◽  
Hatching Rate ◽  
High Survival ◽  
Hollow Fibres ◽  
Group 2 ◽  
Bovine Oocytes ◽  
Group 1

A prospective method of vitrification in triacetate cellulose hollow fibres (HF) introduced by Matsunari et al. (2012 J. Reprod. Dev. 58, 599-608) allowed significant simplification and standardization of vitrification/warming procedures and was successfully used for group cryopreservation of various pre-implantation mammalian embryos. The goal of the current work was to evaluate the effectiveness of the HF vitrification method for cryopreservation of in vitro-matured bovine oocytes. The base medium for all the vitrification and rewarming solutions was calcium-free TBP-like protein-HEPES supplied with 20% of fetal bovine serum. Groups of 15 morphologically normal in vitro-matured bovine oocytes were equilibrated with 3% (vol/vol) ethylene glycol for 15min, loaded into HF, and transferred into vitrification solution containing 30% ethylene glycol and either 0.5M (Group 1) or 1.0M (Group 2) sucrose. Hollow fibres were incubated for either 60s (Group 1) or 30s (Group 2) and immediately plunged into LN. Rewarming was conducted at 39°C. Oocytes within HF were placed in decreasing concentrations of sucrose solutions to remove cryoprotectants. Then, oocytes were subjected to IVF. Non-vitrified denuded oocytes were used as a control. Survival rates were evaluated at 21h post-rewarming. Part of the presumptive zygotes were fixed and stained with acetolacmoid for fertilization rate. The remaining zygotes were cultured for 10 days. Developmental rates were evaluated at 44h and 7 and 10 days post-IVF. All results are presented as mean percentage±standard deviation. Data were analysed using Mann-Whitney U test. Significance was set at P<0.05. Survival rate was significantly lower in Group 1 (79.0±8.0%) and Group 2 (75.0±5.0%) compared with the control group (97.0±4.0%). Fertilization rate in Group 1 differed significantly from the control (80.5±18.3% v. 95.5±9.1%). Cleavage rates in Groups 1 and 2 did not differ significantly from the control (42.5±15.7% v. 60.7±11.1% v. 63.0±15.8%, respectively). Blastocyst yields at 7 days post-IVF were 0.9±2.3 (1/116) and 9.6±5.4% (6/65) in Groups 1 and 2, respectively. The former was significantly lower than in the control group (17.0±10.3%, 23/154). It should be noted that hatching in the control group started at 8 days post-IVF and was delayed in Groups 1 and 2 for at least 24h. Day 10 blastocyst yields were 3.0±3.3 (P<0.05), 20.9±13.8, and 30.4±9.6% in Groups 1 and 2 and the control group, respectively. All obtained Day 10 blastocysts (3/116) in Group 1 hatched. Hatching rate in Group 2 was significantly lower than in the control group. Both Groups 1 and 2 showed relatively high survival and fertilization rates, but embryo development rates in both groups had a tendency to be lower than in the control. However, the obtained results indicate that the modifications of the protocol may increase the effectiveness of HF vitrification. The HF vitrification method remains a prospective option for simultaneous cryopreservation of a group of bovine oocytes.

234 HORMONAL FOLLICLE STIMULATION IN HOLSTEIN COWS FOR IN VITRO EMBRYO PRODUCTION USING SPERM SORTED BY FLOW CYTOMETRY

2016 ◽  
Vol 28 (2) ◽  
pp. 248 ◽  
Author(s):  
L. Ferré ◽  
Y. Bogliotti ◽  
J. Chitwood ◽  
M. Kjelland ◽  
P. Ross
Keyword(s):  
Transvaginal Ultrasound ◽  
Prostaglandin F2α ◽  
Holstein Cows ◽  
Control Group ◽  
Hatching Rate ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
Group 2 ◽  
Group 1

Transvaginal ultrasound needle-guided ovum pick-up (OPU) and in vitro embryo production (IVP) offer a reliable alternative to conventional embryo transfer to produce offspring. The success of OPU/IVP greatly depends on the number and quality of retrieved oocytes. The aim of this study was to compare OPU/IVP performance from stimulated Holstein cows. Holstein (Bos taurus) >8-year-old pluriparous open dry cows (n = 28) were used for OPU as oocyte donors. Follicular waves in all groups were synchronized by gonadotropin-releasing hormone (GnRH), prostaglandin F2α (PGF), and CIDR administrated on Day 0, followed by stimulation treatments 48 h later. No pre-synch was used. Total hormone dosage were administrated as follows: Group 1: pFSH = 180 mg (Folltropin, Bioniche, Belleville, ON, Canada; n = 7), Group 2: pFSH/LH = 500 IU (Pluset, Calier, Barcelona, Spain; n = 7), Group 3: eCG = 1500 IU (eCG, Biogénesis-Bagó, Buenos Aires, Argentina; n = 7) and Group 4: Control (n = 7), no stimulation. All injections were performed intramuscularly (i.m.) twice a day, during three days. OPU was performed 48 (Group 1) or 24 h (Group 2 and 3) after the last injection. The control group received saline solution i.m. Follicles were classified according to diameter in 4 categories: small (2–5 mm); medium (6–9 mm); large (10–14 mm) and extra large (>15 mm). A Mindray DP-30 Vet (Mindray Medical, Shenzhen, China) was equipped with a micro-convex transducer 5.0- to 8.5-MHz probe along with a disposable 21G needle. The OPU flow rate was 15 mL min–1. Retrieved oocytes were classified according to IETS guidelines as viable (grade 1 + 2) and non-viable (grade 3 + 4). The IVP protocol was according to that in Reprod. Fertil. Devel. (2004, 16, 253). Fertilization (Day 0) was carried out using female sex-sorted semen selected with a discontinuous density gradient (PureSperm, Nidacon, Mölndal, Sweden) and diluted to 1 × 106 sperm mL–1. ANOVA was used for comparisons of mean values and a chi-squared test was used for proportions. Results are presented in the Table 1. In conclusion, pFSH stimulation before ovum pick-up in Holstein cows increased the number of collected and viable oocytes, cleavage, embryo development, and hatching rates in comparison to other follicle stimulation hormones and non-stimulation. A cost-benefit analysis of these methods could be valuable in order to inform whether or not a stimulation protocol is necessary for a commercial IVP operation. Table 1.Numbers of follicles, collected and viable oocytes, cleavage rate, blastocysts and hatching rate

scholarly journals Comparison of different irrigation and agitation methods for the removal of two types of calcium hydroxide medicaments from the root canal wall: An in-vitro study

2017 ◽  
Vol 90 (3) ◽  
pp. 327-332 ◽  
Author(s):  
Deepak Singh Kirar ◽  
Pradeep Jain ◽  
Pallav Patni
Keyword(s):  
Calcium Hydroxide ◽  
Root Canal ◽  
Positive Control ◽  
Positive Control Group ◽  
Control Group ◽  
Passive Ultrasonic Irrigation ◽  
Ultrasonic Irrigation ◽  
Group 2 ◽  
Group 1

Background and aim: Comparison of different irrigation and agitation methods for the removal of two types of calcium hydroxide medicaments from the root canal walls.Methods: Fifty extracted single rooted teeth were selected for this study. After decoronation, the root canals of these teeth were prepared to the size F3 (30 no.) using rotary ProTaper file system. These samples were randomly divided into four groups. Group 1 (n=20) were filled completely with water based calcium hydroxide (CH), Group 2 (n=20) were filled with oil based CH using lentulo spiral, Group 3 (n=5) - the positive control group received the CH as intracanal medication, but no subsequent removal, Group 4 (n=5) - the negative control did not receive CH placement. Further on, Group 1 and Group 2 were divided into four sub-groups (n=5). In sub-group A we performed conventional syringe irrigation with side-vented needle sub-group B) manual dynamic agitation, sub-group C sonic agitation using endoactivator, sub-group D passive ultrasonic irrigation (PUI). Roots were split longitudinally into mesial and distal halves. Digital images of the root canal walls were acquired by a Dental Operating Microscope (DOM) and assessed by using a scoring criteria at different thirds (coronal, middle and apical) of the root canal as follows: score 1, score 2, score 3, and score 4. Data were analyzed applying one-way analysis of variance (ANOVA) and Tukey’s multiple comparison tests at a 95% confidence interval (P < 0.05).Results: Statistically significant differences were not found between the experimental groups and the negative group in any one third of the root canal (P>0.05). However, a difference did exist between the experimental groups and the positive control group (P<0.05). None of the experimental groups totally removed CH substances from root canal walls.Conclusion: Among all experimental groups, removal of CH was best achieved by sonic agitation using endoactivator followed by passive ultrasonic irrigation (PUI), manual dynamic agitation and conventional syringe irrigation with side-vented needle.

scholarly journals Effect of Different Irrigation Solutions on the Diffusion of MTA Cement into the Root Canal Dentin

Materials ◽  
2020 ◽  
Vol 13 (23) ◽  
pp. 5472
Author(s):  
José Pedro Martinho ◽  
Sara França ◽  
Siri Paulo ◽  
Anabela Baptista Paula ◽  
Ana Sofia Coelho ◽  
...  
Keyword(s):  
Root Canal ◽  
Ethylenediaminetetraacetic Acid ◽  
Treatment Success ◽  
Confocal Laser ◽  
Control Group ◽  
Middle Section ◽  
Group 2 ◽  
Root Canal Dentin ◽  
Group 1

(1) Aim: This study aims to analyze the in vitro infiltration of a silicate root canal sealer into dentinal tubules after using different endodontic irrigating solutions. (2) Methods: Twenty-nine teeth with single roots were separated into three groups according to the final irrigation protocol: G1 n = 10) = 17% EDTA (ethylenediaminetetraacetic acid) + 3.0% sodium hypochlorite (NaOCl), G2 (n = 10) = 17% EDTA + 2.0% chlorhexidine and G3 (Control group, n = 9) = 17% EDTA + saline solution. Root canals were filled using cold lateral compaction technique with MTA Fillapex sealer and gutta-percha. The sealer was labeled with rhodamine B. The teeth were segmented at the middle and third apical sections, which were visualized using 10× confocal laser microscopy to determine the sealer penetration percentage. (3) Results: In the apical section, no statistically significant differences were found between the groups regarding sealer penetration. In the middle section, Group 1 obtained the highest percentage, and Group 2 the lowest (p = 0.004). Group 1 also presented statistically significant differences in the Control Group (p = 0.031) and had close sealer penetration values. Meanwhile, the Control Group (p = 0.023) and Group 2 (p = 0.029) revealed a significant decrease of sealer penetration between the apical and middle sections. (4) Conclusion: The obtained results support that final irrigation with NaOCl promoted similar sealer penetration in the apical and middle sections. On the other hand, a significant decrease in the sealer penetration of the middle section was observed for the chlorhexidine and saline groups. Compared to other irrigant solutions, NaOCl promotes more uniform sealer penetration, which can correlate with better sealing and, consequently, higher endodontic treatment success.

47 A New Device and Method for Successful Vitrification of In Vitro-Produced Ovine Embryos

2018 ◽  
Vol 30 (1) ◽  
pp. 163
Author(s):  
S. Ledda ◽  
J. M. Kelly ◽  
S. K. Walker ◽  
Y. Natan ◽  
A. Arav
Keyword(s):  
Survival Rate ◽  
Embryo Transfer ◽  
Blastocyst Stage ◽  
Embryo Cryopreservation ◽  
Control Group ◽  
Hatching Rate ◽  
High Survival ◽  
Embryo Survival ◽  
New Device

To advance the use of embryo vitrification technology in veterinary practice, we developed a system in which embryo vitrification, warming, and dilution can be performed within a straw. An in-straw embryo cryopreservation method reduces the need for equipment and technical skills and can facilitate direct embryo transfer to the uterus. This study proposes the use of a new device named “Sarah” that is designed to permit all in-straw embryo cryopreservation procedures. Ovine in vitro-produced (IVP) embryos were vitrified at either early blastocyst stage (EB, n = 65, 6 days post-IVF) or fully expanded blastocyst stage (FB, n = 168, 7 days post-IVF). The vitrification procedure using Sarah constituted a 0.25-mL straw with a capsule having 50-µm pores inserted at one end. Embryos at each stage (EB and FB) were divided into 2 subgroups and vitrified by 1 of 2 methods: (1) multi-step (MS) group-a straw containing 2 embryos was sequentially loaded vertically into 1.5-mL tubes containing 6 different vitrification solutions: 10, 20, 40, 60, 80, or 100% ES (with 100% ES being 7.5% DMSO +7.5% EG + 20% FCS in TCM-199; 90 s each step) followed by 30 s each in 75 and 100% VS (100% VS being 18% DMSO +18% EG + 0.5 M trehalose + BSA in TCM-199); and (2) two-step (TS) group-the straw (2 embryos/straw) was loaded with 100% of ES (5 min), followed by 100% VS solution for 30 s. For both methods, at the end of the preparation steps, the straws were plunged directly into liquid N2. Non-vitrified embryos were maintained in in vitro culture as a control group (n = 102). The warming procedure consisted of placing the straws directly into 5-mL tubes containing 100, 50, 25% WS (WS = 1 M sucrose in TCM-199+ 20% FCS) at 38.6°C (for first solution) and at room temperature for all the rest (5 min each), before being placed into the holding medium. Embryos were recovered from the straws, incubated at 38.6 C in 5% CO2 in air in TCM 199 + 5% FCS, and evaluated for blastocoel re-expansion, embryo survival, and hatching rate at 2, 14, 48 h post-warming. Blastocyst re-expansion (2 h) after warming increased as the developmental stage progressed and was not affected by the vitrification method. In fact, it was significantly (P < 0.05) higher for FB vitrified in the MS and TS methods (77.90% and 71.25%, respectively) compared with the EB method (62.5% and 48.50%, respectively). At 24 h, survival rate of vitrified FB was significantly higher (P < 0.05) in the MS system (95.35%) compared with those in TS (86.25%). Survival rates of FB embryos for both methods (MS and TS) were significantly higher (P < 0.001) than EB embryos vitrified in MS (56.25%) and TS (56.55) methods. After 48 h of culture, the hatching rate for FB vitrified in the MS system (87.21%) was comparable with TS (77.5%) and control (85.3%) groups but significantly higher (P < 0.001) than vitrified EB in MS (43.75%) and TS (36.36%). In conclusion, we showed that a high survival rate of IVP embryos can be achieved by this new in-straw vitrification and warming device (“Sarah”), with hatching rates in vitro comparable with that of control fresh embryos. This method has the potential for use in direct embryo transfer in field conditions.

scholarly journals Effect of Different Finishing and Polishing Methods for Color Stability of the E-Max Dental Ceramics: In vitro Study

2021 ◽  
Vol 14 (4) ◽  
pp. 1508-1513
Author(s):  
Ibraheem F Alshiddi
Keyword(s):  
Surface Brightness ◽  
Color Stability ◽  
Color Measurement ◽  
Control Group ◽  
Group 4 ◽  
Significant Difference ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

In order to assess the influence of finishing and polishing on the surface brightness and color stability of the ceramic veneer, fifty specimens were fabricated with 10 mm diameter and 2 mm thickness using IPS E-Max Ceramic. After glazing, 10 specimens were untouched as control group, and the other 40 specimens were abraded using 125µm diamond bur to create surface roughness. Forty specimens were divided into four groups (n=10), in group 1: specimens were finished using diamond point, in group 2 specimens’ surface was polished with a polishing kit, Group 3: Each specimen surface was polished with the polishing kit as in protocol 2 and was polished a polishing past and group 4 Each specimen was glazed by heating at 621℃ for 3 minutes followed by a temperature increase of 83℃/min up to 918℃ for 30 seconds. Color measurement was performed using spectrophotometer. Color stability data were analyzed using two-way ANOVA and Tukey’s HSD test (α=0.05). For Ra values, paired-samples t-tests were used to analyze the data and compare groups. The change in L and E showed a significant difference among the study groups; (group 1, group 2, group 3 and group 4) with respect to three variables L, a and b. A significant difference was noted when compared each group with the control; however, only group 2 showed a significant difference from group 4; the remaining groups demonstrated similar findings for all three variables. The study displayed a significant impact of the finishing and polishing technique on the surface brightness and color stability of ceramic restoration. However, it was evident that combination of two or three polishing techniques which includes polish kit and glaze enhances the surface finish and adds color stability by alternating the yellow – blue axis (increase in b) and red- green axis (decrease in a).

scholarly journals Re-evaluation of the value of sperm morphology in classical in vitro fertilization in a Northeastern Chinese population

2019 ◽  
Vol 47 (9) ◽  
pp. 4134-4142 ◽  
Author(s):  
Dong-liang Zhu ◽  
Hong-guo Zhang ◽  
Rui-xue Wang ◽  
Yu-ting Jiang ◽  
Rui-zhi Liu
Keyword(s):  
In Vitro Fertilization ◽  
Chinese Population ◽  
Sperm Morphology ◽  
Semen Analysis ◽  
Fertilization Rate ◽  
Vitro Fertilization ◽  
Group 2 ◽  
Normal Fertilization ◽  
Group 1

Objective This study aimed to re-evaluate the clinical value of a 4% cut-off threshold of sperm morphology in in vitro fertilization (IVF) in a cohort of a Northeastern Chinese population. Methods A total of 375 IVF cycles that met strict inclusion criteria were included. These cycles were conducted with semen analysis and oocyte fertilization. A total of 188 embryo-transferred cycles proceeded. According to sperm morphology, 375 cycles were divided into group 1 (329 cycles, <4% normal sperm morphology rate [NSMR]) and group 2 (46 cycles, ≥4% NSMR), and 188 transferred cycles into group A (151 cycles, < 4% NSMR) and group B (37 cycles, ≥4% NSMR). Results The fertilization and normal fertilization rates were significantly lower in group 1 than in group 2. The normal fertilization rate was significantly correlated with an NSMR < 4% or ≥4%, but the fertilization rate was not significantly correlated with the NSMR. No significant differences were found in pregnancy outcomes between groups A and B. Conclusions This study suggests that infertile patients with an NSMR < 4% are more likely to have a poor normal fertilization status in IVF.

75 ADDITION OF HYALURONAN TO A VITRIFICATION SOLUTION FOR IMMATURE BOVINE OOCYTES SELECTED BY BRILLIANT CRESYL BLUE

2013 ◽  
Vol 25 (1) ◽  
pp. 185
Author(s):  
P. Rodriguez Villamil ◽  
F. Ongaratto ◽  
M. Fernandez Taranco ◽  
G. A. Bó
Keyword(s):  
Ethylene Glycol ◽  
Solid Phase ◽  
Animal Health ◽  
Survival Rates ◽  
Control Group ◽  
Cresyl Blue ◽  
Development Rates ◽  
Vitrification Solution ◽  
Bovine Oocytes

An experiment was designed to evaluate the effect of brilliant cresyl blue (BCB) selection of immature oocytes and the addition of sodium hyaluronate (HA) to the vitrification solution on survival rates of bovine oocytes vitrified using solid-phase vitrification. Bovine cumulus–oocyte complexes (COC; n = 716) obtained from slaughterhouse ovaries were used in 6 replicates. Cumulus–oocyte complexes were washed in tissue culture medium 199 (TCM-199) and randomly allocated to 2 groups to be exposed to BCB stain (Sigma Chemical Company, St. Louis, MO, USA) for 90 min as described by Alm et al. (2005 Theriogenology 63, 2194–2205) or (control) maintained in Vigro holding medium (Bioniche Animal Health, Belleville, Canada) for 90 min (n = 220). Cumulus–oocyte complexes in the BCB group were selected based on their response to BCB as BCB+ (colored, n = 248) or BCB– (colorless, n = 248), whereas those in the control group were selected morphologically as described by Rodríguez-González et al. (2002 Theriogenology 57, 1397–1409). Oocytes from both BCB groups and 100 oocytes in the control group were vitrified by solid-phase vitrification as previously described by Rodriguez et al. (2012 Reprod. Fertil. Dev. 24, 132). The remaining 120 oocytes in the control group were not vitrified and were matured, fertilized, and cultured in vitro (in SOFaa in a controlled atmosphere) for 7 days. Vitrified oocytes were exposed to 10% ethylene glycol for 10 min, and 20% ethylene glycol + 0.2-M trehalose for 30 s, and then were subdivided to be exposed to 30% ethylene glycol + 0.5-M trehalose with or without 0.1 mg mL–1 HA (MAP 5, Bioniche Animal Health). Vitrified oocytes were stored in liquid nitrogen for at least one week and then placed directly into a 0.5-M sucrose solution (in TCM 199) at 37°C for 5 min, 0.25 M of sucrose for another 5 min, and finally TCM-199 and matured, fertilized, and cultured. Development rates (i.e. proportion of blastocysts) were examined on Day 7 after fertilization. Proportional data were first transformed by square root and then analyzed by ANOVA to detect the effect of replicate, type of oocyte (BCB+, BCB–, controls), and vitrified with or without HA or not vitrified as main effects, using the software Infostat (UNC, Argentina, 2010). There was a significant effect of oocyte type on blastocyst rate (P < 0.01) following vitrification (BCB+, 6.4 ± 0.4%. v. BCB–, 1.6 ± 0.6%). Control oocytes (not exposed to BCB) resulted in 3.0 ± 2.0% blastocysts following vitrification, which was lower to that obtained with the BCB+ oocytes. Vitrification also influenced development rates (3.0 ± 2.0 v. 32.0 ± 1.3%) for blastocysts produced from vitrified v. nonvitrified oocytes, respectively (P < 0.01). Furthermore, the use of HA in the vitrification solutions did not have a significant effect on development rates (4.7 ± 0.9 v. 3.3 ± 0.9%, for blastocysts obtained from vitrified oocytes with or without HA, respectively). In conclusion, the selection of oocytes by BCB increased the in vitro development rates of vitrified immature oocytes, whereas the use of HA in the vitrification solution did not improve the survival rates of vitrified oocytes.

97 PRELIMINARY DESCRIPTIVE STUDY OF EQUINE PLACENTA GENERATED AFTER TRANSFER OF IN VIVO- AND IN VITRO-PRODUCED EMBRYOS

2017 ◽  
Vol 29 (1) ◽  
pp. 156 ◽  
Author(s):  
A. Lanci ◽  
J. Mariella ◽  
B. Merlo ◽  
C. Castagnetti ◽  
E. Iacono
Keyword(s):  
Embryo Transfer ◽  
Intracytoplasmic Sperm Injection ◽  
Reproductive Technologies ◽  
Control Group ◽  
Placental Development ◽  
Group 3 ◽  
Group 2 ◽  
Group 1

Placental changes associated with artificial reproductive technologies have been described in several species, but little information is available in horses. Joy et al. (2012) reported that human placentas from intracytoplasmic sperm injection derived embryos were heavier and thicker than those produced after natural conception. Despite the most growing interest and efficiency of artificial reproductive technologies in equine species, only recently, Pozor et al. (2016) described placental abnormalities in pregnancies generated by somatic cell NT, but there are no studies on equine placenta generated by intracytoplasmic sperm injection and traditional embryo transfer. In the present preliminary study, macroscopic differences of placentas generated after transfer of in vitro- or in vivo-produced embryos were registered. Twelve Standardbred recipient mares with pregnancy generated after transfer of in vivo-derived (Group 1) and in vitro-derived (Group 2) embryos were enrolled; 10 Standardbred mares with pregnancy derived by traditional AI were included as control (Group 3). All pregnancies were physiological, and newborn foals were healthy. Mare age, parity, length of pregnancy, gross evaluation and weight of placenta, total length of umbilical cord (UC), length of UC, number of UC coils, foal sex, and weight at birth were registered. Collected data are listed in Table 1 and are expressed as mean ± standard deviation. Differences between groups were evaluated by 1-way ANOVA, and the difference in proportion of overweight placentas was evaluated with the Fisher test. The gross evaluation of placenta revealed 8/12 placentas (2/4 Group 1; 6/8 Group 2) were heavier than 11% (Madigan, 1997) due to oedema of the chorioallantois. No overweight placentas were registered in Group 3. In Group 1, 1/4 placentas had villous hypoplasia, and in Group 2, 1/8 placentas had cystic pouches on the UC. There were no significant differences among groups. However, the proportion of overweight placentas between Group 2 (6/8) and Group 3 (0/10) approached significance (P = 0.06). Although preliminary, the results of the present study suggest that production of equine embryos in vitro may lead to alterations in placental development. Several studies in cattle and sheep have suggested that alterations in the placentas of pregnancies derived from in vitro-produced embryos are related to effects of culture on epigenetic regulation. Less is known in the horse about the effects of in vitro embryo production on placental development; thus, further research in this area is necessary. Table 1. Characteristics of full-term placentas derived from AI or embryo transfer with in vivo- and in vitro-produced embryos

scholarly journals Effect of the number of thermocycles on microleakage of resin composite restorations

2003 ◽  
Vol 17 (4) ◽  
pp. 337-341 ◽  
Author(s):  
Flávia Bittencourt Pazinatto ◽  
Bruno Barbosa Campos ◽  
Leonardo César Costa ◽  
Maria Teresa Atta
Keyword(s):  
Resin Composite ◽  
Control Group ◽  
Group 4 ◽  
Thermal Changes ◽  
Group 3 ◽  
Group 5 ◽  
Group 2 ◽  
Number Of Cycles ◽  
Group 1

Thermocycling simulates, in vitro, thermal changes that occur in the oral cavity. The aim of this study was to evaluate the influence of the number of cycles on microleakage. Class V cavities (1.5 mm deep, 3 mm in height and 3 mm in width) were prepared in bovine teeth, restored with a Single Bond/Z250 restorative system (3M/ESPE) and then divided into five groups of ten teeth each: group 1 was not thermocycled (control group), and groups 2, 3, 4 and 5 were thermocycled 500, 1,000, 2,500 and 5,000 times, respectively (5º-55º ± 2ºC, 15 s dwell time). The teeth were immersed in 0.5% basic fuchsin aqueous solution for 24 h, sectioned and the sections with the highest degree of microleakage were selected, scanned and the extent of dye penetration was measured by the ImageTool program. The results submitted to one-way ANOVA showed no significant differences between the groups (p > 0.05). The averages of microleakage values in millimeters were: group 1 (3.92); group 2 (3.13); group 3 (4.48); group 4 (4.33) and group 5 (3.42). Thus, it was concluded that there is no relation between the increase of the number of cycles and the increase in microleakage.

106 EFFECT OF GLYCEROL AND ETHYLENE GLYCOL ON THE DEVELOPMENT OF IN VITRO BOVINE EMBRYOS

2006 ◽  
Vol 18 (2) ◽  
pp. 161
Author(s):  
A. C. Nicacio ◽  
R. Simões ◽  
M. A. Peres ◽  
J. S. A. Gonçalves ◽  
M. E. O. D'Ávila Assumpção ◽  
...  
Keyword(s):  
Ethylene Glycol ◽  
Cell Monolayer ◽  
Control Group ◽  
Hatching Rate ◽  
Bovine Embryos ◽  
Embryo Viability ◽  
Chi Square ◽  
Chi Square Test ◽  
Hatching Rates

The aim of this study was to evaluate the viability of in vitro-produced bovine embryos after exposure to different cryoprotectant solutions and cryopreservation. Bovine ovaries were collected at slaughterhouse and oocytes were matured, fertilized, and cultured in vitro. The embryos were co-cultured on a granulosa cell monolayer in SOF + 5% FCS and nonessential amino acids. In Experiment 1, expanded blastocysts were exposed to 10% ethylene glycol (EG) solution for 10 min (Group EG) or to 10% EG solution for 10 min and to 20% EG + 20% glycerol (Gly) solution for 30 s (Group EG/Gly). Cryoprotectants were diluted with PBS + 0.2% BSA + 0.3 M sucrose and PBS + 0.2% BSA solutions, both for 3 min, and the hatching rate was evaluated after culture. In Experiment 2, after exposure, EG Group was cryopreserved by slow freezing procedure (1.2�C/min) and EG/Gly Group was vitrified on nitrogen vapor for 2 min. After thawing, cryoprotectants were diluted using PBS + 0.2% BSA + 0.3 M sucrose and PBS + 0.2% BSA solutions, both for 3 min; hatching rate was evaluated after culture. As a control group for both experiments, non exposed embryos were cultured and evaluated for hatching rate. In Experiment 1, the hatching rates were 59.72% (43/72) for control, 62.38% (63/101) for EG, and 69.00% (69/100) for EG/Gly groups. In Experiment 2, hatching rates were 59.72% (43/72) for control, 15.22% (7/46) for EG, and 0.00% (0/46) for EG/Gly groups. Results were analyzed by chi-square test. In Experiment 1, no differences were observed among groups (P > 0.05) and in Experiment 2, differences were observed among control, EG, and EG/Gly groups (P < 0.05). In conclusion, the cryoprotectants were not deleterious to the development of in vitro bovine embryos until hatching, but the cryopreservation procedures decreased embryo viability. This work was supported by FAPESP 04/05335-1.

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