Photobiomodulation combined with adipose-derived stem cells encapsulated in methacrylated gelatin hydrogels enhances in vivo bone regeneration.

Reconstruction of bone defects is still a significant challenge. The aim of this study was to evaluate the effect of application of photobiomodulation (PBM) to enhance in vivo bone regeneration and osteogenic differentiation potential of adipose-derived stem cells (ADSCs) encapsulated in methacrylated gelatin (GEL-MA) hydrogels. Thirty-six Sprague-Dawley rats were randomly separated into 3 experimental groups (n = 12 each). The groups were control/blank defect (I), GEL-MA hydrogel (II), and ADSC-loaded GEL-MA (GEL-MA+ADSC) hydrogel (III). Biparietal critical sized bone defects (6 mm in size) are created in each animal. Half of the animals from each group (n = 6 each) were randomly selected for PBM application using polychromatic light in the near infrared region, 600-1200 nm. PBM was administered from 10 cm distance cranially in 48 h interval. The calvaria were harvested at the 20th week, and macroscopic, microtomographic, and histologic evaluation were performed for further analysis. Microtomographic evaluation demonstrated the highest result for mineralized matrix formation (MMF) in group III. PBM receiving samples of group III showed mean MMF of 79.93±3.41%, whereas the non-PBM receiving samples revealed mean MMF of 60.62±6.34 % (p=0.002). In terms of histologic evaluation of bone defect repair, the higher scores were obtained in the groups II and III when compared to the control group (2.0 for both PBM receiving and non-receiving specimens; p<0.001). ADSC-loaded microwave-induced GEL-MA hydrogels and periodic application of photobiomodulation with polychromatic light appear to have beneficial effect on bone regeneration and can stimulate ADSCs for osteogenic differentiation.

Fertilization and early embryonic development of in vitro matured metaphase I oocytes in patients with unexpected low oocyte maturity rate.

To determine the fertilization and embryonic potential of immature metaphase I (MI) oocytes in patients with low oocyte maturity rate in whom the percentage of mature oocytes obtained was less than 75% of the total retrieved ones. In vivo matured metaphase II (MII) oocytes (MII-ICSI, n = 244), and in vitro matured MI oocytes (MI-MII-ICSI, n = 202) underwent an intracytoplasmic sperm injection (ICSI) procedure. Maturation rate, fertilization rate and early embryonic development were compared in both groups. In total, 683 oocytes were collected from 117 ICSI cycles of 117 patients. Among them, 244 (35.7%) were mature MII and 259 (37.9%) were MI after the denudation process. Of those 259 MI oocytes, 202 (77.9%) progressed to MII oocytes after an incubation period of 18-24 h. The maturation rate was 77.9%. Fertilization rate was found to be significantly higher in the rescued in vitro matured MI oocyte group when compared with the in vivo matured MII oocyte group (41.6% vs 25.8%; P = 0.0006). However, no significant difference was observed in terms of cleavage rates on days 2 and 3 between the groups (P = 0.9126 and P = 0.5031, respectively). There may be unidentified in vivo factors on the oocyte maturation causing low developmental capacity in spite of high fertilization rates in the group of patients with low oocyte maturity rate. Furthermore, studies are needed to determine the appropriate culture characteristics as well as culture period and ICSI timing of these oocytes.

The relationship between good quality embryo rates and IVF outcomes/embryo transfer policies in extended embryo culture.

This study aims to investigate which parameters affect the change in good quality embryo rates during the cleavage stage and whether they have any effect on embryo transfer policies and IVF results. We analysed changes in good quality embryo (grades 1 and 2) rates during the period on days 2, 3 and 5; patients with five or fewer embryos (group 1), 6-10 embryos (group 2) and more than 10 embryos (group 3). The good quality embryo rates decreased in all groups on day 5. When the infertility reasons are studied among all of the groups, ovulatory dysfunction is found to be significantly higher in group 2 compared to group 1 and unexplained infertility was found to be significantly higher in group 2 compared to group 1 and group 3. Total antral follicle, mature oocyte and total oocyte counts were found to be significantly lower in group 1. However, there is no significant difference found among all of the groups for ß-HCG levels and clinical pregnancies. Changes in good quality embryo rates at the cleavage stage in extended embryo culture do not have an impact on IVF results.IMPACT STATEMENTWhat is already known on this subject? The number and quality of embryos in the cleavage stage are important parameters affecting the embryo transfer decision on day 5. There is still insufficient knowledge concerning changes in the percentage of increased good quality embryo transfers associated with IVF outcomes during the second to the third day, and the third to the fifth day.What do the results of this study add? Day 5 embryo transfer is possible in patients with a low number of embryos, according to our results. The good quality embryo rates of patients with a low number of embryos at the cleavage stage are more promising compared to patients having more than five embryos.What are the implications of these findings for clinical practice and/or further research? An extended embryo culture option can be used on patients with a low number of embryos for clinical practice.

Photobiomodulation with polychromatic light increases zone 4 survival of transverse rectus abdominis musculocutaneous flap.

OBJECTIVE: The aim of this study was to evaluate the effect of relatively novel approach of application of polychromatic light waves on flap survival of experimental musculocutaneous flap model and to investigate efficacy of this modality as a delay procedure to increase vascularization of zone 4 of transverse rectus abdominis musculocutaneous (TRAM) flap. METHODS: Twenty-one Wistar rats were randomized and divided into 3 experimental groups (n = 7 each). In group 1 (control group), after being raised, the TRAM flap was sutured back to its bed without any further intervention. In group 2 (delay group), photobiomodulation (PBM) was applied for 7 days as a delay procedure, before elevation of the flap. In group 3 (PBM group), the TRAM flap was elevated, and PBM was administered immediately after the flap was sutured back to its bed for therapeutic purpose. PBM was applied in 48 hours interval from 10 cm. distance to the whole abdominal wall both in groups 2 and 3 for one week. After 7 days of postoperative follow-up, as the demarcation of necrosis of the skin paddle was obvious, skin flap survival was further evaluated by macroscopic, histological and microangiographic analysis. RESULTS: The mean percentage of skin flap necrosis was 56.17 ± 23.68 for group 1, 30.92 ± 17.46 for group 2 and 22.73 ± 12.98 for group 3 PBM receiving groups 2 and 3 revealed less necrosis when compared to control group and this difference was statistically significant. Vascularization in zone 4 of PBM applied groups 2 and 3 was higher compared to group 1 (P = 0.001). Acute inflammation in zone 4 of group 1 was significantly higher compared to groups 2 and 3 (P = 0.025). Similarly, evaluation of zone 1 of the flaps reveled more inflammation and less vascularization among the samples of the control group (P = 0.006 and P = 0.007, respectively). Comparison of PBM receiving two groups did not demonstrate further difference in means of vascularization and inflammation density (P = 0.259). CONCLUSION: Application of PBM in polychromatic fashion enhances skin flap survival in experimental TRAM flap model both on preoperative basis as a delay procedure or as a therapeutic approach. Lasers Surg. 51:538-549, 2019. © 2019 Wiley Periodicals, Inc.

Effects of Systemic and Local Caffeine on Vessel Diameter, Anastomosis Patency, and Intimal Hyperplasia in the Rat.

BACKGROUND: The use of caffeine is not recommended prior to elective microsurgery due to its demonstrated negative effects on vessel anastomosis by the presumed sympathomimetic induction of vasoconstriction. In this study, we aimed to elucidate the systemic and local effects of caffeine on vessel diameter, anastomosis patency, and degree of intimal hyperplasia during the healing process. METHODS: Twenty-five rats were randomly assigned to five groups: (1) negative control, (2) preoperative systemic caffeine, (3) postoperative systemic caffeine, (4) perioperative systemic caffeine, and (5) a local caffeine group. Both the right and left femoral arteries were used. Ten anastomoses were performed per group. The arterial diameter was measured by micrometer, anastomosis patency was assessed surgically and histologically, and the histological examination was conducted 3 weeks postoperatively to determine intimal hyperplasia. RESULTS: The overall patency rate was 96%. Mild vasoconstriction was observed in the systemic caffeine groups (statistically insignificant); however, there were no negative effects on anastomosis patency. Local caffeine irrigation resulted in significant vasodilatation in the local caffeine group (p = 0.001); a similar effect was not observed in the other groups. There was a significant decrease in the intima/media ratio in the local caffeine group (p < 0.01), when compared with the control and systemic caffeine groups. No other intima/media ratio differences were observed among other comparison groups. CONCLUSION: The systemic administration of caffeine, although statistically insignificant, has an observable effect on vasoconstriction. However, it does not appear to have negative effects on anastomosis patency regardless of its application period (pre-, post-, or perioperatively). The local application of caffeine resulted in considerable vasodilatation as opposed to the vasoconstriction effect in the systemic caffeine groups. Decreased intimal hyperplasia at the anastomosis edge, and antifibrotic properties in the surgical field were also observed in this group. Histologically, the local caffeine group demonstrated an additional beneficial effect on anastomosis remodeling.

Retrospective comparison of intracytoplasmic sperm injection outcomes of sperm retrieved from a testicular biopsy and freshly ejaculated semen in oligozoospermia / Comparación retrospectiva de los resultados de inyección intracitoplasmática de espermatozoides obtenidos de una biopsia testicular y semen recién eyaculado en oligozoospermia

Objective: The aim of this study was to compare fertilisation, pregnancy rates and perinatal outcomes in patients undergoing intracytoplasmic sperm injection (ICSI) due to oligozoospermia. Methods: A total of 166 patients with oligozoospermia who underwent an ICSI procedure were included in the study. The subjects were divided into two groups according to the sperm retrieval technique used: group 1, ejaculated semen (n=111); group 2, surgical sperm retrieval (n=55). Results: Although the clinical pregnancy rate was lower in group 2, the difference was not statistically significant (36.4% vs. 42.3%, p=0.460). The difference between fertilisation and take-home baby rates of the groups were not significantly different, either (p=0.486, p=0.419, consecutively). Conclusion: Two different sperm retrieval techniques used for ICSI had no statistically significant difference on intracytoplasmic sperm injection outcomes in oligozoospermic patients

Objetivo: El objetivo de este estudio fue comparar la fertilización, las tasas de embarazo y los resultados perinatales en pacientes sometidos a inyección intracitoplasmática de espermatozoides (ICSI) por oligozoospermia. Métodos: Un total de 166 pacientes con oligozoospermia que se sometieron a un procedimiento ICSI se incluyeron en el estudio. Los sujetos se dividieron en dos grupos según la técnica de recuperación de espermatozoides utilizada: grupo 1, semen eyaculado (n=111); Grupo 2, recuperación quirúrgica de espermatozoides (n=55). Resultados: Aunque la tasa de embarazos clínicos fue menor en el grupo 2, la diferencia no fue estadísticamente significativa (36.4% vs. 42.3%, p=0,460). La diferencia entre las tasas de fecundación y de bebés a domicilio de los grupos tampoco fue significativamente diferente (p=0.486, p=0.419, consecutivamente). Conclusión: Dos diferentes técnicas de recuperación de espermatozoides utilizados para la ICSI no tenía ninguna diferencia estadísticamente significativa en la inyección de espermatozoides intracitoplasmáticos resultados en pacientes oligozoospermicos

Retrospective comparison of intracytoplasmic sperm injection outcomes of sperm retrieved from a testicular biopsy and freshly ejaculated semen in oligozoospermia.

OBJECTIVE: The aim of this study was to compare fertilisation, pregnancy rates and perinatal outcomes in patients undergoing intracytoplasmic sperm injection (ICSI) due to oligozoospermia. METHODS: A total of 166 patients with oligozoospermia who underwent an ICSI procedure were included in the study. The subjects were divided into two groups according to the sperm retrieval technique used: group 1, ejaculated semen (n=111); group 2, surgical sperm retrieval (n=55). RESULTS: Although the clinical pregnancy rate was lower in group 2, the difference was not statistically significant (36.4% vs. 42.3%, p=0.460). The difference between fertilisation and take-home baby rates of the groups were not significantly different, either (p=0.486, p=0.419, consecutively). CONCLUSION: Two different sperm retrieval techniques used for ICSI had no statistically significant difference on intracytoplasmic sperm injection outcomes in oligozoospermic patients.

Ribosome biogenesis mediates antitumor activity of flavopiridol in CD44+/CD24- breast cancer stem cells.

Flavopiridol is a synthetically produced flavonoid that potently inhibits the proliferation of human tumor cell lines. Flavopiridol exerts strong antitumor activity via several mechanisms, including the induction of cell cycle arrest and apoptosis, and the modulation of transcriptional regulation. The aim of the present study was to determine the effect of flavopiridol on a subpopulation of cluster of differentiation (CD)44+/CD24- human breast cancer MCF7 stem cells. The CD44+/CD24- cells were isolated from the MCF7 cell line by fluorescence-activated cell sorting and treated with 100, 300, 500, 750 and 1,000 nM flavopiridol for 24, 48 and 72 h. Cell viability and proliferation assays were performed to determine the inhibitory effect of flavopiridol. Gene expression profiling was analyzed using Illumina Human HT-12 v4 Expression BeadChip microarray. According to the results, the half maximal inhibitory concentration (IC50) value of flavopiridol was 500 nM in monolayer cells. Flavopiridol induced growth inhibition and cytotoxicity in breast cancer stem cells (BCSCs) at the IC50 dose. The present study revealed several differentially regulated genes between flavopiridol-treated and untreated cells. The result of the pathway analysis revealed that flavopiridol serves an important role in translation, the ribosome biogenesis pathway, oxidative phosphorylation, the electron transport chain pathway, carbon metabolism and cell cycle. A notable result from the present study is that ribosome-associated gene expression is significantly affected by flavopiridol treatment. The data of the present study indicate that flavopiridol exhibits antitumor activity against CD44+/CD24- MCF7 BCSCs through different mechanisms, mainly by inhibiting translation and the ribosome biogenesis pathway, and could be an effective chemotherapeutic molecule to target and kill BCSCs.

Follicular fluid total antioxidant capacity levels in PCOS.

In this study, our aim was to assess total antioxidant capacity (TAC) levels in follicular fluid (FF) and their relationship to clinical pregnancy rates in PCOS patients undergoing assisted reproduction (ART). Twenty-two women with polycystic ovary syndrome (PCOS) (Group 1) and 41 women without PCOS (Group 2) were included in this study. Clinical and laboratory parameters and FF TAC levels were investigated. No statistically significant differences were found between the groups with regard to age and baseline parameters. Although we could not demonstrate a significant difference in FF TAC levels between the two groups (p=0.469), there was a significant positive correlation between FF TAC and clinical pregnancy rates, BMI, and the duration of infertility for the entire group (r=0.254, p=0.048; r=0.312, p=0.013; r=0.259, p=0.040; respectively). Owing to the correlation between FF TAC and the clinical pregnancy rates, further studies evaluating the impact of FF TAC levels on ART outcomes in patients with PCOS and other etiologies of infertility are needed.

Total fertilization failure: is it the end of the story?

PURPOSE: To study parameters that could predict in-vitro fertilization (IVF) success in patients who experienced total fertilization failure (TFF) with intracytoplasmic sperm injection (ICSI) in their previous cycles. METHODS: Cycle characteristics of patients with TFF (Group I, n = 136 cycles), cycles resulting in embryo transfer (ET) following TFF (Group II, n = 36 cycles) and recurrent TFF (Group III, n = 25 cycles) and were studied retrospectively. Demographic features, cycle characteristics of three groups were compared. RESULTS: Follicle count measuring 15-17 mm was significantly higher in group II when compared to group I (p = 0.02). Total number of retrieved oocytes and mature oocytes were significantly higher in group II when compared to groups I and III (p = 0.001). Estradiol level at oocyte pick up (OPU) day was significantly higher in group II when compared to group I (p = 0.02). When the characteristics of ET cycles and preceding TFF cycles of the same patient were compared, total number of retrieved oocytes (5.11 ± 0.72 (95% CI 3.69-6.52) vs. 11.44 ± 1.60 (95% CI 5.29-17.59)) and mature oocytes (3.26 ± 3.66 (95% CI 2.04-4.47) vs. 6.92 ± 5.61 (95% CI 5.09-8.75)) were found to be significantly lower in TFF cycles (p = 0.001). Five biochemical and 5 clinical pregnancies occurred while only 2 healthy babies were born, corresponding to a live birth rate 5.5%. CONCLUSIONS: Increasing the number of retrieved and mature oocytes may increase the success of fertilization in patients with a history of previous failed fertilization. However, live birth rate is still low in embryo transfer cycles.

Is there a detrimental effect of higher gonadotrophin dose on clinical pregnancy rate in normo-responders undergoing ART with long protocol?

BACKGROUND: In recent years, it has become evident that ovarian stimulation, although a central component of in vitro fertilization (IVF), may itself has detrimental effects on oogenesis, embryo quality, endometrial receptivity, and perhaps also perinatal outcomes. OBJECTIVE: To evaluate the effect of higher gonadotrophin dose on clinical pregnancy rate in normo-responder ICSI cycles with long protocol. METHODS: A retrospective study was planned in the Department of Reproductive Endocrinology of Zekai Tahir Burak Women's Health Education and Research Hospital. 362 normo-responders undergoing ICSI cycles with long protocol were included in the study. Group 1 (n = 260): Total gonadotrophin dose <2198 IU and Group 2 (n = 102): Total gonadotrophin dose >2198 IU. Laboratory IVF outcome, clinical pregnancy rate were evaluated. RESULT(S): There was no statistically significant difference between peak estradiol levels, endometrial thickness, fertilization rates among the Group 1 versus Group 2 (p > 0.05). But there was a statistically significant difference in age, baseline FSH, oocyte number, 2PN, and clinical pregnancy among the Group 1 versus Group 2. Clinical pregnancy rate were significantly higher in Group 1 compared with Group 2 (p < 0.001). Lower gonadotrophin dose, 2PN was an independent positive predictor of clinical pregnancy (OR 2.65 for gonadotrophin dose, OR 1.1 for 2PN) CONCLUSION(S): Higher total gonadotrophin dose adversely affect clinical pregnancy in normo-responder patients undergoing ICSI cycles with long protocol.

Nitric oxide synthase inhibitors protect cerebellar Purkinje cells from zinc-induced cell loss in adult rat.

Zinc is an important trace element in biological systems; however, excessive extracellular zinc could lead to neuronal cell death following ischemia, seizures, and brain trauma. In this study, we investigated whether the intracortical injection of zinc sulphate (200 µg/kg, i.c.) changes total number of Purkinje cells in the cerebellum and whether different types nitric oxide synthase inhibitors, N-(G)-nitro-L-arginine methyl ester (L-NAME), N(omega)-nitro-L-arginine (L-NNA), aminoguanidine and 7-nitroindazole (7-NI), have protective effects against zinc neurotoxicity in Wistar albino rats. Animals were divided into 6 groups: control, zinc, zinc+L-NAME (100 mg/kg, i.p.), zinc+L-NNA (100 mg/kg, i.p.), zinc+7-NI (100 mg/kg, i.p.) and zinc+aminoguanidine (100 mg/kg, i.p.) groups. Total number of Purkinje cells in the cerebellum was estimated using unbiased stereological technique as 318,947 ± 20,549, 123,483 ± 23,762, 206,537 ± 43,128, 178,135 ± 26,635, 193,148 ± 46,104 and 212,910 ± 26,399 in the control, zinc, zinc+L-NAME, zinc+L-NNA, zinc+7-NI and zinc+aminoguanidine groups, respectively (mean ± SD). The number of Purkinje cells in zinc group was significantly lower than that of the other groups (P<0.001). It was found that the nitric oxide synthase inhibitors have neuroprotective effect against zinc neurotoxicity on Purkinje cells. These data show that the inhibition of the nitric oxide synthase could prevent some of the deleterious effects of zinc on Purkinje cells.