RESUMO
Tissue cryopreservation has allowed long term banking of biomaterials in medicine. Ovarian tissue cryopreservation in particular helps patients by extending their fertility window. However, protection against tissue injury during the thawing process has proven to be challenging. This is mainly due to the heterogenous and slow distribution of the thermal energy across the vitrified tissue during a conventional warming process. Nanowarming is a technique that utilizes hyperthermia of magnetic nanoparticles to accelerate this process. Herein, hyperthermia of synthesized PEGylated silica-coated iron oxide nanoparticles was used to deter the injury of cryopreserved ovarian tissue in a sheep model. When compared to the conventional technique, our findings suggest that follicular development and gene expression in tissues warmed by the proposed technique have been improved. In addition, Nanowarming prevented cellular apoptosis and oxidative stress. We therefore conclude that Nanowarming is a potential complementary candidate to increase efficiency in the ovarian cryopreservation field.
RESUMO
Background: Enhanced sperm motility is necessary for the successful journey of sperm inside the female genital tract, successful fertilization, and the increased chance of pregnancy. Objective: We investigated the impact of red and near-infrared (NIR) ranges of photobiomodulation (PBM) alone and together on fresh human sperm to validate an optimized PBM protocol that would maximize sperm motility and viability in vitro. Methods: We randomly divided 30 normal human semen samples into 3 different PBM protocols (red, NIR, and red+NIR lasers). Each sample was divided into four subparts, one control group sample and three experimental group samples. Each experimental group received one of the PBM protocols (red, NIR, or red+NIR). Each protocol was adjusted to three energy densities (0.6, 1.2, and 2.4 J/cm2). After exposure to the selected protocol, we determined the percentage of either viable or progressive sperm motility (PSM) and measured the DNA Fragmentation Index (DFI). Results: The NIR and red+NIR lasers at 2.4 J/cm2 energy density significantly increased PSM after 60 min compared with the control groups [least significant difference (LSD) test, p = 0.023 and p = 0.04, respectively]. Samples treated with the red laser at 0.6 J/cm2 had significantly decreased viability compared with the control group (LSD test, p = 0.003). Samples treated with the red+NIR lasers had significantly decreased viability at 0.6 J/cm2 (p = 0.003), 1.2 J/cm2 (p = 0.001), and 2.4 J/cm2 (p = 0.04) energy densities when compared with the control groups. The NIR laser resulted in no significant difference in sperm viability between the control and experimental groups. At 120 min after exposure, treatment with the red+NIR and red lasers at 2.4 J/cm2 density significantly increased DFI compared to the control groups (LSD test, p = 0.000, p = 0.007). Conclusions: In this study, sperm motility, viability, and DFI data confirmed the superiority of the NIR laser at 0.6 J/cm2 energy density compared with the red and red+NIR PBM protocols.
Assuntos
Terapia com Luz de Baixa Intensidade , Motilidade dos Espermatozoides/efeitos da radiação , Espermatozoides/efeitos da radiação , Adulto , Técnicas de Cultura de Células , Sobrevivência Celular/efeitos da radiação , Fragmentação do DNA/efeitos da radiação , Humanos , Irã (Geográfico) , MasculinoRESUMO
The probable positive effects of photobiomodulation therapy (PBMT) and oxytocin (OT) treatments together or alone were evaluated on cell viability along with the changes in the gene expression of Osteocalcin (OC), Osteoprotegerin (OPG), and Runt-related transcription factor 2 (Runx2) levels of sham (healthy)-Bone marrow mesenchymal stem cell(BMMSC) and ovariectomy-induced osteoporosis (OVX)-BMMSC. BMMSC was harvested from healthy and OVX rats and was cultured in osteogenic induction medium (OIM). There were five groups of BMMSCs: (1) sham -BMMSCs; (2) control -OVX-BMMSCs; (3) OT-treated-OVX-BMMSCs; (4) PBMT-treated-OVX-BMMSCs, and (5) OT + PBMT-OVX-BMMSCs. In all 5 groups, BMMSC viability and proliferation as well as gene expression of OC, OPG, and RUNX2 were evaluated. PBMT and PBMT + OT treatments showed a promising effect on the increased viability of OVX-BMMSC (ANOVA test; LSD test, p = 0.01, p = 0.002). The results of gene expression analysis revealed that the sham- BMMSCs responded optimally to OT treatment. It was also found that OVX-BMMSCs responded optimally to PBMT + OT and PBMT treatments at early and middle stages of osteogenic induction process. Nevertheless, they responded optimally to PBMT + OT and OT especially at the late stage of osteogenic induction process. PBMT and PBMT + OT treatments significantly increased viability of OVX-BMMSC in OIM in vitro. Both PBMT and PBMT + OT treatments could promote mineralization of OVX-BMMSC in the culture medium at early and middle stages of osteogenic induction process. Both OT and PBMT + OT treatments could promote mineralization of OVX-BMMSC in vitro at late stages of osteogenic induction process.
Assuntos
Calcificação Fisiológica/efeitos dos fármacos , Calcificação Fisiológica/efeitos da radiação , Terapia com Luz de Baixa Intensidade , Células-Tronco Mesenquimais/citologia , Osteoporose/patologia , Osteoporose/fisiopatologia , Ocitocina/farmacologia , Animais , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/efeitos da radiação , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Terapia Combinada , Feminino , Células-Tronco Mesenquimais/efeitos dos fármacos , Células-Tronco Mesenquimais/patologia , Células-Tronco Mesenquimais/efeitos da radiação , Osteoporose/tratamento farmacológico , Osteoporose/radioterapia , Ocitocina/uso terapêutico , RatosRESUMO
Background: Numerous people suffer from diabetes mellitus (DM) and resultant diabetic foot ulcers (DFU), which lack effective treatment. Photobiomodulation (PBM) has accelerated wound healing in diabetic animals and patients in some studies. However, there is scant information on the number and activation state of skin mast cells (MCs) in PBM-treated diabetic wounds. Objective: We intend to assess the influence of the number of MCs and degranulation in the remodeling step of an infected wound model on wound strength and its microbial flora in a type 1 DM (T1DM) rat model by administration of PBM, condition medium (CM) derived from human bone marrow mesenchymal stem cells (hBMMSCs), and the combination of PBM+CM. Methods: We prepared CM by culturing hBMMSCs. T1DM was induced in 72 rats and, after 1 month, we created one excisional wound in each rat. All wounds were infected with methicillin-resistant Staphylococcus aureus (MRSA). We divided the rats into four groups: (n = 18): (i) control; (ii) PBM; (iii) CM, and (iv) PBM+CM. On days 4, 7, and 15, we conducted microbiological, tensiometrical, and stereological analyses. The type of MCs (T1MCs, T2MCs, or T3MCs) and total number of MCs (TOMCs) were counted by light microscopy. Results: On day 15, the PBM+CM, PBM, and CM groups had significantly increased wound strength compared with the control group. There was a significant decrease in colony-forming units (CFU) at all time points in the PBM+CM and PBM groups. The PBM+CM and PBM groups had more stable MCs (T1MCs), less significant degranulated MCs (T2MCs), less significant disintegrated MCs (T3MCs), and less significant TOMCs compared with the control group at all time points. Conclusions: PBM+CM and PBM treatments significantly increased the healing process in an ischemic and MRSA-infected wound model of T1DM rats. PBM+CM and PBM significantly decreased both TOMCs and their degranulation, and significantly decreased CFU.
Assuntos
Degranulação Celular/efeitos da radiação , Terapia com Luz de Baixa Intensidade , Mastócitos/metabolismo , Cicatrização/efeitos da radiação , Infecção dos Ferimentos/radioterapia , Animais , Contagem de Células , Meios de Cultivo Condicionados , Diabetes Mellitus Experimental , Modelos Animais de Doenças , Humanos , Mastócitos/efeitos da radiação , Transplante de Células-Tronco Mesenquimais , Staphylococcus aureus Resistente à Meticilina , Microscopia , Ratos Wistar , Infecções Estafilocócicas/radioterapia , Infecção dos Ferimentos/microbiologiaRESUMO
OBJECTIVE: Epididymo-orchitis (EO) potentially results in reduced fertility in up to 60% of affected patients. The anti-inflammatory effects of Korean red ginseng (KRG) and its ability to act as an immunoenhancer in parallel with the beneficial effects of this ancient herbal medicine on the reproductive systems of animals and humans led us to evaluate its protective effects against acute EO. MATERIALS AND METHODS: This animal experimental study was conducted in the Department of Anatomical Sciences, Faculty of Medicine, Zanjan University of Medical Sciences (ZUMS), Zanjan, Iran during 2013-2015. We divided 50 Wistar rats into five following groups (n=10 per group): i. Control-intact animals, ii. Vehicle-phosphate buffered saline (PBS) injection into the vas deferens, iii. KRG-an intraperitoneal (IP) injection of KRG, iv. EO-an injection of uropathogenic Escherichia coli (UPEC) strain M39 into the vas defer- ens, and v. EO/ KRG-injections of both UPEC strain M39 and KRG. The treatment lasted seven days. We then evaluated sperm parameters, number of germ cell layers, Johnson's criteria, germ cell apoptosis, body weight and relative sex organs weight. RESULTS: Acute EO increased the relative weight of prostate and seminal vesicles (P≤0.05). It also reduced sperm quality such as total motility, sperm concentration (P≤0.01), and the percentage of normal sperm (P≤0.001). Moreover, acute EO decreased Miller's (P≤0.05) and Johnsen's scores and increased apoptotic indexes of spermatogenic cells (P≤0.001). KRG treatment decreased prostate weight gain (P≤0.05) and improved the percentage of sperm with normal morphology, total motility (P≤0.01), and progressive motility (P≤0.05). The apoptotic indexes of spermatogenic cells reduced (P≤0.001), whereas both Johnsen's (P≤0.01) and Miller's criteria increased in the KRG-treated EO testis (P≤0.05). CONCLUSION: Consequently, KRG ameliorated the devastating effects of EO on the sperm retrieved from either epididymis or testicle in rats.
RESUMO
Feature outcome of hippocampus and extra-hippocampal cortices was evaluated in melatonin treated lithium-pilocarpine epileptic rats during early and chronic phases of temporal lobe epilepsy (TLE). After status epilepticus (SE) induction, 5 and 20 mg/kg melatonin were administered for 14 days or 60 days. All animals were killed 60 days post SE induction and the histological features of the rosrto-caudal axis of the dorsal hippocampus, piriform and entorhinal cortices were evaluated utilizing Nissl, Timm, and synapsin I immunoflorescent staining. Melatonin (20 mg/kg) effect on CA1 and CA3 neurons showed a region-specific pattern along the rostro-caudal axis of the dorsal hippocampus. The number of counted granular cells by melatonin (20 mg/kg) treatment increased along the rostro-caudal axis of the dorsal hippocampus in comparison to the untreated epileptic group. The density of Timm granules in the inner molecular layer of the dentate gyrus decreased significantly in all melatonin treated groups in comparison to the untreated epileptic animals. The increased density of synapsin I immunoreactivity in the outer molecular layer of the dentate gyrus of untreated epileptic rats showed a profound decrease following melatonin treatment. There was no neuronal protection in the piriform and entorhinal cortices whatever the melatonin treatment. Long-term melatonin administration as a co-adjuvant probably could reduce the post-lesion histological consequences of TLE in a region-specific pattern along the rostro-caudal axis of the dorsal hippocampus.