Impact of fetal brain tissue derived mesenchymal stem cell and fibrin glue on facial nerve crash injury

Background/aim: To evaluate the clinical and histopathological effects of fetal brain tissue derived mesenchymal stem cells (FBTMSC) and fibrin glue (FG) on the facial nerve (FN) regeneration in rats with traumatic FN injury. Materials and methods: Twenty-eight Sprague Dawley rats were included in the study and divided into 4 groups. Traumatic FN injury (FP) was created by a surgical clamp compression to the main trunk of left FN in all groups. In the control group (group 1) no treatment was applied, in group 2 (FBTMSC group) 2 × 106 FBTMSC was injected, in group 3 (FG group) only FG was applied, in group 4 (FBTMSC and FG groups) both FBTMSC and FG were applied to the injured section of the nerve. The FN functions were evaluated clinically, immediately after the procedure and at 3rd, 5th, and 8th weeks postoperatively. The FNs of all subjects were excised after the 8th week; then the rats were sacrificed. The presence of stem cells in the injured zone was assessed using bromo-deoxyuridine (BrdU), and apoptosis was determined by the TUNEL method. Results: After the damage, total FP was observed in all subjects. Statistically significant functional improvement was observed in group 4 compared to all other groups (P < 0.005). TUNEL-positive cell count was statistically significantly higher in the control group than the other groups (P < 0.001). TUNEL-positive cell count was statistically significantly lower in group 4 than the other groups. The proportion of BrdU-stained cells in group 4 (5%) was higher than group 2 (2%). Conclusion: Clinically and histopathologically FBTMSC applied with FG may play a promising role as a regenerative treatment in posttraumatic FP.

Effects of uterus derived mesenchymal stem cells and their exosomes on asherman's syndrome.

Asherman's syndrome has become a growing problem with the incidence of cesarean and endometrial surgical procedures. A surgical procedure that can damage to the basal layer of the endometrium is formed as intrauterine adhesion and can cause asherman's syndrome. Mesenchymal stem cells (MSCs) are characterized by some characteristics such as non-immunogenic, angiogenic, antifibrotic, antiapoptotic and antiinflammatory properties, also they support tissue repair by secretion of various factors and chemokines in cellular therapy. Exosomes are active paracrine components with a great potential for repairing damaged tissue. Exosomes include many paracrine factors responsible for regeneration and angiogenesis. In this study, 10 newborn Wistar rats were used to obtain MSCs. A total of 24 adult Wistar rats were also used. The rats were divided into 4 groups: untreated control group; asherman control group; asherman + uterine-derived MSCs group; asherman + uterine-derived MSCs-exosomes group. At the end of the experiment, uterine tissues were evaluated by histochemical and immunohistochemical. As a result of MSCs and exosomes treatments, proliferation and vascularization in uterine tissue was increased. It was also shown to reduce fibrosis with masson's trichrome staining. MMP-2 and MMP-9 expression was enhanced by MSC and exosomal therapy; in addition, TIMP-2 expression was decreased. In our study, it was shown that proliferation and vascularization increased and fibrosis decreased in uterus as a result of MSC and exosome treatments. Our results indicate that the exosomal treatment restored the damage of asherman's syndrome at tissue at a shorter time than the MSCs group.

Sutureless approach with vein grafts and mesenchymal stem cells in primary nerve repair: Functional and immunohistological results.

PURPOSE: The aim of this study was to define a sutureless peripheral nerve repair technique with a vein graft and bone marrow-derived stem cells (BMSC) and compare it to epineural repair. MATERIALS AND METHODS: Thirty Wistar Albino rats were divided into five groups evenly. In the control group (C), epineural repair was performed. In the SV (suture + vein) and MSV (BMSC + suture + vein) groups, epineural repair was wrapped with a vein graft. In the V (vein) and MV (vein + BMSC) groups, sutureless repair using a vein graft was performed by taking sutures away from the regeneration site. Rats were evaluated with pinprick, toe spread tests and sciatic nerve index (SFI) at 4th, 8th, and 12th weeks. They were sacrificed at 12th week, repair sites were harvested and evaluated immunohistochemically. RESULTS: There was no difference in pinprick and toe spread tests between the groups at 12th week. The mean SFI was -76.5 ± 3.7, -65.2 ± 11.7, -46.2 ± 19.4, -68.8 ± 9.8, -56 ± 8.8 in the C, SV, MSV, V, MV groups, respectively. The MSV group showed significantly the best SFI results (P < .05). NF-H immunostaining scores were as C; 1 ± 0.18, SV; 2.5 ± 0.36, MSV; 4 ± 0.49, V; 1.56 ± 0.54, MV; 3 ± 0.39, whereas GAP-43 scores were as C; 1 ± 0.31, SV; 2.66 ± 0.56, MSV; 4.50 ± 0.23, V; 2 ± 0.23, MV; 3 ± 0.6. The best nerve regeneration according to immunostaining results was observed in the MSV group (P < .05). The mean fibrosis area was 221.5 ± 25.9, 101.6 ± 7.1, 121.3 ± 18.8, 150.3 ± 12.1, 152.4 ± 11.8 µm2 in the above groups, respectively. SV and MSV groups showed the significantly less fibrosis area (P < .05). CONCLUSION: Epineural suture repair combined with vein wrapping and BMSCs (MSV) showed the best SFI, GAP-43, and NF-H immunostaining results.

Histopathological effects of mesenchymal stem cells in rats with bladder and posterior urethral injuries

Background/aim: To evaluate the effects of mesenchymal stem cell (MSC) therapy in an experimental bladder and posterior urethral injury model. Materials and methods: The study subjects consisted of 40 male Wistar albino rats that were divided into four groups: control group (n = 10) (the bladder was only surgically opened and closed), sham group (n = 10) (surgical procedure), IVMSC group (n= 10) (surgical procedure and intravenous MSC treatment), and LMSC group (n = 10) (surgical procedure and local MSC treatment). Histopathological evaluation was performed for the degree of fibrosis and inflammation and the extent and intensity of staining of vascular endothelial growth factor (VEGF) and endoglin (CD105). Results: There were no significant differences between the control and LMSC groups with respect to fibrosis (P = 0.070) or inflammation (P = 0.048). Fibrosis and inflammation were significantly lower in the IVMSC (P = 0.034 for fibrosis, P = 0.080 for inflammation) and LMSC (P = 0.01 for fibrosis, P = 0.013 for inflammation) groups when compared with the sham group. No significant differences regarding fibrosis and inflammation were observed between the IVMSC and LMSC groups (P = 0.198 for fibrosis, P = 0.248 for inflammation). A significant difference was noted between the sham and LMSC groups concerning VEGF staining intensity (P = 0.017). However, no significant difference was found among the groups with regard to the extent or intensity of CD105 staining (P > 0.05). Conclusion: MSC treatment significantly decreased the development of fibrosis in a uroepithelial injury model.

Effects of commonly used intravitreal anti-vascular endothelial growth factor drugs on mesenchymal stem cells derived from the limbus and ciliary body.

BACKGROUND: To investigate the effects of commonly used intravitreal anti-vascular endothelial growth factor (anti-VEGF) antibodies on proliferation index and viability of mesenchymal stem cells derived from ciliary body and limbus (CB-MSC and LMSC). METHODS: CB-MSCs and LMSCs were isolated from newborn rats' eyes, and they were expanded in medium by the explant method. Intravitreally used anti-VEGF drugs, aflibercept, bevacizumab and ranibizumab were tested into the 16-well plates, respectively, at four different concentrations. After keeping them for 48 h, the proliferation indexes and viabilities of CB-MSCs and LMSCs were compared separately by Real-Time Cell Analyzer and Methylthiazoltetrazoli (MTT) test. RESULTS: Anti-VEGFs used at 5-times and 10-times of the standard clinical dosage caused statistically significant negative effects on proliferation indexes of CB-MSCs and LMSCs at the 24th hour compared to control group. Only the anti-VEGF group that had 10-times dosage of those used clinically had a statistically significant negative effect on the viabilties of CB-MSCs and LMSCs. CONCLUSION: Administrations of high doses or repeated standard doses of intravitreal anti-VEGF agents may affect the proliferation indexes and viabilities of CB-MSCs and LMSCs adversely. These novel findings deserve further in vivo investigations.

The effect of mesenchymal stem cells and sildenafil on flap viability in perforator-based flaps for ischemia/reperfusion injury: An experimental study.

PURPOSE: In this study, we evaluated the individual and combined effects of mesenchymal stem cells (MSCs) and sildenafil citrate (SC) on the viability of pedicled perforator flaps in which ischemia/reperfusion injury developed after induction of primary ischemia. MATERIALS AND METHODS: Seven Sprague-Dawley rats were used as donors of cells. Rectangular flaps (7 × 7 cm2 ) were created featuring the right second epigastric musculocutaneous perforator in 63 male Sprague-Dawley rats. The animals were randomly divided into two experimental groups (based on the ischemia time of 4 or 8 hours) and a control group. Each of the experimental group was further divided into four subgroups with no treatment, subcutaneous administration of MSCs after termination of ischemia, intraperitoneal administration of SC after termination of ischemia, and combined MSCs and SC treatments at the end of the period of ischemia (n = 7 for each subgroup). A sham group with no-ischemia to flap was used as the control (n = 7). On day 7, viable areas on the flaps were calculated from photographs. The levels of the antioxidative enzymes, such as superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX), were analyzed in tissue samples obtained from the most distal regions of the flap prior to ischemia and on day 7 after induction of ischemia. RESULTS: No difference was detected between the no-ischemia group and 4-hours SC-treated subgroup, 4-hours combined MSC and SC treated subgroup, 8-hours MSC-treated subgroup, or 8-hours SC-treated subgroup (P > 0.05). In 4-hours ischemia group, the viable flap area of combined MSC and SC treated subgroup was significantly greater than that of the ischemia subgroup (17.17 ± 12.56 cm2 vs. 7.24 ± 7.17 cm2 ; P = 0.015). However, in 8-hours ischemia group, the viable flap area of MSC- treated subgroup was significantly greater than that of the ischemia subgroup (2.69 ± 3.71 cm2 vs. 14.52 ± 8.57 cm2 ; P = 0.004). There were no significant differences in SOD, CAT, and GPX levels detected between no-ischemia group and any of the treated subgroups in 4- and 8-hours ischemia groups (P > 0.05). However, SOD, CAT, and GPX levels in the no-ischemia group were lower than that in 4-hours ischemia control subgroup or 8-hours ischemia control subgroup (P < 0.05). CONCLUSION: In this rat pedicled perforator based abdominal flap, we found that after primary ischemia, application of MSCs and SC, either individually or in combined form, significantly enhanced antioxidant enzyme levels compared with those in the control group, and provided protection against ischemia/reperfusion injury. The two treatments acted synergistically to protect against damage after 4 hours of ischemia, but either treatment alone more effectively enhanced viable flap area after 8 hours of ischemia, although some flap damage was apparent. © 2015 Wiley Periodicals, Inc. Microsurgery 36:402-409, 2016.