ABSTRACT
OBJECTIVES@#To explore the effect of hydrogel loaded with exosomes from Wharton's Jelly-derived mesenchymal stem cell (WJMSC) on wound healing.@*METHODS@#Exosomes were extracted from WJMSC, and the morphology and size of WJMSC-derived exosomes (WEX) were analyzed by transmission electron microscopy and nanoparticle size analyzer, respectively. The surface markers CD9, CD81, and Calnexin of WEX were detected by Western blotting. Exosome-loaded alginate hydrogel (WEX-gel) was prepared; its morphology was studied by scanning electron microscope, and its rheological behavior was examined by a rheometer. The in vitro drug release performance of WEX-gel was investigated by BCA method. RAW264.7 cells were treated with alginate hydrogel, WEX and WEX-gel, respectively; and the expression of CD86 and CD206 in macrophages was detected by flow cytometry. A full-thickness skin wound model was established in mice; the model mice were randomly divided into blank control group, WEX control group and WEX-gel group, and PBS, WEX and WEX-gel were applied to the wound area of mice, respectively. On day 3, the skin tissue of mice was excised, and the antibacterial effect of WEX hydrogel was evaluated by plate counting. On day 15, the mice were euthanized and the percentage of residual wounds was calculated. The histological changes of the skin wound were observed after hematoxylin and eosin (HE) and Masson stainings. The expression of CD86, CD206, CD31 and vascular endothelial growth factor (VEGF) in the skin wound tissue was detected by immunohistochemistry.@*RESULTS@#Exosomes were successfully extracted from WJMSC. WEX-gel presented a regular three-dimensional network structure, good rheology and controlled drug release performance. WEX-gel promoted the polarization of RAW264.7 cells from the M1 phenotype to M2 phenotype in vitro. The residual wound percentage in blank control group, WEX control group and WEX-gel group were (27.5±3.4)%, (15.3±1.2)% and (7.6±1.1)%, respectively (P<0.05). The antibacterial property of WEX-gel is better than that of WEX (P<0.05). The dermis thickness, the number of new hair follicles, and the rate of collagen deposition in the WEX-gel group were significantly higher than those in the other two groups (all P<0.05). The expression of CD206, CD31 and VEGF in skin wound tissue was higher and the expression of CD86 was lower in WEX-gel group than those in other two groups (all P<0.05).@*CONCLUSIONS@#WEX-gel can significantly promote wound healing in mice by regulating the polarization of macrophages.
Subject(s)
Mice , Animals , Vascular Endothelial Growth Factor A , Wharton Jelly , Exosomes , Hydrogels , Wound Healing/physiology , Mesenchymal Stem Cells , Anti-Bacterial Agents , AlginatesABSTRACT
Objective:To investigate the effect of monosialotetrahexosylganglioside sodium treatment on neurological function, inflammatory factor, and blood coagulation function in patients with traumatic brain injury.Methods:The clinical data of 90 patients with traumatic brain injury who received treatment in Taizhou Central Hospital from February 2018 to May 2020 were retrospectively analyzed. These patients were divided into a control group ( n = 46) and an observation group ( n = 44) according to different treatment methods. The control group was given routine symptomatic treatment and the observation group was given monosialotetrahexosylganglioside sodium treatment based on routine symptomatic treatment. Remission rate, inflammatory factor level, the National Institutes of Health Stroke Scale score, Glasgow Outcome Scale score, and coagulation function were compared between the two groups at each time point. Results:At 3 days and 2 weeks post-surgery, neuropeptide Y in the observation group was (121.13 ± 12.68) ng/L and (68.52 ± 10.21) ng/L, tumor necrosis factor α was (96.15 ± 8.16) ng/L and (46.68 ± 5.95) ng/L, interleukin-6 was (231.26 ± 9.41) ng/L and (126.74 ± 12.23) ng/L, C-reactive protein was (47.52 ± 4.32) μg/L and (18.65 ± 1.32) μg/L, the National Institutes of Health Stroke Scale score was (20.12 ± 2.22) points and (17.67 ± 1.31) points. They were significantly lower than those in the control group [neuropeptide Y: (135.69 ± 15.42) ng/L, (79.36 ± 11.15) ng/L; tumor necrosis factor-α: (108.56 ± 10.13) ng/L, (69.33 ± 6.42) ng/L; interleukin-6: (264.13 ± 10.24) ng/L and (157.89 ± 12.13) ng/L; C-reactive protein: (65.19 ± 5.17) μg/L and (24.39 ± 3.45) μg/L; the National Institutes of Health Stroke Scale score: (24.56 ± 2.54) points and (20.39 ± 2.55) points] ( t3 days post-surgery = 4.88, 6.38, 15.83, 17.55, 8.81; t2 weeks post-surgery= 4.80, 17.33, 12.12, 10.33, 6.32, all P < 0.001). At 3 days and 2 weeks post-surgery, the Glasgow Outcome Scale score in the observation group was (3.65 ± 0.35) points and (4.65 ± 0.26) points, respectively, which was significantly higher than (3.15 ± 0.10) points and (4.11 ± 0.11) points in the control group ( t = 9.30, 12.93, both P < 0.05). At 3 days and 2 weeks post-surgery, fibrinogen in the observation group was (4.52 ± 0.39) g/L and (3.12 ± 0.10) g/L, thrombin time was (18.46 ± 2.95) seconds and (21.79 ± 2.45) seconds, prothrombin time was (12.42 ± 1.33) seconds and (15.79 ± 2.36) seconds, activated partial thromboplastin time was (34.59 ± 2.64) seconds and (38.98 ± 2.78) seconds, which were significantly superior to those in the control group [fibrinogen: (5.02 ± 0.13) g/L and (4.29 ± 0.16) g/L; thrombin time: (17.36 ± 1.56) seconds and (19.63 ± 1.62) seconds; prothrombin time: (10.69 ± 1.21) seconds and (13.26 ± 1.78) seconds; activated partial thromboplastin time: (32.16 ± 2.59) seconds and (35.69 ± 2.91) seconds] ( t3 days post-surgery = 8.23, 2.22, 6.46, 4.40; t2 weeks post-surgery = 41.38, 4.95, 5.75, 5.48, all P < 0.001). At 1 and 2 weeks post-surgery, the remission rate in the observation group was significantly higher than that in the control group ( χ2 = 4.75, 4.44, both P < 0.05). Conclusion:Monosialotetrahexosylganglioside sodium treatment for a traumatic brain injury can inhibit inflammatory reactions, improve blood coagulation and protect brain tissue.