Hypoxia-conditioned extracellular vesicles from human amniotic membrane stem cells attenuate D-galactose-induced aging in mice

Figure 1 Characteristics of extracellular vesicles obtained from amniotic membrane stem cells. A: Size distribution of extracellular vesicle particles from nanoparticle-tracking analysis; B: Concentrations of growth factors and neurotrophic factors in normoxic (white bars) and hypoxic (black bars) culture extracellular vesicles. ^aP < 0.05, compared with each normoxic culture extracellular vesicles. BDNF: Brain-derived neurotrophic factor; NGF: Nerve growth factor; PDGF: Platelet-derived growth factor; IGF: Insulin-like growth factor; VEGF: Vascular endothelial growth factor.

Figure 2 Body weights of D-galactose-induced aging rats treated with extracellular vesicles. D-galactose (300 mg/kg) was subcutaneously administered every day for 6 weeks, and extracellular vesicles (6 × 10⁸ particles/body) were intravenously injected at 2-week intervals (on days 0, 13, 26, and 41). EVs: Extracellular vesicles.

Figure 3 Effects of extracellular vesicles on the Rota-rod performance of D-galactose-induced aging rats on days 1, 7, 21, and 35. D-galactose (300 mg/kg) was subcutaneously administered every day for 6 weeks, and extracellular vesicles (6 × 10⁸ particles/body) were intravenously injected at 2-week intervals (on days 0, 13, 26, and 41). ^aP < 0.05, compared with normal control; ^bP < 0.05, compared with D-galactose alone. EVs: Extracellular vesicles; D-Gal: D-galactose.

Figure 4 Effects of extracellular vesicles on the maximum swimming time of D-galactose-induced aging rats on days 2, 8, 22, and 36. D-galactose (300 mg/kg) was subcutaneously administered every day for 6 weeks, and extracellular vesicles (6 × 10⁸ particles/body) were intravenously injected at 2-week intervals (on days 0, 13, 26, and 41). ^aP < 0.05, compared with normal control; ^bP < 0.05, compared with D-galactose alone. EVs: Extracellular vesicles; D-Gal: D-galactose.

Figure 5 Effects of extracellular vesicles on the passive avoidance performance of D-galactose-induced aging rats on day 43. D-galactose (300 mg/kg) was subcutaneously administered every day for 6 weeks, and extracellular vesicles (6 × 10⁸ particles/body) were intravenously injected at 2-week intervals (on days 0, 13, 26, and 41). ^aP < 0.05, compared with normal control; ^bP < 0.05, compared with D-galactose alone. EVs: Extracellular vesicles.

Figure 6 Effects of extracellular vesicles on the brain acetylcholine, brain brain-derived neurotrophic factor, muscular brain-derived neurotrophic factor, and brain sirtuin 1 in D-galactose-induced aging rats on day 44. A: Effects of extracellular vesicles (EVs) on the brain acetylcholine in D-galactose (D-Gal)-induced aging rats on day 44; B: Effects of EVs on the brain brain-derived neurotrophic factor in D-Gal-induced aging rats on day 44; C: Effects of EVs on the muscular brain-derived neurotrophic factor in D-Gal-induced aging rats on day 44; D: Effects of EVs on the brain sirtuin 1 in D-Gal-induced aging rats on day 44. D-Gal (300 mg/kg) was subcutaneously administered every day for 6 weeks, and EVs (6 × 10⁸ particles/body) were intravenously injected at 2-week intervals (on days 0, 13, 26, and 41). ^aP < 0.05, compared with normal control; ^bP < 0.05, compared with D-galactose alone. ACh: Acetylcholine; BDNF: Brain-derived neurotrophic factor; SIRT1: Sirtuin 1; EVs: Extracellular vesicles; D-Gal: D-galactose.

Figure 7 Effects of extracellular vesicles on the brain glial fibrillary acidic protein, muscular glycogen, and muscular thiobarbituric acid-reactive substances in D-galactose-induced aging rats on day 44. A: Effects of extracellular vesicles (EVs) on the brain glial fibrillary acidic protein in D-galactose (D-Gal)-induced aging rats on day 44; B: Effects of EVs on the muscular glycogen in D-Gal-induced aging rats on day 44; C: Effects of EVs on the muscular thiobarbituric acid-reactive substances in D-Gal-induced aging rats on day 44. D-Gal (300 mg/kg) was subcutaneously administered every day for 6 weeks, and EVs (6 × 10⁸ particles/body) were intravenously injected at 2-week intervals (on days 0, 13, 26, and 41). ^aP < 0.05, compared with normal control; ^bP < 0.05, compared with D-galactose alone. GFAP: Glial fibrillary acidic protein; TBARS: Thiobarbituric acid-reactive substances; EVs: Extracellular vesicles; D-Gal: D-galactose.