Application of temperature-sensitive chitosan hydrogel encapsulated exosomes in ischemic diseases

doi:10.3969/j.issn.2095-4344.3130

Abstract

Abstract: BACKGROUND: Exosomes derived from mesenchymal stem cells can reduce myocardial ischemia and reperfusion injury, but there are some problems such as short half-life, fast clearance and low targeting.
OBJECTIVE: To modify and encapsulate exosomes with temperature-sensitive chitosan hydrogel to increase the retention rate of exosomes in the body, and to achieve better therapeutic effect.
METHODS: Cell transfection method was used to knock down piR823 in exosomes derived from umbilical cord mesenchymal stem cells, and the effect of knockdown of piR823 exosomes on the proliferation and apoptosis of C2C12 cells was detected. Chitosan/β-sodium glycerophosphate temperature-sensitive hydrogel was prepared and mixed directly with exosomes to prepare chitosan/β-sodium glycerophosphate temperature-sensitive hydrogel encapsulated with exosomes. The gel-forming properties, rheology and in vitro sustained release properties of the hydrogel after encapsulation of exosomes were tested. Thirty C57BL/6J mice were taken to establish hind limb ischemia models, and randomly divided into five groups. The gastrocnemius of group A was injected with PBS; group B was injected with chitosan/β-sodium glycerophosphate temperature-sensitive hydrogel; and group C was injected with exosomes-encapsulated chitosan/β-sodium glycerophosphate temperature-sensitive hydrogel; group D was injected with exosomes; group E was injected with exosomes knocking down piR823. Limb function and recovery, blood flow, grip strength, exercise endurance and muscle regeneration were detected in each group.
RESULTS AND CONCLUSION: (1) Exosomes knocking down piR823 inhibited the proliferation of C2C12 cells; normal exosomes inhibited hydrogen peroxide-induced apoptosis of C2C12 cells. The inhibitory effect of exosomes on apoptosis was weakened after piR823 was knocked down. (2) The hydrogel encapsulating exosomes had gel-forming properties, but the gel-forming time was shortened, and it could slowly and continuously release exosomes for more than 30 days. (3) After 28 days of hindlimb ischemia, the blood flow recovery of the left limb in group C was better than that in groups B and D (P < 0.05); it in group D was better than group E (P < 0.05); the grip strength, endurance, running time and distance of group C were better than those of groups D and B (P < 0.05), and above indexes in group D were better than in group E (P < 0.05). Muscle regeneration was better in group C than in groups B and D (P < 0.05), and it in group D was better than group E (P < 0.05). (4) The results showed that encapsulation of exosomes by chitosan/β-sodium glycerophosphate temperature-sensitive hydrogel prolonged the residence time of exosomes in vivo, significantly enhanced blood perfusion and recovery of tissue function after ischemia, and the treatment effect was more significant.

Key words: material, exosomes, chitosan, temperature-sensitive hydrogel, mesenchymal stem cells, hindlimb ischemia, animal mode, piRNA

CLC Number:

Liu Feng, Zhang Yu, Wang Yanli, Luo Wei, Han Chaoshan, Li Yangxin. Application of temperature-sensitive chitosan hydrogel encapsulated exosomes in ischemic diseases[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(16): 2479-2487.

Figures/Tables 9

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