Dextran/poly(lactic-co-glycolic acid) microspheres combined with three growth factors promote neovascularization in ischemic lower limbs of rats

doi:10.3969/j.issn.2095-4344.1353

Abstract

Abstract:

BACKGROUND: The dextran/poly(lactic-co-glycolic acid) (PLGA) microspheres as a sustained release drug delivery system enables the drug to maintain an effective concentration at the site of action.

OBJECTIVE: To investigate the effects of dextran/PLGA microspheres carrying granulocyte colony-stimulating factor, erythropoietin and vascular endothelial growth factor on the neovascularization of ischemic lower limb of rats.

METHODS: Dextran/PLGA microspheres carrying granulocyte colony-stimulating factor, erythropoietin, and vascular endothelial growth factor were prepared and their surface morphology was observed by scanning electron microscopy. The encapsulation efficiency and cumulative release rate were detected by ELISA method. Dextran/PLGA microspheres carrying three growth factors were mixed with fibrin glue to prepare a growth factor dextran/PLGA/fibrin glue complex. Right lower limb ischemia models were produced in 24 male Sprague-Dawley rats (Laboratory Animal Center, Jiangsu University of Traditional Chinese Medicine, China). These animal models were randomly divided into two groups (n = 12/group). In the experimental group, dextran/PLGA/fibrin glue complex carrying three growth factors was injected into the inner thigh muscle at 5 points. In the control group, dextran/PLGA/fibrin glue complex without growth factors was identically injected. At 1 week after surgery, the muscle tissue around the injection site was harvested for immunohistochemistry to detect the expression of proliferating cell nuclear antigen, Bcl-2, stromal cell-derived factor 1, and C-X-C chemokine receptor type 4. At 4 weeks after surgery, the muscle tissue around the injection site was harvested for histological and immunohistochemical examination to measure capillary density. This study was approved by Animal Ethics Committee of Nanchang University School of Medicine, China.

RESULTS AND CONCLUSION: The dextran/PLGA microspheres were spherical and had a smooth surface with a diameter of 40-120 μm. The encapsulation efficiency of dextran/PLGA microspheres carrying granulocyte colony-stimulating factor, erythropoietin, and vascular endothelial growth factor was 84%, 85%, and 82%, respectively. The 4-week cumulative release rate was 89.5%, 90.3%, 91.2%, respectively. The expression of proliferating cell nuclear antigen, Bcl-2, stromal cell-derived factor 1, and C-X-C chemokine receptor type 4 in the experimental group was significantly higher than that in the control group. Capillary density and α-smooth muscle actin-positive vascular density in the experimental group were significantly higher than those in the control group (P < 0.05). These results suggest that dextran/PLGA microspheres carrying multiple growth factors for treatment of lower limb ischemia is a promising therapeutic method.

Key words: poly(lactic-co-glycolic acid), granulocyte colony-stimulating factor, erythropoietin, vascular endothelial growth factor, microsphere, ischemia, dextran, angiogenesis

CLC Number:

Zhu Xianhua, Chen Feng, Liu Chongdong, Zhou Wei, Tang Xinhua . Dextran/poly(lactic-co-glycolic acid) microspheres combined with three growth factors promote neovascularization in ischemic lower limbs of rats[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(26): 4142-4147.

Figures/Tables 9

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