Effects of bone marrow mesenchymal stem cells and human umbilical vein endothelial cells combined with hydroxyapatite-tricalcium phosphate scaffolds on early angiogenesis in skull defect repair in rats

doi:10.3969/j.issn.2095-4344.3207

Abstract

Abstract: BACKGROUND: Tissue-engineered bone provides a new direction for the repair of large bone defects, but the survival of tissue-engineered bone grafts in vivo can be guaranteed only if the capillary network is formed in the early stage. Therefore, vascularization of early tissue-engineered bone is the key to its clinical application.
OBJECTIVE: To explore the effect of bone marrow mesenchymal stem cells and human umbilical vein endothelial cells combined with hydroxyapatite-tricalcium phosphate scaffolds on early angiogenesis in skull defect repair in rats.
METHODS: (1) The hydroxyapatite-tricalcium phosphate scaffold was divided into four groups: Group A was added with DMEM culture medium and extracellular matrix culture medium with volume ratio of 1:1. Group B was added with bone marrow mesenchymal stem cells suspension and human umbilical vein endothelial cells suspension with a volume ratio of 1:1. Group C was cultured with DMEM culture medium and human umbilical vein endothelial cells suspension with a volume ratio of 1:1. Group D was cultured with bone marrow mesenchymal stem cells suspension and extracellular matrix medium with a volume ratio of 1:1 for 21 days for in vivo implantation experiment. (2) Bone defects with a diameter of 1 cm were made in the skull of adult SD rats, and materials of groups A, B, C and D were implanted respectively. Skull micro-CT and skull scaffold complex gross, histomorphology, CD31/CD34 immunohistochemical staining, CD31/CD34 molecular content, and vascular endothelial growth factor A protein were detected respectively 4, 8 and 12 weeks after the operation. Animal experiment was approved by the Animal Ethics Committee of Southwest Medical University.
RESULTS AND CONCLUSION: (1) Micro-CT showed that 8 weeks after operation, the mass of new bone in group B was higher than that in the other three groups, and the mass of new bone in groups C and D was higher than that in group A. At 12 weeks after surgery, the new bone mass was from high to low: group B>group C>group D>group A. (2) Gross observation showed that the bone crawling area on the scaffold of group B was higher than that of the other three groups, and that of groups C and D was higher than that of group A. At the same time point, the amount of bloody exudate was from most to least: group B>group C>group D>group A. (3) Histological observation showed that at 4 weeks after the operation, the bone regeneration amount from high to low was: group B>group C>group D>group A. At other time points, the bone regeneration amount from high to low was: group B>group D>group C>group A. (4) Immunohistochemical staining showed that at the same time point, the microvascular density of CD31/CD34 was from high to low successively: group B>group C>group D>group A. Except for groups C and D, there was no significant difference (P > 0.05), and the difference between the other groups was significant (P < 0.05). (5) Western blot assay showed that at the same time point, the protein expression of vascular endothelial growth factor A was in descending order from high to low: group B>group C>group D >group A, except for no significant difference between groups C and D (P > 0.05), and the difference between the other groups was significant (P < 0.05). (6) The results showed that bone marrow mesenchymal stem cells and human umbilical vein endothelial cells combined with hydroxyapatite-tricalcium phosphate scaffold materials had strong early vascularization ability in repairing skull defects in rats.

Key words: bone, materials, scaffold, tissue engineering bone, bone defect repair, stem cells, human umbilical vein endothelial cells, early angiogenesis

CLC Number:

Chen Siqi, Xian Debin, Xu Rongsheng, Qin Zhongjie, Zhang Lei, Xia Delin. Effects of bone marrow mesenchymal stem cells and human umbilical vein endothelial cells combined with hydroxyapatite-tricalcium phosphate scaffolds on early angiogenesis in skull defect repair in rats[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(22): 3458-3465.

Figures/Tables 11

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