Evaluation of Vascularization of Porous Calcium Phosphate by Chick Chorioallantoic Membrane Model ex vivo

doi:10.15541/jim20160535

Journal of Inorganic Materials ›› 2017, Vol. 32 ›› Issue (6): 649-654.DOI: 10.15541/jim20160535

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Evaluation of Vascularization of Porous Calcium Phosphate by Chick Chorioallantoic Membrane Model ex vivo

XIAO Wen¹, LIU Yu-Mei¹, REN Kai-Ge², SHI Feng¹, LI Yan², ZHI Wei¹, WENG Jie¹, QU Shu-Xin¹

(1. Key Lab of Advanced Technologies of Materials, Ministry of Education, School of Material Science and Engineering, Southwest Jiaotong University, Chengdu 610031, China; 2. Department of Stomatology, Chengdu Army General Hospital, Chengdu 610083, China)

Received:2016-09-22 Revised:2016-11-02 Published:2017-06-20 Online:2017-05-27
About author:XIAO Wen. E-mail: 18728428596@163.com
Supported by:
National Basic Research Program of China (973 Program, 2012CB933602);National Natural Science Foundation of China (51372210);Basic Research Foundation Key Project of Sichuan Province (2016JY0011);Research Fund for the Doctoral Program of Higher Education of China (20130184110023);Fundamental Research Funds for the Central Universities (2682016YXZT11);The Construction Program for Innovative Research Team of University in Sichuan Province (14TD0050)

Abstract

Abstract:

In this study, ex vivo chick chorioallantoic membrane (CAM) model with abundant blood vessels was developed to investigate the vascularization of porous calcium phosphate. Porous hydroxyapatite ceramic (HA) and calcium phosphate cement with segmental porous structure (CPCs) were implanted into CAM, respectively. Stereomicroscope and scanning electron microscope (SEM) were employed to observe the angiogenesis on the surface of CPCs. The vascular density, diameters and numbers were quantified by Image-Pro Plus and Nano Measurer. Furthermore, undecalcified histological staining was conducted to observe the inner angiogenesis of porous HA. The vascular density and number in the porous part of CPCs both increased significantly with the increase of culture time, while the angiogenesis was tardily in the part of CPCs without pores. Most of the blood vessels less than 50 μm on the porous part of CPCs grew in budding manner to form vascular network. SEM images showed that blood vessels grew tightly on the surface of CPCs. Histological staining demonstrated that some blood vessels grew into the inside of porous HA. These results suggest that three-dimensional porous structure may promote the angiogenesis of porous calcium phosphate and the present CAM model may provide a convenient, efficient and with low-cost approach to evaluate vascularization of porous biomaterials.

Key words: chick chorioallantoic membrane, porous calcium phosphate materials, vascularization, ex vivo

CLC Number:

TB321

XIAO Wen, LIU Yu-Mei, REN Kai-Ge, SHI Feng, LI Yan, ZHI Wei, WENG Jie, QU Shu-Xin. Evaluation of Vascularization of Porous Calcium Phosphate by Chick Chorioallantoic Membrane Model ex vivo[J]. Journal of Inorganic Materials, 2017, 32(6): 649-654.

Figures/Tables 6

Fig. 1 Vascular network of CAM after pre-incubated for 8 d (a), CAM after implanted CPCs (b) and HA (c)

Fig. 2 Angiogensis on the surface of CPCs after implanted in CAM model for 2 d (a), 6 d (b) and 8 d (c). Angiogensis of different parts (1 (d), 2 (e) and 3 (f)) on the surface of CPCs after implanted for 8 d

Fig. 3 Total density of blood vessels on CPCs after implanted in CAM model for 2, 6 and 8 d (a). The percentage of vascular densities of CPCs, with pores and without pores (b)**p<0.01, n=3

Fig. 4 The distribution of vascular diameters (a), numbers of different vascular diameters (Φ<50 μm, 50 μm<Φ<100 μm and Φ>100 μm) (b) and the percentage of different diameters in different parts (with pores and without pores) (c) on the surface of CPCs after implanted for 2, 6 and 8 d*p<0.05, **p<0.01, n=3

Fig. 5 SEM images of the surface on CPCs after implanted in CAM model for 8 d

Fig. 6 H&E staining images of porous hydroxyapatite ceramic after implanted for 4 d in CAM modelBlack arrow: red blood cell grew into the inner pore of HA, white arrow: blood vessel grew into the inner pore of HA

References 20

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Evaluation of Vascularization of Porous Calcium Phosphate by Chick Chorioallantoic Membrane Model ex vivo

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