The Relationship between Osteoinduction and Vascularization: Comparing the Ectopic Bone Formation of Five Different Calcium Phosphate Biomaterials

Yun He; Yu Peng; Lishuang Liu; Sha Hou; Junyu Mu; Liang Lan; Lijia Cheng; Zheng Shi

doi:10.3390/ma15103440

The Relationship between Osteoinduction and Vascularization: Comparing the Ectopic Bone Formation of Five Different Calcium Phosphate Biomaterials

Materials (Basel). 2022 May 10;15(10):3440. doi: 10.3390/ma15103440.

Authors

Yun He¹, Yu Peng¹, Lishuang Liu¹, Sha Hou¹, Junyu Mu¹, Liang Lan¹, Lijia Cheng¹, Zheng Shi¹

Affiliation

¹ College of Basic Medicine, Clinical Medical College, Affiliated Hospital of Chengdu University, Chengdu 610106, China.

Abstract

Objective: The objective of this study is to compare the bone induction of five kinds of calcium phosphate (Ca-P) biomaterials implanted in mice and explore the vascularization and particle-size-related osteoinductive mechanism. Methods: The following five kinds of Ca-P biomaterials including hydroxyapatite (HA) and/or tricalcium phosphate (TCP) were implanted in the muscle of 30 BALB/c mice (n = 6): 20 nm HA (20HA), 60 nm HA (60HA), 12 µm HA (12HA), 100 nm TCP (100TCP) and 12 µm HA + 100 nm TCP (HATCP). Then, all animals were put on a treadmill to run 30 min at a 6 m/h speed each day. Five and ten weeks later, three mice of each group were killed, and the samples were harvested to assess the osteoinductive effects by hematoxylin eosin (HE), Masson’s trichrome and safranine−fast green stainings, and the immunohistochemistry of the angiogenesis and osteogenesis markers CD31 and type I collagen (ColI). Results: The numbers of blood vessels were 139 ± 29, 118 ± 25, 78 ± 15, 65 ± 14 in groups HATCP, 100TCP, 60HA and 20HA, respectively, which were significantly higher than that of group 12HA (12 ± 5) in week 5 (p < 0.05). The area percentages of new bone tissue were (7.33 ± 1.26)% and (8.49 ± 1.38)% in groups 100TCP and HATCP, respectively, which were significantly higher than those in groups 20HA (3.27 ± 0.38)% and 60HA (3.43 ± 0.27)% (p < 0.05); however, no bone tissue was found in group 12HA 10 weeks after transplantation. The expression of CD31 was positive in new blood vessels, and the expression of ColI was positive in new bone tissue. Conclusions: Nanoscale Ca-P biomaterials could induce osteogenesis in mice muscle, and the osteoinductive effects of TCP were about 124% higher than those of 20HA and 114% higher than those of 60HA. The particle size of the biomaterials affected angiogenesis and osteogenesis. There was a positive correlation between the number of blood vessels and the area percentage of new bone tissue; therefore, osteoinduction is closely related to vascularization. Our results provide an experimental basis for the synthesis of calcium−phosphorus matrix composites and for further exploration of the osteoinductive mechanism.

Keywords: hydroxyapatite; nanoscale; osteoinduction; tricalcium phosphate; vascularization.

Grants and funding

This research was funded by The Key Laboratory of Medical Clinical Genetics of Sichuan Province, China (ZD-01-05), Medical Scientific Research Project of Chengdu City, China (2021043); CC National Creative Space Innovation and Entrepreneurship Education Special topic of Chengdu University (ccyg202101008); Higher Education Talent Training Quality and Teaching Reform Project of Education Department of Sichuan Province, China (JG2021-1102); College Student Innovation and Entrepreneurship Training Program of Sichuan province, China (S202111079028, S202111079043X, S202111079095, S202111079124X, S202111079041X).