AAS ›› 2013, Vol. 44 ›› Issue (3 ): 386-392.doi: 10.3969/j.issn.0529-1356.2013.03.019

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Nano-hydroxyapatite-collegen-chitosan substance compound with bone marrow mesenchymal stem cells to repair tibia defect in mice

LI Yang FANG Yan BAI Shu-ling* TIAN Xiao-hong TONG Hao HOU Wei-jian   

  1. Department of Tissue Engineering,China Medical University, Shenyang 110001,China
  • Received:2012-05-15 Revised:2012-07-08 Online:2013-06-06 Published:2013-07-16

Abstract:

Objective To use nano-hydroxyapatite-collegen-chitosan compounded with bone marrow mesenchymal stem cells (BMSCs)
to repair tibia bulk defect in mice, and to investigate the application effectiveness of the composite to the long bone defect.
Methods BMSCs of mice were isolated and cultured. Biological characteristics of the stem cells were detected. The
nano-hydroxyapatite-collegen-chitosan was prepared. The structure, poriness and degradation rate of the composite were observed
under a scanning electronic microscope(SEM). Male mice with a 5 mm tibia defect were randomly divided into three groups:
group A treated with the nano-hydroxyapatite-collegen-chitosan compound with BMSCs; group B with the nano-hydroxyapatite-collegen-chitosan
and group C with the nanoscale hydroxyapatite compound with BMSCs. At 8 weeks after restoration, the mice were anesthetize
and samples were collected. HE staining and X-ray were used to observe the osteogenesis and healing process in the defect
area. Results The BMSCs presented fibroblast morphology and expressed the surface markers of mesenchymal stem cells which were
of the biological characteristics of stem cells. The nano-hydroxyapatite-collegen-chitosan had abundant pores and poriness
with a relatively even diameter of 100-200μm and an appearance ratio of 72.3%-92.7% (an average of about 82.5%). The degradation
rate was(60.3±5.4)% . There were extensive communications inside the scaffold. At the 8th week after transplantation, there
was no obvious inflammatory response but mass mature osteoid structure, and the scaffold degraded mostly in group A. X-ray
showed that the defect area had new callus,increased new bone density and the bridge grafting in group A. The above featrues
were less obvious in groups B and C than that in group A(P<0.01). Conclusion Nano-hydroxyapatite-collegen-chitosan compounded
with BMSCs works well for repairing the tibia bulk defect in mice.

Key words: Bone marrow mesenchymal mesenchymal stem cell, Hydroxyapatite, Collegen, Chitosan, Bone defect, Scanning electronic microscopy, Mouse