Compatibility of bone marrow mesenchymal stem cells with nano-hydroxyapatite/collagen

Yonghui HUANG; Jin TIAN; Qing XIA; Tiecheng SHEN; Wenrong XU

ABSTRACT

BACKGROUND:

The microcosmic and submicroscopic organizations of tissue engineering scaffold matedals’superficial structure have all important effect on the eell adhesion and growth.By means of nano.Technique and three-dimensional porous technique,the resultant nano-hydroxyapatite/collagen(n-HAC)call imitate the component and microstructure of natural bone.

OBJECTIVE:

To observe the biocompatibility of human bone m arrow mesenchymal stem cells(MSCs)cultured in vitro with nHAC.DESIGN,TIME AND

SETTING:

Single samples observation was performed in the Experimental Center of School ofMedical Technology,Jiangsu University from September 2005 to December 2006. MATERIALDnHAc was provided by the Material Science and Engineering Department of Tsinghua University.Humanbone marrow mesenchymal stem cells were derived from healthy adult volunteers.All the subiects signed the informedconsents.

METHODS:

Whole bone marrow culture and successive adherence method was used to culture MSCs in vitro,and the cells were then induced to differentiate into the phenotype of osteoblasts by the revulsants(methylprednisolone,vitamin C,β-glycerophosphate and basic fibroblast growth factor).MSCs at passage 3 were co-cultured with nHACfor 14 days.MAIN OUTCOME

MEASURES:

The cytological characteristics of the osteoblast were identified throue,alkalinephosphatase immunohistochemistry method and Von Kossa stain.The growth condition with or without nHAC wasevaluated through invert microscope and scanning electron microscope,respectively.

RESULTS:

The cultured MSCs proliferated into uniform fibroblast-like cells rapidly.MSCs reached confluence and started to form multilayers averaging from 10 to 12 days,passaged stably as well.Then the MSCs passaged from 7 to 9 days.Cytochemistry evaluation showed that MSCs in induced culture were positive for alkaline phosphatase and Von Kossa stain,and deposited calcified matrix.It showed a typical ostcoblast feature in morphology and biology.In coculture model ofMSCs with nHAC,cells would attach to the inner surface of nHAC.At 8 days,the osteoblasts proliferated in the nHAC and the secretion of the matrix was observed.Lots ofcells adheredon the surfaceand pores of nHAC at 14 days.There wereextensive prominent connections among cells.

CONCLUSION:

THE nHAC is suitable for MSCs to adhere,grow and proliferate,with a good compatibility.