The Research On The Biological Characteristic Of Seed Cells And Constructed Periosteum In Vitro

Objectives To investigate the practicability of periosteum construction in vitro by small intestinal submucosa(SIS) as a tissue scaffold with bone marrow mesenchymal stem cells(BMSC) and osteoblast induced from BMSC respectively. There is also a studying of biological characteristics of seed cells.Methods The bone marrow mesenchymal stem cells were cultivated by the conventional methods in vitro, and BMSC were induced osteoblast under conditional culture medium and then co-cultivated with SIS respectively. The morphological feature of BMSC and induced osteoblast were observed by drawing the growth curve and assaying the cell cycle and phenotype with flow cytometer respectively. The morphological feature of the complex material, the extracellular matrices, the adhesion and proliferation of BMSC and induced osteoblast on SIS were observed by optical microscope, scanning electronic microscope, Western Blot respectively.Results Bone marrow mesenchymal stem cells and induced osteoblast adhered and proliferated on SIS, and secreted a great deal of extracellular matrices with active cellular function, acting like a biological periosteum. The expression of Type I collagens and alkaline phosphatase(ALP) and osteocalcin(OC) confirmed that they had osteogenic potential. There were some differences between BMSC and induced osteoblast on SIS in the expression of Type I collagens and ALP, OC. BMSC and induced osteoblast on SIS had similar grow tendency to those without combining with SIS.Conclusions Combined culture of BMSC and osteoblast induced from BMSC with SIS respectively in vitro can construct a tissue engineering periosteum similar to the natural one. BMSC and osteoblast induced from BMSC showed good compatibility on the surface of SIS, while they maintained their normal physiological function. Furthermore, tissue engineering periosteum combined with osteoblast induced from BMSC showed more dominance than that combined with BMSC. All of this provided a basis for further study of osteogenic regeneration in vivo with tissue engineering periosteum.