In vitro culture of bone marrow stromal cell and ceramic xenogenic bone: Observation of bone marrow stromal cell on morphology property, growth, adhesion and proliferation in rabbits

Yanlin LI; Rui HAN; He HUANG; Shihe LI; Caiming ZENG; Hongbang WANG

ABSTRACT

BACKGROUND:

Proper physiochemical method can alleviate or eliminate the antigenicity of natural xenogenic bone and preserve its natural porous reticular system so that its structure and components are in conformity with physiological requirements. It is expected that xenogenic bone becomes a kind of substitutive material in bone transplantation.

OBJECTIVE:

It was to adopt generative culture of marrow stromal cell and complex culture of ceramic xenogenic bone in vitro to understand the influence of ceramic xenogenic bone on morphology, growth, adhesion and proliferation of marrow stromal cell in rabbits.

DESIGN:

Blank, control experiment was designed.

SETTING:

Center Laboratory of First Hospital Affiliated to Kunming Medical College.MATERIALS Five Japanese big ear white rabbits of 3-month old were employed, body mass varied from 1.5 to 2.0 kg, of either sex.

INTERVENTIONS:

The experiment was performed in Center Laboratory of First Hospital Affiliated to Kunming Medical College from June to October 2003. With physiochemical management, the fresh pig rib bought from market was prepared into ceramic xenogenic bone and was verified with Xray diffractometry to be mainly composed of hydroxyapatite. It was verified with scanning electron microscopic observation that such material presented porous meshwork system as osteogenie tissue. Five Japanese big ear white rabbits were employed to collect 1mL marrow for each. PRMI 1640culture solution was used to dilute it into suspension of marrow stromal cell and the cells of upper layer were collected for culture. With 1×106 cells/bottle, they were vaccinated to 8 culture bottles of 25 mL and generated at the ratio of 11. The prepared ceramic xenogenic bone was placed into culture bottles, 5 pieces in each bottle, totally for 5 bottles, taken as experimental group and the other 3 bottles (without xenogenic bone ) were taken as the control. The culture ended in 2 weeks and a part of cultured material was used for observation with phase microscope and scanning electron microscope.MAIN OUTCOME

MEASURES:

Influence of ceramic xenogenie bone on morphology, growth, adhesion and proliferation of marrow stromal cell in rabbits were observed with phase microscope and scanning electron microscope.to 14 days after vaccination, cell proliferation was connected to be monolayer meshwork. Generative culture in 24 hours, adhesion was accomplished and the cells grew well in both experimental group and the cortrol.In the experimental group, fibroblastic cells were arranged more closely rabbits with scanning electron microscope in experimental group, it was visible in 2 weeks of generative culture that some counected reticular fibroblastic cells adhered with the inner wall of reticular pores and porous base of ceramic xenogenic bone as well as the surface of bone trabecula.The cells were mostly fusiform, triangular or multiangular.

CONCLUSION:

Ceramic xenogenic bone is mainly composed of hydroxyapatite and presents reticular porous structural system of osteogenic tissue, which will not produce harmful effects on morphology, growth, adhesion and proliferation of marrow stromal cell and it is expected to be used in complex graft for repair of bone defects with bone marrow.