lyophilization primer drying at 32 u00b0 c 2 pa for 12 hours second drying at 32 u00b0 c 0 pa for 12 hours the mineralized allograft was frittered into cubic or round shape pieces the superficies of the allograft pieces was one square centimeter and their height was 5 millimeters thus prepared allografts were incubated in protein solution at 4 u00b0 c overnight the used protein solutions were 10 20 u03bcg ml fibronectin and 10 20 albumin of human serum origin furthermore fetal calf serum and 1 5 collagen type i was derived from pig the proteins were also used in combination like fibronectin with albumin fibronectin with collagen type i these bones were used either directly for the u201c without drying u201d experiments or subsequently the incubated bones were lyophilized at 32 u00b0 c at 1 pa for 24 hours the coated allografts were subjected to uv irradiation for decontamination for 30 minutes the seeded mscs were labeled with the fluorescent membrane dye vybrant did excitation emission 644 665 nm for 30 minutes at 37 u00b0 c did labeled mscs were suspended in dmem culture medium and dropped with pipette to the surface of the coated allografts 60 000 cells scaffold were used in all the experiments the mscs were expanded on the allograft in vitro for 18 days the seeded did labeled msc 39 s proliferation was observed with confocal microscopy zeiss lsm 510 meta carl zeiss jena germany on the coated allograft individual areas.