| OBJECTIVETo study the effects of platelet-rich plasma (PRP) on the proliferation and Osteoblast differentiation activity of the bone marrow stromal cells (BMSCs) cultured in vitro and to provide the experimental evidence for the application of PRP on the bone tissue projects.METHODSThe bone marrow was taken from the Greater trochanter of femur in 8 rabbits. The rabbit's BMSCs were separated and cultivated by the density gradient centrifugation and the passage bolting of the adherence cells at extraorgan. The growth character of the BMSCs was observed. All the third generation of the BMSCs were divided into two groups by own control: the experimental group and the control group .In the experimental group, the BMSCs were interfused with 1%PRP , while in the control group, the BMSCs were interfused with the dexamethasone 10-6 mol/l, theβ-glycerophosphate(β-GP)10-3mol/l, and the vitamin C 50 mg/l. The shape and proliferation of the cultivated cells were observed using the inverted phase contrast microscope. The biological character of the cultivated cells was studied by the inverted phase contrast microscope, the alkaline phosphatase(ALP) determination and the calcium node alizarin bordeaux staining.RESULTSAfter the primary cells were inoculated, plenty of rounded cells could be found ,most of which were in the same shape. 24-36 hours after the adherence ,most of the cells were polygon ellipse or spindle-shaped. 48-72 hours later, they were spindle-shaped. 4 days later ,cell colony increased, but each cell's adumbration was not clear in the center and with small volume. 6 days later ,most of the adhered cells took on the spindle-shaped,when cell- conjunctions were close and the adumbrations were not clear. Some regions were fused. 10-12 days later, the cells fused into single deck, taking on the typical spindle-shaped, and arranged as the cluster. As the time went,cell-islands were formed,and they turned into calcified scobinations 1 month later. (2) 2 dayes after passaging ,cells in both experimental group and control group adhered ,and grew as clusters.On the 3rd day ,they became spindle-shaped ,arranging as radiatiform. On the 6th day ,most of the cells fused, and arranging as fishes. On the 10th day , cells increased rapidly ,and turned from spindle-shaped to polygon or squared.(3)The ALP levels in both two groups achieved highest on the 10th day,and then started to step down. By statistical analysis,it could be concluded that the ALP level in the experimental group was higher than the one in the control group(P< 0. 05). The difference was statistically significant. After the alizarin bordeaux staining ,calcified scobinations could be observed clearly under the microscopes. The alkaline phosphatase(ALP) determination and the calcium node alizarin bordeaux staining showed that the rabbit osteobalasts cultured in vitro grew well by this method and the biochemical indexes were stable.They also had the same morphological and biological characters as the osteoblasts.CONCLUSION1 The platelet-rich plasma (PRP) had well effects on the proliferation and Osteoblast differentiation of the bone marrow stromal cells (BMSCs ).Compared with dexamethasone. theβ-glycerophosphate(β-GP)and the vitamin C , this effection was more significant.2 Because the platelet-rich plasma (PRP) could be collected easily,and had a better ability differentiating to the Osteoblasts ,which was cheaper as well ,it could be a good inductor used in the bone tissue projects.
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