The Experimental Study On The Biocompatibility Between HAMSCs And Porcine Cancellous Bone Scaffold Modified With P-15

Objective: To study whether the Porcine Cancellous Bone Scaffold appear better biocompatibility when modified with P-15.Methods:(1)Bone scaffolds were treated by physical and chemical methods to deproteinization and degreasing.(2) Passaging h AMSCs to the 5th generation(3) The scaffolds were divided into experimental group and control group, each group has 36 materials.Experimental group was coated by P-15, control group was coated by PBS. Both groups were dried in the room temperature.Every scaffold was put in 250μl of cell suspension with 1×106/ml cell density. The cell adhesion rates were observed by counting the number of cell which was digested by trypsin at the 6, 12, 24, 48 hours after mixed-culture. The cell proliferation were observed by counting the number of cell which was digested by trypsin on the 3,5,7,9 days after mixed-culture.The cell differentiation were observed by testing the ALP levels on the 3,7,14,21 days after mixed-culture.Results:(1) After deproteinization and degreasing, the bone scaffold material was white, light gray, like porous spongy, no obvious changed in hardness, but the brittleness increased.(2) Primary h AMSCs were spindle, polygonal or star.The cell line grew with adherence.After continuous cell culture,they cloned like fibroblast,arranged radially or spirally by spindle shape.The cell adhesion rates were observed. There was no significant difference between the two groups(P>0.05) at 6 hour,experimental group(20.0±2.6,%)>control group(19.3±1.5,%). There was significant difference between the two groups(P<0.05) at 12,24,48 hours and the result showed that experimental group[(40.7±1.5,%)、(45.3±2.5,%)、(52.7±1.2,%)] > control group[(24.3±4.1,%)、(31.0±1.7,%)、(37.3±4.7,%)].The cell proliferation was significant different between the two groups(P<0.05)on the3,5,7,9 days, experimental group[(20.7±2.5,×104)、(30.3±2.3,×104)、(42.3±3.5,×104)、(41.3±6.5,×104)]>control group[(14.7±0.6,×104)、(20.7±4.0,×104)、(29.7±1.1,×104)、(33.0±5.6,×104)].There was difference between the two groups( P<0.05) on cell differentiation at every time point,experimental group>control group.The OD of experimental group[(0.362±0.052)、(0.551±0.011)、(0.785±0.034)、(0.812±0.125)].The OD of control group[(0.171±0.027)、(0.383±0.055)、(0.445±0.043)、(0.580±0.662)].Conclusions: When combined with P-15,the Porcine Cancellous Bone Scaffold provide a better environment for h AMSCs to adhere and proliferate.P-15 can promote the process of bone formation.