Bone marrow-derived mesenchymal stem cells transfected with adenovirus vector containing bone morphogenetic protein 2 encapsulated by hyaluronic acid hydrogel and its effect on proliferation and expression of target protein

English Keywords:Hyaluronic acid hydrogels  BMP-2  BMSCs  Tissue engineering nucleus pulp

English Abstract:

  【Abstract】 Objectives： To observe the proliferative ability of bone marrow-derived mesenchymal stem cells(BMSCs) transfected with the adenovirus vector containing bone morphogenetic protein 2(BMP-2) in the three-dimensional condition of hyaluronic acid hydrogel(HAH) and its effect on the expresseion of BMP-2 originated from BMSCs. Methods: BMSCs were isolated, extracted, identified in vitro and transfected with the constructed adenovirus vector containing BMP-2(Ad-BMP-2) in advance, then BMSCs were encapsulated by HAH; the three-dimensional shape of Ad-BMP-2 BMSCs in HAH was observed by laser scanning confocal microscope. Four groups in the experiments(all groups no difference in culture conditions) were classified as followings: group A, untransfected BMSCs; group B, HAH+untransfected BMSCs; group C, transfected BMSCs; group D, HAH+transfected BMSCs. CCK-8 method was used to detect each OD value for evaluating cell proliferation of transfected cells encapsulated by HAH at different time point(1d, 2d, 3d……7d). ELISA method was used to detect the quantity of each BMP-2 expression(pg/ml). Results: In HAH by used confocal laser 3d reconstruction of BMSCs morphology could be observed that BMSCs rendered arbitrary planar spreading with plenty of growth space. BMSCs proliferation activitis detection showed the extension of incubation time as, cell proliferation activities increased, OD value of group A and group C after 4d was obviously higher than that in 1d(P<0.05); OD value of group B and group D after 5d was obviously higher than that in 1d(P<0.05). At 7d, the OD value of group A was 1.283±0.061, group B was 1.844±0.075, group C was 1.570±0.099, group D was 1.976±0.142 and encapsulated HAH OD value of group B and group D were better than that in group A and group C without encapsulated HAH(P<0.05). Weak expression of BMP-2 were in group A and group B, there were no significant difference between different time points(P>0.05). At various time points, the expression of BMP-2 in both group C and group D were obviously higher than that in group A and group B(P<0.05). Since 4d, group C and group D of BMP-2 expression significantly increased(compared to 1d, P<0.05); the expression of BMP-2 in group D was significantly better than that in group C, and the difference between the two group was statistically significant(P<0.05). Conclusions: Ad-BMP-2 BMSCs not only have strong proliferationpotency, but also can promote the expression of BMP-2 in three-dimensional environment(encapsulated by HAH).