目的观察rhBMP2(重组人骨形成蛋白-2)与rhVEGF(重组人表皮生长因子)联合用药对体外培养SD大鼠成骨细胞不同成熟阶段的增殖、分化及胎鼠跖骨生长的影响。方法分离新生大鼠颅盖骨的细胞培养获得前成骨细胞(preosteoblast,pOB),再诱导分化为成骨细胞(osteoblast,OB),以rhBMP2和低(2μg/L)、中(4μg/L)及高(8μg/L)剂量rhVEGF干预48h,比较乳酸脱氢酶(LDH)活性、细胞相对增殖率(RGR)和碱性磷酸酶(AKP)活性。另取胎鼠跖骨,以rhBMP2和rhVEGF干预7d和14d,比较其纵向骨生长率(10ngitudinalbonegrowthrate,LBDR)。结果①LDH组间差异无统计学意义。②RGR:pOB的中、高剂量联用组较同剂量rhBMP2组高。OB的中、高剂量联用组较同剂量rhBMP2及rhVEGF组高。③AKP:pOB的中、高剂量联用组较同剂量rhBMP2及rhVEGF组高。OB的组间差异无统计学意义。④LBDR;7d联用组较rhBMP2及rhVEGF组高,但14d联用组差异无统计学意义。结论适量联用rhBMP2与rhVEGF对骨再生诱导及骨组织早期生长有协同促进效应。
Objectives To observe the effect of rhBMP2 (recombinate human bone morphogenetic protein-2) combined with rhVEGF (recombinate human vascular endothelial growth factor) on the osteoblast proliferation arid differentiation and the metatarsal growth in SD rats in vitro. Methods Cells were isolated enzymatically from the cranium of newborn rats. The pre-osteoblasts (pOBs) were cultured and then induced into the osteoblasts (OBs). Then, the cells were cultured with rhBMP-2 (300 μg'L-1 ) and/or rhVEGF in low dose (2 μg'L-1), middle dose (4μg.L-1), and high dose (8 μg.L-1) for 48h, respectively. The lactate dehydrogenase (LDH) activity, cell relative growth rate (RGR), and alkaline phosphatase (AKP) activity were determined. The metatarsal collected from the SD fetal rats were intervened by rhBMP-2 and/or rhVEGF for 7 days or 14 days. The longitudinal bone growth rate (LBDR) was determined. Results No significant difference of LDH activity among groups was observed. RGR in the middle and high-dose combined medication groups increased compared with that in the same dose or rhBMP2 group for pOBs. The results were the same in the middle and high-dose combined medication groups compared with that in the same dose of rhBMP2 group and rhVEGF group for OBs. The activity of AKP increased in the middle and high-dose combined medication groups compared with that in the same dose of rhBMP2 group and rhVEGF group for pOBs, but no significant difference was observed for OBs. LBDR increased significantly in combined medication group of 7-day culturing compared with that in the rhBMP2 and rhVEGF group, but no significant difference was observed in 14-day culturing. Conclusion The combination of proper dose of rhBMP2 and rhVEGF can promote osteoblastic generation and bone tissue growth at the early stage in a synergic way.