Effect Of Ectogenous Nerve Growth Factor On Proliferation,Differentiation And Mineralization Of Osteoblast In Vitro

ObjectiveIn clinical practice,a large number of immature callus was often found in limb fracture,when these patients got paraplegia and brain injuries,even the heterotopic ossification could be found in muscles.At the same time,much more bone was found in patients with neurological diseases.So,we can consider the nervous system involved in the fracture healing process.It is not clear that how NGF which is very important in nervous system effects on osteoblast which is the most important cell in bone.So,the influencing of ectogenous nerve growth factor on proliferation, differentiation and mineralization of osteoblast in vitro.was going to be investigated.MethodCranium of newborn Wistar rat within 24h was obtained,and other tissue around the cranium was removed.The cranium was cut into pieces.Improved explant culture and enzymatic digestion for culturing osteoblast are taken.Different concentration NGF(1pg/ml,10pg/ml,100pg/ml,500pg/ml,1000pg/ml)were added into the 3rd generation osteoblast.Proliferation of the cells are detected by MTT on the 7th day,and activity of ALP was measured by ALP kit,and capacity of the typeⅠcollagen was assayed by Immunohistostaining,and the mineralized matrix was also observed and calculated by Dahl McGec-Russll's alizarin red stain.ResultsThe effect on osteoblast proliferation:low concentration NGF's(＜100 pg/ml) OD value is more high than control group(P＜0.05).there is significant differences in statistics;The effect on osteoblast differentiation:these three group 500 pg/ml,100 pg/ml,10 pg/ml has more high ALP activity(OD value)(P＜0.05),and more typeⅠ collagen,comparing with the control group(P＜0.05).The effect on osteoblast mineralization:Each group has no significant difference.ConclusionsIn vitro,ectogenous nerve growth factor could significantly stimulate the osteoblast proliferation and differentiation,enhance the activity of ALP,promote the secretion of typeⅠcollagen,but they couldn't stimulate the osteoblast mineralization obviously.