Promoted Proloferation And Differentiate Into Osteoblast Of Human Adipose Derived Stem Cells And The Biocompatibility Of HADSCs With A Scaffold In Vitro

OBJECT:â‘ To improve the method for preparing platelet-rieh plasma(PRP).To investigate the effect of platelet-rich plasma (PRP) on human adipose derived stem cells(hADSCs)when it differentiate into osteoblast, and the biocompatibility of hADSCs with a scaffold-- nano-HAp/Collagen.â‘¡Assessing the effect of different concentrations of EMPs on Beagle dog adipose tissue-derived stromal cells proliferationMETHODS:â‘ Whole blood was collected from 3 healthy human and then PRP was separated by both the second routine method and the third routine method. hADSCs were obtained from legal abortion fetal's by enzyme digestion, and 3th passage were cultured in DMEM with 10% FBS, Osteoblasts induced medium and Osteoblasts induced medium with PRP respectively, investigate the effect of PRP on hADSCs'differentiate by Von Kossa. hADSCs were coculture with nano-HAp/Collagen,cell attachment was observed by environmental scanning electron microscope after 7 days.â‘¡dADSCs were exposed to various concentrations of EMPs (400 mg/L,200 mg/L,100 mg/L,50 mg/L) and DMEM(negative control), respectively. After 48,72 and 96 hours, Proliferation of the ADSCs cell proliferation was evaluated by thiazoly blue(MTT).Statistical analysis was performed using one- way ANOVA and Student's test in SPSS10.0. RESULTS:â‘ The the third routine method has a higher platelet count in PRP than that of the second routine method. hADSCs can obtain by enzyme digestion successfully, With the contribution of PRP, hADSCs can differentiate into osteoblast more effectively with more calcified nodules which are fine but dense. hADSCs can adhere to and fully extend on nano-HAp/Collagen scaffold material under environmental scanning electron microscope.â‘¡The proliferation of dADSCs was significantly stimulated by EMP of 50mg /L and100mg /L from 48h over the experiment. There was no significant change between EMP of 200mg /L and400mg /L. EMPs at a concentration of 100 mg /L significantly enhanced dADSCs proliferation(P<0.05).CONCLUSION:â‘ The third routine method is an effetive method in preparation of PRP.hADSCs isolation and differentiation successfully offered a new chioce of seed cell of periodontal tissue engineering,PRP was a effective cell factor for hADSCs'differentiation into osteoblasts and nano-HAp/Collagen could be used as a potential scaffold material in periodontal tissue regeneration.â‘¡EMPs could promote the proliferating ability of human hADSCs, EMPs at a concentration of 100 mg /L significantly enhanced dADSCs proliferation from 48h over the experiment.