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Objective:To investigate the effect of concentrated growth factor (CGF) on proliferation and differentiation in Beagle adipose-derived stem cells (ADSCs).Methods:ADSCs were isolated from adipose tissue of healthy Beagles and cultured.The multidirectional differentiation potential of ADSCs was identified.The ADSCs were assigned to a CGF group and a control group.The rate of proliferation was analyzed by CCK-8 assay.The osteogenic differentiation capability was detected by ALP staining after the osteoinduction.Bone formationrelated gene expression was detected by RT-PCR.Results:CGF promoted the proliferation of ADSCs in vitro.ADSCs in the CGF group showed higher level of ALP activity than that in the control group (P＜0.05).CGF stimulated the expression of the genes associated with osteogenesis,such as Col-I and Runx2.Conclusion:CGF can promote the proliferation and osteogenic differentiation in ADSCs in vitro.
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Objective · To explore the biological characteristics of platelet-rich fibrin (PRF) and its effects on proliferation and differentiation of adipose derived stem cells (ADSCs). Methods · The whole blood was collected from the forelimb vein of healthy beagles to prepare the PRF membrane, which were observed with optical microscope and scanning electron microscope. ADSCs were collected from the inguinaladipose tissue and were isolated and cultured. Identification of multi-directionaldifferentiation potential was performed. ADSCs were assigned to the PRF group and the control group, the former was treated with PRF in vitro. Cell proliferation wasmeasured with CCK-8. Osteogenesis induction was performed for two groups and the expression of genes associated with osteogenesis, including osteocalcin (OCN), osteopontin (OPN) and collagen I (Col- Ⅰ ), was measured with RT-PCR before induction and 4 and 7 days after induction. The alkaline phosphatase (ALP) activitywas measured 7 days after induction. Results · PRF is a milk white fibrin glue with elasticity and toughness. PRF can form loose and porous three dimensional network structure, which harbors lots of platelets and leucocytes. The cell proliferation activity was significantly higher in the PRF group than in the control group. After osteogenesis induction, the ALP activity and the mRNA levels of OCN, OPN, and Col- Ⅰ were significantly increased. Conclusion · PRF is a fibrin glue with three dimension network structure and contains lots of platelets, which can slowly release growth factors. PRF can promote the proliferation and osteogenic differentiation of ADSCs.
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Objective Recent studies have indicated that early brain injury is the leading cause of death in patients with subarachnoid hemorrhage ( SAH) .Our study investigated the role of aminoguanidine ( AG) in early brain injury after SAH . Methods Sixty-eight male SD rats were equally randomized into four groups of equal number :control, sham, SAH, and AG.The animals in the sham group were injected with isotonic saline solution , while those of the latter two groups with femoral artery blood ( FAB) and FAB+AG, respectively, into the pre-chiasmatic cistern to induce SAH. At 24 hours after modeling , all the rats were killed for HE staining , obtainment of behavioral neurological assessment ( BNA ) scores by Garcia, measurement of the apoptosis of neurons by TUNEL , and de-termination of the expressions of the iNOS and NSE proteins by West-ern blot. Results The results of HE staining showed the presence of more red blood cells in the subarachnoid cavity of the rats in the SAH group, with a significantly decreased BNA score ( 14.47 ± 0.62) as compared with those in the control (17.94 ±0.24), sham (17.59 ±0.51), and AG group (15.71 ±0.47) (P<0.05). The rate of positive cells was remarkably higher in the SAH group ([42.38 ±2.38]%) than in the control ([6.35 ±0.94]%), sham ([6.85 ±0.69]%), and AG group ([30.48 ±2.89]%) ( P<0.01), with significant differences among the latter three groups (P<0.05).The expressions of iNOS and NSE were markedly higher in the SAH group ([3.86 ±0.07] and [1.59 ±0.06]) than in the control (0 and[0.35 ±0.09]), sham ([2.96 ±0.34] and [0.38 ±0.08]), and AG group ([3.41 ±0.04] and [0.70 ±0.12]) ( P<0.05).Both the expression levels of iNOS and NSE were positively correlated with the rate of positive cells (r=0 .879 and 0.935, P<0.01). Conclusion AG can alleviate early brain injury after SAH in SD rats by improving the neuro-ethologic function , suppressing the apoptosis of neurons , and reducing the expressions of iNOS and NSE .