R, the biological activity of compounded MSCs was determined and compared with uncompounded MSCs. The results of EdU incorporation and WST-8 assays showed that MSCs from the MSC-fibrin glue complex have a high proliferation rate similar as uncompounded MSCs resulting from the expression of HGF transgene (Figure 2A-C). The expression level of secreted HGF from transgenic MSCs was assayed using ELISA at 24 h after MSCs released from fibrin glue. 6-(Thiophen-3-yl)pyridin-3-amine The results showed that transgenic MSCsexpressed similar level of Methyl 3-amino-2,4-dichloro-5-fluorobenzoate secreted HGF no matter ever compounded with or without fibrin glue (Figure 2D). Moreover, MSCs from fibrin glue composite exhibited similar osteogenic differentiation ability as uncompounded MSCs, 4-Dichlorophenoxy)propanoic acid N-Methylhydrazinecarboximidamide hydroiodide Tembotrione Methyl 1-hydroxycyclopropanecarboxylate (2R which was showed by ALP expression (Figure 2E) and alizarin red (ARS) staining (Figure 2F, G). The above results suggested that short-term compounding with fibrin glue will not change the biological activities of MSCs in osteogenic differentiation and responsive sensitivity to HGF stimulation.Fibrin glue composite improved the therapeutic effects of HGF-transgenic MSCs on ANFHAfter the ANFH PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/11834444 model was established, the animals were given MSCs or MSCs + FG treatment. Similar as previous report, there is no significant difference could be observedFigure 2 Proliferation, HGF expression and osteoblast differentiation of MSCs compounded with fibrin glue. Cell proliferation and viability were assayed using EdU incorporation (A, B) and CCK-8 method (C). HGF expression was assayed using ELISA (D). Osteoblast differentiation of MSCs compounded with or without fibrin glue was assayed using ALP staining (E) and ARS staining (F, G) after treatment with osteogenic culture medium.Wen et al. Journal of Translational Medicine 2014, 12:114 http://www.translational-medicine.com/content/12/1/Page 5 ofbetween fibrin glue-compounded and uncompounded groups while obvious and similar therapeutic effects were observed in both treatment groups within 4 weeks [21]. However, the MSCs + FG group exhibited more significant improvement in recovery from ANFH when the observation period was extended to 8 weeks. Results of HE staining indicated that hematopoietic cells in the MSCs + FG group increased PubMed ID:https://www.ncbi.nlm.nih.gov/pubmed/7780048 more significantly than that of the MSCs group and the number of empty lacuna was dramatically decreased in the MSCs + FG group compared with the MSCs group (P < 0.05) (Figure 3). The above results suggested better angiogenesis and formation of osteogenic microenvironment in MSCs + FG group during tissue repairing. As a specific marker of bone formation during bone turnover, OCN could be used to characterize the initial osteogenic differentiation of MSCs [24,25]. In addition, OCN was mainly expressed in perivascular vessels and part of osteocytes in normal tissues. In ANFH model, expression of OCN was almost lost completely. Such loss could be rescued by transplantation of Ad-HGF transfected MSCs regardless of the presence of fibrin glue in short-term treatment. However, the significant differences only could be clearly observed at the end of 8 weeks post treatment, OCN levels in the MSCs + FGgroup was significantly higher than that in the MSCs group (P < 0.05) (Figure 4A, B). As expected, transplantation of HGF transgenic MSCs restored the vascularization of the femoral head indicated by the expression of CD105, the marker of newly-formed blood vessels [26,27]. Notably, the level of CD105 in the MSCs + FG group was also higher than that in the MSCs group (P < 0.05) at 8 weeks.
R, the biological activity of compounded MSCs was determined and compared
