ABSTRACT
Decellularized extracellular matrix (dECM) has been widely used as a scaffold for regenerative medicine due to its high biomimetic and excellent biocompatibility. As a functional polymer material with high water content and controlled fluidity, hydrogel is very promising for some minimally invasive surgery in clinical practice. In recent years, with the rapid development of hydrogel theory and technology, dECM hydrogel has gradually become a research hotspot in the field of regenerative medicine. In this paper, the related researches in recent years are reviewed regarding the preparation of dECM hydrogel and its preclinical application. The future clinical use is also prospected.
ABSTRACT
Objective To observe the effects of a new immunosuppressive agent, FFY720, on rat glomerular mesangial cell (GMC) apoptosis and on gene expression profiles of cell cycle regulatory proteins. Methods Rat GMCs were cultured with 20 μmol/L FTY720 for 6 h, 12 h, 24 h and 48 h, and then were evaluated for proliferation through MTT method, and for apoptosis by flow cytometry and fluorescence stainig with Hoechst33258 and PI, and DNA fragmentation analysis. The gene expression profile of cell cycle regulatory proteins was characterized in rat GMCs before and after FTY720 treatment by SuperArray real-time PCR microarray analysis. Results After incubation with FTY720 for 6 h, apoptotic sub-G1 peak was identified in GMC through flow cytometry. After incubation with FTY720 for 12 h, not only apoptosis bodies of GMC were observed by fluorescence staining with Hoechst33258 and PI, but also typical morphological changes of apoptosis were found in GMC. After incubation with FRY720 for 24 h, typical DNA ladder pattern was identified. The percentage of FTY720-iuduced GMC apoptosis gradually increased with the extension of incubation time. SuperArray real-time PCRmicroarray analysis revealed that FTY720 could respectively up-regulate the expression of Dnajc2, LOC688900 and RGDi562436_predicted genes to 41.6, 38 and 16 folds. Conclusion FTY720 can induce GMC apoptosis in a time-dependent manner, probably through influencing gene expression of cell cycle regulatory proteins.
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Objective To observe the effects of myriocin(ISP-1) on improvement of glomerular messangial cell(GMC) apoptosis and on gene expression profiles of cell cycle regulatory proteins in GMC.Methods Rat GMC was cultured with 100 ?mol/L ISP-1 for 6,12,24,and 48 h,apoptosis was evaluated through flow cytometry,Hoechst33258/PI fluorescence stainig and DNA fragmentation analysis was carried out under Sepharose electrophoresis to observe the changes of apoptosis and DNA fragmentation.Gene expression profiles of cell cycle regulatory proteins were detected by SuperArray Real-Time PCR microarray analysis.The expression of Bax and Bcl-2 proteins was investigated by Western blotting.Results ISP-1 could significantly induce GMC apoptosis in a time-dependent manner,which was the most significant after 48 h.Apoptosis bodys of GMC were observed by Hoechst/PI fluorescence staining,and a typical ladder pattern was identified in DNA electrophoresis.SuperArray Real-Time PCR microarray analysis revealed that ISP-1 could up-regulate the expression of genes involved in DNA damage,apoptosis and cell cycle regulation,such as Rad51,Atm,Brcal,caspases,cyclinA2,cyclinC,chek1,cyclinB1,cyclinB2,cyclinD2,cyclinF,Cdc25a,and P27.Western blotting showed that ISP-1 could significantly up-regulate Bax protein expression and down-regulate Bcl-2 protein expression,respectively.ConclusionISP-1 could induce GMC apoptosis in a time-dependent manner,propabably through influencing gene expression of cell cycle regulatory proteins and apoptosis proteins.