ABSTRACT
Objective To explored the therapeutic benefit and safety of acute kidney injury ( AKI) mice induced by ischemia/reperfusion ( I/R ) treating by thermosensitive chitosan chloride ( CSCI ) hydrogel, an injectable scaffold for adipose-derived MSCs ( ADMSCs ) delivery.Methods After establishing the rat model of acute kidney injury, all rats were randomly divided into 4 groups: ADMSCs/PBS group, ADMSCs/CSCI group, PBS group and CSCI group.Live/Dead staining was used to evaluate the Cytocompatibility between CSCI hydrogel and ADMSCs, dihydroethidium ( DHE) staining was used to detect the number of ROS in vivo, and bioluminescence imaging ( BLI) was used to track the retention and survival of ( fluc-mrfp) labeled ADMSCs in the rat kidney, respectively.The biocompatibility and biodegradability of CSCI hydrogel in kidney tissue were detected by HE staining.The improvement of renal function was evaluated by detecting the level of serum creatinine and urea nitrogen.Results Live/Dead staining manifested an satisfactory Cytocompatibility between CSCI hydrogel and ADMSCs.CSCI hydrogel can improve the micro-environment of AKI kidney tissue by lowering the level of ROS.In PBS and CSCI groups, the positive rate of DHE staining was 68.8%±8.5% and 38.5%±5.8%( P <0.05 ) , respectively.These suggested that CSCI hydrogels could improve the retention and survival of grafted ADMSCs.In ADMSCs /CSCI hydrogel group, BLI fluorescent signal can be detected on the seventh day and was undetectable on the 21st day.ROI value of BLI signal changed from 38 ×105 p/(s? cm2? sr) on the first day to 8.5 ×105 p/(s? cm2? sr) on the 14th day.In ADMSCs /PBS group, BLI fluorescent signal only can be detected on the seventh day and was undetectable on the 14th day.ROI value of BLI signal changed from 17 ×105 p/( s?cm2? sr) on the first day to 3.3 ×105 p/( s? cm2? sr) on the 14th day.Results detected on the 3th and 28th day suggested that ADMSCs/CSCI hydrogel group , ADMSCs/PBS group and CSCI hydrogel group have a better self-repairing capability than the PBS injection group ( P<0.05 ) .In addition, compared with the ADMSCs /PBS group and CSCI hydrogel group, ADMSCs /CSCI hydrogel group got a lower tissue injury scores (P<0.05).At the 4th week, significant improvement of the renal function was observed in CSCI hydrogels with ADMSCs groups.Conclusions CSCI hydrogel is a very promising cell carrier for the treatment of AKI.
ABSTRACT
Objective To study the mechanism of adipose-derived mesenchymal stem cell (ADMSC) in treating rats ischemia/reperfusion (I/R) induced acute kidney injury (AKI).Methods From June 2011 to March 2012,AKI was induced in 40 male SD rats (weight 180-220 g) by clamping bilateral renal pedicles for 40 minutes.Another 8 SD rats (weight 60-80 g) were used to obtain the primary ADMSC from inguinal fat tissue.After being transferred by lentivirus,those cells were cultured for transplantation until passage two.Animals with AKI were then randomly treated by intraparenchymally injecting ADMSC/PBS solutions (ADMSC/PBS group,n =20) or PBS solutions only (PBS group,n =20) (2× 106 cells,100 μl).During injection,the solutions were injected into the upper,middle and lower part of left kidney.The HE staining from 5 rats in each group was used to detect the histological injury at 3 days and 4 weeks after injection,respectively.The apoptosis and proliferation of host renal cells were evaluated using TUNEL staining and PCNA staining.The serum levels of SCr and BUN in animals were measured at day 1,3,14 and 28 after injection.Results HE staining showed ADMSC/PBS group got a lower injury score compared with that in PBS group at 3 days and 4 weeks,respectively (2.0 vs.3.4,1.3 vs.2.6,P＜0.05).In the result of TUNEL staining,the mean number of apoptosis cells was 30 in ADMSC/PBS group and 55 in PBS group.In terms of PCNA staining,the mean number of proliferative cells was 35 in ADMSC/PBS group and 10 in PBS group.All those results suggested that ADMSC could significantly reduce apoptosis and increase proliferation of renal cell (P＜0.05,repectively).The levels of serum SCr in ADMSC/PBS group were lower than those in PBS groups at day 1,3,14 and 28,after injection,respectively (40 vs.70 μmol/L,32 vs.58 μmol/L,26 vs.38 μ mol/L,26 vs.37 μmol/L; P＜0.05).The similar results were shown in the levels of serum BUN between the 2 groups (15 vs.24 mmol/L,13 vs.20 mmol/L,10 vs.13 mmol/L,7 vs.10 mmol/L; P＜0.05).Conclusion ADMSC could repair AKI after acute I/R injury.
ABSTRACT
Objective To detect whether adipose-derived mesenchymal stem cells (ADMCSs)can differentiate into renal tubular epithelial cells and be used for treatment of acute kidney injury (AKI).Methods ADMSCs were separated and cultured in vitro,and were transfected with lentivirus carrying reporter genes of firefly luciferase and monomeric red fluorescent protein (fluc-mrfp).Transfected ADMCSs (2 × l06) were directly injected into renal parenchyma after establishment of AKI models induced by ischemic reperfusion injury.Distribution and differentiation of ADMSCs in rats were detected by bioluminescence imaging (BLI) and immunofluorescent staining,respectively.Results ADMSCs had positive expressions of CD29 and CD90,rather than expressions of CD34,CD45 and CD31.Lentivirus-transduced ADMSCs had stable expressions of mrfp and fluc reporter genes.BLI showed persistent fluorescence signal in vivo even at day 14 after ADMSCs transplantation.Immunofluorescence staining further indicated that the transplanted ADMSCs could differentiate into tubular epithelial cells.Conclusion ADMSCs in vivo can differentiate into renal tubular epithelial cells,which provides an experimental basis for more researches on ADMSCs transplantation for treatment of AKI.