| Objective:When the intravenous concentration of USPIO is 20 mg/kg,explore the rabbitbone marrow mesenchymal stem cells(BMSCs)symbol rates,and the effect of USPIO for rabbit bone marrow mesenchymal stem cells hyperplastic and multi-directional differentiation capacity,make use of magnetic resonance imaging(MRI)to trace the process and signal strength of repairation of rabbit articular cartilage defect after the bone marrow mesenchymal stem cells transplantation intravitally.Provide experimental basis for further experiments to determine the intravenous concentration settings of USPIO.Methods:(1)The six New Zealand white rabbits completely randomly divided into 2 groups,each group of three and a group for the unit,experimental group injects 20 mg/kg of USPIO by ear marginal vein,the control group does not do any process.Executed each rabbit after 48 h,rabbit BMSCs were isolated and cultured to the third generation as experiment object;(2)Prussian blue stain was performed to show intracellular iron nanoparticles,enhanced CCK-8 Cell Counting Kit was applied to evaluate proliferation of labeled cells;osteoplastic,adipogenic and chondrogenic differentiation potential of labled cells were identified using Alizarin red staining,Oil red O stain and Sarranine O staining;(3)36 New Zealand white rabbit completely randomly divided into 4 groups,each group of nine,prepare the rabbit model of double knee joint cartilage defect,experimental group was injected USPIO tag rabbit BMSCs-sodium alginate gel complex,positive control group was injected unmarked rabbit BMSCs-sodium alginate gel complex,negative control group was injected pure sodium alginate gel,blank group doesn’t do any deal.In postoperative 1 month,2 months,3 months for gross observation,MRI scans and histologic study.Results:(1)After the rabbit ear margin vein was injected USPIO of 20 mg/kg,isolated and cultured the rabbit BMSCs,then compared with the control group in rabbit BMSCs,cell morphology is basically same.(2)Prussian blue staining showed the rabbit BMSCs marked with USPIO can be seen much blue dye in the cytoplasm and USPIO has been swallowed by rabbit BMSCs;Enhanced CCK-8 Cell Counting Kit,automatic enzyme standard instrument measured absorbance value and proliferation curve drawing showed USPIO concentration of 20 mg/kg does not affect the proliferation activity of rabbit BMSCs;Multidirectional differentiation experiments showed USPIO concentration of 20 mg/kg does not affect the multi-directional differentiation ability of rabbit BMSCs.(3)BMSCs transplanted in the rabbit knee joint cartilage defect showed marked cell transplantation group and unmarked cell transplantation group were gradually repaired,otherwise pure sodium alginate transplantation group and blank group were slowly repaired.(4)Statistical analysis showed after BMSCs were transplanted in rabbit knee joint cartilage defect,marked cell transplantation group and unmarked cell transplantation group did not see the obvious difference of the repair effect at the same time(P>0.05),compared with pure sodium alginate transplantation group and the blank group repair effect was statistically significant(P<0.05);Marked cell transplantation group and unmarked cell transplantation group did not see the significantly statistical differences of SI at the same time(P>0.05).Conclusion:(1)When the USPIO intravenous concentration is 20 mg/kg,rabbit BMSCs could be marked with USPIO.(2)This injection concentration does not affect the physiological activity of rabbit BMSCs.(3)When the USPIO intravenous concentration is 20 mg/kg,the marked rabbit BMSCs can be used for transplantation of rabbit knee joint cartilage defect,but failed to be clearly traced using MRI. |
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