| Objective:Study the effect of percutaneous micro electrical stimulation of the right foot sole of rats in the treatment of femoral fractures,Exploring the relationship between Wnt3 a changes and fracture healing.Methods:1.Twenty SD rats were randomly divided into a blank group(4 rats),a control group(8 rats),and an experimental group(8 rats)using computer randomization.The model control group and experimental group were used to construct a right femoral fracture model.2.Percutaneous micro electrical stimulation therapy: Starting from the second day after modeling,transcutaneous micro electrical stimulation therapy(parameter:Intermediate frequency 3KHz,wavelength 167 us,low frequency 0.125-150Hz)should be performed once every 2 days,with each treatment lasting for 30 minutes.Both the experimental group and the control group attached electrode pads to the right foot of the rat;The experimental group received electrical stimulation treatment,while the control group did not receive electrical stimulation;The blank group of rats did not receive any treatment.3.Sample and data acquisition: Blood samples were collected from the heart every week before and after modeling,and serum was extracted and stored in a-80 ℃refrigerator for testing;Gait analysis was conducted at the 1st and 5th weeks after modeling(using a gait analysis platform to collect the footprints of the hind legs of experimental rats,measure the foot spacing(cm),and collect data for statistical analysis);Observe the DR images of the right femur in the experimental group and control group at 0,4,and 6 weeks after modeling,and determine the fracture healing status;At the 6th week of treatment,the rats were euthanized and the right femur and small intestine of the rats were harvested.4.Detection:(1)Using ELISA method to detect the Wnt3 a OD values in serum and small intestine plasma,and calculate the concentration of Wnt3a;(2)Extract m RNA of Wnt3 a 、β-catenin and T cell factor 4/lymphoid enhancer factor 1(TCF4/LEF1),measure their expression using q PCR,(3)Using Masson staining to analyze the proportion of callus area in femoral bone slices(%),(4)Immunohistochemical staining analysis of Wnt3 a protein in bone tissue was performed.5.Statistical analysis;The experimental results were analyzed using SPSS 27.0software;Bone tissue pathological sections were analyzed using image j software;Use Graph Pad Prism 9.0.0 software to create statistical charts.Two independent samples were compared using t-test,and multiple samples were compared using one-way analysis of variance,with significant differences indicated by P<0.05.Results:1.The DR films of fracture healing in the experimental group and the control group indicate that the experimental group has better healing than the control group.2.The numerical changes in foot spacing between the experimental group and the control group(1 week,5 weeks)were significantly different from those of the blank control group(P<0.05),while there was no difference in foot spacing between the experimental group and the control group(1 week,5 weeks)(P>0.05).3.There was no difference in the ELISA detection concentration of Wnt3 a in the small intestine and serum between the experimental group,control group,and blank group(P>0.05),there was no difference in the numerical changes in q PCR detection results of femoral bone tissue Wnt3a、β-catenin、TCF4 and LEF1(P>0.05)..4.The immunohistochemical expression of Wnt3 a protein in the experimental group was higher than that in the control group(P<0.05).The proportion of callus area(%)stained by Masson in the bone tissue of the experimental group was smaller than that of the control group(P<0.05).Conclusions:Percutaneous micro electrical stimulation of the right foot sole of rats with femoral fractures can improve their gait,promote femoral fracture healing,and increase the expression of Wnt3 a protein at the femoral fracture site.But it does not affect the concentration of Wnt3 a in the small intestine and serum. |
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