| ObjectiveThe study of naringin promotes the osteoporotic fracture healing of rat femur through X-ray,histological, bone mineral density(BMD)and biomechanical testing.Methods1. The establishment of the osteoporosis model:50adult-female SD rats are selected, divided into ovariectomized group (OVX) and sham-operation group(SHAM) randomly, the former has40rats, the latter has the rest10. In the OVX group, the rats’ bilateral ovaries are excised from dorsal part under aseptic conditions.In SHAM group, when their ovaries are found, do not excise them, just excise the same weight of fat tissue in the same position.2. The establishment of the osteoporotic fracture model:under aseptic conditions, fracture modeling is builton all rats’midpiece backbone of the left femur, and bones is cut transversely on the midpiece of the femurs, then transverse fracture model is made, finally, it is fixed by kirschnerintra medullary needle.3. The medication and dose:the40rats of the fracture model were divided into four groups randomly,the first group is osteoporotic model group (OVX), the other three are experimental groups (OVX+L/M/H).The three experimental groups are given a gavage of naringin respectively (low dose group,1mg/kg; mediate dose group,10mg/kg, high dose group,100mg/kg), control group and model group are given a gavage of double evaporate water in the same volume, do it for8weeks.4. The observational index of the experiment4.1Bone mineral density check:Raise them for12weeks after operation, use dual-energy X-ray to check the density of their bones, verify the established osteoporotic model.4.2Imaging observation:X-ray check is conducted with C shape arm machine right away,4weeks and8weeks afte r OPF, the location of kirschner intramedullary needleand the situation of fracture healingwereobserved.4.3Callus histological examination:8weeks after OPF, allthe rats are put to death, then get the callus histology of the fracture,(including fracture end and bone tissue about3cm long), fixed the tissue-decalcify-embedding-section-HE stain, histological observation under the lightmicroscope.4.4Biomechanical measurement:the three point bending experiment is conducted by using biological materials experiment instrument.Results1. Bone mineral density:12weeks after the oophorectomy, bone mineral density is checked, the BMD of the OVX group is inferior to that ofthe Sham group (P<0.05), the weight is significantly higher than Sham group, the differences are statistically significant (P<0.05), this suggests that osteoporotic model is set up。2. The imaging observation:X-raycheck is conducted right away after OPF, counterpoint to the line in the broken end of the femoral fracture of all groups is good, no obvious difference.4weeks after OPF, the X-ray show that fracture line in SHAM group is slightly fuzzy,bright line still existedat the broken ends of fractured bone, and there are a lot of callus;The two groups, OVX+M and OVX+H, fracture lineswere clearwith a small amount of callus;fracture line of Group OVX and group OVX+L are clear with lesscallus.8weeks after OPF,X-ray show that: SHAME group hadfuzzy fracture line and there are continuous callus through the fracture end;OVX+H, M, L,these three groups’fracture line is slightly blurred, and there is a lot of callus, the generated callus size from large to small is as follows:H> M>L; OVX+N group’s fracture end still had a bright line, callus of fracture end is small.3. Callus histologic examination:8weeks after operation, a large number of bone trabeculas are formedin SHAM group, it is coarser and its arrange is compact and orderly, the number waslarge, and the orientation is the same with the direction of principal stress;OVX+M group’s bone trabecular is of small number, uneven and disordered, the orientation isn’t the same with the principal stress direction, they are staggered into mesh with each other;the trabecular bone of group OVX+L and group OVX+M are fine, the clearance is big, the orientation is uneven, osteoblast and bone cell surrounding the bone trabecularwas of large number;chondrocytes around the OVX+N group’s soft callus are mostly degenerated and necrotic, and there is a small amount of original trabecular bone, osteoblast near the original trabecular bone is of small number, the original bone trabecular belonged to connective tissue.4. Biomechanical test:8weeks postoperatively, three point bending load in the left femur of experimental group are higher than in OVX+N group, but lower than that of SHAM group, in detail:SHAM group> OVX+H> OVX+M> OVX+L> OVX+N, differences are statistically significant (P<0.05).Conclusion1. Naringin has the effect of promoting fracture healing for OPF of ovariectomized rat.2. The bilateral ovariectomy for manufacturing osteoporotic model and the observation time are scientific and reliable. |
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