[Research of You Gui Yin and MSCs interventional therapy on early avascular necrosis of the femoral head].
OBJECTIVE
To observe the effect of You Gui Yin and MSCs interventional therapy on the early stage of avascular necrosis of the femoral head (ANFH) and its role of improving revascularization and reossification.


METHODS
Twenty-four adult Beagle dogs weighing (10.0 +/- 0.5) kg were divided into 4 groups (n=6): group A (model group), group B (You Gui Yin group), group C (MSCs intervention group), group D(You Gui Yin and MSCs intervention group). The model of ANFH at early stage was established by liquid-nitrogen cryopreservation method, and MSCs were isolated, cultured and labeled by BrdU. Three weeks after modeling, groups C and D received 1 mL MSCs with artery perfusion [(0.5-1.0) x 10(6)/mL)], groups B and D received intragastric administration of 100 mL You Gui Yin per day, groups A and C received intragastric administration of 100 mL distilled water. Gross observation on femoral head was conducted 4 and 8 weeks after continuous treatment. Meanwhile, DSA and MRI were adopted to observe the quantity and the diameter of femoral head blood vessel, histology and immunohistochemistry staining were performed to observe the expression of VEGF and BrdU, and the expression of VEGF mRNA was detected by real-time fluorescence quantitative RT-PCR.


RESULTS
At 4 and 8 weeks after treatment, the figuration of the femoral head in group A was flat and mushroom-shaped, while it was relatively normal in groups B, C and D. DSA observation: the number and the diameter of blood vessel in groups C and D were increased, and the obstructed blood vessel was open. At 4 and 8 weeks after treatment, significant differences between group C and group D were evident in the number and the diameter of blood vessel (P < 0.05); compared with before treatment, the diameter of blood vessel in two groups were significantly improved (P < 0.05) and the number of blood vessel in group D was significantly increased (P < 0.05). MRI observation: compared with group A, groups B, C and D were obviously improved, especially group D, T1W showed lower signal, T2W and STIR showed no abnormal changes of signal. Histopathology and immunohistochemistry staining: compared with group A, the structure of groups B, C and D were obviously improved, the positive expression of VEGF in group D was significantly higher than that of other groups (P < 0.05), the positive rate of BrdU, the number of positive osteoblast and the number of positive blood vessel in group D was obviously higher than that of group C (P < 0.05). Real-time fluorescence quantitative RT-PCR detection: the expression of VEGF mRNA in group D was significantly higher than that of other groups (P < 0.05), the expression of VEGF in groups B, C and D was higher than that of group A (P < 0.05).


CONCLUSIONS
The combination of You Gui Yin and MSCs interventional treatment has significant therapeutic effects on the early-stage ANFH, can improve the blood supply of the necrotic femoral head, promote repair and prevent collapse.
Tong
Peijian
P
Department of Orthopaedics, the First Hospital Affiliated to Zhejiang University of Traditional Chinese Medicine, Hangzhou Zhejiang, PR China. tongpeijian@163.com
Jin
Hongtin
H
Shen
Yan
Y
He
Bangjian
B
Xiao
Luwei
L
Zhao
Hongchang
H
Ma
Zhenchuan
Z
D000818:Animals; D002478:Cells, Cultured; D004195:Disease Models, Animal; D004285:Dogs; D005271:Femur Head Necrosis / Q000188:drug therapy / Q000628:therapy*; D008297:Male; D008516:Medicine, Chinese Traditional; D045164:Mesenchymal Stem Cell Transplantation