1.Effects of Electroacupuncture on Activation of Microglia in Peri-infarct Cortex of Cerebral Ischemia-reperfusion Injury Rats
Xian WANG ; Jia HUANG ; Weilin LIU ; Hao SHANGGUAN ; Yi ZHENG ; Lulu WANG ; Yunjiao LIN ; Jing TAO ; Lidian CHEN
Chinese Journal of Rehabilitation Theory and Practice 2015;(11):1251-1255
Objective To explore the effect of electroacupuncture on activation of microglia in peri-infarct cortex after cerebral isch-emia-reperfusion in rats. Methods 36 male Sprague-Dawley rats were randomly divided into sham group (n=12), model group (n=12) and electroacupuncture group (n=12). The latter two groups were occluded the left middle cerebral arteries with modified Longa's method for 2 hours and reperfused. The electroacupuncture group received electroacupuncture at Quchi (LI11) and Zusanli (ST36) acupoints for 3 days. The nerve cell damage in peri-infarct cortex was observed with HE staining, while the expression of ED1 was determined with immunohisto-chemical staining, and the expression of tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and IL-6 were determined with Western blotting. Results The neurological deficits score improved significantly in the electroacupuncture group (P<0.05), with less nerve cell dam-age, less number of ED1 positive microglia (P<0.05) and less levels of TNF-α, IL-1βand IL-6 (P<0.05), compared with the model group. Conclusion The electroacupuncture at Quchi (LI11) and Zusanli (ST36) acupoints can protect brain from ischemia-reperfusion injury, which might be associated with inhibiting the microglial activation and proinflammatory response in peri-infarct cortex.
2.Effects of Electroacupuncture at Quchi (LI11) and Zusanli (ST36) on Neuronal Apoptosis Induced by Mitochondria- caspase-3 Pathway in Rats with Cerebral Ischemia-reperfusion Injury
Hao SHANGGUAN ; Weilin LIU ; Wenlie CHEN ; Yi ZHENG ; Xian WANG ; Yunjiao LIN ; Lulu WANG ; Lidian CHEN ; Jing TAO
Chinese Journal of Rehabilitation Theory and Practice 2015;21(8):900-904
Objective To investigate the effects of electroacupuncture (EA) at Quchi (LI11) and Zusanli (ST36) acupoints on the ultrastructural structure of cortical neurons in peripheral area and the protein expression of caspase- 3, Bcl- 2, Bax in rats with cerebral ischemia- reperfusion injury. Methods 36 male adult Sprague-Dawley rats were randomly divided into sham operation group, model group, and electroacupuncture group, with 12 rats in each group. The model group and electroacupuncture group were performed with left middle cerebral artery occlusion (MCAO) according to the modified Longa' methods. The electroacupuncture group received electroacupuncture at Quchi (LI11) and Zusanli (ST36) on the paralyzed limb, for 30 minute. The neurobehavioral scores were recorded before and after treatment. The ultrastructural structure of cortical neurons was observed with transmission electron microscope (TEM). The protein expression of caspase- 3, Bcl-2 and Bax were detected by Western blotting technique. Results The neurobehavioral score was lower in the electroacupuncture group than in the control group (P<0.05). Compared with the model group, the chromatin of neurons was even relatively, and the number of mitochondria increased. The expression of Bcl-2 was higher and the expression of caspase-3 and Bax was lower in the electroacupuncture group than in the model group (P<0.05). Conclusion Electroacupuncture at Quchi (LI11) and Zusanli (ST36) acupoints can inhibit the neurons apoptosis in peripheral area through mitochondria-caspase-3 pathway.
3.The promotion of bone formation at the tendon-bone interface after ACL reconstruction with De-BMSCs transplantation and its mechanism
Kai TIE ; Jinghang CAI ; Jun QIN ; Hao XIAO ; Yangfan SHANGGUAN ; Liaobin CHEN
Chinese Journal of Orthopaedics 2022;42(8):519-529
Objective:This study aimed to investigate the effect of differentiation osteogenic bone marrow mesenchymal stem cells (De-BMSCs) transplantation on the promotion of bone formation at the tendon-bone interface after anterior cruciate ligament reconstruction (ACLR), and further explored the molecular mechanism of the enhanced osteogenic effect of De-BMSCs.Methods:BMSCs from femur and tibia of New Zealand White rabbit were subjected to osteogenic induction and then cultured in no osteogenic factor medium; the obtained cell population was termed De-BMSCs. De-BMSCs were induced into osteo-, chondro-and adipo-differentiation in vitro to examine the characteristics of primitive stem cells. ACLR model with a semitendinosus tendon were performed in 48 adult rabbits, three groups were established: control group with alginate gel injectionat the tendon-bone interface, BMSCs group with the injection of alginate gel containing BMSCs, De-BMSCs group with the injection of alginate gel containing De-BMSCs. At 4 and 12 weeks after surgery, rabbits in each group were sacrificed to evaluate tendon-bone healing by histologic staining, micro-CT examination, and biomechanical test. During osteogenic differentiation of De-BMSCs, si-RNA of nuclear factor of activated T cells 2 (NFATc2) si-RNA of nuclear factor of activated T cells 1 (NFATc1) were used to verify the molecular mechanism of enhanced osteogenic effect of De-BMSCs.Results:De-BMSCs exhibited some properties similar to BMSCs including multiple differentiation potential and cell surface marker. At 4 weeks after surgery, the BV/TV value of the De-BMSCs group 0.36±0.01 was significantly higher than that of the control group 0.24±0.03 and BMSCs group 0.30±0.02 (all P<0.05), and the maximum load 40.34±1.19 N and stiffness 20.67±2.14 N/mm were significantly higher than those in the control group 14.88±2.74N, 8.67±2.19 N/mm and the BMSCs group 26.31±1.76 N, 13.81±2.14 N/mm (all P<0.05). At 12 weeks after surgery, the BV/TV value of the De-BMSCs transplantation group 0.47±0.02 was significantly higher than that of the control group 0.30±0.02 and the BMSCs group 0.35±0.03 (all P<0.05), and the maximum load 64.46±6.69 N and stiffness 25.18±3.11 N/mm were significantly higher than those in the control group 41.01±6.12 N, 11.59±2.54 N/mm and the BMSCs group 48.21±4.12 N, 15.89±2.94 N/mm (all P<0.05). During the osteogenic differentiation of De-BMSCs, the expressions of Nanog and NFATc1 were synergistically increased which promoted interaction of NFATc1 and Osterix ( P< 0.05), resulting in the increased expression of osteoblast marker genessuch as COL1A, OCN, OPN (all P< 0.05). Conclusion:De-BMSCs transplantation could promote bone formation at the tendon-bone interface after ACLR,Nanog/NFATc1/Osterix signaling pathway mediated the enhancement of the osteogenic differentiation effect of De-BMSCs.
4.Effects of hyperbaric oxygen on the expression of osteoprotegerin and integrin β1 in rabbit's mandibular distraction gap
Wensong SHANGGUAN ; Chunbing HU ; Hao DAI ; Shu WANG ; Guoping WU
Chinese Journal of Medical Aesthetics and Cosmetology 2019;25(3):174-178
Objective To investigate the effect of hyperbaric-oxygen therapy on osteoprotegerin (OPG) and integrin β1 (Itgβ1 ) expression during mandibular distraction osteogenesis .Methods Forty New-Zealand rabbits were used .The animals were randomly divided into two groups (experimental group and control group) with 20 animals each ;after accomplished osteotomy and implant distraction devices on mandible bilaterally for 3 days of latency period ,the device was activated at the rate of 0 .8 mm per day for 10 days .All animals in the experimental group were subjected to hyperbaric oxy -gen for 90 minutes once a day since the beginning of distraction ,and lasted for four weeks .In control group ,all the distractors were activated following the same distraction protocol as the experimental group ,but without hyperbaric oxygen therapy .Five animals of each group were sacrificed at 10th day after distraction ,7th ,14th and 28th day of consolidation , respectively .The lengthened mandibles were harvested and processed for immunohistochemical examinations to detect OPG and Itg β1 expres-sion in the distraction gap .Semi-quantitative analysis was carried out by image analysis software . Results OPG staining was mainly located in the membrane and cytoplasm of osteoprogenitor cells and osteoblasts in the distraction zones .The expression of OPG increased after distraction accomplished and reached to the peak at 7th day of consolidation ,and then decreased gradually .At every time point , the level of expression of OPG in the experimental group was remarkably higher than those in control group .There were significant differences between the experimental group and control group ( P <0 .01) .Itgβ1 mainly located in actively proliferating osteoblasts ,fibroblasts and mesenchymal cells . The expression of Itgβ1 decreased significantly after reached to the peak at the 10th day distraction .At 10th day distraction ,7th and 14th day of consolidation ,Itgβ1 expressed more strongly than that in the experimental group ,which was remarkably higher than those in control group .There were significant differences between experimental group and control group (P < 0 .05) .At 28th day of consolidation , Itgβ1 expressed weakly ; there was no significant difference between the two groups ( P > 0 .05 ) . Conclusions Hyperbaric oxygen therapy can up-regulate the expression of OPG and Itgβ1 in the dis-traction gap ,which may promote osteoblast differentiation ,proliferation ,enhance osteoblast func-tion ,and new bone formation in distraction gap .
5.Influence of different costal cartilage harvest techniques on donor site pain in rhinoplasty
Wensong SHANGGUAN ; Guoping WU ; Hao DAI ; Chao HU ; Shu WANG ; Qiaofeng JIANG
Chinese Journal of Medical Aesthetics and Cosmetology 2020;26(4):272-275
Objective:To seek a minimally invasive method for costal cartilage harvest by using two different costal cartilage harvest techniques in rhinoplasty and to compare their influence on donor site pain.Methods:Fourty-three female patients who underwent costal cartilage harvest for rhinoplasty from Dec. 2016 to Dec. 2017 were randomly divided into two groups. We harvested the right side seventh costal cartilage in both groups. In control group, we harvested a full thickness segment of costal cartilage in each patient, whereas we harvested a split thickness segment of costal cartilage by preserving the superior strip in experimental group. Donor site pain was evaluated via visual analogue scale (VAS) 6 h, 24 h, 48 h and 72 h after surgery.Results:The VAS scores (mean±standard deviation) at different time points in the control group were (5.515±1.085), (5.250±1.302), (5.315±1.117) and (4.895±1.042). And in the experimental group, they were (2.665±0.713), (2.261±0.642), (1.609±0.398) and (1.383±0.514), respectively. The VAS scores at different time points were significantly higher in the control group than that in the experimental group ( P<0.05). Conclusions:The superior strip preserved costal cartilage harvest technique significantly reduces postoperative donor site pain.
6. The expression of β-catenin in callus of distraction gap during mandibular distraction osteogenesis in rabbits
Hao DAI ; Chunli ZHANG ; Wensong SHANGGUAN ; Chunbing HU ; Shu WANG ; Guoping WU
Chinese Journal of Plastic Surgery 2019;35(11):1128-1134
Objective:
To explore the role of β-catenin in distraction osteogenesis of new bone formation, the expression of β-catenin in the distraction gap callus was detected during rabbit mandibular distraction osteogenesis.
Methods:
26 New-Zealand rabbits were randomly divided into 3 groups. Group A is a normal control group with 2 rabbits, group B is the mandibular defect control group, and group C is the distraction group. Group B and C with 12 rabbits, respectively. The two rabbits in group A without surgery, their mandibles are normal control. In group B, vertical osteotomy was performed between the first molar and the mental foramen on the mandibles bilaterally, followed by rigid internal fixation with titanium plates and screw with 5 mm gap immediately. In group C, after the same osteotomy was performed, the fragments of mandibles were reduced and fixed with mandibular distractors bilaterally. On the fourth day postoperatively, the distraction started at a rate of 0.8 mm/d and lasted for 7 days, followed by consolidation period. Two rabbits of group B and C were sacrificed at 6th, 10th, 17th, 24th, 31st, 38th day postoperatively, respectively. The newly formed callus in the distraction gap of mandibles was harvested for Western blotting and immunohistochemistry examination to detect the distribution and expression of β-catenin. The experimental data were analyzed by SPSS 22.0 statistical software using Spearman function.
Results:
The result of Western blotting showed that the expression of β-catenin gradually increased at distraction period(6-10 days after surgery) and reached the peak at the end of the distraction period(10th day postoperatively), it gradually decreased during the consolidation period. However, the expression of β-catenin in group C was higher than that of group B. Immunohistochemistry stain showed that the expression of β-catenin mainly located in inflammatory cells(eg. monocyte), fibroblast of the granulation tissue, the osteoblasts, osteocyte on the surface of new formed trabecular, and the connective tissues surrounding the new bone in the new formed callus. Cytoplasmic and nuclear staining were positive. In group C, the expression of β-catenin was strong (3.245 8±0.132 3) after distraction (6th day postoperatively), and reached a peak (4.602 8±0.021 9) on the 10th day postoperatively. With the disappearance of the distraction stress, the expression of β-catenin gradually decreased since17th day postoperatively(3.639 8±0.125 5), but the staining was still positive. In group B, the strong positive staining of β-catenin on the 6th day after surgery (2.734 0±0.134 7), the strongest staining on the 10th day after surgery (3.101 3±0.104 8), and the expression of β-catenin on the 17th day after surgery (2.542 8±0.211 1) was weaker than that on the 10th day after surgery. At each time point, the expression of β-catenin in group C was significantly higher than that in group B, and the difference was statistically significant (