Li Wen Yuan, Tian Yong Hui, Gao Yu Hai, Zhu Bao Ying, Xi Hui Rong, Chen Ke Ming
Institute of Orthopaedics,the 940th Hospital of Joint Logistic Support Force of Chinese People's Liberation Army,Lanzhou 730050,China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2019 Feb 28;41(1):11-20. doi: 10.3881/j.issn.1000-503X.10471.
Objective To compare the effects of 50-Hz 0.6-mT low-frequency pulsed electromagnetic fields(PEMFs) and 50-Hz 1.8-mT sinusoidal alternating electromagnetic fields(SEMFs) in preventing bone loss in tail-suspended rats,with an attempt to improve the prevention and treatment of bone loss caused by weightlessness.Methods Tail-suspension rat models were used to simulate microgravity on the ground. Forty rats were randomly divided into four groups[control group,hindlimb-suspended(HLS) group,HLS+PEMFs group,and HLS+SEMFs group],with 10 rats in each group. In the PEMFs treatment group and SEMFs treatment group,the intervention was 90 min per day. Rats were sacrificed after four weeks. Bone mineral density(BMD) of femur and vertebra was measured by dual-energy X-ray absorptiometry and biomechanical strength by AG-IS biomechanical instrument. Serum osteocalcin(OC),tartrate-resistant acid phosphatase 5b(Tracp 5b),parathyroid hormone(PTH),and cyclic adenosine monophosphate(cAMP) were detected by ELISA. The microstructure of bone tissue was observed by Micro-CT and HE staining.Results The BMD of the femur(P=0.000) and vertebrae(P=0.001) in the HLS group was significantly lower than in the control group;the BMD of the femurs(P=0.001) and vertebrae(P=0.039) in the HLS+PEMFs group was significantly higher than in the HLS group;the BMD of the femurs in the HLS+SEMFs group was significantly higher than in the HLS group(P=0.003),but the BMD of the vertebrae showed no significant difference(P=0.130). There was no significant difference in the BMD of the femur(P=0.818) and vertebrae(P=0.614) between the HLS+PEMFs group and the HLS+SEMFs group. The maximum load(P=0.000,P=0.009) and elastic modulus(P=0.015,P=0.009) of the femurs and vertebrae in the HLS group were significantly lower than those in the control group;the maximum load of the femur(P=0.038) and vertebrae(P=0.087) in the HLS+PEMFs group was significantly higher than that in the HLS group,but the elastic modulus was not significantly different from that in the HLS group(P=0.324,P=0.091). The maximum load(P=0.190,P=0.222) and elastic modulus(P=0.512,P=0.437) of femurs and vertebrae in the HLS+SEMFs group were not significantly different from those in the HLS group. There were no significant differences in the maximum load and elastic modulus of femurs(P=0.585,P=0.948) and vertebrae(P=0.668,P=0.349) between the HLS+PEMFs group and the HLS+SEMFs group. The serum OC level in the HLS group was significantly lower than that in the control group(P=0.000),and the OC level in HLS+PEMFs group(P=0.000) and HLS+SEMFs group(P=0.006) were significantly higher than that in the HLS group. The serum Tracp 5b concentration in the HLS group was significantly higher than that in the control group(P=0.011). There was no significant difference between the HLS+PEMFs group(P=0.459) and the HLS+SEMFs group(P=0.469) compared with the control group.Serum Tracp 5b concentrations in the HLS+PEMFs group(P=0.056) and the HLS+SEMFs group(P=0.054) were not significantly different from those in the HLS group. The PTH(P=0.000) and cAMP concentrations(P=0.000) in the HLS group were significantly lower than those in the control group. The PTH(P=0.000,P=0.000) and cAMP concentrations(P=0.000,P=0.000) in the HLS+PEMFs group and the HLS+SEMFs group were significantly higher than in the HLS group. The femoral cancellous bone of the HLS group was very sparse and small compared with the control group. The density and volume of the cancellous bone were similar among the control group,HLS+PEMFs group,and HLS+SEMFs group. Compared with the control group,the HLS group had lower BMD(P=0.000),bone volume (BV)/tissue volume(TV)(P=0.000),number of trabecular bone (Tb.N)(P=0.000),and trabecular thickness(Tb.Th)(P=0.000) and higher trabecular bone dispersion(Tb.Sp)(P=0.000) and bone surface area(BS)/BV(P=0.000). Compared with the HLS group,the HLS+PEMFs group and the HLS+SEMFs group had significantly lower Tb.Sp(P=0.000,P=0.000) and BS/BV(P=0.000,P=0.000) and significantly increased BMD(P=0.000,P=0.000),BV/TV(P=0.001,P=0.004),Tb.Th(P=0.000,P=0.001),and Tb.N(P=0.000,P=0.001). The trabecular thickness significantly differed between the HLS+PEMFs group and the HLS+SEMFs group(P=0.024). The HLS group(P=0.000),HLS+PEMFs group(P=0.000),and HLS+SEMFs group(P=0.000) had the significantly lower osteoblast density on the trabecular bone surface than the control group;however,it was significantly higher in the HLS+SEMFs group(P=0.000) and the HLS+PEMFs group(P=0.000) than in the HLS group. The HLS group had significantly lower density of osteoblasts in the endothelium than the control group(P=0.000);however,the density of osteoblasts was significantly higher in HLS+PEMFs group(P=0.000) and HLS+SEMFs group(P=0.000) than HLS group and was significantly higher in HLS+PEMFs group than in HLS+SEMFs group(P=0.041). Compared with the control group,a large number of fatty cavities were produced in the bone marrow cavity in the HLS group,but the fat globules remarkably decreased in the treatment groups,showing no significant difference from the control group. The number of adipose cells per mm bone marrow in the HLS group was 4 times that of the control group(P=0.000);it was significantly smaller in the HLS+PEMFs group(P=0.000) and HLS+SEMFs group(P=0.000) than in the HLS group,whereas the difference between the HLS+PEMFs group and the HLS+SEMFs group was not statistically significant(P=0.086). Conclusions 50-Hz 0.6-mT PEMFs and 50-Hz 1.8-mT SEMFs can effectively increase bone mineral density and biomechanical values in tail-suspended rats,increase the concentration of bone formation markers in rat blood,activate the cAMP pathway by affecting PTH levels,and thus further increase the content of osteoblasts to prevent the deterioration of bone micro-structure. In particular,PEMFs can prevent the reduction of bone mineral density and maximum load value by about 50% and increase the bone mass of tail-suspended rats by promoting bone formation.
目的 比较50Hz 0.6mT低频脉冲电磁场(PEMFs)和50Hz 1.8mT正弦交变电磁场(SEMFs)对尾悬吊大鼠骨质流失的预防作用,以期改善失重导致的骨质流失的防治效果。方法 采用尾悬吊大鼠模型模拟地面微重力环境。40只大鼠随机分为4组[对照组、后肢悬吊(HLS)组、HLS+PEMFs组和HLS+SEMFs组],每组10只。PEMFs治疗组和SEMFs治疗组每天干预90分钟。4周后处死大鼠。采用双能X线吸收法测量股骨和椎体的骨密度(BMD),用AG-IS生物力学仪测量生物力学强度。采用酶联免疫吸附测定法(ELISA)检测血清骨钙素(OC)、抗酒石酸酸性磷酸酶5b(Tracp 5b)、甲状旁腺激素(PTH)和环磷酸腺苷(cAMP)。通过Micro-CT和苏木精-伊红(HE)染色观察骨组织微观结构。结果 HLS组股骨(P=0.000)和椎体(P=0.001)的BMD显著低于对照组;HLS+PEMFs组股骨(P=0.001)和椎体(P=0.039)的BMD显著高于HLS组;HLS+SEMFs组股骨的BMD显著高于HLS组(P=0.003),但椎体BMD差异无统计学意义(P=0.130)。HLS+PEMFs组和HLS+SEMFs组股骨(P=0.818)和椎体(P=0.614)的BMD差异无统计学意义。HLS组股骨和椎体的最大负荷(P=0.000,P=0.009)和弹性模量(P=0.015,P=0.009)显著低于对照组;HLS+PEMFs组股骨(P=0.038)和椎体(P=0.087)的最大负荷显著高于HLS组,但弹性模量与HLS组差异无统计学意义(P=0.324,P=0.091)。HLS+SEMFs组股骨和椎体的最大负荷(P=0.190,P=0.222)和弹性模量(P=0.512,P=0.437)与HLS组差异无统计学意义。HLS+PEMFs组和HLS+SEMFs组股骨(P=0.585,P=0.948)和椎体(P=0.668,P=0.349)的最大负荷和弹性模量差异无统计学意义。HLS组血清OC水平显著低于对照组(P=0.000),HLS+PEMFs组(P=0.000)和HLS+SEMFs组(P=0.006)的OC水平显著高于HLS组。HLS组血清Tracp 5b浓度显著高于对照组(P=0.011)。HLS+PEMFs组(P=0.459)和HLS+SEMFs组(P=0.469)与对照组相比差异无统计学意义。HLS+PEMFs组(P=0.056)和HLS+SEMFs组(P=0.054)血清Tracp 5b浓度与HLS组差异无统计学意义。HLS组PTH(P=0.000)和cAMP浓度(P=0.000)显著低于对照组。HLS+PEMFs组和HLS+SEMFs组PTH(P=0.000,P=0.000)和cAMP浓度(P=0.000,P=0.000)显著高于HLS组。与对照组相比,HLS组股骨松质骨非常稀疏且细小。对照组、HLS+PEMFs组和HLS+SEMFs组松质骨的密度和体积相似。与对照组相比,HLS组BMD(P=0.000)、骨体积(BV)/组织体积(TV)(P=0.000)、骨小梁数量(Tb.N)(P=0.000)和骨小梁厚度(Tb.Th)(P=0.000)较低,骨小梁离散度(Tb.Sp)(P=0.000)和骨表面积(BS)/BV(P=0.000)较高。与HLS组相比,HLS+PEMFs组和HLS+SEMFs组Tb.Sp(P=0.000,P=0.000)和BS/BV(P=0.000,P=0.000)显著降低,BMD(P=0.000,P=0.000)、BV/TV(P=0.001,P=0.004)、Tb.Th(P=0.000,P=0.001)和Tb.N(P=0.000,P=0.001)显著增加。HLS+PEMFs组和HLS+SEMFs组骨小梁厚度差异有统计学意义(P=0.024)。HLS组(P=0.000)、HLS+PEMFs组(P=0.000)和HLS+SEMFs组(P=0.000)骨小梁表面的成骨细胞密度显著低于对照组;然而,HLS+SEMFs组(P=0.000)和HLS+PEMFs组(P=0.000)的成骨细胞密度显著高于HLS组。HLS组内皮中成骨细胞密度显著低于对照组(P=0.000);然而,HLS+PEMFs组(P=0.000)和HLS+SEMFs组(P=0.000)的成骨细胞密度显著高于HLS组,且HLS+PEMFs组显著高于HLS+SEMFs组(P=0.041)。与对照组相比,HLS组骨髓腔内产生大量脂肪空泡,但治疗组脂肪球明显减少,与对照组差异无统计学意义。HLS组每毫米骨髓中脂肪细胞数量是对照组的4倍(P=0.000);HLS+PEMFs组(P=0.000)和HLS+SEMFs组(P=0.000)明显少于HLS组,而HLS+PEMFs组和HLS+SEMFs组之间差异无统计学意义(P=0.086)。结论 50Hz 0.6mT的PEMFs和50Hz 1.8mT的SEMFs可有效提高尾悬吊大鼠的骨密度和生物力学值,提高大鼠血液中骨形成标志物的浓度,通过影响PTH水平激活cAMP通路,进而增加成骨细胞含量,防止骨微观结构恶化。特别是,PEMFs可预防骨密度和最大负荷值降低约50%,并通过促进骨形成增加尾悬吊大鼠的骨量。