重组Tet-on COLIA1基因腺病毒骨组织靶向表达对大鼠骨质疏松骨折愈合生物力学影响的实验研究  

Effect of targeted expression of recombinant Tet-on COLIA1 gene adenovirus in bone tissue on biomechanics of osteoporotic fracture healing in rats: an experimental study

作　　者：吴元刚[1] 曾羿[1] 李明阳 刘渊[1] 裴福兴[1] 沈彬[1] WU Yuangang;ZENG Yi;LI Mingyang;LIU Yuan;PEI Fuxing;SHEN Bin(Department of Orthopaedics,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院骨科,成都610041

出　　处：《中华骨与关节外科杂志》2022年第3期170-175,共6页Chinese Journal of Bone and Joint Surgery

基　　金：国家自然科学基金(81974347);中国博士后基金(2021M702351)。

摘　　要：目的:研究重组Tet-on COLIA1基因腺病毒骨组织靶向表达对大鼠骨质疏松骨折愈合生物力学的影响。方法:40只健康雌性大鼠按照随机数字表法分为手术组(n=36)和假手术组(n=4)。手术组大鼠切除双侧卵巢,建立骨质疏松动物模型。假手术组大鼠仅切开皮肤,不切除双侧卵巢。手术组36只大鼠切除双侧卵巢3个月后,随机选取4只并处死,同时处死假手术组4只大鼠。收集其胫骨标本,采用双能X线测量假手术组和手术组大鼠的骨密度,以证实大鼠骨质疏松模型建立成功。随后,将剩余的32只手术组大鼠,采用薄电锯片从胫骨中段横行切断骨干,克氏针固定骨折断端,以构建大鼠骨质疏松骨折模型。同时按照随机数字表法分为4组:A组,重组Tet-on COLIA1基因腺病毒+强力霉素组;B组,重组Tet-on COLIA1基因腺病毒;C组,Tet-on空白腺病毒+强力霉素组;D组,空白对照组。分别于注射腺病毒6、8周后在电子万能材料试验机上进行胫骨三点弯曲试验。根据载荷-变形曲线计算骨折标本的生物力学参数,包括最大负荷、弹性模量、屈服点最大应力,同时采用micro-CT检测骨折愈合情况。结果:术后3个月手术组大鼠胫骨骨密度低于假手术组大鼠,且差异有统计学意义(P=0.005),表明大鼠骨质疏松模型制备成功。胫骨三点弯曲试验发现A组大鼠的最大负荷、弹性模量、屈服点最大应力,在腺病毒转染6、8周后,均高于B、C、D三组大鼠,且差异均有统计学意义(P<0.05)。Micro-CT显示,在转染6、8周后,A组大鼠骨折断端更加模糊,骨皮质较连续,骨折断端愈合更好。结论:经重组Tet-on COLIA1基因腺病毒转染的骨质疏松骨折大鼠,在强力霉素的作用下,能增加大鼠的最大载荷、弹性模量、屈服点最大应力,改善骨折愈合的能力,缩短骨折愈合的时间。Objective: To study the effect of targeted expression of recombinant Tet-on COLIA1 gene adenovirus in bone tissue on biomechanics of osteoporotic fracture healing in rats. Methods: A total of 40 healthy female rats were randomly divided into an operation group(n=36) and a sham operation group(n=4). Bilateral ovaries of rats in the operation group were removed to establish an animal model of osteoporosis. In the sham operation group, an incision was made on rats’ skin and sutured. After 3 months, 4 rats randomly selected from the operation group and 4 rats in the sham operation group were sacrificed. Tibial specimens were collected, and dual-energy X-rays were used to measure the bone mineral density of the rats. A rat osteoporotic fracture model was established in the remaining 32 rats in the operation group by cutting off the shaft of the tibia and fixing with Kirschner wires.Then, they were randomly divided into 4 subgroups: group A(recombinant Tet-on COLIA1 gene adenovirus + doxycycline group), group B(recombinant Tet-on COLIA1 gene adenovirus group), group C(Tet-on blank adenovirus + doxycycline group), and group D(control group). Tibial three-point bending test was performed on an electronic universal material testing machine 6 and 8 weeks after adenovirus injection. The biomechanical parameters of fracture specimens were calculated according to the load-deformation curve, including maximum load, elastic modulus, and maximum stress at the yield point. Micro-CT was used to detect fracture healing. Results: Three months after operation, the bone mineral density of the tibia in the operation group was significantly lower than that in the sham operation group(P=0.005), indicating that the rat osteoporosis model was successfully prepared. The tibia three-point bending test found that the maximum load, elastic modulus and maximum stress at the yield point in group A were significantly higher than those in group B, C and D after 6 and 8 weeks of adenovirus transfection(P<0.05). Micro-CT showed that, after 6 and 8

关 键 词：重组Tet-on COLIA1基因腺病毒 骨质疏松 骨折 强力霉素 生物力学 大鼠 动物模型 基因治疗 

分 类 号：R683[医药卫生—骨科学] R589.5[医药卫生—外科学]