载异烟肼、利福平纳米羟基磷灰石-硫酸钙-壳聚糖人工骨在兔脊柱结核模型中的释药研究

Release of isoniazid and rifampicin ano-hydroxyapatite-calcium-sulfate-chitosan artificial bone in rabbit spinal tuberculosis model

中文关键词:  脊柱结核  异烟肼  利福平  人工骨  体内缓释

英文关键词:Spinal tuberculosis  Isoniazid  Rifampin  Artificial bone  Slow-release in vivo

中文摘要:

  【摘要】 目的：研究载异烟肼（isoniazid，INH）、利福平（rifampicin，RFP）纳米羟基磷灰石（nHA）-硫酸钙（CSH）-壳聚糖（CTS）人工骨在兔脊柱结核模型病灶中的释药特点。方法：用结核分枝杆菌H37Rv标准株，采用侧腹入路的手术方法建立兔脊柱结核模型27只，其中雌性15只，雄性12只，3个月龄，平均体重为2.50±0.25kg，每只模型兔均在行结核病灶清除术后置入载INH、RFP纳米羟基磷灰石-硫酸钙-壳聚糖人工骨，按照时相点分为24h、72h和1周、2周、4周、6周、8周、10周、12周共9组，每组3只。采用高效液相色谱法（high performance liquid chromatography，HPLC）测定各时相点动物模型病灶中椎体及椎旁肌肉中的INH及RFP的药物浓度，并研究其浓度与时间的关系。结果：人工骨中的INH在兔脊柱结核模型的病灶椎体中24h、72h和1周、2周、4周、6周、8周的药物浓度分别为75.66±1.95μg/g、48.46±2.34μg/g、30.69±2.74μg/g、20.34±1.63μg/g、9.36±1.17μg/g、5.19±1.40μg/g、2.73±1.19μg/g，在第10周以后未检测到药物浓度，INH在病灶椎旁肌肉中的各时相点浓度为39.51±2.25μg/g、23.65±1.55μg/g、16.39±2.10μg/g、10.38±1.44μg/g、3.66±0.79μg/g、1.89±0.73μg/g、0.26±0.10μg/g，在第10周以后未检测到药物浓度。对INH在同一时间的椎体和椎旁肌两种不同组织中的药物浓度进行两两比较，均有统计学意义（P＜0.05）；RFP在动物模型的病灶椎体中24h、72h和1周、2周、4周、6周、8周、10周、12周的药物浓度分别为10.85±2.45μg/g、22.47±1.94μg/g、38.32±1.73μg/g、24.22±1.45μg/g、17.85±1.50μg/g、9.81±1.30μg/g、6.35±1.30μg/g、5.11±0.53μg/g、1.32±0.33μg/g，RFP在病灶椎旁肌肉中的药物浓度分别为5.39±1.50μg/g、20.66±1.29μg/g、48.72±2.24μg/g、32.27±1.63μg/g、15.58±1.88μg/g、8.69±0.79μg/g、3.43±0.39μg/g，在第10周后未检测到药物浓度。RFP在椎体和椎旁肌两个部位的药物浓度除72h、4周和6周外，其余时间点的差异均有统计学意义（P<0.05）。结论：载INH、RFP纳米羟基磷灰石-硫酸钙-壳聚糖人工骨在兔脊柱结核模型病灶中可持续、长效的释放抗结核药物。

英文摘要:

  【Abstract】 Objectives: To study drug release characteristics of isoniazid(INH) and rifampicin(RFP) nano-hydroxyapatite(nHA)-calcium sulfate(CSH)-chitosan(CTS) artificial bone in rabbit spinal tuberculosis model. Methods: By using standard strain of Mycobacterium tuberculosis H37Rv, 27 rabbit spinal tuberculosis models were established by lateral ventral approach, 15 females and 12 males, 3 months old, with an average body weight of 2.50±0.25kg, each model rabbit was implanted with artificial bone materials after spinal tuberculosis debridement. According to the time points, the models were divided into 24h, 72h and 1, 2, 4, 6, 8, 10, 12 weeks, a total of nine groups, with 3 rabbits in each group. High performance liquid chromatography(HPLC) was used to determine the concentration of INH and RFP in focal bone tissue and focal muscle tissue of animal models at different time points, and the relationship between concentration and time was studied. Results: The concentration of INH in focal bone tissue at 24h, 72h and 1 week, 2 weeks, 4 weeks, 6 weeks and 8 weeks was 75.66±1.95μg/g, 48.46±2.34μg/g, 30.69±2.74μg/g, 20.34±1.63μg/g, 9.36±1.17μg/g, 5.19±1.40μg/g and 2.73±1.19μg/g respectively， no drug concentration was detected after 10 weeks, the concentration of INH in focal muscle tissue was 39.51±2.25μg/g, 23.65±1.55μg/g, 16.39±2.10μg/g, 10.38±1.44μg/g, 3.66±0.79μg/g, 1.89±0.73μg/g, 0.26±0.10μg/g respectively， no drug concentration was detected after 10 weeks. Comparison of drug concentrations of INH in two different tissues at the same time showed statistical significance(P<0.05). At 24h, 72h and 1 week, 2 weeks, 4 weeks, 6 weeks, 8 weeks and 10 weeks, the drug concentration of RFP in focal bone tissue was 10.85±2.45μg/g, 22.47±1.94μg/g, 38.32±1.73μg/g, 24.22±1.45μg/g, 17.85±1.50μg/g, 9.81±1.30μg/g, 6.35±1.30μg/g, 5.11±0.53μg/g, 1.32±0.33μg/g respectively. The drug concentration of RFP in focal muscle tissue was 5.39±1.50μg/g, 20.66±1.29μg/g, 48.72±2.24μg/g, 32.27±1.63μg/g, 15.58±1.88μg/g, 8.69±0.79μg/g, 3.43±0.39μg/g respectively. No drug concentration was detected after 10 weeks. The concentrations of RFP in two different tissues were significantly different at other time points except 72h, 4 weeks and 6 weeks(P<0.05). Conclusions: INH-RFP nano hydroxyapatite-calcium sulfate-chitosan artificial bone can sustainably and effectively release anti-tuberculosis drugs in the focal tissues of rabbit spinal tuberculosis model.