载三联抗结核药物硫酸钙/聚氨基酸人工材料体外缓释性能的观察

目的:探讨载三联抗结核药物硫酸钙/聚氨基酸人工材料在模拟体液中的药物缓释性能。方法:避光环境下以100:3:3:12的比例称取硫酸钙/氨基酸复合材料500mg、异烟肼(isoniazid,INH)15mg、利福平(rifampicin,RFP)15mg、吡嗪酰胺(pyrazinamide,PZA)60mg制备载药人工缓释材料,将其置于模拟体液中,分别于浸泡3h、12h、24h、36h、48h、60h、72h、1~14周时取材料浸提液,应用高效液相色谱法(HPLC)检测其中INH、RFP、PZA三种药物的浓度,并据其计算单位时间段内药物释出质量。结果:载三联抗结核药的硫酸钙/聚氨基酸人工材料在模拟体液中浸泡3h时浸提液中释出INH、RFP、PZA的浓度分别达到152.96±1.32μg/ml、92.90±2.17μg/ml和334.90±12.3μg/ml,在8周前各时间点的浸提液中,3种药物浓度均较高;至8周时PZA的释出浓度、10周时RFP释出浓度、11周时INH的释出浓度仍高于其10倍的最小抑菌浓度,之后逐渐降低;未载药硫酸钙/聚氨基酸人工材料在有效检测时间内药物出峰时间处未见有意义杂质峰出现。结论:载三联抗结核药硫酸钙/聚氨基酸人工材料具有较为平稳、持续时间较长的有效缓释性能,三种药物在模拟体液中释出药物的浓度均可达到体内杀死结核分枝杆菌的浓度。     

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

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(P0.05);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(P0.05)。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。     

载异烟肼、利福平白蛋白纳米粒在兔体内的药代动力学研究

目的 :研究载异烟肼(Isoniazid,INH)、利福平(Rifampicin,RFP)牛血清白蛋白(Bovine serum albumin,BSA)纳米粒经新西兰大白兔静脉给药后的体内药代动力学特征及其相关参数。方法 :16只新西兰大白兔随机分为2组:纳米粒组8只,INH、RPP对照组8只。经耳缘静脉分别滴注等计量的载INH、RFP白蛋白纳米粒和普通对照品(INH 11.43mg/kg、RFP 12mg/kg)。采用高效液相色谱分析方法测定各自INH、RFP的血浆药物浓度。对比研究静脉给药后两种剂型药物在不同时间(0、0.5、1、1.5、2、2.5、3、4、6、8、12、24、36、48、72、96、120h)兔体内的血浆药代动力学参数,包括最大血药浓度(Cmax)、生物半衰期(T1/2)、血药浓度时间曲线下的面积(AUC)及体内平均驻留时间(MRT),以DAS 2.0药动学程序进行血药浓度数据拟合分析。结果:给药后两组药物的血药浓度-时间曲线显示:纳米粒药组INH、RFP血药浓度相对稳定,药物浓度持续缓慢下降,给药后96h INH仍能检测到,RFP则为48h。INH的Cmax=9.79±1.38(mg/L),T1/2=28.10±6.69(h),AUC=22.34±8.81(mg/L·h),MRT=15.13±5.48(h);RFP的Cmax=16.79±3.74(mg/L),T1/2=14.13±3.78(h),AUC=116.64±66.26(mg/L·h),MRT=15.89±2.49(h)。对照组中INH的Cmax=9.52±2.19(mg/L),T1/2=1.92±0.80(h),AUC=20.97±3.11(mg/L·h),MRT=2.03±0.62(h);RFP的Cmax=12.50±1.58(mg/L),T1/2=2.87±0.78(h),AUC=50.05±14.50(mg/L·h),MRT=3.70±0.99(h)。对照组INH、RFP血药浓度快速下降,分别在给药后24h和12h检测不到。结论 :载异烟肼、利福平白蛋白纳米粒在兔体内的药代动力学过程符合双隔室模型(权重系数为1/cc)。白蛋白纳米粒可有效改变异烟肼和利福平的药动学行为并具有显著的缓释效果。     

异烟肼聚乳酸缓释体的制备及体内外释药特性

目的探讨异烟肼聚乳酸微球的制备方法和体内外释药特性.方法用复乳法制备异烟肼聚乳酸微球,扫描电子显微镜观察微球的特征,高效液相色谱法测定其包封率、载药率;用溶出法、高效液相色谱法测定其体外释药特性.观测第3、6、9、12、15、18、25、32、39、42、49、56天的释药情况,并计算其日均释药率、累计释药率将异烟肼聚乳酸微球、异烟肼(INH)分别置入24只新西兰大白兔的左右股骨粗隆间髓腔内,测定微球在体内3、7、14、28、42、56d时的释药情况;实验组将第42、49、56天时的异烟肼聚乳酸微球洗涤液(空白组用磷酸缓冲液)与结核菌、钛块一起共同培养7d,扫描电子显微镜下观察结核菌生长情况.结果异烟肼聚乳酸微球呈比较完整的球形且分散性好,无明显聚集现象;微球平均直径为59.4±2.9μm,包封率为(67.51±0.57)%,载药率为(32.82±0.65)%.体外释药结果显示,最初14d累计释药达3.784±0.359mg,约占总量的30%,日平均释药0.270±0.024mg;随后42d累计释药5.328±0.203mg,约占总量的50%,日平均释药0.126±0.013mg,第56天时释药0.032±0.009mg.体内释药实验显示,3d时右侧股骨粗隆间骨质内INH浓度明显高于左侧(P＜0.01);7～56d右侧INH浓度明显降低并低于左侧(P＜0.01);7～56d左侧股骨粗隆间骨质内INH浓度持续在20±g/s以上.空白组钛块表面有大量结核菌生长,实验组钛块表面几乎无结核菌生长.结论异烟肼聚乳酸微球具有缓慢释放药物的特性.可以作为持久抗结核治疗的缓释体置入骨结核病灶清除术后病灶局部.     

脊柱结核病灶中抗痨药物浓度的测定

目的研究脊柱结核病椎各部位抗结核药物的分布特点,为脊柱结核化疗方案与手术治疗方法的改进提供理论依据。方法24例应用2SHRZ/2.5H2R2Z2(4.5个月)方案化疗结合手术治疗的脊柱结核患者,按有无病椎骨硬化分为硬化组和非硬化组。化疗第4周手术,依据手术取材时间分为术晨服药后120~130、180~190min两个时相点,取硬化组的病椎硬化壁、硬化壁外“亚正常骨”、硬化壁内结核病变组织和非硬化组的病椎结核病变组织、病变组织外“亚正常骨”以及两组患者的血浆、髂骨,采用高效液相色谱法测定上述样本的药物浓度。结果(1)吡嗪酰胺(PZA)和异烟肼(INH)各时相血药浓度与文献报道健康人单次服药的药时数据相近,利福平(RFP)则低25%,各药血药浓度高于髂骨及病椎组织中的药物浓度。(2)两组中“亚正常骨”与髂骨内的INH和RFP均达到了各自的杀菌浓度水平,PZA超过其细胞内酸性条件下MIC的5倍。三种药物各自在“亚正常骨”与自身对照的髂骨之间浓度相近。(3)硬化组中硬化壁的药物浓度仅为各药的最小抑菌浓度水平(MICs),远低于壁外围“亚正常骨”,壁内的结核病变组织中未检出上述三种药。(4)非硬化组结核坏死、干酪组织中RFP及PZA的浓度相当于各自的MICs,远低于其他组分,而INH却达到细胞内杀菌浓度。结论INH、RFP和PZA在脊柱结     

利福平硫酸钙植入剂修复动物骨缺损的体内实验研究

目的探讨利福平硫酸钙植入剂（RCPPs）在体内的生物相容性、降解情况及成骨特点。方法将利福平硫酸钙植入剂（RCPPs）随机植入成年新西兰白兔1．5cm的桡骨缺损内．并设立不植入任何材料的空白对照。观察利福平硫酸钙植入剂（RCPPs）植入后动物的局部反应；于术后6、12周取材．作X线、大体标本及组织形态学观察，分析不同时期组织反应、骨缺损修复及材料降解情况。结果利福平硫酸钙植入剂（RCPPs）植入后无明显的局部不良反应。X线检查、大体标本及组织形态学观察显示利福平硫酸钙植入剂（RCPPs）的成骨方式主要是骨传导成骨，至术后12周时骨缺损基本修复，RCPPs与宿主骨结合紧密。而空白对照骨缺损断端仅有少量骨修复，形成骨不连。利福平硫酸钙植入剂（RCPPs）植入后即开始其降解过程，12周以后材料周围出现较多吞噬有材料颗粒的巨噬细胞和多核口细胞，仍有部分材料未降解吸收。结论利福平硫酸钙植入剂（RCPPs）具有持续的成骨能力、良好的生物相容性。     

载三联抗结核药硫酸钙/氨基酸聚合物人工骨体内缓释实验研究

目的:观察载异烟肼、利福平、吡嗪酰胺三联抗结核药硫酸钙/氨基酸聚合物人工骨在大鼠体内的缓释效果.方法:取6～7周龄SD大鼠80只,随机分为实验组与对照组,实验组载三联抗结核药人工骨植入大鼠骶脊肌内,40只;对照组植入空白未载药人工骨于大鼠骶脊肌内,40只;术后1d、3d、1周、2周、4周、6周、8周、12周处死大鼠,每次处死实验组、对照组大鼠各5只,应用高效液相色谱法检测大鼠植入人工骨局部周边组织及血液中三种药物浓度,2周及6周时HE染色观察大鼠肝、肾病理组织学变化.结果:植入载三联抗痨药人工骨局部周边1cm肌肉组织中,术后1d、3d、1周、2周、4周、6周、8周、12周时,高效液相色谱法检测异烟肼为4.09 ±0.56μg/ml、2.45 ±1.33μg/ml、2.12±1.56μg/ml、1.58 ±4.12μg/ml、2.09 ±2.35μg/ml、2.31 ±1.44μg/ml、4.26±2.17 μg/ml、3.79 ±0.49 μg/ml;利福平为4.02±1.14 μg/ml、1.90±0.11 μg/ml、1.88±0.90μg/ml、0.79±1.08 μg/ml、0.86±0.44μ g/ml、0.89±0.98μg/ml、3.92± 1.09μ g/ml、3.57±0.22μg/ml;吡嗪酰胺为460.87±1.41 μg/ml、440.91±1.69 μg/ml、430.21 ±0.86μg/ml、340.73± 1.45μg/ml 、320.85±2.0μg/ml、270.61±1.0μg/ml、230.38±0.48μg/ml;12周时植骨块局部组织,周边1cm肌肉组织异烟肼、利福平、吡嗪酰胺能达到10倍最低抑菌浓度,静脉血三种药物平均浓度为2.79μg/ml、2.02μg/ml、4.38 μg/ml低于植骨块局部周边浓度;肝、肾组织组织病理学检查未见损伤.结论:载三联抗结核药硫酸钙/氨基酸聚合物人工骨体内释放平稳,可实现体内局部用药缓释;其可直接植入体内,对肝、肾组织无明显药物损伤.     

生物活性复合人工骨的制备与理化性能研究

目的探讨硫酸钙/骨基质明胶生物活性复合人工骨的制备方法及其理化性能。方法分别制备硫酸钙、骨基质明胶颗粒,按1：1、2：1、3：1、1：0不同质量比例制备复合人工骨。经扫描电镜观察、生物力学测试及体外降解试验,研究不同配比复合人工骨的结构特征、力学强度及降解速率。结果硫酸钙与骨基质明胶呈均匀混合分布,材料内见较多孔径为100～500μm的微孔结构,孔隙间相互交通,随着骨粒含量的增加,孔径逐渐增大。质量比为1：1、2：1、3：1、1：0的复合人工骨的抗压强度分别为(3．53±0．62)、(6．74±0．78)、(13．60±1．01)、(39．85±2．33)MPa,100%体外降解时间分别为8、10、12、12周。结论硫酸钙/骨基质明胶复合人工骨具有利于新骨长入的微孔结构,随着骨粒含量的增高,材料的力学强度逐渐减低,降解时间加快,不同配比的复合材料可适用于不同的植骨需要。     

载异烟肼及利福平白蛋白纳米粒治疗兔脊柱结核的实验观察

目的 :观察载异烟肼(isoniazid,INH)、利福平(rifampicin,RFP)白蛋白纳米粒治疗兔脊柱结核的临床疗效。方法:以结核分枝杆菌H37Rv标准株构建新西兰大白兔脊柱结核模型,选取造模成功的新西兰大白兔35只。随机分为3组,实验组(A组)和对照组(B组)各14只,空白对照组(C组)7只。A组耳缘静脉滴注载INH、RFP白蛋白纳米粒(INH 3mg/kg,RFP 4mg/kg,一周一次,共12周)。B组耳缘静脉滴注INH和RFP普通注射液(INH 3mg/kg,RFP 4mg/kg,一天一次,共12周)。C组耳缘静脉滴注生理盐水(7mg/kg,一天一次,共12周)。观察三组给药后6周、12周各组脊柱结核兔的体重及病灶CT变化,并在给药的6周和12周每组各处死3只兔留取病灶椎体组织标本病理检查,采用高效液相色谱法测定血液、病灶椎体、正常椎体及肌肉组织中两种药物浓度,评估载INH、RFP白蛋白纳米粒对于兔脊柱结核模型的临床疗效。结果:A组14只模型兔经治疗后全部成活且体重均增加,治疗6周、12周后纳米粒组兔平均体重为2.94±0.11kg、3.04±0.18kg,与治疗前比较差异有统计学差异(P0.05)。B组14例模型兔经治疗后均存活,4例出现后腿瘫痪的神经症状。治疗6周、12周后平均体重维持在2.5±0.2kg,与治疗前相比无统计学差异(P0.05)。A组治疗6周时组织病理观察到椎体呈现骨质破坏的炎症修复背景,大量淋巴细胞和嗜酸性粒细胞,还有一些浆细胞存在,治疗12周时后组织病理可见大量的成骨细胞和已经修复完成的骨质结构骨小梁形成,影像学显示模型病灶处碎片骨组织消失,周围脓肿和坏死被正常骨组织替代,病椎基本治愈。B组治疗6周时病灶椎体组织病理提示椎体骨质破坏炎症反应重,大量的坏死物及纤维素性渗出物。CT扫描提示病椎有部分修复,但仍有碎片征。治疗12周时病灶椎体组织病理可见椎体大量破骨细胞、间充质细胞、毛细血管,显示病灶处于修复期。CT显示病椎为不全修复,碎片征基本消失,但椎体内仍然存在小的空洞,周边硬化带形成。治疗6周时,A、B两组INH、RFP血浆药物浓度差异有统计学意义(P0.05),病灶椎体INH、RFP有效骨组织药物浓度差异有统计学意义(P0.05),肌肉组织INH、RFP药物浓度差异无统计学意义(P0.05),治疗12周时两组INH、RFP血浆药物浓度差异有统计学意义(P0.05),病灶椎体INH、RFP有效骨组织药物浓度较差异有统计学意义(P0.05),肌肉组织INH、RFP有效组织药物浓度差异无统计学意义(P0.05)。C组分别在给予安慰剂生理盐水耳缘静脉滴注45~60d内死亡。结论:与INH、RFP普通制剂相比,载INH、RFP白蛋白纳米粒具有良好的体内缓释特性以及病灶骨组织中的有效生物分布,在兔脊柱结核模型中的治疗效果显著优于普通制剂。     

硫酸钙/聚氨基酸复合三联抗痨药人工缓释材料的制备及物理性能测定

[目的]通过压缩强度的测定和扫描电镜的观察,评价本研究制备的载三联抗痨药HRZ硫酸钙/聚氨基酸人工缓释材料的物理性能,从而制定构成缓释材料各成分的优化配比方案。[方法]实验组分为A1、A2、A3、A4、A5五个亚组,避光环境下分别根据五种不同比例的配比方案,称取适量硫酸钙/聚氨基酸、异烟肼(INH,H)、利福平(RFP,R)、吡嗪酰胺(PZA,Z)制备成直径9 mm、高6 mm圆柱状载药人工缓释材料,每一亚组制备8块,干燥。对照组B组为不含任何药物的人工缓释材料,以同法制备8块。将制备好的人工缓释材料在电子力学万能材料试验机上测定压缩强度;并在扫描电子显微镜下观察显微结构。[结果](1)实验组各亚组药占比重量A19.09%、A212.28%、A313.79%、A416.67%、A528.57%时,压缩强度分别依次为(11.63±0.18)MPa、(10.74±0.25)MPa、(10.11±0.24)MPa、(3.48±0.22)MPa、(1.18±0.18)MPa;对照组不含药物,压缩强度为(18.62±0.16)MPa;6组数据分别进行组间比较,存在差异,具有统计学意义(F=7 141.068,P=0.000);(2)硫酸钙/聚氨基酸人工缓释材料晶体之间存在大量的空隙,为承载药物提供了丰富的空间。[结论]载三联抗痨药HRZ硫酸钙/聚氨基酸人工缓释材料随着抗痨药物药占比的增加,压缩强度逐渐降低;但在一定药物含量范围内,人工材料的强度变化并不明显;硫酸钙晶体间的空隙是该人工材料承载抗痨药物的空间。     

Presenteeism and Absenteeism Before and After Total Hip and Knee Arthroplasty

BackgroundAbsenteeism is costly, yet evidence suggests that presenteeism—illness-related reduced productivity at work—is costlier. We quantified employed patients’ presenteeism and absenteeism before and after total joint arthroplasty (TJA).MethodsWe measured presenteeism (0-100 scale, 100 full performance) and absenteeism using the World Health Organization’s Health and Work Performance Questionnaire before and after TJA among a convenience sample of employed patients. We captured detailed information about employment and job characteristics and evaluated how and among whom presenteeism and absenteeism improved.ResultsIn total, 636 primary, unilateral TJA patients responded to an enrollment email, confirmed employment, and completed a preoperative survey (mean age: 62.1 years, 55.3% women). Full at-work performance was reported by 19.7%. Among 520 (81.8%) who responded to a 1-year follow-up, 473 (91.0%) were still employed, and 461 (88.7%) had resumed working. Among patients reporting at baseline and 1 year, average at-work performance improved from 80.7 to 89.4. A Wilcoxon signed-rank test indicated that postoperative performance was significantly higher than preoperative performance (P < .0001). The percentage of patients who reported full at-work performance increased from 20.9% to 36.8% (delta = 15.9%, 95% confidence interval = [10.0%, 21.9%], P < .0001). Presenteeism gains were concentrated among patients who reported declining work performance leading up to surgery. Average changes in absences were relatively small. Combined, the average monthly value lost by employers to presenteeism declined from 15.3% to 8.3% and to absenteeism from 16.9% to 15.5% (ie, mitigated loss of 8.4% of monthly value).ConclusionAmong employed patients before TJA, presenteeism and absenteeism were similarly costly. After, employed patients reported increased performance, concentrated among those with declining performance leading up to surgery.     

The German emergency and disaster medicine and management systemdhistory and present

As well for optimized emergency management in individual cases as for optimized mass medicine in disaster management, the principle of the medical doctors approaching the patient directly and timely, even close to the site of the incident, is a long-standing marker for quality of care and patient survival in Germany. Professional rescue and emergency forces, including medical services, are the “Golden Standard” of emergency management systems. Regulative laws, proper organization of resources, equipment, training and adequate delivery of medical measures are key factors in systematic approaches to manage emergencies and disasters alike and thus save lives. During disasters command, communication, coordination and cooperation are essential to cope with extreme situations, even more so in a globalized world. In this article, we describe the major historical milestones, the current state of the German system in emergency and disaster management and its integration into the broader European approach.     

Differences in Osteocyte Density and Bone Histomorphometry Between Men and Women and Between Healthy and Osteoporotic Subjects

Bone defects related to osteoporosis develop with increasing age and differ between males and females. It is currently thought that the bone remodeling process is supervised by osteocytes in a strain-dependent manner. We have shown an altered response of osteocytes from osteoporotic patients to mechanical loading, and osteocyte density is reduced in osteoporotic patients, which might relate to imperfect bone remodeling, leading to lack of bone mass and strength. Hence, information on osteocyte density will contribute to a better understanding of bone biology in males and females and to the assessment of osteoporosis. Osteocyte density as well as conventional histomorphometric parameters of trabecular bone were determined in cancellous iliac crest bone of healthy postmenopausal women and men and of osteoporotic women and men. Osteocyte density was higher in healthy females than in healthy males and lower in osteoporotic females than in healthy females. Bone mass was reduced in osteoporotic patients, both male and female. In females, trabecular number was reduced, whereas in males, trabecular thickness was reduced and eroded surface was increased. There were no correlations between the parameter groups bone architecture, bone formation, bone resorption, and osteocyte density. These results are consistent with impaired osteoblast function in osteoporotic patients and with a different mechanism of bone loss between men and women, in which osteocyte density might play a role. The reduced osteocyte numbers in female osteoporotic patients might relate to imperfect bone remodeling leading to lack of bone mass and strength. M. G. Mullender and S. D. Tan contributed equally to this work.     

Tendon and Ligament Healing and Current Approaches to Tendon and Ligament Regeneration

Ligament and tendon injuries are common problems in orthopedics. There is a need for treatments that can expedite nonoperative healing or improve the efficacy of surgical repair or reconstruction of ligaments and tendons. Successful biologically-based attempts at repair and reconstruction would require a thorough understanding of normal tendon and ligament healing. The inflammatory, proliferative, and remodeling phases, and the cells involved in tendon and ligament healing will be reviewed. Then, current research efforts focusing on biologically-based treatments of ligament and tendon injuries will be summarized, with a focus on stem cells endogenous to tendons and ligaments. Statement of clinical significance: This paper details mechanisms of ligament and tendon healing, as well as attempts to apply stem cells to ligament and tendon healing. Understanding of these topics could lead to more efficacious therapies to treat ligament and tendon injuries. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 38:7–12, 2020