磷酸钙骨水泥/纤维蛋白胶复合材料填充桡骨缺损的生物相容性
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摘要
背景:磷酸钙骨水泥与人体自身骨矿物的化学组成和结构相似,能够填充因骨折造成的骨质塌陷,具有诱导成骨作用,但其降解速度较慢。目的:评估磷酸钙骨水泥/纤维蛋白胶复合材料的生物相容性及修复桡骨缺损的可行性。方法:(1)体外细胞毒性实验:分别以磷酸钙骨水泥/纤维蛋白胶材料浸提液、苯酚溶液,含体积分数为10%胎牛血清RPMI1640培养液培养小鼠成纤维细胞,检测细胞毒性分级;(2)血液相容性实验:在兔抗凝血中分别加入磷酸钙骨水泥/纤维蛋白胶材料浸提液、生理盐水及双蒸水,检测溶血率;(3)取45只新西兰大白兔,制备双侧桡骨缺损模型,随机分3组,空白对照组不进行任何干预,实验组与骨缺损处植入磷酸钙骨水泥/纤维蛋白胶复合材料,对照组植入自体桡骨。术后4,8,16周,进行X射线、组织学、骨密度及生物力学检测。结果与结论:(1)细胞毒性:磷酸钙骨水泥/纤维蛋白胶材料浸提液的毒性为0至1级;(2)血液相容性:磷酸钙骨水泥/纤维蛋白胶材料的溶血率为3.15%;(3)体内修复实验:术后16周,X射线显示,实验组和对照组骨折线完全消失,髓腔复通,塑性完全;组织学显示,对照组骨小梁重建较明显,板层骨较成熟,髓腔再通,实验组有大量新生编织骨成网格状长入材料中,材料降解明显,降解与骨长入同步;实验组与对照组骨密度、最大负荷、最大应力和破坏能量均明显高于空白对照组(P<0.05),实验组与对照组各指标比较差异无显著性意义;(4)结果表明:磷酸钙骨水泥/纤维蛋白胶复合材料具有良好的生物相容性,修复骨缺损可促进骨组织再生,获得与自体骨移植基本相当的治疗效果。
BACKGROUND: The chemical compositions and structure of calcium phosphate bone cement are similar to those of human bone, which can fill the bone collapse caused by fracture and induce osteogenesis, but its degradation rate is slow. OBJECTIVE: To evaluate the biocompatibility of the calcium phosphate cement/fibrin glue and the feasibility of repairing radius defects. METHODS: In vitro cytotoxicity experiment: Mouse fibroblasts were cultured in the calcium phosphate bone cement/fibrin glue extracts, phenol solution, and RPMI 1640 culture medium containing 10% fetal bovine serum, respectively, to detect the cytotoxicity grade. Hemocompatibility experiment: Calcium phosphate bone cement/fibrin glue extracts, normal saline and distilled water were respectively added into the rabbit anticoagulation, to detect the hemolytic rate. Forty-five New Zealand white rabbits were enrolled and modeled into bilateral radius defects, followed by randomly allotted into three groups: blank control group without any intervention, experimental and control groups were given the implantation with calcium phosphate bone cement/fibrin glue and autologous radius, respectively. X-ray, histology, bone mineral density and biomechanical test were performed at postoperative 4, 8 and 16 weeks. RESULTS AND CONCLUSION: The toxicity grade of the calcium phosphate cement/fibrin glue was 0 to 1. The hemolytic rate of the calcium phosphate cement/fibrin glue was 3.15%. At 16 weeks postoperatively, X-ray showed that in the experimental and control groups, the fracture line disappeared completely, pulp cavity was recanalized, and in plastic completely. Histology showed that the reconstructed bone trabecular was obvious, plate layer of bone was mature, and medullary cavity recanalization appeared in the control group; there were a large number of new grid-shaped woven bone tissues growing into the material in the experimental group, with overt degradation, and degradation rate was in parallel to bone ingrowth. The bone density, the maximum load, maximum stress and failure energy in the experimental and control groups were significantly higher than those in the blank control group(P < 0.05), and all above indicators showed no significant differences between the experimental and control groups. These results manifest that the calcium phosphate bone cement/fiber protein glue composite material holding a good biocompatibility can promote bone tissue regeneration for bone defect repair, achieving similar curative effect with autologous bone transplantation.
BACKGROUND: The chemical compositions and structure of calcium phosphate bone cement are similar to those of human bone, which can fill the bone collapse caused by fracture and induce osteogenesis, but its degradation rate is slow. OBJECTIVE: To evaluate the biocompatibility of the calcium phosphate cement/fibrin glue and the feasibility of repairing radius defects. METHODS: In vitro cytotoxicity experiment: Mouse fibroblasts were cultured in the calcium phosphate bone cement/fibrin glue extracts, phenol solution, and RPMI 1640 culture medium containing 10% fetal bovine serum, respectively, to detect the cytotoxicity grade. Hemocompatibility experiment: Calcium phosphate bone cement/fibrin glue extracts, normal saline and distilled water were respectively added into the rabbit anticoagulation, to detect the hemolytic rate. Forty-five New Zealand white rabbits were enrolled and modeled into bilateral radius defects, followed by randomly allotted into three groups: blank control group without any intervention, experimental and control groups were given the implantation with calcium phosphate bone cement/fibrin glue and autologous radius, respectively. X-ray, histology, bone mineral density and biomechanical test were performed at postoperative 4, 8 and 16 weeks. RESULTS AND CONCLUSION: The toxicity grade of the calcium phosphate cement/fibrin glue was 0 to 1. The hemolytic rate of the calcium phosphate cement/fibrin glue was 3.15%. At 16 weeks postoperatively, X-ray showed that in the experimental and control groups, the fracture line disappeared completely, pulp cavity was recanalized, and in plastic completely. Histology showed that the reconstructed bone trabecular was obvious, plate layer of bone was mature, and medullary cavity recanalization appeared in the control group; there were a large number of new grid-shaped woven bone tissues growing into the material in the experimental group, with overt degradation, and degradation rate was in parallel to bone ingrowth. The bone density, the maximum load, maximum stress and failure energy in the experimental and control groups were significantly higher than those in the blank control group(P < 0.05), and all above indicators showed no significant differences between the experimental and control groups. These results manifest that the calcium phosphate bone cement/fiber protein glue composite material holding a good biocompatibility can promote bone tissue regeneration for bone defect repair, achieving similar curative effect with autologous bone transplantation.
引文
[1]周磊,闫景龙,胡春杰,等.聚磷酸钙纤维/磷酸钙骨水泥复合材料修复骨缺损的实验研究[J].中国脊柱脊髓杂志,2006,16(11):851-855.
[2]Tyllianakis M,Giannikas D,Panagopoulos A,et al.Use of injectable calcium phosphate in the treatment of intra-articular distal radius fractures.Orthopedics.2002;25(3):311-315.
[3]高俊,张曦,盛永华,等.外固定支架结合注射用磷酸钙骨水泥治疗桡骨远端不稳定骨折[J].中国骨与关节损伤杂志,2008,23(5):407-408.
[4]黄程川,马川,黄家骏,等.磷酸钙骨水泥与自体骨植骨治疗C2型桡骨远端骨折的疗效比较[J].四川医学,2012,33(3):505-507.
[5]胡运生,范清宇,马保安,等.磷酸钙/纤维蛋白胶复合支架材料的结构及力学性能分析[J].功能材料,2006,37(4):607-610.
[6]Tobe M,Mizutani K,Tsubuku Y.Treatment of Distal Radius Fracture With the Use of Calcium Phosphate Bone Cement as a Filler.Tech Hand Up Extrem Surg.2004;8(2):95-101.
[7]Iida K,Sudo A,Ishiguro S,et al.Clinical and radiological results of calcium phosphate cement-assisted balloon osteoplasty for Colles'fractures in osteoporotic senile female patients.J Orthop Sci.2010;15(2):204-209.
[8]Bohner M,Baumgart F.Theoretical model to determine the effects of geometrical factors on the resorption of calcium phosphate bone substitutes.Biomaterials.2004;25(17):3569-3582.
[9]代宇.磷酸钙骨水泥填充治疗桡骨远端骨质疏松骨折的实验研究[D].遵义医学院,2010.
[10]唐理英,刘运辉,谢诗涓,等.经皮注射磷酸钙骨水泥治疗桡骨远端骨质疏松性骨折的临床应用[J].临床医学工程,2015,22(6):709-710.
[11]Tobe M,Mizutani K,Tsubuku Y.Treatment of distal radius fracture with the use of calcium phosphate bone cement as a filler.Tech Hand Up Extrem Surg.2004;8(2):95.
[12]Leroux T,Perez-Ordonez B,von-Schroeder HP,et al.Osteolysis after the use of a silicon-stabilized tricalcium phosphate-based bone substitute in a radius fracture:a case report.J Hand Surg Am.2007;32(4):497-500.
[13]代宇,何慕舜,叶俊强,等.磷酸钙骨水泥治疗老年骨质疏松性桡骨远端骨折的疗效观察[J].中国骨与关节损伤杂志,2010,25(3):246-247.
[14]夏长所,叶发刚,洪光祥,等.磷酸钙人工骨结合骨髓基质干细胞移植修复骨缺损的实验研究[J].中华手外科杂志,2004,20(3):174-176.
[15]张才龙,夏长所,叶发刚,等.磷酸钙人工骨复合骨髓基质干细胞移植修复骨缺损的实验[J].中国临床康复,2005,9(46):140-142.
[16]Ishiguro S,Oota Y,Sudo A,et al.Calcium phosphate cement-assisted balloon osteoplasty for a Colles'fracture on arteriovenous fistula forearm of a maintenance hemodialysis patient.J Hand Surg Am.2007;32(6):821-826.
[17]邹国耀,江峰.自体游离骨膜骨髓自固化磷酸钙复合移植治疗低负重或非负重部位节段性骨缺损的实验研究[J].中国临床康复,2004,8(11):2152-2153.
[18]Cross KJ,Huq NL,Stanton DP,et al.NMR studies of a novel calcium,phosphate and fluoride delivery vehicle-alpha(S1)-casein(59-79)by stabilized amorphous calcium fluoride phosphate nanocomplexes.Biomaterials.2004;25(20):5061-5069.
[19]immermann R,Gabl M,Lutz M,et al.Injectable calcium phosphate bone cement Norian SRS for the treatment of intra-articular compression fractures of the distal radius in osteoporotic women.Arch Orthop Trauma Surg.2003;123(1):22-27.
[20]张志鹏,闫冰,闫景龙,等.同种异体微小颗粒骨/磷酸钙骨水泥复合物修复骨缺损的实验研究[J].中国康复医学杂志,2005,20(7):514-516.
[21]Abramo A,Geijer M,Kopylov P,et al.Osteotomy of distal radius fracture malunion using a fast remodeling bone substitute consisting of calcium sulphate and calcium phosphate.J Biomed Mater Res B Appl Biomater.2010;92B(1):281-286.
[22]王剑龙,米雷,候光辉,等.新型组织工程化骨磷酸钙骨水泥/聚乳酸-聚羟基乙酸复合骨髓间充质干细胞修复兔桡骨缺损[J].中国组织工程研究与临床康复,2008,12(41):8001-8005.
[23]廖红兴,刘展亮,邹学农.磷酸钙骨水泥复合骨形态发生蛋白6和血管内皮生长因子修复骨缺损[J].中国组织工程研究,2014,18(8):1155-1160.
[24]李光宇,董洲,刘鑫,等.明胶-壳聚糖-羟基磷灰石-米诺环素仿生纳米复合材料修复桡骨缺损[J].中国组织工程研究,2015,19(30):4757-4763.
[25]陈盈君,王南翔,周磊,等.丝素纤维/磷酸钙骨水泥复合材料修复兔桡骨骨缺损的实验研究[J].中华创伤骨科杂志,2014,16(1):62-66.
[26]郝伟,姜明,王新,等.重组骨形态发生蛋白2、碱性成纤维细胞生长子共转染骨髓间充质干细胞复合纳米羟基磷灰石/重组类人胶原基/聚乳酸复合支架材料支架修复桡骨缺损研究[J].中华实验外科杂志,2013,30(5):1016-1019.
[27]马佳滨,周磊,闫景龙,等.磷酸钙骨水泥/聚磷酸钙纤维复合不同配比的颗粒骨修复兔桡骨缺损[J].中华创伤杂志,2011,27(8):737-741.
[28]梁建平,刘兴炎,葛宝丰,等.磷酸钙骨水泥复合微小颗粒骨修复兔桡骨缺损[J].中国矫形外科杂志,2010,18(6):486-488.
[29]刘洪,臧小方,赵自平,等.磷酸钙骨水泥与外源性神经生长因子复合移植修复免桡骨缺损[J].中国组织工程研究与临床康复,2008,12(41):8037-8041.
[30]Hidaka N,Yamano Y,Kadoya Y,et al.Calcium phosphate bone cement for treatment of distal radius fractures:a preliminary report.J Orthop Sci.2002;7(2):182-187.
[31]Seeherman HJ,Li XJ,Smith E,et al.rhB MP-2/calcium phosphate matrix induces bone formation while limiting transient bone resorption in a nonhuman primate core defect model.J Bone Joint Surg Am.2012;94/A(19):1765-1776.
[2]Tyllianakis M,Giannikas D,Panagopoulos A,et al.Use of injectable calcium phosphate in the treatment of intra-articular distal radius fractures.Orthopedics.2002;25(3):311-315.
[3]高俊,张曦,盛永华,等.外固定支架结合注射用磷酸钙骨水泥治疗桡骨远端不稳定骨折[J].中国骨与关节损伤杂志,2008,23(5):407-408.
[4]黄程川,马川,黄家骏,等.磷酸钙骨水泥与自体骨植骨治疗C2型桡骨远端骨折的疗效比较[J].四川医学,2012,33(3):505-507.
[5]胡运生,范清宇,马保安,等.磷酸钙/纤维蛋白胶复合支架材料的结构及力学性能分析[J].功能材料,2006,37(4):607-610.
[6]Tobe M,Mizutani K,Tsubuku Y.Treatment of Distal Radius Fracture With the Use of Calcium Phosphate Bone Cement as a Filler.Tech Hand Up Extrem Surg.2004;8(2):95-101.
[7]Iida K,Sudo A,Ishiguro S,et al.Clinical and radiological results of calcium phosphate cement-assisted balloon osteoplasty for Colles'fractures in osteoporotic senile female patients.J Orthop Sci.2010;15(2):204-209.
[8]Bohner M,Baumgart F.Theoretical model to determine the effects of geometrical factors on the resorption of calcium phosphate bone substitutes.Biomaterials.2004;25(17):3569-3582.
[9]代宇.磷酸钙骨水泥填充治疗桡骨远端骨质疏松骨折的实验研究[D].遵义医学院,2010.
[10]唐理英,刘运辉,谢诗涓,等.经皮注射磷酸钙骨水泥治疗桡骨远端骨质疏松性骨折的临床应用[J].临床医学工程,2015,22(6):709-710.
[11]Tobe M,Mizutani K,Tsubuku Y.Treatment of distal radius fracture with the use of calcium phosphate bone cement as a filler.Tech Hand Up Extrem Surg.2004;8(2):95.
[12]Leroux T,Perez-Ordonez B,von-Schroeder HP,et al.Osteolysis after the use of a silicon-stabilized tricalcium phosphate-based bone substitute in a radius fracture:a case report.J Hand Surg Am.2007;32(4):497-500.
[13]代宇,何慕舜,叶俊强,等.磷酸钙骨水泥治疗老年骨质疏松性桡骨远端骨折的疗效观察[J].中国骨与关节损伤杂志,2010,25(3):246-247.
[14]夏长所,叶发刚,洪光祥,等.磷酸钙人工骨结合骨髓基质干细胞移植修复骨缺损的实验研究[J].中华手外科杂志,2004,20(3):174-176.
[15]张才龙,夏长所,叶发刚,等.磷酸钙人工骨复合骨髓基质干细胞移植修复骨缺损的实验[J].中国临床康复,2005,9(46):140-142.
[16]Ishiguro S,Oota Y,Sudo A,et al.Calcium phosphate cement-assisted balloon osteoplasty for a Colles'fracture on arteriovenous fistula forearm of a maintenance hemodialysis patient.J Hand Surg Am.2007;32(6):821-826.
[17]邹国耀,江峰.自体游离骨膜骨髓自固化磷酸钙复合移植治疗低负重或非负重部位节段性骨缺损的实验研究[J].中国临床康复,2004,8(11):2152-2153.
[18]Cross KJ,Huq NL,Stanton DP,et al.NMR studies of a novel calcium,phosphate and fluoride delivery vehicle-alpha(S1)-casein(59-79)by stabilized amorphous calcium fluoride phosphate nanocomplexes.Biomaterials.2004;25(20):5061-5069.
[19]immermann R,Gabl M,Lutz M,et al.Injectable calcium phosphate bone cement Norian SRS for the treatment of intra-articular compression fractures of the distal radius in osteoporotic women.Arch Orthop Trauma Surg.2003;123(1):22-27.
[20]张志鹏,闫冰,闫景龙,等.同种异体微小颗粒骨/磷酸钙骨水泥复合物修复骨缺损的实验研究[J].中国康复医学杂志,2005,20(7):514-516.
[21]Abramo A,Geijer M,Kopylov P,et al.Osteotomy of distal radius fracture malunion using a fast remodeling bone substitute consisting of calcium sulphate and calcium phosphate.J Biomed Mater Res B Appl Biomater.2010;92B(1):281-286.
[22]王剑龙,米雷,候光辉,等.新型组织工程化骨磷酸钙骨水泥/聚乳酸-聚羟基乙酸复合骨髓间充质干细胞修复兔桡骨缺损[J].中国组织工程研究与临床康复,2008,12(41):8001-8005.
[23]廖红兴,刘展亮,邹学农.磷酸钙骨水泥复合骨形态发生蛋白6和血管内皮生长因子修复骨缺损[J].中国组织工程研究,2014,18(8):1155-1160.
[24]李光宇,董洲,刘鑫,等.明胶-壳聚糖-羟基磷灰石-米诺环素仿生纳米复合材料修复桡骨缺损[J].中国组织工程研究,2015,19(30):4757-4763.
[25]陈盈君,王南翔,周磊,等.丝素纤维/磷酸钙骨水泥复合材料修复兔桡骨骨缺损的实验研究[J].中华创伤骨科杂志,2014,16(1):62-66.
[26]郝伟,姜明,王新,等.重组骨形态发生蛋白2、碱性成纤维细胞生长子共转染骨髓间充质干细胞复合纳米羟基磷灰石/重组类人胶原基/聚乳酸复合支架材料支架修复桡骨缺损研究[J].中华实验外科杂志,2013,30(5):1016-1019.
[27]马佳滨,周磊,闫景龙,等.磷酸钙骨水泥/聚磷酸钙纤维复合不同配比的颗粒骨修复兔桡骨缺损[J].中华创伤杂志,2011,27(8):737-741.
[28]梁建平,刘兴炎,葛宝丰,等.磷酸钙骨水泥复合微小颗粒骨修复兔桡骨缺损[J].中国矫形外科杂志,2010,18(6):486-488.
[29]刘洪,臧小方,赵自平,等.磷酸钙骨水泥与外源性神经生长因子复合移植修复免桡骨缺损[J].中国组织工程研究与临床康复,2008,12(41):8037-8041.
[30]Hidaka N,Yamano Y,Kadoya Y,et al.Calcium phosphate bone cement for treatment of distal radius fractures:a preliminary report.J Orthop Sci.2002;7(2):182-187.
[31]Seeherman HJ,Li XJ,Smith E,et al.rhB MP-2/calcium phosphate matrix induces bone formation while limiting transient bone resorption in a nonhuman primate core defect model.J Bone Joint Surg Am.2012;94/A(19):1765-1776.