壳聚糖复合骨髓间充质干细胞修复兔骨缺损的研究

目的 观察壳聚糖复合骨髓间充质干细胞(MSCs)修复兔骨缺损的作用.方法 将45只骨缺损模型兔随机分为3组.A组:骨缺损不填充任何材料;B组:骨缺损填充单纯壳聚糖微球;C组:骨缺损填充壳聚糖微球复合MSCs.分别在治疗后第4、8和12周对3组动物模型的骨缺损部位行影像学和组织学观察,并检测骨形成蛋白-2 (BMP-2)的表达.结果 影像学和组织学显示,C组复合材料有良好的诱导成骨能力,治疗第4周即有明显的成骨反应和新骨形成,治疗第12周基本修复骨缺损;B组修复能力较C组差,但B组和C组均优于A组;C组毛细血管数、管径及骨陷窝空缺百分比依次为(7.8±1.4)、(9.0±1.7) μm和(24.8±5.6)%,与A、B两组比较差异均有统计学意义(P<0.05);各时期C组BMP-2的表达水平依次为(9.8±1.5)%、(11.2±2.0)%和(16.7±2.5)%,与A、B两组同时期比较均明显升高(P<0.05).B组和C组对壳聚糖无明显异物反应.结论 壳聚糖是修复兔骨缺损的良好移植材料,复合MSCs后能促进骨缺损的修复.

Abstract：

Objective To investigate the effects of chitosan combined with bone marrow mesenchymal stem cells (MSCs) on the repair of rabbit with bone defect. Methods 45 rabbits with bone defect were randomly divided into 3 groups.Group A: defect was not filled with any implants;Group B: defect was filled with chitosan; Group C: defect was filled with chitosan combined with bone MSCs.The expression of bone morphogenetic protein-2 (BMP-2) in the 3 groups were observed by imaging and histology in the 4, 8 and 12 week after treatment. Results Group C had a better osteogenesis ability than group A and B. Group B had a better osteogenesis ability than group A.New bones and osteogenesis were obviously observed in group C in the 4 week, and the defect areas in group C were almost repaired 12 weeks after operation;The number and diameter of capillaries,percentage of vacant lacunae in group C were (7.8±1.4), (9.0±1.7) μm and (24.8±5.6)%, compared with the group A and B, it improved significantly (P<0.05);The expression of BMP-2 in group C every time were (9.8±1.5)%, (11.2±2.0)% and (16.7±2.5)%, compared with the group A and B,it improved significantly (P<0.05) during all periods.There was no significant foreign body reaction to chitosan in group B and C. Conclusion Chitosan is a superior material for repairing bone defect, and chitosan combined with MSCs have potential applications in treating bone defect.     

多孔双相陶瓷复合重组人骨形态发生蛋白-2和碱性成纤维细胞生长因子缓释微球的异位成骨活性

目的 观察复合重组人骨形态发生蛋白-2(rhBMP-2)和碱性成纤维细胞生长因子(bFGF)缓释微球的多孔双相陶瓷(BCP)异位成骨活性.方法 A组(rhBMP-2+bFGF缓释微球/BCP)、B组(BCP)、C组(bFGF缓释微球/BCP)、D组(rhBMP-2缓释微球/BCP),其中bFGF和rhBMP的浓度各为5、15μg.规格均为3mm×8mm×28mm.将材料植入兔背部肌袋内,术后4、8、12周分别取材,行大体、组织学观察,测量新骨面积和血管生成.结果 8、12周A组成骨活性较其他组优,差异有统计学意义 (P<0.05).A组4周材料中有较多间充质细胞,并有少量毛细血管的生长;8周时可见散在分布的不成熟新生骨组织,材料基本降解;12周时出现成熟度较低的编织骨,为膜内成骨.结论 BCP复合rhBMP-2和bFGF缓释微球具有良好的异位成骨活性.

Abstract：

Objective To investigate the heterotopic bone formation ability of biphasic ceramics phosphate (BCP) combined with recombinant human bone morphogenetic protein-2 (rhBMP-2) and basic fibroblast growth factor (bFGF) microspheres.Methods The rabbits were divided into group A (rhBMP-2+bFGF/BCP), group B (BCP), group C (bFGF/BCP), and group D (rhBMP-2/BCP). The concentrations of bFGF and rhBMP were 5 μg and 15 μg, respectively. All the samples were 3 mm×8 mm×28 mm in size. The muscle pouches of the rabbits were implanted with the samples. The ectopic bone formation was evaluated in the following aspects: histology, osteogenic area, and blood capillary number at 4th, 8th and 12th week after operation.Results At any specified time point, the value of the heterotopic bone formation was significantly higher in group A than other groups (P< 0.05). The histological analysis showed that group A had more mesenchymal (MES) cells and fewer blood capillaries at 4th week after operation. At 8th week, the composite was degradated and diffusely distributed, and the immature new bone was found. There were low-grade mature woven bones at 12th week, and the membranous bone formation occurred.Conclusion BCP combined with rhBMP-2 and bFGF microspheres has good heterotopic bone formation ability.     

新型多孔可吸收骨替代材料修复兔股骨缺损的实验研究

Objective To test the osteogenetic ability and biocompatibility of a new type of porous morsel blocks of resorbable bioglass bone graft substitute. Methods Thirty healthy New Zealand adult white rabbits were selected to establish bone defect models and randomly assigned into 3 even groups(with 20hind legs in each group)for 3 methods of repairing the defects.The 6 mm×10 to 12 mm defeets were made at the bilateral fenloral condyles of each rabbit.In group A,a porous morsel block of resorbable bioactive glass bone substitute was implanted;in group B,commonly used bioactive glass was implanted;in group C,nothing was implanted.Specimens were collected at 6 and 12 weeks after operation to evaluate the biodegradation and osteogenic properties by gross observation, radiography, Micro-CT and histopathological examinations.ResulIs No local adverse reaction was observed in each group.At 6 weeks.gradual growth of the new bone was observed inside the implants in groups A and B.The amount of new bone in group A was significantly higher than in group B.In group A,the porous morsel material was almost completely degraded,bone defects were completely repaired,and bone trabecula moulding reconstruction was completed.In group B,a great amount of material was not completely degraded in the central area of the graft,while bone defects were not healed in the group C.The percentages of new bone area were 0.30±0.02 in group A,0.17±0.03 in group B and 0.06±0.01 in group C,with significant differences(F=374.202,P<0.001).At 12weeks.the percentages of new bone area were 0.53 ±0.05 in group A,0.39±0.05 in group B and 0.07±0.01 in group C,with significant differences(F=317.243,P<0.001). Conclusion New porous morsel blocks of resorbable bioglass bone graft substitute show superior osteogenetic properties and biodegradability because of their good three dimensional structure.     

Repair of radius defect with bone-morphogenetic-protein loaded hydroxyapatite／collagen-poly（ L-lactic acid）composite

Objective: To explore the method to repair bone defect with bone-morphogenetic-protein loaded hydroxyapatite/collagen-poly ( L-lactic acid) composite. Methods: 18 adult beagle dogs were randomly divided into 3 groups. In Group A, bone-morphogeneticprotein ( BMP ) loaded hydroxyapatite/collagen-poly ( L-lactic acid) (HAC-PLA) scaffold was implanted in a 2 cm diaphyseal defect in the radius. In Group B, unloaded pure HAC-PLA scaffold was implanted in the defects. No material was implanted in Group C (control group). The dogs were sacrificed 6 months postoperatively. Features of biocompatibility, biodegradability and osteoinduction were evaluated with histological, radiological examinations and bone mineral density (BMD) measurements. Results: In Group A, the radius defect healed after the treatment with BMP loaded HAC-PLA. BMD at the site of the defect was higher than that of the contralateral radius. Fibrous union developed in the animals of the control group. Conclusions: BMP not only promotes osteogenesis but also accelerates degradation of the biomaterials.Optimized design parameters of a three-dimensional porous biomaterial would give full scope to the role of BMP as an osteoinductive growth factor.     

新型多孔可吸收骨替代材料修复兔股骨缺损的实验研究

目的 探讨新型多孔块状可吸收生物活性玻璃骨替代材料的骨缺损修复能力和生物相容性.方法选取30只健康成年新西兰大白兔,建立舣侧股骨髁直径为6 mm、深10～12 mm的骨缺损模型,根据骨缺损中植入的材料不同随机分为3组,每组10只20侧:实验组(A组)植人多孔块状可吸收生物活性玻璃骨替代材料,材料对照组(B组)植入常用非多孔生物活性玻璃骨替代材料,空白对照组(C组)不植入任何材料.术后6、12周取材,通过大体观察、X线片、显微CT和病理组织切片检测评价新型多孔块状可吸收生物活性玻璃骨替代材料的成骨能力和生物降解性能.结果植入后6周A、B组可见新生骨逐渐向材料内部生长,A组新生骨的数量明显多于B组.12周时A组植入材料几乎完全降解,骨缺损完全修复,成熟的骨小梁塑形改造;B组植骨中心区仍有较多材料尚未完全降解;C组骨缺损末愈合.组织切片定量分析显示:6周时A、B、C组新生骨占骨缺损的面积比平均分别为0.30±0.02、0.17±0.03、0.06±0.01,差异有统计学意义(F=374.202,P<0.001),组问两两比较差异均有统计学意义(P<0.05);12周时A、B、C组新生骨占骨缺损的面积比平均分别为0.53 ±0.05、0.39±0.05、0.07±0.01,差异有统计学意义(F=317.243,P<0.001),组间两两比较差异均有统计学意义(P<0.05).A组与B组6、12周时残留材料占骨缺损面积比差异均有统计学意义(P<0.05).结论新型多孔块状可吸收生物活性玻璃骨替代材料因具有良好的三维结构而表现出更好的成骨能力和生物降解性能.

Abstract：

Objective To test the osteogenetic ability and biocompatibility of a new type of porous morsel blocks of resorbable bioglass bone graft substitute. Methods Thirty healthy New Zealand adult white rabbits were selected to establish bone defect models and randomly assigned into 3 even groups(with 20hind legs in each group)for 3 methods of repairing the defects.The 6 mm×10 to 12 mm defeets were made at the bilateral fenloral condyles of each rabbit.In group A,a porous morsel block of resorbable bioactive glass bone substitute was implanted;in group B,commonly used bioactive glass was implanted;in group C,nothing was implanted.Specimens were collected at 6 and 12 weeks after operation to evaluate the biodegradation and osteogenic properties by gross observation, radiography, Micro-CT and histopathological examinations.ResulIs No local adverse reaction was observed in each group.At 6 weeks.gradual growth of the new bone was observed inside the implants in groups A and B.The amount of new bone in group A was significantly higher than in group B.In group A,the porous morsel material was almost completely degraded,bone defects were completely repaired,and bone trabecula moulding reconstruction was completed.In group B,a great amount of material was not completely degraded in the central area of the graft,while bone defects were not healed in the group C.The percentages of new bone area were 0.30±0.02 in group A,0.17±0.03 in group B and 0.06±0.01 in group C,with significant differences(F=374.202,P<0.001).At 12weeks.the percentages of new bone area were 0.53 ±0.05 in group A,0.39±0.05 in group B and 0.07±0.01 in group C,with significant differences(F=317.243,P<0.001). Conclusion New porous morsel blocks of resorbable bioglass bone graft substitute show superior osteogenetic properties and biodegradability because of their good three dimensional structure.     

Biplane osteotomy and bone transport combined external locking plating for sequential treatment of massive tibial bone defects北大核心CSCD

Objective To evaluate biplane osteotomy and bone transport combined external locking plating for sequential treatment of massive tibial bone defects. Methods Twenty-eight patients with massive tibial bone defects were reviewed who had been treated at Department of Repair and Reconstruction, Wuhan Puai Hospital from October 2013 to October 2019. They were divided into a bone transport group and a combined bone transport group (bone transport combined external locking plating) according to their surgical methods. In the bone transport group of 14 cases, there were 10 males and 4 females with an age of (38. 6 ±3. 2) years and a bone defect of (8. 0 ± 0. 5) cm; in the combined bone transport group of 14 cases, there were 9 males and 5 females with an age of (39. 1 ± 3. 9) years and a bone defect of (8. 3 ± 0. 3) cm. The time for wearing external fixator, fracture healing time, dock-in-site healing time, postoperative function assessment and complications were observed and compared between the 2 groups. Results There was no significant difference between the 2 groups in their preoperative general data, showing they were comparable (P > 0. 05). The bone transport group were followed up for 12 to 28 months (average, 18. 4 months) and the combined bone transport group for 12 to 26 months (average, 16. 8 months) . The time for wearing external fixator in the combined bone transport group [(8.4±0.7) months] was significantly shorter than that [(13.3±1.4) months ] in the bone transport group (P< 0. 05). No significant difference was observed between the 2 groups in either the fracture healing time [ (8. 4 土1.3) months versus (7. 4 ± 1. 2) months] or the dock-in-site healing time [(210. 2 ±9.1) months versus (206. 2 ± 9. 8) months ] (P > 0. 05). By the Association for the Study and Application of the Method of Ilizarov (ASAMI) scoring, the bone healing was excellent in 6, good in 5 and fair in 3 cases in the bone transport group while excellent in 8, good in 4 and fair in 2 cases in the combined bone transport group, showing no significant difference between the 2 groups in the excellent to good rate (P > 0. 05). By the postoperative functional assessment of the lower extremity, there were 7 excellent, 3 good, 3 fair and one poor cases in the bone transport group while 8 excellent, 5 good and one poor cases in the combined bone transport group, showing no significant difference between the 2 groups in the excellent to good rate (P > 0. 05). In the bone transport group, there were 3 cases of pin track infection, one case of dock-in-site nonunion, 2 cases of poor alignment of lower extremities, 2 cases of skin depression, one case of nail loosening, 5 cases of joint stiffness and 3 cases of delayed union of the distracted bone; in the combined bone transport group, there were one case of pin track infection, 2 cases of poor alignment of lower extremities, 3 cases of skin depression, 3 cases of joint stiffness, 2 cases of delayed union of the distracted bone and one case of refracture. Conclusion In the sequential treatment of massive tibial bone defects, biplane osteotomy and bone transport combined external locking plating can reduce the time for wearing external fixator and increase the satisfaction of patients. © The Author(s) 2022.     

RGD多肽接枝聚复合导管桥接神经缺损的实验研究

Objective To investigate the effect of RGD peptide conjugated poly[ LA-(Glc-Lys) ]/βTCP/PLA nerve conduit for bridging peripheral nerve regeneration defect. Methods Forty-five male Wister rots were randomly divided into 3 groups, with 15 rats each. A 10 mm defect was created in the right sciatic nerve. In group A the gap was bridged by PLA tube. In group B RGD peptide conjugated poly[ LA-(Glc-Lys) ]/β-TCP/PLA nerve conduit was used to repair the defect. Autologous nerve graft was done in group C which served as control. Twelve weeks postoperatively nerve regeneration was evaluated by gross observation,electrophysiology, muscle weight and muscle morphometry of triceps surae, and ultrastructure of the regenerating nerve. Results Twelve weeks after the operation, nerve conduction velocity and muscle weight recovery of group B were better than those of group A. The differences were statistically significant( P < 0.05). There was no significant difference between group B and group C ( P > 0. 05). The results of histology and ultrastructure showed that nerve regeneration in group B and group C was significantly superior to that in group A.Conclusion RGD peptide conjugated poly[LA-(Glc-Lys)]/β-TCP/PLA conduit can achieve similar results in repairing sciatic nerve defect to that of autogentic nerve graft. It may be an ideal material to repair nerve defect.     

RGD多肽接枝聚复合导管桥接神经缺损的实验研究

Objective To investigate the effect of RGD peptide conjugated poly[ LA-(Glc-Lys) ]/βTCP/PLA nerve conduit for bridging peripheral nerve regeneration defect. Methods Forty-five male Wister rots were randomly divided into 3 groups, with 15 rats each. A 10 mm defect was created in the right sciatic nerve. In group A the gap was bridged by PLA tube. In group B RGD peptide conjugated poly[ LA-(Glc-Lys) ]/β-TCP/PLA nerve conduit was used to repair the defect. Autologous nerve graft was done in group C which served as control. Twelve weeks postoperatively nerve regeneration was evaluated by gross observation,electrophysiology, muscle weight and muscle morphometry of triceps surae, and ultrastructure of the regenerating nerve. Results Twelve weeks after the operation, nerve conduction velocity and muscle weight recovery of group B were better than those of group A. The differences were statistically significant( P < 0.05). There was no significant difference between group B and group C ( P > 0. 05). The results of histology and ultrastructure showed that nerve regeneration in group B and group C was significantly superior to that in group A.Conclusion RGD peptide conjugated poly[LA-(Glc-Lys)]/β-TCP/PLA conduit can achieve similar results in repairing sciatic nerve defect to that of autogentic nerve graft. It may be an ideal material to repair nerve defect.     

RGD多肽接枝聚复合导管桥接神经缺损的实验研究

Objective To investigate the effect of RGD peptide conjugated poly[ LA-(Glc-Lys) ]/βTCP/PLA nerve conduit for bridging peripheral nerve regeneration defect. Methods Forty-five male Wister rots were randomly divided into 3 groups, with 15 rats each. A 10 mm defect was created in the right sciatic nerve. In group A the gap was bridged by PLA tube. In group B RGD peptide conjugated poly[ LA-(Glc-Lys) ]/β-TCP/PLA nerve conduit was used to repair the defect. Autologous nerve graft was done in group C which served as control. Twelve weeks postoperatively nerve regeneration was evaluated by gross observation,electrophysiology, muscle weight and muscle morphometry of triceps surae, and ultrastructure of the regenerating nerve. Results Twelve weeks after the operation, nerve conduction velocity and muscle weight recovery of group B were better than those of group A. The differences were statistically significant( P < 0.05). There was no significant difference between group B and group C ( P > 0. 05). The results of histology and ultrastructure showed that nerve regeneration in group B and group C was significantly superior to that in group A.Conclusion RGD peptide conjugated poly[LA-(Glc-Lys)]/β-TCP/PLA conduit can achieve similar results in repairing sciatic nerve defect to that of autogentic nerve graft. It may be an ideal material to repair nerve defect.     

RGD多肽接枝聚复合导管桥接神经缺损的实验研究

Objective To investigate the effect of RGD peptide conjugated poly[ LA-(Glc-Lys) ]/βTCP/PLA nerve conduit for bridging peripheral nerve regeneration defect. Methods Forty-five male Wister rots were randomly divided into 3 groups, with 15 rats each. A 10 mm defect was created in the right sciatic nerve. In group A the gap was bridged by PLA tube. In group B RGD peptide conjugated poly[ LA-(Glc-Lys) ]/β-TCP/PLA nerve conduit was used to repair the defect. Autologous nerve graft was done in group C which served as control. Twelve weeks postoperatively nerve regeneration was evaluated by gross observation,electrophysiology, muscle weight and muscle morphometry of triceps surae, and ultrastructure of the regenerating nerve. Results Twelve weeks after the operation, nerve conduction velocity and muscle weight recovery of group B were better than those of group A. The differences were statistically significant( P < 0.05). There was no significant difference between group B and group C ( P > 0. 05). The results of histology and ultrastructure showed that nerve regeneration in group B and group C was significantly superior to that in group A.Conclusion RGD peptide conjugated poly[LA-(Glc-Lys)]/β-TCP/PLA conduit can achieve similar results in repairing sciatic nerve defect to that of autogentic nerve graft. It may be an ideal material to repair nerve defect.     

自体骨髓间充质干细胞复合壳聚糖/羟基磷灰石支架修复兔膝骨软骨缺损

目的 探讨骨髓间充质干细胞(bone mesenehymal stem cells,BMSCs)复合壳聚糖(chitosan,CS)/羟基磷灰石(hydmxyapatite,HA)支架修复兔膝关节局部骨软骨缺损.方法 选健康日本大耳白兔36只,2～3月龄,体重1.7～2.0 kg,每只抽取自体骨髓4～6ml,体外分离培养BMSCs后以2×107/ml密度植于CS/HA支架上体外培养10 h,制成BMSCs-CS/HA支架复合物.将36只实验动物手术制成右膝股骨外侧髁负重区骨缺损模型后,随机分成A、B、C 3组,每组12只.A组植入BMSCs-CS/HA复合物,B组植入单纯CS/HA支架;C组不作任何植入,为空白对照组.分别于术后6周、12周各处死6只动物,取材后进行大体、组织学观察6根据改良Wakitani评分标准进行评分,评估软骨组织的修复情况,并行成组设计方差分析.结果 A组术后6周即可重建关节软骨缺损;修复软骨在观察期内逐渐变厚,软骨下骨有少量骨修复;术后12周透明软骨样修复,表面光整,与周围软骨色泽相近,软骨下骨有部分修复.而B组和C组12周时缺损区仍为纤维软骨样纤维组织修复,色泽浅黄.术后6、12周各组组织学半定量评分显示:股骨髁负重区修复A组评分明显优于B、C组(F=27.26,P＜0.05).结论 自体BMSCs复合CS/HA支架在体内环境下可形成透明软骨修复兔膝关节负重区骨软骨缺损.     

带血供脂肪筋膜瓣对骺板缺损骨桥形成的预防作用

目的 制作未成年兔的股骨远端中心型骺板缺损模型,观察带血供脂肪筋膜瓣充填骺板缺损后对骨桥形成的预防作用.方法 用3.5mm克氏针造成兔一侧股骨远端中心型骺板缺损,另一侧作正常参照.实验动物分为3组,A组用带血供脂肪筋膜瓣填塞骺板缺损;B组用游离脂肪筋膜瓣填塞;C组不做任何填塞,为单纯缺损组.20周后,取3组手术侧股骨长度、膝关节外翻角与健侧的差值进行比较,评价股骨的畸形.并通过组织学的方法观察骨桥形成及骺板自身修复情况.结果 3组的手术侧股骨长度较对侧正常股骨短,A、B、C组短缩的程度分别为(1.73±1.37)、(3.89±1.49)、(6.52±1.62) mm,3组之间差异有统计学意义(P＜0.05);3组手术侧股骨与健侧对比均有不同程度的外翻畸形,外翻角的差异分别为A组(3.73±3.41)°,B组(6.34±6.07)°,C组( 18.38±10.65)°,C组外翻畸形明显,而A、B两组之间无统计学意义(P＞0.05).组织学观察,A组脂肪筋膜瓣未被吸收,并有较规则的骺板再生;B组脂肪筋膜瓣大部分吸收,与骺板之间有骨桥形成,骺板再生不规则;C组有明显骨桥形成.结论 带血供脂肪筋膜瓣对中心型骺板缺损后的骨桥形成有明显的预防作用,且能减小骺板缺损后股骨的畸形程度.     

骨髓基质干细胞联合血管束植入构筑血管化组织工程骨修复兔大段骨缺损

目的 研究兔骨髓基质干细胞(BMSCs)联合动静脉血管束植入异种脱蛋白松质骨(XDCB)构筑血管化组织工程骨修复兔桡骨中远段完全骨膜骨缺损的能力. 方法 从兔髂嵴捕骨髓培养制备兔BMSCs,将第5代BMSCs种植于多孔XDCB,并进行成骨诱导2周制备组织工程骨,手术中分离兔桡动、静脉血管束.动物模型为制备24只兔舣侧桡骨中远段完全骨膜骨缺损1.5 cm共48侧,分4组修复(n=12),A组为空白未治疗组,B组为单纯材料+血管束植入组(XDCB+VB),C组为组织工程骨组(XDCB+BMSCs),D组为组织工程骨+血管束植入组(XDCB+BMSCs+VB),符组交叉配对.分别于术后4、8、12周行X线片、大体解剖、组织切片、生物力学等检查,观察各组骨缺损修复效能及移植物血管化情况.结果 D组骨缺损修复效能(术后12周新骨面积比2.02%±0.16%)及血管化情况(术后12周血管面积比6.89%±0.32%)优于C组(1.50%±0.28%和3.17%±0.19%),而C组又优于B组(1.59%±0.19%和6.52%±0.23%),A组骨缺损未修复,各组结果差异有统计学意义(P<0.05).结论 BMSCs联合动静脉血管束植入构筑的血管化组织工程骨能促进成骨过程和新生骨的血管化,显著提高组织工程骨修复大段骨缺损的能力.     

双相磷酸钙组织工程骨块重建股骨头内骨缺损的实验研究

目的 观察在体外构建的具有仿生结构的双相磷酸钙（biphasic calcium phosphate，BCP）组织工程骨块植入犬股骨头缺损内的骨再生情况及预防股骨头塌陷的效果。方法 以犬股骨头的松质骨样本Micro—CT（micro—computed tomography）图像为基础，提取其中的图像信息，利用三维凝胶叠层成形法制备出具有仿骨小梁结构的陶瓷支架。在体外利用诱导分化的自体骨髓间充质细胞与仿生BCP支架复合，构建组织工程骨块，将其植入10只犬的股骨头负重区骨缺损内；另取10只火作为对照组，在股骨头骨缺损区内打球植入自体松质骨粒，对比观察仿生BCP骨块的植入效果。结果 制备出的股骨头仿生BCP支架具有良好的三维空间结构，支架小梁具有一定的方向性，呈板状模型；细胞在支架表面大量生长。植入动物体内30周后，实验组火股骨头外形基本完整，新生骨质包绕支架小梁并沿支架表面生长，而对照组犬股坩头均出现不同程度的塌陷。结论 仿生BCP组织工程骨块具有良好的生物相容性，植入犬股骨头负曩区骨缺损后，能在一定程度上防止股骨头塌陷。     

多孔双相陶瓷复合重组人骨形态发生蛋白-2和碱性成纤维细胞生长因子缓释微球的生物相容性

目的 观察复合重组人骨形态发生蛋白-2(rhBMP-2)和碱性成纤维细胞生长因子(bFGF)缓释微球的多孔双相陶瓷(BCP)组织相容性.方法 A组(rhBMP-2+bFGF缓释微球/BCP)、B组(单纯BCP)、C组(bFGF缓释微球/BCP)、D组(rhBMP-2缓释微球/BCP)和E组(不含BCP).其中bFGF、dlBMP的浓度各为5、15μg,材料均为5 mm×5 mm×1mm.采用体外细胞培养的方法测定细胞黏附能力、浸提液对于兔骨髓基质细胞(BMSCs)增殖和成骨分化的影响及体外诱导BMSCs成骨分化的能力.结果 细胞在材料上黏附、生长良好.各组噻唑蓝(MTT)吸光度值、碱性磷酸酶(ALP)值均随培养时间的延长而增大.A组与B、D和E组比较对MSC有显著的促增殖作用,但低于C组(P<0.05).A组ALP活性显著高于B、C和E组,但低于D组(P<0.05).结论 复合rhBMP-2和bFGF缓释微球的BCP具有良好的生物相容性.     

髓芯减压联合自体骨髓干细胞移植治疗股骨头坏死的早期临床观察

目的 观察髓芯减压植骨联合自体骨髓干细胞移植治疗早期股骨头坏死的临床疗效.方法 根据国际骨循环研究学会(Association Research Circulation Osseous,ARCO)骨坏死分期标准,选取Ⅰ、Ⅱ期股骨头坏死32例(39髋),治疗组18例(24髋),对照组14例(15髋).治疗组经过自体骨髓干细胞采集、分离、髓芯减压后,骨髓干细胞和松质骨混合植入;对照组行髓芯减压松质骨植入.结果 所有患者均获18个月的随访,行Harris评分和影像学检查.(1)髋关节Harris评分:治疗组由(55.90±9.71)分升至(81.86±7.95)分,优良率为87.5%;而对照组由(56.69±8.32)分升至(68.14±8.65)分,优良率为60.0%,两组比较差异有统计学意义(P<0.01);(2)髋关节影像学检查:治疗组仅1例(1髋)发展为Ⅲ期塌陷,而对照组2例(2髋)发展为Ⅲ期塌陷;MRI测得坏死面积百分比,治疗组由31.88%±7.59%降至13.20%±9.56%,而对照组由32.64%±6.32%降至21.18%±8.83%,两组比较差异有统计学意义(P<0.05).结论 髓芯减压植骨联合自体骨髓干细胞移植治疗早期股骨头坏死是一种安全、有效的方法,但还需要大样本临床试验及长期随访以进一步验证.     

双相生物钙磷陶瓷生物相容性及异位骨诱导的实验研究

目的检测运用羟基磷灰石及β-磷酸三钙制备的双相钙磷陶瓷的生物相容性及其异位骨诱导效率。方法采用化学共沉淀法及过氧化氢发泡法,将羟基磷灰石及β-磷酸三钙以6∶4的比例在1 100 ℃条件下烧结3 h获得双相钙磷陶瓷,利用X线衍射评估材料组成成分。分离SD大鼠骨髓间充质干细胞,接种于双相钙磷陶瓷,通过扫描电镜、鬼笔环肽及DAPI染色观察细胞的黏附,CCK8法评估细胞增殖,碱性磷酸酶测定法评估骨髓间充质干细胞的碱性磷酸酶表达活性。将不含骨髓间充质干细胞的双相钙磷陶瓷置入比格犬竖脊肌内,于4、8、12周对样本行大体检测、组织染色,测算新骨生成率,从而评估双相钙磷陶瓷的异位骨诱导效率。结果成功制备双相钙磷陶瓷,X线衍射分析可见羟基磷灰石及β-磷酸三钙特异性的衍射峰。扫描电镜可见双相钙磷陶瓷表面广泛分布大孔及连通孔,孔壁粗糙不平,孔内可见均匀分布的微孔。鬼笔环肽及DAPI染色显示,骨髓充质干细胞在材料表面伸展黏附,共培养后逐渐从不规则形转变为均一的长梭形。CCK8法提示共培养后第1天,细胞活力降低,而第3、4、5、7天,细胞增殖活力逐渐增加。碱性磷酸酶活性检测提示,与对照组相比,共培养后第1、7天,双相钙磷陶瓷组的骨髓间充质干细胞可分泌更多的碱性磷酸酶。双相钙磷陶瓷顺利置入比格犬竖脊肌内,术后8周材料孔隙内可见骨样组织沉积,术后12周大孔成骨比例为0.77±0.11,孔内成骨面积比例为0.71±0.14。结论双相钙磷陶瓷具有良好的生物相容性及异位骨诱导效率。     

BMSCs来源成骨细胞和内皮细胞复合壳聚糖-羟基磷灰石多孔支架构建血管化组织工程骨研究

目的探讨大鼠BMSCs来源的成骨细胞和内皮细胞复合壳聚糖-羟基磷灰石多孔支架植入大鼠桡骨缺损处的成骨作用和成血管作用。方法取分离培养至第3代的SD大鼠BMSCs行成骨和成内皮细胞诱导并鉴定。分别将内皮细胞(A组)、成骨细胞(B组)、混合细胞(成骨细胞和内皮细胞比例为1∶1,C组)均匀滴加于壳聚糖-羟基磷灰石多孔支架上制备3组细胞-支架复合物,MTT检测支架内细胞增殖活性。取2月龄雄性SD大鼠30只,制作大鼠桡骨5 mm长缺损模型并分别植入3组细胞-支架复合物(n=10)。术后4、8、12周分别取移植物行HE染色观察,CD34免疫组织化学染色计数微血管密度,RT-PCR法检测骨桥蛋白(osteopontin,OPN)和骨保护素(osteoprotegrin,OPG)mRNA表达。结果 BMSCs成骨诱导7 d后ALP染色可见细胞质内蓝染颗粒,细胞核呈红染;内皮细胞诱导14 d后,CD34免疫细胞化学染色可见细胞内棕色颗粒。MTT检测示3组细胞活性随时间延长逐渐升高。HE染色示,术后12周A组未见明显类骨质形成,而有较密集的微血管结构及较多纤维组织形成;B、C组可见均质的类骨质,呈条索状和岛状分布,可见大量成骨样细胞存在。术后各时间点A、C组微血管密度均显著高于B组(P<0.05);A组术后12周微血管密度高于C组(P<0.05),其余2个时间点A、C组间差异无统计学意义(P>0.05)。A组3个时间点OPN和OPG mRNA表达水平均较低,与B、C组比较差异有统计学意义(P<0.05);B、C组分别于术后8、12周OPN mRNA表达达峰值,4周时OPG mRNA表达达峰值。结论 BMSCs来源的成骨细胞和内皮细胞按1∶1比例共培养于壳聚糖-羟基磷灰石多孔支架作为组织工程骨移植物,可以促进大鼠桡骨缺损部位骨的形成和血管化,促进骨缺损愈合。