骨膜成骨细胞修复骨缺损的实验研究

为了进一步验证骨膜成骨细胞培养建立“成骨细胞库”用于修复骨缺损的可能性，取８只家兔胫前骨膜进行成骨细胞分离培养，体外繁殖后，以明胶海绵为载体，回植于自体的桡骨缺损部。对侧以明胶海绵吸附Ｈａｎｋｓ液为对照。分别在细胞植入后第２，４，８和１２周拍摄Ｘ线片，按Ｌａｎｅ方法计分，评价其骨缺损修复愈合的能力。结果表明，植入的成骨细胞因自身的成骨能力而促进骨缺损的愈合。提示自体“成骨细胞库”的建立和移植在骨缺损的修复中是一种可以继续探索的方法     

含孔磷酸钙陶瓷复合自体成骨细胞骨组织工程大鼠模型的构建

目的 构建封闭群动物SD大鼠骨组织工程模型。研究含孔磷酸钙陶瓷为支架复合自体成骨细胞的异位骨形成和极量骨缺损修复能力。方法 成年雄性SD大鼠20只，全麻及无菌条件下取其左侧股骨及胫骨，将骨髓冲入75ml培养瓶中并在诱导培养条件下进行体外扩增分化；培养14d后将已具成骨细胞表型的细胞消化接种于经纤连蛋白表面修饰的含孔磷酸钙陶瓷上继续培养15d，再对应地将细胞-陶瓷复合体种植于大鼠皮下，肌肉内并用于颅骨极量缺损修复，分别于2-20周处死动物，从形态学及组织学角度观察比较异位骨形成及骨缺损修复情况。结果 细胞-陶瓷复合体无论种植于皮下或肌肉内均表现明显的异位骨形成能力；颅骨极量缺损修复也优于同期仅植入经表面修饰但无细胞的陶瓷对照组。结论 应用封闭群动物SD大鼠构建的骨组织工程模型避免了采用免疫缺陷动物及同源近交系动物价格昂贵，近交衰退等缺点，其结果接近于临床，因而更具有说服力。     

骨-关节软骨复合组织块构建的实验研究

目的探讨采用组织工程技术构建骨．关节软骨复合组织块的可行性。方法将分离、培养的第2代软骨细胞和成骨细胞分别接种于磷酸三钙支架上，培养2周后通过物理方法将两者连接成一个整体，然后接种于裸鼠皮下。术后8周取材，行组织学观察。结果复合组织块在体内分别形成软骨组织和成骨组织，而且两者界面整合良好。结论以软骨细胞和成骨细胞为种子细胞、以磷酸三钙为支架体外构建的组织工程软骨和骨，可在体内形成组织工程骨．关节软骨复合组织块，有望用于关节软骨缺损的修复。     

间充质干细胞体内成骨的实验研究

[目的]探讨间充质干细胞在体内的成骨能力。[方法]从羊髂嵴处抽取10～15ml骨髓组织，用Pereoll分离液按梯度离心法分离出间充质干细胞，培养、增殖后种植在多孔β-磷酸三钙上，构建组织工程化骨，植入8只羊的左后肢跖骨缺损区（长21mm）作为实验组；单纯植入多孔β-磷酸三钙陶瓷材料的8只羊作为对照组；分别在术后6、12、24周处死动物行放射学、组织学和生物力学检测。[结果]放射学和组织学检测，术后6周实验组即可见有新骨生成，对照组则无明显新骨生成；骨缺损部位新生骨样组织、编织骨和板状骨出现的时间实验组也都较对照组早，并且不经软骨介导即能直接成骨，而对照组从两端以“爬行替代”方式成骨。术后24周，放射学和生物力学检测显示实验组骨缺损几乎完全修复，对照组只有部分愈合。[结论]间充质干细胞不仅在体外具有良好的增殖能力，回植人体内后仍然具有良好的成骨能力，不需经过软骨过程就能直接形成新骨，显示了间充质干细胞作为骨组织工程种子细胞良好的应用前景。     

自体软骨细胞修复关节软骨缺损的机制探讨

目的:观察团块样自体软骨细胞植入关节软骨缺损后的病理变化,探讨自体软骨细胞移植修复关节软骨缺损的病理生理机制。方法:24只3.0kg以上4～6月龄新西兰大白兔,雌雄不限,随机分为两组:实验组和对照组。实验组12只,20%的速眠新(1mg/kg)肌肉注射麻醉后取肩关节软骨组织,0.2%Ⅱ型胶原酶消化分离软骨细胞,体外单层培养,细胞长成肉眼可见的膜状后收集固体的组织样细胞团,动物再次麻醉制造双膝股骨滑车4.0mm×6.0mm方形缺损,植入细胞团块,骨膜覆盖,缝合骨膜于双股骨髁上。对照组12只,同实验组手术方法进行缺损单纯骨膜移植。1、3、12、24周两组各3只动物空气栓塞处死取材,观察细胞团块变化和缺损修复情况。结果:1周时软骨细胞朝向关节面部分细胞变大变圆,产生大量基质;3周时此种变化更加明显,但骨膜与细胞团块已然不能分开;12周时缺损为类透明软骨组织修复;24周时修复组织为透明软骨样组织,对照组为纤维软骨组织修复。结论:关节软骨细胞体外聚集培养形成的细胞团块内的细胞有迁移生长能力;细胞团块移植方法植入的细胞数量大,表型好,细胞在缺损内不会流失;关节软骨缺损修复是由植入的细胞团块生长分化而来;自体关节软骨细胞团块植入关节缺损内后,在关节应力的影响下,先从朝向关节面的一侧逐渐发生细胞成熟分化和软骨基质产生,逐渐完成缺损的修复。     

细胞复合β-磷酸三钙生物陶瓷修复软骨缺损的实验研究

[目的]通过将骨髓间充质干细胞（MCSc）诱导的具有软骨细胞、成骨细胞表型的细胞接种到三维多孔β-磷酸三钙（β-TCP）生物陶瓷支架材料上，体外构建骨软骨复合体，探讨以β-TCP为载体建造组织工程化软骨修复骨软骨缺损的可行性。[方法]将β-TCP多孔陶瓷加工成圆柱状，并将其作为构建人工软骨的细胞支架。在支架材料上分别接种从犬骨髓干细胞培养成的具有软骨细胞、成骨细胞表型的细胞，将细胞-支架复合体共同培养1周后，移植到犬关节软骨缺损处。植入后12、16周末取材，进行大体观察、组织学及组织化学等观察。[结果]复合体体内移植后，在犬关节软骨缺损处有新生软骨形成，形成的软骨基本保持了支架材料原有形态。[结论]β-TCP多孔陶瓷可作为支架材料，复合细胞后具有修复软骨缺损的作用。     

骨膜移植修复骨缺损的实验研究及临床应用

为探寻大块骨缺损修复方法的新途径，进行了自体骨膜游离移植修复骨缺损的实验及临床研究。实验用４２只兔，于双侧胫骨作人工骨缺损模型（６ｍｍ×１８ｍｍ×５ｍｍ）。一侧随机植入自体游离骨膜片，另一侧不植入，作为对照。以组织学、Ｘ线和放射性同位素为观察指标，研究成骨过程。结果表明，骨膜植入侧骨缺损的愈合比对照侧缺损的愈合快一倍。原因可能是骨膜提供了大量成骨细胞并直接呈膜内成骨而非软骨内化骨。在此基础上，为２１例骨缺损患者应用自体胫骨骨膜片植入治疗骨缺损，面积最大１０．５ｃｍ×４ｃｍ×４ｃｍ，最小２ｃｍ×２ｃｍ×２ｃｍ；其中１７例为良性骨肿瘤，４例为骨慢性感染。骨缺损均得到修复，关节功能恢复满意。为腔洞性骨缺损的修复提供了一种新方法。     

骨膜成骨细胞分离培养及其BMP－2cDNA基因转染的初步研究

目的：为骨膜成骨细胞作为骨缺损和骨坏死基因治疗中的治疗性细胞提供依据。方法：兔胫骨骨膜分离培养其成骨细胞。利用脂质体转染技术把荧光素酶基因和ＢＭＰ－２ｃＤＮＡ基因转入骨膜成骨细胞内，观察其导入基因表达情况。结果：荧光素酶基因和ＢＭＰ－２ｃＤＮＡ基因均可以在成骨细胞内表达，结论：培养的骨膜成骨细胞可以作为受体细胞用于骨缺损和骨坏死的基因治疗。     

骨髓间质干细胞复合生物陶瓷构建组织工程化人工软骨

目的 探索以多孔β-磷酸三钙(β-TCP)生物陶瓷材料为支架，经体外诱导的骨髓间质干细胞(MSCs)为种子细胞，构建组织工程化软骨修复羊关节软骨缺损的可行性。方法将28只中国美利奴绵羊分为三组。实验组(n＝12)：分离培养羊自体第7代MSCs，经转化生长因子-β诱导后接种到顶制的β-TCP多孔生物陶瓷材料上，细胞—材料复合体经体外孵育后，无菌条件下植入预制的羊单例肋骨近端关节面缺损处；单纯材料组(n＝12)：采用单纯β-TCP材料修复羊关节软骨缺损；空白对照组(n＝4)：制备的羊关节软骨缺损区未做任何修复。术后12周和24周分别取材，进行组织学、组织化学和免疫组织化学分折。结果 实验组术后12周羊关节软骨缺损处肉眼可见透明软骨祥组织形成，组织学检查发现，材料降解明显，未降解吸收的材料孔洞内广泛分布着新生软骨组织，软骨细胞外基质丰富，Ⅱ型胶原染色阳性；术后24周，支架材料几乎完全降解，缺损区被新生软骨组织所取代。单纯材料组术后12周，缺损区边缘有新生软骨组织向支架材料内长入，支架材料吸收明显；术后24周，见从缺损区边缘长入到支架材料内的新生软骨组织逐渐增多，但材料的中心部位未发现新生软骨形成。空白对照组至术后24周，仅见少量软骨组织从缺损区边缘向缺损区内长入，缺损中央大部分区域仍未得到修复。结论 以β-TCP多孔生物陶瓷作为支架材料，以自体MSCs作为种子细胞构建组织工程化软骨修复关节软骨缺损是可行的。     

骨膜间质干细胞移植时机的实验和临床研究

目的 研究骨膜间质干细胞异体移植时机。方法 根据骨缺损内纤维组织生长时间的不同，将从大鼠骨膜分离培养的细胞分别分批植入，在植入的第2、3、4和8周时动态观察其成骨量，即骨小梁体积比；另将幼儿骨膜间质干细胞悬液，移植于26例长管状骨干骨折2周后的缺损内，观察骨折临床愈合的时间。结果 大鼠骨缺损后2周以内移植干细胞其骨小梁体积比大，愈合快，2周以后与对照组比无显著差异；临床应用9个月内病人骨折缺损愈合15例(57．7％)，骨不愈合者11例(42．3％)。结论 大鼠骨缺损2周以内移植干细胞其疗效好，大鼠实验和临床上骨缺损2周以后移植骨膜间质干细胞不能明显促进骨缺损愈合。     

不同应力环境对兔骨髓间充质干细胞修复关节软骨缺损的影响

目的探讨不同应力环境对骨髓间充质干细胞(MSCs)修复关节软骨缺损的影响. 方法将日本大耳白兔15只制成髌骨外侧脱位动物模型,平均分成3组,每组5只:即单纯载体脱位组(对照组)、移植物正常应力组及移植物脱位组.对兔MSCs进行分离、培养,以兔MSCs为种子细胞构建自体组织工程移植物修复关节软骨缺损.6周后处死动物,观察修复组织的成分和结构. 结果术后6周,移植物正常应力组修复组织浅层为软骨组织,甲苯胺蓝染色接近正常关节软骨;深层为软骨下骨,与正常关节软骨结构相似.移植物脱位组为骨组织所修复,缺损周围的正常关节软骨变薄,软骨下血管侵入正常关节软骨内,遗留在股骨髁滑车槽内的移植物在滑车槽正常关节软骨表面形成新生类透明软骨组织.单纯载体脱位组为纤维组织修复. 结论 MSCs修复关节软骨缺损,只有在正常应力状态下修复效果最佳;提示维持负重关节正常的应力刺激,对组织工程软骨修复组织的形成和维持必不可少.     

Osteochondral progenitor cells in acute and chronic canine nonunions.

This study examined the ability of cells isolated from early healing segmental defects and from tissue from chronic nonunions to support bone and cartilage formation in vivo and their response to transforming growth factor-beta1 in vitro. Ostectomies (3 mm) were created in the radial diaphysis of four dogs. The dogs were splinted 3-5 days postoperatively and then allowed to bear full weight. At 7 days, tissue in the defect was removed and any periosteum was discarded; cells in the defect tissue were released by enzymatic digestion. The dogs were splinted again and allowed to bear full weight for 12 weeks. Radiographs confirmed a persistent nonunion in each dog. Defect tissue was again removed, any periosteum was discarded, and cells were isolated. Cells were also obtained from the defect tissue by nonenzymatic means with use of explant cultures. One-half of the tissue and one-half of any preconfluent, first-passage cultures were shipped to Cleveland by overnight carrier. At second passage, cells were loaded into ceramic cubes and implanted into immunocompromised mice for 3 or 6 weeks. Harvested cubes were examined histologically for cartilage and bone with use of a semiquantitative scoring system. Confluent fourth-passage cultures of 7 and 84-day defect tissue cells were cultured with 0.03-0.88 ng/ml transforming growth factor-beta1 for 24 hours, and [3H]thymidine incorporation and alkaline phosphatase specific activity were determined. Donor-dependent differences were noted in the rate at which defect cells achieved confluence; in general, cells from 7-day tissue divided most rapidly. Seven-day defect cells formed less bone and at a slower rate than was seen in the ceramic cubes containing samples from day 84. Cells derived enzymatically behaved similarly to those from explant cultures. Ceramic cubes contained fibrous connective tissue, cartilage, bone, and fat, indicating that multipotent cells were present. Stimulation of [3H]thymidine incorporation in response to transforming growth factor-beta1 was donor dependent and variable; only two of six separate isolates of cells exposed to it had measurable alkaline phosphatase activity (which was relatively low), and none of the cultures exhibited an increase in response to transforming growth factor-beta1 for 24 hours. This indicates that mesenchymal progenitor cells are present in the healing defect tissue at 7 and 84 days and that the relative proportion of osteochondroprogenitor cells is greater at the later time. The response to transforming growth factor-beta1 is typical of multipotent mesenchymal cells but not of committed chondrocytes or osteoblasts, indicating that these committed and differentiated cells are not present in early stages of healing and suggesting that their differentiation is inhibited in chronic nonunion.     

骨髓基质细胞源性软骨细胞修复兔全层关节软骨缺损

目的观察体外诱导骨髓基质细胞(MSCs)源性软骨细胞在兔股骨滑车关节面全层软骨缺损修复中的作用. 方法高密度传代培养第3代诱导MSCs分化为软骨细胞,以酸溶性Ⅰ型胶原为载体,两者混合后形成凝胶样植入物(细胞浓度为5×106/ml).于36只新西兰大耳白兔一侧股骨滑车关节面造成3 mm×5 mm全层关节软骨缺损,凝胶样植入为实验侧;另一侧分别为单纯胶原植入组(18个膝关节)和空白对照组(18个膝关节).术后4、8、12、24、32和48周取材观察缺损修复情况及新生组织的类型.参照Pineda标准对新生组织评分. 结果实验侧术后4周,植入细胞类似软骨细胞,周围有异染基质,形成透明软骨样组织;8周,深层有软骨下骨形成,软骨细胞层较正常关节软骨厚;12周,新生软骨厚度减小,与正常软骨相近,细胞呈柱状排列,结构与正常关节软骨相似,软骨下骨形成,潮线恢复;24周,新生软骨厚度较正常薄,约占55%,表面平整,潮线附近仍有肥大的软骨细胞;32周,潮线附近无肥大软骨细胞;48周,组织结构与32周时基本相同,为类透明软骨.Pineda评分24、32和48周间无差异,与4周比较有统计学意义(P＜0.05).实验组2～48周期间关节功能良好.单纯胶原组与空白对照组缺损无修复,48周时软骨下骨外露,关节退变;关节功能逐渐减退,动度受限. 结论 MSCs源性软骨细胞移植体内可形成透明样软骨组织,24周后新生软骨特性稳定,48周时为透明样软骨,能维持良好的关节功能.     

组织工程化骨修复猕猴长段骨缺损的实验研究

目的 探讨用生物衍生骨材料和骨髓基质干细胞(MSCs)复合后构成的组织工程化骨对异体猕猴长段骨缺损的修复作用。方法 于猕猴胫骨结节抽取MSCs并使诱导分化为成骨样细胞，用5-溴脱氧尿嘧啶核苷(BrdU)标记，培养后与人源生物衍生骨材料体外复合构成组织工程化骨，植入15只异体猕猴修复桡骨2．5cm长段骨缺损作为实验组；用单纯生物衍生骨材料修复对侧同样骨缺损作为对照组；另取2只猕猴双侧桡骨同样部位和大小骨缺损旷置作为空白组。术后1、2、3、6和12周时各处死3只动物取材，空白组12周取材，行大体观察、组织学和免疫组织化学检测。结果 实验组和对照组术后1、2和3周移植物周围组织反应较明显，6周后明显减轻，12周时基本消失。实验组标记成骨样细胞于术后6周仍存在，术后12周基本消失；骨缺损部位骨样组织、软骨、编织骨和板层骨出现时间均较对照组早，且骨愈合时间提前3～6周。实验组骨缺损以多点方式直接成骨，对照组则从两端以“爬行替代”方式成骨。空白组术后12周骨缺损均无愈合。结论 生物衍生骨材料和MSCs复合构建的组织工程化骨异体植入修复猕猴长段骨缺损可超越骨段移植的“爬行替代”过程，使骨缺损能较快愈合。生物衍生骨材料和同种异体MSCs复合组织工程化骨可作为构建骨组织工程的一种较好选择方式。     

Use of a guanidine extract of demineralized bone in the treatment of osteochondral defects of articular cartilage

In order to evaluate the ability of a guanidine extract of demineralized bone to repair osteochondral defects in articular cartilage, plugs made of this extract were implanted into defects in rabbit knees. The repair tissue was examined macroscopically, histologically, and immunohistochemically at 4, 8, 12 and 30 weeks. Controls (defects that were left empty) showed no cartilage formation. Four weeks after implantation of a guanidine extract plug, histological examination showed a nonhomogeneous metachromatically stained region extending from the surface of the repair tissue down to cancellous bone. This region also was labeled by an anti-type-II collagen antibody, indicating that cartilage-like tissue had been induced. At 8 weeks, the newly formed cartilage in the subchondral and cancellous bone had been partially replaced by bone. At 12 weeks, the thickness of the newly formed cartilage layer had decreased, and most of the newly formed cartilage in the subchondral and cancellous bone had been replaced by bone. In addition, a tidemark was observed. At 30 weeks, the repair tissue was a mixture of cartilage and fibrocartilage, and there was severe degeneration of the cartilage surrounding the repaired defects. These findings indicate that osteochondral defects of articular cartilage can be partially repaired by the implantation of a guanidine extract and that the newly formed cartilage-like tissue is not permanent.     

骨髓间质干细胞复合多孔磷酸钙陶瓷修复兔颅骨缺损的研究

目的探讨以骨髓间质干细胞(MSCs)作为种子细胞、β-磷酸三钙(β-TCP)多孔生物陶瓷作为支架材料构建组织工程化人工骨，修复兔实验性颅骨标准缺损的可行性。方法选择5月龄新西兰大白兔34只，制作颅骨标准缺损后分成3组。A组(n=20)：分离培养兔同胎异体MSCs。体外扩增后接种到预制的β-TCP多孔生物陶瓷材料上，细胞-材料复合体经体外孵育后，无菌条件下植入颅骨缺损；B组(n=10)：采用单纯β-TCP材料修复兔颅骨缺损；C组(n=4)：兔实验性颅骨标准缺损区未做骨修复。术后6周和12周分别处死动物、取材，进行缺损区大体、组织学、组织化学和免疫组织化学分析。结果颅骨缺损处A组术后6周表面肉眼可见骨样组织形成，组织学提示材料部分降解，未降解吸收的材料孔洞内广泛分布着新生骨组织，成骨细胞外基质丰富，I型胶原染色阳性；术后12周，支架材料几乎完全降解，缺损区被新生骨组织所取代。B组术后6周，可见从缺损区边缘有新生骨组织向支架材料内长入，支架材料部分吸收；术后12周，可见从缺损区边缘长入到支架材料内的新生骨组织逐渐增多，但材料的中心部位未发现新生骨形成。C组术后12周，仅见少量骨组织从缺损区边缘向缺损区内长入，缺损中央大部分区域未得到修复。结论体外培养扩增的兔MSCs在不添加外源性生长因子的情况下，与pTCP复合植入后可以在体内诱导、分化为成骨细胞，并能够对标准的颅骨缺损进行有效的修复，可进一步推广应用。     

Characterization of Matrix-Induced Osteogenesis in Rat Calvarial Bone Defects: II. Origins of Bone-Forming Cells

Two experimental models that separated demineralized bone matrix (DBM) implants from the host bone were utilized to identify the origins of bone-forming cells in the repair of calvarial defects in rats. Rat DBM, Guanadine-HCl (Gdn-HCl) extracted insoluble residue of DBM, and Gdn-HCl extracted insoluble DBM to which the dialyzed Gdn-HCl extract was added back, were implanted in the two models which prevented cells of the adjacent host bone from participating in the repair. In addition, cells in the dura and in the subcutaneous tissue overlying the calvarial defect were locally labeled with ³H-thymidine to identify the origins of those cells that were stimulated to divide and differentiate to osteoblasts. Histological studies of the temporal events that occurred during the healing process in these defect models, combined with ³H-thymidine labeling demonstrated that the osteoblasts induced by DBM were initially derived from undifferentiated mesenchymal stem cells of the dura and later augmented by cells in the overlying connective tissue covering the defect, and not from cells in the cranial bone surrounding the circular defect. The cells of both dura and subcutaneous tissue were stimulated to proliferate and differentiate principally to osteoblasts and to a very much lesser extent to chondroblasts by DBM and by reconstituted components of DBM after Gdn-HCl extraction. Gdn-HCl-extracted insoluble DBM failed to induce bone or cartilage. These results indicate that the cytokines or other factors present in DBM are required to induce bone-forming cells derived from the dura and the overlying connective tissue for the repair of the calvarial defect. Received: 1 January 1999 / Accepted: 13 August 1999     

以藻酸钙为载体的可注射组织工程骨行隆鼻术的实验研究

目的：研究可注射组织工程骨做为隆鼻术植入材料的可行性。方法：从兔髂骨中获取骨髓基质成骨细胞，将骨髓基质成骨细胞与25g/L藻酸钠溶胶混合形成藻酸钠/骨髓基质成骨细胞复合物，取其2mL与0.17g硫酸钙粉末混合均匀，注射于新西兰兔鼻背部骨膜下，观察成骨情况。结果：藻酸钙/骨髓基质成骨细胞复合物植入兔鼻背部皮下4周后有类软骨样组织形成，8周时有骨小梁、骨髓腔等骨组织结构。结论：以藻酸钙为载体的可注射性组织工程骨可用于隆鼻术植入材料。     

Bone regeneration by implantation of purified,culture-expanded human mesenchymal stem cells

Bone marrow contains a population of rare progenitor cells capable of differentiating into bone, cartilage, tendon, and other connective tissues. These cells, referred to as mesenchymal stem cells, can be purified and culture-expanded from animals and humans and have been shown to regenerate functional tissue when delivered to the site of musculoskeletal defects in experimental animals. To test the ability of purified human mesenchymal stem cells to heal a clinically significant bone defect, mesenchymal stem cells isolated from normal human bone marrow were culture-expanded, loaded onto a ceramic carrier, and implanted into critical-sized segmental defects in the femurs of adult athymic rats. For comparison, cell-free ceramics were implanted in the contralateral limb. The animals were elthanized at 4, 8, or 12 weeks, and healing bone defects were compared by high-resolution radiography, immunohistochemistry, quantitative histomorphometry, and biomechanical testing. In mesenchymal stem cell-loaded samples, radiographic and histologic evidence of new bone was apparent by 8 weeks and histomorphometry demonstrated increasing bone formation through 12 weeks. Biomechanical evaluation confirmed that femurs implanted with mesenchymal stem cell-loaded ceramics were significantly stronger than those that received cell-free ceramics. These studies demonstrate that human mesenchymal stem cells can regenerate bone in a clinically significant osseous defect and may therefore provide an alternative to autogenous bone grafts.     

运用组织工程原理结合纳米技术构建骨组织的实验研究

目的 探索表面有纳米级沟槽的冻干脱钙骨基质（freeze-dried demineralized bone matrix with nanoscale topography，nFDBM）的制备方法，并探讨其构建组织工程骨的可行性。方法取犬趾皮质骨按改良Urist法制备FDBM，经掺钕钇铝石榴石（Nd：YAG）激光处理后，与犬自体骨髓间充质干细胞诱导分化来的成骨细胞体外构建nFDBM-细胞复合物，原子力显微镜及扫描电镜观察材料表面不同形态特征对细胞行为的影响;同时将片状nFDBM复合物植入右侧犬背深筋膜袋内（实验组A），短管状nFDBM植入右侧趾骨节段性缺损处（实验组C），以片状FDBM-成骨细胞复合物（对照组B）及短管状FDBM（对照组D）植入同一犬体内左侧相对应位置作为对照，术后第4、8、12周行X线片、组织学及扫描电镜观察。结果FDBM经Nd：YAG激光处理后表面形成了规则的纳米级沟槽状三维结构（深150nm，宽600-800nm）。成骨细胞在nFDBM上的黏附密度及分泌基质量均高于FDBM。实验组复合物植入体内12周时，HE染色、扫描电镜发现材料纳米沟槽一侧、材料内有新生骨组织，X线片检查呈部分钙化，而对照组几无成骨表现，其中X线片表现评分差异有统计学意义（P〈0．05）。结论Nd：YAG激光可在FDBM表面形成纳米沟槽结构，该结构有利于成骨细胞的黏附、生长和基质分泌。以组织工程方法结合纳米技术构建的nFDBM，成骨细胞复合物植入动物模型体内表现出一定的成骨能力。