长骨节段性缺损修复重建的理论及应用进展

背景：长骨节段性骨缺损修复重建方法众多,各有优缺点,限制了其广泛应用,仍是一个未完美解决的骨科难题。 目的：综述近年国内外长骨节段性缺损的修复重建进展研究。 方法：第一作者应用计算机检索1990年1月至2012年12月PubMed数据库、中国期刊全文数据库、维普数据库有关长骨节段性缺损修复重建的文章,英文检索词“bone defect, long bone reconstruction, tissue engineering of bone,scaffolds,bone reconstruction,bone graft,bone tumor; tumor resection,musculoskeletal tumors,regeneration,autografts”;中文检索词“大段,骨缺损,骨肿瘤,软组织肿瘤,瘤段切除,组织工程,骨移植”。共检索到104篇相关文献,纳入52篇文献进一步研究。 结果与结论：传统长骨节段性骨缺损的修复方法包括自体骨移植、异体骨移植、骨延长、人工假体置换等。研制适用于儿童保肢需要,符合儿童生长发育特点的假体成为儿童保肢的一个挑战课题。为了解决这一问题,人们研制出可以不断延长的假体,用于儿童骨肿瘤的保肢重建。随着医疗技术的进步,各种人工骨,尤其是骨组织工程和基因治疗在骨缺损修复中的逐步应用将进一步提高骨缺损的修复效果,是未来的研究方向。     

丝素蛋白-壳聚糖支架复合骨髓间充质干细胞体内构建组织工程化软骨的生物相容性

文题释义： 生物相容性：是指生命体组织对非活性材料产生的一种性能,一般是指材料与宿主之间的相容性,包括组织相容性和血液相容性。 检测相容性的方法：是将支架材料与种子细胞在体外共培养,检测支架毒性、细胞活性、细胞增殖及细胞与支架的黏附情况等指标,该方法具有客观性强、可重复性强、影响因素相对简单及敏感性高等特点。 背景：课题组前期的研究中发现,丝素蛋白-壳聚糖支架材料复合诱导后骨髓间充质干细胞在兔体内能修复缺损的软骨组织,但对于该组织工程化软骨组织的生物相容性还未进一步研究。 目的：研究丝素蛋白-壳聚糖支架材料复合骨髓间充质干细胞在体内构建组织工程化软骨的生物相容性。 方法：使用丝素蛋白-壳聚糖按1∶1比例混合制备三维支架材料,提取兔骨髓间充质干细胞,将诱导后的骨髓间充质干细胞与丝素蛋白-壳聚糖支架构建修复体,再将修复体移植到兔关节软骨缺损模型中修复软骨组织。实验分为3组,实验组植入诱导后骨髓间充质干细胞+丝素蛋白-壳聚糖支架,对照组植入丝素蛋白-壳聚糖支架干预,空白组未植入修复体。 结果与结论：①实验成功制备丝素蛋白-壳聚糖三维支架材料及提取骨髓间充质干细胞,并构建软骨缺损的修复体,将修复体植入兔体内能成功修复缺损的软骨组织;②建模后2,4,8,12周,3组血常规、降钙素原、血沉、C-反应蛋白结果提示无明显的全身感染征象,3组血常规及肝肾功能各时间段比较差异无显著性意义(P > 0.05);③一般观察、苏木精-伊红染色及扫描电镜观察：建模后12周,相比其他两组,实验组软骨缺损已修复,支架材料已吸收,修复组织周围未见炎性细胞,修复组织已正常组织整合良好;④结果证实,丝素蛋白-壳聚糖支架复合骨髓间充质干细胞在体内构建的组织工程化软骨具有良好的生物相容性。 ORCID: 0000-0002-8139-1175(佘荣峰) 中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程     

A model for tissue engineering applications: femoral critical size defect in immunodeficient mice

Animal models for preclinical functionality assays lie midway between in vitro systems such as cell culture and actual clinical trials. We have developed a novel external fixation device for femoral critical size defect (CSD) in the femurs of immunodeficient mice as an experimental model for studying bone regeneration and bone tissue engineering. The external fixation device comprises four pointed rods and dental acrylic paste. A segmental bone defect (2 mm) was created in the midshaft of the mouse femur. The CSD in the femur of the mice were either left untreated or treated with a bone allograft, a cell-scaffold construct, or a scaffold-only construct. The repair and healing processes of the CSD were monitored by digital x-ray radiography, microcomputed tomography, and histology. Repair of the femoral CSD was achieved with the bone allografts, and partial repair of the femoral CSD was achieved with the cell scaffold and the scaffold-only constructs. No repair of the nongrafted femoral CSD was observed. Our results establish the feasibility of this new mouse femoral model for CSD repair of segmental bone using a simple stabilized external fixation device. The model should prove especially useful for in vivo preclinical proof-of-concept studies that involve cell therapy-based technologies for bone tissue engineering applications in humans.     

Repair of canine mandibular bone defects with bone marrow stromal cells and porous beta-tricalcium phosphate

Tissue engineering has become a new approach for repairing bone defects. Previous studies have been limited to the use of slow-degradable scaffolds with bone marrow stromal cells (BMSCs) in mandibular reconstruction. In this study, a 30 mm long mandibular segmental defect was repaired by engineered bone graft using osteogenically induced autologous BMSCs seeded on porous beta-tricalcium phosphate (beta-TCP, n=5). The repair of defects was compared with those treated with beta-TCP alone (n=6) or with autologous mandibular segment (n=4). In the BMSCs/beta-TCP group, new bone formation was observed from 4 weeks post-operation, and bony-union was achieved after 32 weeks, which was detected by radiographic and histological examination. In contrast, minimal bone formation with almost fibrous connection was observed in the group treated with beta-TCP alone. More importantly, the engineered bone with BMSCs/beta-TCP achieved a satisfactory biomechanical property in terms of bending load strength, bending displacement, bending stress and Young's modulus at 32 weeks post-operation, which was very close to those of contralateral edentulous mandible and autograft bone (p>0.05). Based on these results, we conclude that engineered bone from osteogenically induced BMSCs and biodegradable beta-TCP can well repair the critical-sized segmental mandibular defects in canines.     

组织工程人工骨材料修复长骨节段性缺损的现状与展望

背景：骨移植、人工替代物置换、骨延长等长骨节段性缺损修复方法虽已在临床上应用,但均有各自的局限性。 目的：就目前骨缺损的治疗加以回顾总结,并对长骨大段性缺损的修复方式作展望性分析。 方法：由第一作者检索PubMed数据库、中文生物医学文献期刊数据库2000-01/2010-12有关长骨大段性缺损修复的文章。英文关键词为“bone defect, bone tissue engineering,artificial bone,bone transplantation,therapy”,中文关键词为“骨缺损,骨组织工程,人工骨,骨移植,治疗”。排除标准陈旧性文章及重复性研究。 结果与结论：保留33篇文献进行文献证据提取。长骨节段性缺损的治疗包括自体骨移植、异体骨移植、骨延长、人工骨替代移植等,以组织工程人工骨的研究、利用为热点和主要发展方向,但以基础性研究和动物实验为多,少见于临床应用。组织工程人工骨在应用于临床长骨缺损治疗之前,尚存诸多问题急需解决,但仍为最有开发前景的骨缺损修复材料。     

Use of a biomimetic strategy to engineer bone

Engineering trabecular-like, three-dimensional bone tissue throughout biodegradable polymer scaffolds is a significant challenge. Using a novel processing technique, we have created a biodegradable scaffold with geometry similar to that of trabecular bone. When seeded with bone-marrow cells, new bone tissue, the geometry of which reflected that of the scaffold, was evident throughout the scaffold volume and to a depth of 10 mm. Preseeded scaffolds implanted in non-healing rabbit segmental bone defects allowed new functional bone formation and bony union to be achieved throughout the defects within 8 weeks. This marks the first report of successful three-dimensional bone-tissue engineering repair using autologous marrow cells without the use of supplementary growth factors. We attribute our success to the novel scaffold morphology.     

Experimental repair of segmental bone defects in rabbits by angiopoietin-1 gene transfected MSCs seeded on porous β-TCP scaffolds

Segmental bone defect repair remains a clinical and experimental challenge in tissue engineering with increasing focus on angiogenesis in the bone substitutes. The objective of this study was to investigate the osteogenic effects of angiopoietin-1 (Ang-1) gene transfected bone marrow-derived mesenchymal stem cells (MSCs) seeded on porous β-TCP scaffolds. This bone substitute (experimental group) and MSCs/β-TCP compounds (control group) were implanted into 15 mm segmental bone defects of the radii of 30 New Zealand white rabbits, with platelet-rich plasma injected at the same time. Bone regeneration and angiogenesis were assessed by Scanning electron microscope (SEM), X-ray, histology, immunohistology, and biomechanical outcome measurements made on the 2nd, 4th, 8th, and 12th week after the operation. In vitro, the amount of proliferation and differentiation of Ang-1 gene transfected MSCs was found to be gross increased than that of the control groups. In vivo, a significantly increased amount of new bone formation accompanied by active capillary vasculature regeneration was observed in the pores of the scaffolds which had been seeded with Ang-1 gene transfected MSCs, as compared with the control groups. The biomechanical test confirmed the failure load of new born bone was close to normal bone. These results suggest that transfer of gene encoding Ang-1 to MSCs increases their osteogenic properties by enhancing capillary regeneration, thus providing a rich blood supply for new bone formation in segmental bone defects.     

脂肪干细胞和生物支架应用于牙槽骨修复

背景：各种生理或病理因素导致的牙槽骨的吸收、缺损是口腔临床医学中的常见问题,但目前较常用的修复缺损牙槽骨的方法不能完全满足临床需要。骨组织工程的出现成为修复骨缺损的研究热点。  目的：就脂肪干细胞的来源和应用价值、生物支架的种类及特性、生物支架对种子细胞的影响及脂肪干细胞复合支架用于动物实验的研究等方面作一总结。 方法：应用计算机检索CNKI和Pubmed数据库中1995年1月至2013年4月关于脂肪干细胞、生物支架及骨修复的文章,在标题和摘要中以“脂肪干细胞,分化、增殖和成骨生物支架,牙槽骨,骨组织工程”或“Adipose stem cells,Differentiation、proliferation and Osteogenesis,Biological scaffold,alveolar bone,bone tissue engineering”为检索词进行检索。选择文章内容与脂肪干细胞和生物支架应用于修复骨缺损有关者,同一领域文献则选择近期发表或发表在权威杂志文章。初检得到163篇文献,根据纳入标准选择关于脂肪干细胞和生物支架应用于修复骨缺损的40篇文献进行综述。 结果与结论：脂肪干细胞具有与骨髓基质干细胞相似的分化潜能,因其来源广、易采集、易培养低衰老,成骨分化好和风险小等特点被广泛关注,尤其和生物支架应用于骨修复表现出更好的成骨效果。随着有关各科学的发展牙槽骨缺损的修复有关问题都可以解决,脂肪干细胞和生物支架构建工程骨将是实现真正意义上牙槽骨再生的发展趋势且具有良好的发展前景。中国组织工程研究杂志出版内容重点：干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程全文链接：     

Hierarchically biomimetic bone scaffold materials: nano-HA/collagen/PLA composite

A bone scaffold material (nano-HA/ collagen/PLA composite) was developed by biomimetic synthesis. It shows some features of natural bone both in main composition and hierarchical microstructure. Nano-hydroxyapatite and collagen assembled into mineralized fibril. The three-dimensional porous scaffold materials mimic the microstructure of cancellous bone. Cell culture and animal model tests showed that the composite material is bioactive. The osteoblasts were separated from the neonatal rat calvaria. Osteoblasts adhered, spread, and proliferated throughout the pores of the scaffold material within a week. A 15-mm segmental defect model in the radius of the rabbit was used to evaluate the bone-remodeling ability of the composite. Combined with 0.5 mg rhBMP-2, the material block was implanted into the defect. The segmental defect was integrated 12 weeks after surgery, and the implanted composite was partially substituted by new bone tissue. This scaffold composite has promise for the clinical repair of large bony defects according to the principles of bone tissue engineering.     

3D bioprinting in orthopedics translational research

Abstract

The repair of critical-size bone defect remains a challenge for orthopedic surgeons. With the advent of an aging society and their accompanying chronic diseases, it is becoming more difficult to treat bone defects, especially large segmental bone defects that are caused by trauma, tumors, infections, and congenital malformations. New materials and technologies need to be developed to address these conditions. 3D bioprinting is a novel technology that bridges the biomaterial and living cells and is an important method in tissue engineering projects. 3D bioprinting has the advantages of replacing or repairing damaged tissue and organs. The progress in material science and 3D printing devices make 3D bioprinting a technology which can be used to create various scaffolds with a large range of advanced material and cell types. However, in regard to the widespread use of bioprinting, biosafety, immunogenicity and rising costs are rising to be concerned. This article reviews the developments and applications of 3D bioprinting and highlights newly applied techniques and materials and the recent achievements in the orthopedic field. This paper also briefly reviews the difference between the methods of 3D bioprinting. The challenges are also elaborated with the aim to research materials, manufacture scaffolds, promote vascularization and maintain cell viability.     

组织工程骨修复兔桡骨节段性缺损血液流变性及局部血流量的变化

目的 观察组织工程骨修复骨缺损过程中，实验动物血液流变性和骨缺损修复区血流量的变化。方法 选择30只新西兰白兔，制作15mm长的桡骨节段性骨缺损模型，根据植入不同移植材料分为实验组和对照组，实验组于动物左侧桡骨缺损区植入组织工程骨，对照组植入部分脱蛋白骨，观察各组动物术后1h、7h、14d血液流变性和术后14d骨缺损修复区血流量的变化。结果 实验组与对照组比较，血液流变学指标和骨缺损修复区血流量差异显著。组织工程骨修复骨缺损，实验动物全身血液黏度降低，骨缺损修复区局部血流量增加。结论 与单纯材料比较，组织工程骨可促进血液流变学及局部血流量的改善。     

骨诱导性磷酸钙陶瓷为支架的组织工程骨修复狗下颌骨箱状缺损*☆◆

背景：应用不外加生长因子或细胞而具有骨诱导性的生物材料,在非骨部位构建骨移植物,即体内组织工程骨,其在修复箱状及节段性骨缺损方面,具有更可行的前景。 目的：采用骨诱导性钙磷陶瓷材料构建体内组织工程化类骨移植物,探索其应用于修复实验动物下颌骨箱状骨缺损的可行性。 方法：以骨诱导性磷酸钙陶瓷材料为支架植入狗肌肉内构建体内组织工程骨,同期在狗自体下颌骨左右两侧各拔除牙弓中段牙2颗,形成约20 mm无牙区。8周后在无牙区形成箱状缺损,同期取出支架即刻移植入一侧自体下颌骨缺损区,对侧骨缺损区直接移植入未经体内构建的磷酸钙陶瓷作为对照。 结果与结论：经肌肉内构建的体内组织工程骨移植物的力学性能较单纯磷酸钙陶瓷有明显提高。颌骨缺损区的核吸收强度明显强于对照区,其移植物内长入的骨组织较多,两者的成骨面积差异有非常显著性意义(P < 0.01)。说明在修复颌骨大范围缺损中,体内组织工程骨移植物较单纯骨磷酸钙陶瓷替代材料表现出明显的力学和生物学优势,修复效果显著,有良好的应用前景。关键词：骨诱导性磷酸钙陶瓷;体内组织工程骨;骨缺损;支架;组织工程 缩略语注释：BCP：biphasic calcium phosphate,双相磷酸钙 doi:10.3969/j.issn.1673-8225.2012.16.001     

人骨形成蛋白7修饰骨髓间充质干细胞复合仿生型支架材料的体内成骨研究

目的 应用携带人骨形成蛋白7(hBMP7)基因的兔骨髓间充质干细胞(BMSC)与仿生型生物玻璃-胶原-透明质酸-磷脂酰丝氨酸(BG-COL-HYA-PS)支架材料复合培养,植入兔桡骨缺损模型中观察其在体内成骨的能力.方法 携带hBMP7基因或增强型绿色荧光蛋白(EGFP)基因的2型重组腺相关病毒(rAAV2)载体在体外分别转染兔BMSC,再将转染后和未转染的兔BMSC分别与BG-COL-HYA-PS支架材料复合培养7 d后植入3组兔桡骨缺损模型,每组6只兔.各组在分别术后8周、12周通过大体标本观察、影像学、组织学等方法观察骨缺损的修复情况.以正常兔桡骨为对照组(n=3),术后12周比较各组骨缺损修复组织生物力学差异.结果 rAAV2-hBMP7转染的兔BMSC与BG-COL-HYA-PS复合支架材料有良好的生物相容性,植入兔桡骨缺损模型内表现出明确的成骨能力和骨修复能力,而形成的新骨最大压力载荷值低于正常桡骨组织[(188.46±12.24)N比(203.25±19.29)N,P＜0.05].结论 用hBMP7修饰BMSC复合仿生型BG-COL-HYA-PS支架材料构建的组织工程骨具有较强的骨修复能力,但形成的新生骨组织与正常骨组织比较仍然有早期生物力学方面的不足.     

Fibroin scaffold repairs critical-size bone defects in vivo supported by human amniotic fluid and dental pulp stem cells

The main aim of this study was the comparative evaluation of fibroin scaffolds combined with human stem cells, such as dental pulp stem cells (hDPSCs) and amniotic fluid stem cells (hAFSCs), used to repair critical-size cranial bone defects in immunocompromised rats. Two symmetric full-thickness cranial defects on each parietal region of rats have been replenished with silk fibroin scaffolds with or without preseeded stem cells addressed toward osteogenic lineage in vitro. Animals were euthanized after 4 weeks postoperatively and cranial tissue samples were taken for histological analysis. The presence of human cells in the new-formed bone was confirmed by confocal analysis with an antibody directed to a human mitochondrial protein. Fibroin scaffolds induced mature bone formation and defect correction, with higher bone amount produced by hAFSC-seeded scaffolds. Our findings demonstrated the strong potential of stem cells/fibroin bioengineered constructs for correcting large cranial defects in animal model and is likely a promising approach for the reconstruction of human large skeletal defects in craniofacial surgery.     

The bone formation in vitro and mandibular defect repair using PLGA porous scaffolds

Highly porous scaffolds of poly(lactide-co-glycolide) (PLGA) were prepared by solution-casting/salt-leaching method. The in vitro degradation behavior of PLGA scaffold was investigated by measuring the change of normalized weight, water absorption, pH, and molecular weight during degradation period. Mesenchymal stem cells (MSCs) were seeded and cultured in three-dimensional PLGA scaffolds to fabricate in vitro tissue engineering bone, which was investigated by cell morphology, cell number and deposition of mineralized matrix. The proliferation of seeded MSCs and their differentiated function were demonstrated by experimental results. To compare the reconstructive functions of different groups, mandibular defect repair of rabbit was made with PLGA/MSCs tissue engineering bone, control PLGA scaffold, and blank group without scaffold. Histopathologic methods were used to estimate the reconstructive functions. The result suggests that it is feasible to regenerate bone tissue in vitro using PLGA foams with pore size ranging from 100-250 microm as scaffolding for the transplantation of MSCs, and the PLGA/MSCs tissue engineering bone can greatly promote cell growth and have better healing functions for mandibular defect repair. The defect can be completely recuperated after 3 months with PLGA/MSCs tissue engineering bone, and the contrastive experiments show that the defects could not be repaired with blank PLGA scaffold. PLGA/MSCs tissue engineering bone has great potential as appropriate replacement for successful repair of bone defect.     

In vitro osteogenic differentiation of human mesenchymal stem cells and in vivo bone formation in composite nanofiber meshes

Tissue engineering has become an alternative method to traditional surgical treatments for the repair of bone defects, and an appropriate scaffold supporting bone formation is a key element in this approach. In the present study, nanofibrous organic and inorganic composite scaffolds containing nano-sized demineralized bone powders (DBPs) with biodegradable poly(L-lactide) (PLA) were developed using an electrospinning process for engineering bone. To assess their biocompatibility, in vitro osteogenic differentiation of human mandible-derived mesenchymal stem cells (hMSCs) cultured on PLA or PLA/DBP composite nanofiber scaffolds were examined. The mineralization of hMSCs cultured with osteogenic supplements on the PLA/DBP nanofiber scaffolds was remarkably greater than on the PLA nanofiber scaffold during the first 14 days of culture but reached the same level after 21 days. The in vivo osteoconductive effect of PLA/DBP nanofibrous scaffolds was further investigated using rats with critical-sized skull defects. Micro-computerized tomography revealed that a greater amount of newly formed bone extended across the defect area in PLA/DBP scaffolds than in the nonimplant and PLA scaffolds 12 weeks after implantation and that the defect size was almost 90% smaller. Therefore, PLA/DBP composite nanofiber scaffolds may serve as a favorable matrix for the regeneration of bone tissue.     

聚左旋乳酸/壳聚糖纳米纤维三维多孔支架复合骨髓间充质干细胞修复兔骨缺损**△

背景：骨髓间充质干细胞具有向多种间质细胞谱系分化的能力,且支架材料的性能对骨缺损的修复有重要影响。 目的：观察聚左旋乳酸/壳聚糖纳米纤维三维多孔支架复合骨髓间充质干细胞治疗骨缺损。 方法：对骨缺损模型兔分别采用空白植入、髂后上棘自体松质骨移植、聚左旋乳酸/壳聚糖纳米纤维多孔支架移植和复合了骨髓间充质干细胞的聚左旋乳酸/壳聚糖纳米纤维多孔支架移植修复缺损部位。 结果与结论：至移植12周,移植复合了骨髓间充质干细胞的聚左旋乳酸/壳聚糖纳米纤维多孔支架的实验兔的缺损处有骨组织生成,支架材料降解,已完成缺损修复,其修复情况接近松质骨组;髂后上棘自体松质骨移植的实验兔的缺损修复完好,新形成的骨组织较规则;只植入聚左旋乳酸/壳聚糖纳米纤维多孔支架的实验兔有少量骨组织形成,材料部分降解;空白植入的实验兔缺损处无新生骨组织生成,主要由纤维结缔组织填充。说明新型的生物支架材料聚左旋乳酸/壳聚糖纳米纤维三维多孔支架与来源于新西兰大白兔的骨髓间充质干细胞复合培养后,植入同种异体兔股骨髁缺损处,使骨缺损的修复速度加快,表现为较好的体内诱导成骨的作用。     

Scaffold translation: barriers between concept and clinic

Translation of scaffold-based bone tissue engineering (BTE) therapies to clinical use remains, bluntly, a failure. This dearth of translated tissue engineering therapies (including scaffolds) remains despite 25 years of research, research funding totaling hundreds of millions of dollars, over 12,000 papers on BTE and over 2000 papers on BTE scaffolds alone in the past 10 years (PubMed search). Enabling scaffold translation requires first an understanding of the challenges, and second, addressing the complete range of these challenges. There are the obvious technical challenges of designing, manufacturing, and functionalizing scaffolds to fill the Form, Fixation, Function, and Formation needs of bone defect repair. However, these technical solutions should be targeted to specific clinical indications (e.g., mandibular defects, spine fusion, long bone defects, etc.). Further, technical solutions should also address business challenges, including the need to obtain regulatory approval, meet specific market needs, and obtain private investment to develop products, again for specific clinical indications. Finally, these business and technical challenges present a much different model than the typical research paradigm, presenting the field with philosophical challenges in terms of publishing and funding priorities that should be addressed as well. In this article, we review in detail the technical, business, and philosophical barriers of translating scaffolds from Concept to Clinic. We argue that envisioning and engineering scaffolds as modular systems with a sliding scale of complexity offers the best path to addressing these translational challenges.     

A novel strategy for prefabrication of large and axially vascularized tissue engineered bone by using an arteriovenous loop

The repair of bone defects remains a major clinical challenge because none available reconstruction methods and biomaterials have been proved completely satisfactory. As a promising approach for bone regeneration, tissue engineered bone has become a technically feasible method to repair small to moderate sized bone defects in clinical practice, but it is difficult to repair large one, particularly when the recipient site is scarred by infection or radiation injury. Construction of large and vascularized tissue engineered bone may overcome the problems since vascularization is an essential prerequisite for the constructs to survive and integrate with existing host tissue. On the other hand, prefabrication large artificial bone in vivo bioreactor and axial vascularization by means of arteriovenous loop model in soft tissue have been proved to be feasible. Therefore, we hypothesize that combination of cells, solid scaffold, growth factors, and arteriovenous loop may eventually generate a large and vascularized tissue engineered bone flap in vivo bioreactor. Like vascularized autologous bone grafts, the new constructs could be transferred to the defect site by using microsurgical techniques. The strategy would facilitate clinical translation in bone tissue engineering and offer new therapeutic strategies for reconstruction of extended bone defects if the hypothesis proved to be practical.