Finite element analysis on the biomechanical stability of open porous titanium scaffolds for large segmental bone defects under physiological load conditions

Repairing large segmental defects in long bones caused by fracture, tumour or infection is still a challenging problem in orthopaedic surgery. Artificial materials, i.e. titanium and its alloys performed well in clinical applications, are plenary available, and can be manufactured in a wide range of scaffold designs. Although the mechanical properties are determined, studies about the biomechanical behaviour under physiological loading conditions are rare. The goal of our numerical study was to determine the suitability of open-porous titanium scaffolds to act as bone scaffolds. Hence, the mechanical stability of fourteen different scaffold designs was characterized under both axial compression and biomechanical loading within a large segmental distal femoral defect of 30 mm. This defect was stabilized with an osteosynthesis plate and physiological hip reaction forces as well as additional muscle forces were implemented to the femoral bone. Material properties of titanium scaffolds were evaluated from experimental testing. Scaffold porosity was varied between 64 and 80%. Furthermore, the amount of material was reduced up to 50%. Uniaxial compression testing revealed a structural modulus for the scaffolds between 3.5 GPa and 19.1 GPa depending on porosity and material consumption. The biomechanical testing showed defect gap alterations between 0.03 mm and 0.22 mm for the applied scaffolds and 0.09 mm for the intact bone. Our results revealed that minimizing the amount of material of the inner core has a smaller influence than increasing the porosity when the scaffolds are loaded under biomechanical loading. Furthermore, an advanced scaffold design was found acting similar as the intact bone.     

Biodegradable composite scaffolds incorporating an intramedullary rod and delivering bone morphogenetic protein-2 for stabilization and bone regeneration in segmental long bone defects

In this study, a two-part bone tissue engineering scaffold was investigated. The scaffold consists of a solid poly(propylene fumarate) (PPF) intramedullary rod for mechanical support surrounded by a porous PPF sleeve for osseointegration and delivery of poly(dl-lactic-co-glycolic acid) (PLGA) microspheres with adsorbed recombinant human bone morphogenetic protein-2 (rhBMP-2). Scaffolds were implanted into critical size rat segmental femoral defects with internal fixation for 12 weeks. Bone formation was assessed throughout the study via radiography, and following euthanasia, via microcomputed tomography and histology. Mechanical stabilization was evaluated further via torsional testing. Experimental implant groups included the PPF rod alone and the rod with a porous PPF sleeve containing PLGA microspheres with 0, 2 or 8 μg of rhBMP-2 adsorbed onto their surface. Results showed that presence of the scaffold increased mechanical stabilization of the defect, as evidenced by the increased torsional stiffness of the femurs by the presence of a rod compared to the empty defect. Although the presence of a rod decreased bone formation, the presence of a sleeve combined with a low or high dose of rhBMP-2 increased the torsional stiffness to 2.06 ± 0.63 and 1.68 ± 0.56 N·mm, respectively, from 0.56 ± 0.24 N·mm for the rod alone. The results indicate that, while scaffolds may provide structural support to regenerating tissues and increase their mechanical properties, the presence of scaffolds within defects may hinder overall bone formation if they interfere with cellular processes.     

Autologous vs. allogenic mesenchymal progenitor cells for the reconstruction of critical sized segmental tibial bone defects in aged sheep

Mesenchymal progenitor cells (MPCs) represent an attractive cell population for bone tissue engineering. Their special immunological characteristics suggest that MPCs may be used in allogenic applications. The objective of this study was to compare the regenerative potential of autologous vs. allogenic MPCs in an ovine critical size segmental defect model. Ovine MPCs were isolated from bone marrow aspirates, expanded and cultured with osteogenic medium for 2 weeks before implantation. Autologous and allogenic transplantation was performed using the cell-seeded scaffolds and unloaded scaffolds, while the application of autologous bone grafts served as a control group (n = 6). Bone healing was assessed 12 weeks after surgery by radiology, microcomputed tomography, biomechanical testing and histology. Radiology, biomechanical testing and histology revealed no significant differences in bone formation between the autologous and allogenic groups. Both cell groups showed more bone formation than the scaffold alone, whereas the biomechanical data showed no significant differences between the cell groups and the unloaded scaffolds. The results of the study suggest that scaffold-based bone tissue engineering using allogenic cells offers the potential for an off-the-shelf product. Thus the results of this study serve as an important baseline for translation of the assessed concepts into clinical applications.     

Segmental bone regeneration using a load-bearing biodegradable carrier of bone morphogenetic protein-2

Segmental defect regeneration has been a clinical challenge. Current tissue-engineering approach using porous biodegradable scaffolds to delivery osteogenic cells and growth factors demonstrated success in facilitating bone regeneration in these cases. However, due to the lack of mechanical property, the porous scaffolds were evaluated in non-load bearing area or were stabilized with stress-shielding devices (bone plate or external fixation). In this paper, we tested a scaffold that does not require a bone plate because it has sufficient biomechanical strength. The tube-shaped scaffolds were manufactured from poly(propylene) fumarate/tricalcium phosphate (PPF/TCP) composites. Dicalcium phosphate dehydrate (DCPD) were used as bone morphogenetic protein-2 (BMP-2) carrier. Twenty-two scaffolds were implanted in 5mm segmental defects in rat femurs stabilized with K-wire for 6 and 15 weeks with and without 10 microg of rhBMP-2. Bridging of the segmental defect was evaluated first radiographically and was confirmed by histology and micro-computer tomography (microCT) imaging. The scaffolds in the BMP group maintained the bone length throughout the duration of the study and allow for bridging. The scaffolds in the control group failed to induce bridging and collapsed at 15 weeks. Peripheral computed tomography (pQCT) showed that BMP-2 does not increase the bone mineral density in the callus. Finally, the scaffold in BMP group was found to restore the mechanical property of the rat femur after 15 weeks. Our results demonstrated that the load-bearing BMP-2 scaffold can maintain bone length and allow successfully regeneration in segmental defects.     

Combination of platelet-rich plasma with polycaprolactone-tricalcium phosphate scaffolds for segmental bone defect repair

Porous scaffold biomaterials may offer a clinical alternative to bone grafts; however, scaffolds alone are typically insufficient to heal large bone defects. Numerous studies have demonstrated that osteoinductive growth factor or gene delivery significantly improves bone repair. However, given the important role of vascularization during bone regeneration, it may also be beneficial to incorporate factors that promote vascular ingrowth into constructs. In this study, a strategy combining structural polycaprolactone-20% tricalcium phosphate (PCL-TCP) composite scaffolds with platelet-rich plasma (PRP) was tested. Following bilateral implantation of constructs into 8 mm rat nonunion femoral defects, 3D vascular and bone ingrowth were quantified at 3 and 12 weeks using contrast-enhanced microcomputed tomography (micro-CT) imaging. At week 3, PRP-treated femurs displayed 70.3% higher vascular volume fraction than control femurs. Interestingly, bone volume fraction (BVF) was significantly higher for the empty scaffold group at the early time point. At 12 weeks, BVF measurements between the two groups were statistically equivalent. However, a greater proportion of PRP-treated femurs (83%) achieved bone union as compared to empty scaffold controls (33%). Consistent with this observation, biomechanical evaluation of functional integration also revealed a significantly higher torsional stiffness observed for PRP-treated defects compared to empty scaffolds. Ultimate torque at failure was not improved, however, perhaps due to the slow resorption profile of the scaffold material. Histological evaluation illustrated infiltration of vascularized connective tissue and bone in both groups. Given that bone ingrowth into untreated defects in this model is minimal, PCL-TCP scaffolds were clearly able to promote bone ingrowth but failed to consistently bridge the defect. The addition of PRP to PCL-TCP scaffolds accelerated early vascular ingrowth and improved longer-term functional integration. Taken together, the results of this study suggest that the use of PRP, alone or in combination with other bioactive components, may be an effective approach to augment the ability of porous biomaterial scaffolds to repair orthotopic defects.     

In vivo monitoring of structural and mechanical changes of tissue scaffolds by multi-modality imaging

Degradable tissue scaffolds are implanted to serve a mechanical role while healing processes occur and putatively assume the physiological load as the scaffold degrades. Mechanical failure during this period can be unpredictable as monitoring of structural degradation and mechanical strength changes at the implant site is not readily achieved in vivo, and non-invasively. To address this need, a multi-modality approach using ultrasound shear wave imaging (USWI) and photoacoustic imaging (PAI) for both mechanical and structural assessment in vivo was demonstrated with degradable poly(ester urethane)urea (PEUU) and polydioxanone (PDO) scaffolds. The fibrous scaffolds were fabricated with wet electrospinning, dyed with indocyanine green (ICG) for optical contrast in PAI, and implanted in the abdominal wall of 36 rats. The scaffolds were monitored monthly using USWI and PAI and were extracted at 0, 4, 8 and 12 wk for mechanical and histological assessment. The change in shear modulus of the constructs in vivo obtained by USWI correlated with the change in average Young's modulus of the constructs ex vivo obtained by compression measurements. The PEUU and PDO scaffolds exhibited distinctly different degradation rates and average PAI signal intensity. The distribution of PAI signal intensity also corresponded well to the remaining scaffolds as seen in explant histology. This evidence using a small animal abdominal wall repair model demonstrates that multi-modality imaging of USWI and PAI may allow tissue engineers to noninvasively evaluate concurrent mechanical stiffness and structural changes of tissue constructs in vivo for a variety of applications.     

股骨头松质骨力学性质实验研究

目的 研究股骨头松质骨力学性质，为临床提供生物力学参数。方法 对正常国人新鲜尸体股骨头松质骨的拉伸、压缩、扭转、剪切、弯曲、冲击力学性能进行实验研究。结果 得出了股骨头松质骨的拉伸、压缩破坏载荷、强度极限、弹性模量，扭转破坏扭矩、扭转剪切强度极限，弯曲破坏载荷、弯曲强度极限、剪切破坏载荷、剪切强度极限，冲击功、冲击韧性等测试指标的实验结果。结论 股骨头松质骨抗压强度大于抗拉强度，压缩弹性模量大于拉伸弹性模量，扭转强度大于剪切强度，抗弯强度与抗压强度接近。     

In vivo evaluation of a bioactive scaffold for bone tissue engineering

Revision cases of total hip implants are complicated by the significant amount of bone loss. New materials and/or approaches are needed to provide stability to the site, stimulate bone formation, and ultimately lead to fully functional bone tissue. Porous bioactive glasses (prepared from 45S5 granules, 45% SiO2, 24.5% Na2O, 24.5% CaO, and 6% P2O5) have been developed as scaffolds for bone tissue engineering and have been studied in vitro. In this study, we investigated the incorporation of tissue-engineered constructs utilizing these scaffolds in large, cortical bone defects in the rat simulating revision conditions. With implantation times of 2, 4, and 12 weeks the results were compared to those using the bioactive ceramic scaffold alone. Two tissue-engineered constructs were studied: osteoprogenitor cells that were either seeded onto the scaffold prior to implantation ("primary") or those that were culture expanded to form bonelike tissue on the scaffold prior to implantation ("hybrid"). Defects treated with the hybrid had the greatest amount of bone in the available pore space of the defect over all other groups at 2 weeks (p < 0.05). For both the primary and hybrid groups, woven and lamellar bone was present along the interface of the scaffold and the host cortex and within the porous space of the scaffold at 2 weeks. By 4 weeks, very uniform, lamellar bone was present throughout the scaffold for both tissue-engineered groups. The amount of bone significantly increased over time for all groups while the bioactive ceramic gradually resorbed by 40% at 12 weeks (p < 0.05). Structural properties of the treated long bones improved over time. Long bones treated with the hybrid had an early return in torsional stiffness by 2 weeks. Both tissue-engineered constructs achieved normal torsional strength and stiffness by 4 weeks as compared to the scaffold alone, which achieved this by 12 weeks. Porous, surface modified bioactive ceramic is a promising scaffold material for tissue-engineered bone repair.     

Substrate stiffness and contractile behaviour modulate the functional maturation of osteoblasts on a collagen–GAG scaffold

Anchorage-dependent cells respond to the mechanical and physical properties of biomaterials. One such cue is the mechanical stiffness of a material. We compared the osteogenic potential of collagen–glycosaminoglycan (CG) scaffolds with varying stiffness for up to 6 weeks in culture. The mechanical stiffness of CG scaffolds were varied by cross-linking by physical (dehydrothermal (DHT)) and chemical (1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDAC) and glutaraldehyde (GLUT)) methods. The results showed that all CG substrates allowed cellular attachment, infiltration and osteogenic differentiation. CG scaffolds treated with EDAC and GLUT were mechanically stiffer, retained their original scaffold structure and resisted cellular contraction. Consequently, they facilitated a 2-fold greater cell number, probably due to the pore architecture being maintained, allowing improved diffusion of nutrients. On the other hand, the less stiff substrates cross-linked with DHT allowed increased cell-mediated scaffold contraction, contracting by 70% following 6 weeks (P < 0.01) of culture. This reduction in scaffold area resulted in cells reaching the centre of the scaffold quicker up to 4 weeks; however, at 6 weeks all scaffolds showed similar levels of cellular infiltration, with higher cell numbers found on the stiffer EDAC- and GLUT-treated scaffolds. Analysis of osteogenesis showed that scaffolds cross-linked with DHT expressed higher levels of the late stage bone formation markers osteopontin and osteocalcin (P < 0.01) and increased levels of mineralisation. In conclusion, the more compliant CG scaffolds allowed cell-mediated contraction and supported a greater level of osteogenic maturation of MC3T3 cells, while the stiffer, non-contractible scaffolds resulted in lower levels of cell maturation, but higher cell numbers on the scaffold. Therefore, we found scaffold stiffness had different effects on differentiation and cell number whereby the increased cell-mediated contraction facilitated by the less stiff scaffolds positively modulated osteoblast differentiation while reducing cell numbers.     

Repair of segmental long bone defect in rabbit femur using bioactive titanium cylindrical mesh cage

A segmental rabbit femur defect was repaired using an empty bioactive titanium (BAT) mesh cage. A 10mm long titanium mesh cage was positioned in the bony defect and reinforced by intramedullary fixation. The BAT surface was prepared by chemical and thermal treatment. Pure titanium cages were used as a control. Torsional stiffness of the BAT group at 4 weeks was approximately equal to, and at 8 weeks twice, that of the intact femur. Differences between the torsional stiffness of the control and BAT groups were significant at both time intervals. Histological examinations showed that woven bone appeared around the cage by 4 weeks and transformed to lamella bone by 8 weeks. New bone bonded to the BAT surface without an intervening layer. The BAT cage enhanced the bone repairing process and achieved faster repair of long bone segmental defects.     

Osteointegration of hydroxyapatite-coated and uncoated titanium screws in long-term ovariectomized sheep.

This study was designed to evaluate the osteointegration of HA-coated and uncoated titanium screws in the cortical bone of long-term (24 months) ovariectomized sheep (OVX group) compared to sham-aged sheep (Control group). At 12 weeks after implantation, the screws were tested biomechanically (extraction torque) and histomorphometrically (affinity index: AI) in both femoral and tibial diaphyses. Cancellous bone status was assessed by iliac crest biopsy. BMD of the L5 vertebra and a histomorphological study of the femoral and tibial shafts were performed to acquire data on cortical bone. A significant difference was found between the OVX and Control groups for BMD (p<0.0005), and a significant reduction in the cancellous bone area was observed in the OVX group. Femoral and tibial cortical bone parameters showed significant differences between the groups. The type of material selected (femurs: p<0.0005; tibiae: p<0.0005) and ovariectomy (femurs: p<0.005; tibiae: p<0.005) had a significant effect on the extraction torque. AI results were related to the presence or absence of ovariectomy (p<0.05) and strictly depended on the material implanted in the femur and tibia (p<0.0005). In conclusion, at implantation OVX sheep showed a significant loss of trabecular and cortical bone versus sham-aged sheep. The biomechanical and histomorphological results achieved suggest employing HA-coated screws in the presence of osteopenic cortical bone.     

Repair of an articular cartilage defect using adipose-derived stem cells loaded on a polyelectrolyte complex scaffold based on poly(l-glutamic acid) and chitosan

As a synthetic polypeptide water-soluble poly(l-glutamic acid) (PLGA) was designed to fabricate scaffolds for cartilage tissue engineering. Chitosan (CHI) has been employed as a physical cross-linking component in the construction of scaffolds. PLGA/CHI scaffolds act as sponges with a swelling ratio of 760 ± 45% (mass%), showing promising biocompatibility and biodegradation. Autologous adipose-derived stem cells (ASCs) were expanded and seeded on PLGA/CHI scaffolds, ASC/scaffold constructs were then subjected to chondrogenic induction in vitro for 2 weeks. The results showed that PLGA/CHI scaffolds could effectively support ASC adherence, proliferation and chondrogenic differentiation. The ASCs/scaffold constructs were then transplanted to repair full thickness articular cartilage defects (4 mm in diameter, to the depth of subchondral bone) created in rabbit femur trochlea. Histological observations found that articular defects were covered with newly formed cartilage 6 weeks post-implantation. After 12 weeks the regenerated cartilage had integrated well with the surrounding native cartilage and subchondral bone. Toluidine blue and immunohistochemical staining confirmed similar accumulation of glycosaminoglycans and type II collagen in engineered cartilage as in native cartilage 12 weeks post-implantation. The result was further supported by quantitative analysis of extracellular matrix deposition. The compressive modulus of the engineered cartilage increased significantly from 30% of that of normal cartilage at 6 weeks to 83% at 12 weeks. Cyto-nanoindentation also showed analogous biomechanical behavior of the engineered cartilage to that of native cartilage. The results of the present study thus demonstrate the potentiality of PLGA/CHI scaffolds in cartilage tissue engineering.     

Novel bilayer bacterial nanocellulose scaffold supports neocartilage formation in vitro and in vivo

Tissue engineering provides a promising alternative therapy to the complex surgical reconstruction of auricular cartilage by using ear-shaped autologous costal cartilage. Bacterial nanocellulose (BNC) is proposed as a promising scaffold material for auricular cartilage reconstruction, as it exhibits excellent biocompatibility and secures tissue integration. Thus, this study evaluates a novel bilayer BNC scaffold for auricular cartilage tissue engineering. Bilayer BNC scaffolds, composed of a dense nanocellulose layer joined with a macroporous composite layer of nanocellulose and alginate, were seeded with human nasoseptal chondrocytes (NC) and cultured in vitro for up to 6 weeks. To scale up for clinical translation, bilayer BNC scaffolds were seeded with a low number of freshly isolated (uncultured) human NCs combined with freshly isolated human mononuclear cells (MNC) from bone marrow in alginate and subcutaneously implanted in nude mice for 8 weeks. 3D morphometric analysis showed that bilayer BNC scaffolds have a porosity of 75% and mean pore size of 50 ± 25 μm. Furthermore, endotoxin analysis and in vitro cytotoxicity testing revealed that the produced bilayer BNC scaffolds were non-pyrogenic (0.15 ± 0.09 EU/ml) and non-cytotoxic (cell viability: 97.8 ± 4.7%). This study demonstrates that bilayer BNC scaffolds offer a good mechanical stability and maintain a structural integrity while providing a porous architecture that supports cell ingrowth. Moreover, bilayer BNC scaffolds provide a suitable environment for culture-expanded NCs as well as a combination of freshly isolated NCs and MNCs to form cartilage in vitro and in vivo as demonstrated by immunohistochemistry, biochemical and biomechanical analyses.     

Small intestinal submucosa versus salt-extracted polyglycolic acid-poly-L-lactic acid: a comparison of neocartilage formed in two scaffold materials

This study sought to compare differences in neocartilage produced over time from two types of resorbable scaffold materials. One material was entirely synthetic and contained a polyglycolic acid-poly-L-lactic acid matrix (PGA-PLLA). The second scaffold material was bioactive and consisted of a four-layered construct of porcine small intestinal submucosa (SIS). Disk-shaped scaffolds were seeded with canine chondrocytes and implanted into athymic mice for periods of 5, 8, 12, and 24 weeks. Constructs were examined microscopically, assayed for hydroxyproline (HP) and glycosaminoglycan (GAG) content, and collagen typed (I or II) at each time period. Creep indentation tests determined aggregate and shear modulus, permeability, and thickness. Results indicated that SIS maintained its thickness through the first 12 weeks, and then doubled by week 24. The 24-week tissue appeared chondroid-like and possessed high GAG content. Tissues derived from PGA-PLLA scaffolds were lower in HP content than SIS-derived tissues, but type II collagen was demonstrated only in PGA-PLLA-derived tissues at 24 weeks. Mechanical properties were not significantly different for any tissue over time (p > 0.05), but aggregate and shear modulus mean values were consistently higher for PGA-PLLA-derived tissues at nearly every time interval. This, coupled with the presence of collagen types I and II, suggested a more congruent solid phase may be forming within the extracellular matrix of tissues derived from PGA-PLLA scaffolds. Future study is necessary to compare these materials under simulated loading conditions.     

Matrix deposition modulates the viscoelastic shear properties of hydrogel-based cartilage grafts

Hydrogel-based scaffolds such as alginate have been extensively investigated for cartilage tissue engineering, largely due to their biocompatibility, ambient gelling conditions, and the ability to support chondrocyte phenotype. While it is well established that the viscoelastic response of articular cartilage is essential for articulation and load bearing, the time-dependent mechanical properties of hydrogel-based cartilage scaffolds have not been extensively studied. Therefore, the objective of this study was to characterize the intrinsic viscoelastic shear properties of chondrocyte-laden alginate scaffolds and determine the effects of seeding density and culturing time on these properties. Specifically, the viscoelastic properties (equilibrium and dynamic shear moduli and dynamic phase shift angle) of these engineered cartilage grafts were measured under torsional shear. In addition, the rapid ramp-step shear stress relaxation of the alginate-based cartilage scaffolds was modeled using the quasi-linear viscoelastic (QLV) theory. It was found that scaffold stiffness increased with both culturing time and cell density, whereas viscosity did not change significantly with cell density (30 vs. 60 million/mL). Similar to native cartilage, the energy dissipation of engineered scaffolds under pure shear is highly correlated to the glycosaminoglycan content. In contrast, collagen content was not strongly correlated to scaffold shear modulus, especially the instantaneous shear modulus predicted by the quasi-linear viscoelastic model. The findings of this study provide new insights into the structure-function relationship of engineered cartilage and design of functional grafts for cartilage repair.     

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.     

通过骨支架的数字化建模分析其力学性能与内部流场分布

目的研究不同骨支架结构的力学性能与内部流场分布,为模型结构的优劣提供直观上的比较和评判,为骨支架结构设计提供有效的指导方法。方法利用Pro/Engineer和MIMICS等软件重建自然结构骨支架、编织状骨支架和球形孔骨支架,并通过有限元方法分析3种支架的有效弹性模量、应力分布和三维灌注培养下支架内部流场分布。结果采用相同材料设计得到的自然结构骨支架表现出更小的有效弹性模量;当3种支架受到相同压力时,自然结构骨支架内部应力峰值更小且应力分布更均匀;初始流速和流体黏度相同时,自然结构骨支架表现出更小的内部流速、壁面剪切应力和壁面压力。结论自然结构骨支架模型具有相对较好的生物力学性能,在3种骨支架中最适合用于骨组织工程中骨支架结构选型。     

The effect of BMP-2 on micro- and macroscale osteointegration of biphasic calcium phosphate scaffolds with multiscale porosity

It is well established that scaffolds for applications in bone tissue engineering require interconnected pores on the order of 100 μm for bone in growth and nutrient and waste transport. As a result, most studies have focused on scaffold macroporosity (>100 μm). More recently researchers have investigated the role of microporosity in calcium phosphate -based scaffolds. Osteointegration into macropores improves when scaffold rods or struts contain micropores, typically defined as pores less than ～50 μm. We recently demonstrated multiscale osteointegration, or growth into both macropores and intra-red micropores (<10 μm), of biphasic calcium phosphate (BCP) scaffolds. The combined effect of BMP-2, a potent osteoinductive growth factor, and multiscale porosity has yet to be investigated. In this study we implanted BCP scaffolds into porcine mandibular defects for 3, 6, 12 and 24 weeks and evaluated the effect of BMP-2 on multiscale osteointegration. The results showed that given this in vivo model BMP-2 influences osteointegration at the microscale, but not at the macroscale, but not at the macroscale. Cell density was higher in the rod micropores for scaffolds containing BMP-2 compared with controls at all time points, but BMP-2 was not required for bone formation in micropores. In contrast, there was essentially no difference in the fraction of bone in macropores for scaffolds with BMP-2 compared with controls. Additionally, bone in macropores seemed to have reached steady-state by 3 weeks. Multiscale osteointegration results in bone-scaffold composites that are fully osteointegrated, with no ‘dead space’. These composites are likely to contain a continuous cell network as well as the potential for enhanced load transfer and improved mechanical properties.     

壳聚糖-磷酸钙/骨形态发生蛋白2复合骨髓间充质干细胞促进兔脊柱的融合

背景：前期实验已经证明壳聚糖-磷酸钙/骨形态发生蛋白2复合材料能够促进兔脊柱融合。 目的：评价壳聚糖-磷酸钙/骨形态发生蛋白2/碱性成纤维细胞生长因子支架材料在兔椎间融合中的应用效果。 方法：制备壳聚糖-磷酸钙/骨形态发生蛋白2/碱性成纤维细胞生长因子支架材料,并与大鼠骨髓间充质干细胞在体外构成组织工程骨。摘除40只新西兰大白兔椎间盘,随机分为4组：空白对照组未植入任何材料,对照组植入自体髂骨,支架材料组植入壳聚糖-磷酸钙/骨形态发生蛋白2/碱性成纤维细胞生长因子复合材料,实验组植入组织工程骨材料。 结果与结论：术后12周：①X射线片：对照组与实验组椎体融合,两组间融合节段生物力学强度大致相同,生物力学强度高于空白对照组与支架材料组(P < 0.05),且支架材料组高于空白对照组(P < 0.05)。②组织学切片：实验组与对照组有编织骨岛和新生毛细血管生成,支架材料组仅观察到壳聚糖支架网络,空白对照组未发现特殊组织结构。表明壳聚糖-磷酸钙/骨形态发生蛋白2/碱性成纤维细胞生长因子复合鼠骨髓间充质干细胞能够明显促进脊柱融合。     

Shaping the micromechanical behavior of multi-phase composites for bone tissue engineering

Mechanical stiffness is a fundamental parameter in the rational design of composites for bone tissue engineering in that it affects both the mechanical stability and the osteo-regeneration process at the fracture site. A mathematical model is presented for predicting the effective Young’s modulus (E) and shear modulus (G) of a multi-phase biocomposite as a function of the geometry, material properties and volume concentration of each individual phase. It is demonstrated that the shape of the reinforcing particles may dramatically affect the mechanical stiffness: E and G can be maximized by employing particles with large geometrical anisotropy, such as thin platelet-like or long fibrillar-like particles. For a porous poly(propylene fumarate) (60% porosity) scaffold reinforced with silicon particles (10% volume concentration) the Young’s (shear) modulus could be increased by more than 10 times by just using thin platelet-like as opposed to classical spherical particles, achieving an effective modulus E ～ 8 GPa (G ～ 3.5 GPa). The mathematical model proposed provides results in good agreement with several experimental test cases and could help in identifying the proper formulation of bone scaffolds, reducing the development time and guiding the experimental testing.