In vivo performance of microstructured calcium phosphate formulated in novel water-free carriers

Osteoinductive calcium phosphate (CaP) ceramics can be combined with polymeric carriers to make shapeable bone substitutes as an alternative to autologous bone; however, carriers containing water may degrade the ceramic surface microstructure, which is crucial to bone formation. In this study five novel tricalcium phosphate (TCP) formulations were designed from water-free polymeric binders and osteoinductive TCP granules of different particle sizes (500–1000 μm for moldable putty forms, and 150–500 μm for flowable paste forms). The performance of these novel TCP formulations was studied and compared with control TCP granules alone (both 150–500 and 500–1000 μm). In vitro the five TCP formulations were characterized by their carrier dissolution times and TCP mineralization kinetic profiles in simulated body fluid. In vivo the formulations were implanted in the dorsal muscle and a unicortical femoral defect (Ø = 5 mm) of dogs for 12 weeks. The TCP formulation based on a xanthan gum–glycerol carrier exhibited fast carrier dissolution (1 h) and TCP mineralization (7 days) in vitro, but induced inflammation and showed little ectopic bone formation. This carrier chemistry was thus found to disrupt the early cellular response related to osteoinduction by microstructured TCP. TCP formulations based on carboxymethyl cellulose–glycerol and Polyoxyl 15-hydroxystearate–Pluronic® F127 allowed the in vitro surface mineralization of TCP by day 7 and produced the highest level of orthotopic bone bridging and ectopic bone formation, which was equivalent to the control. These results demonstrate that water-free carriers can preserve the chemistry, microstructure, and performance of osteoinductive CaP ceramics.     

The osteoinductive potential of printable, cell-laden hydrogel-ceramic composites

Hydrogels used as injectables or in organ printing often lack the appropriate stimuli to direct osteogenic differentiation of embedded multipotent stromal cells (MSCs), resulting in limited bone formation in these matrices. Addition of calcium phosphate (CaP) particles to the printing mixture is hypothesized to overcome this drawback. In this study we have investigated the effect of CaP particles on the osteoinductive potential of cell-laden hydrogel-CaP composite matrices. To this end, apatitic nanoparticles have been included in Matrigel constructs where after the viability of embedded progenitor cells was assessed in vitro. In addition, the osteoinductive potential of cell-laden Matrigel containing apatitic nanoparticles was investigated in vivo and compared with composites containing osteoinductive biphasic calcium phosphate (BCP) microparticles after subcutaneous implantation in immunodeficient mice. Histological and immunohistochemical analysis of the tissue response as well as in vivo bone formation revealed that apatitic nanoparticles were osteoinductive and induced osteoclast activation, but without bone formation. The BCP particles were more effective in inducing elaborate bone formation at the ectopic location.     

Liposomal clodronate inhibition of osteoclastogenesis and osteoinduction by submicrostructured beta-tricalcium phosphate

Bone graft substitutes such as calcium phosphates are subject to the innate inflammatory reaction, which may bear important consequences for bone regeneration. We speculate that the surface architecture of osteoinductive β-tricalcium phosphate (TCP) stimulates the differentiation of invading monocyte/macrophages into osteoclasts, and that these cells may be essential to ectopic bone formation. To test this, porous TCP cubes with either submicron-scale surface architecture known to induce ectopic bone formation (TCPs, positive control) or micron-scale, non-osteoinductive surface architecture (TCPb, negative control) were subcutaneously implanted on the backs of FVB strain mice for 12 weeks. Additional TCPs samples received local, weekly injections of liposome-encapsulated clodronate (TCPs + LipClod) to deplete invading monocyte/macrophages. TCPs induced osteoclast formation, evident by positive tartrate resistant acid phosphatase (TRAP) cytochemical staining and negative macrophage membrane marker F4/80 immunostaining. No TRAP positive cells were found in TCPb or TCPs + LipClod, only F4/80 positive macrophages and foreign body giant cells. TCPs stimulated subcutaneous bone formation in all implants, while no bone could be found in TCPb or TCPs + LipClod. In agreement, expression of bone and osteoclast gene markers was upregulated in TCPs versus both TCPb and TCPs + LipClod, which were equivalent. In summary, submicron-scale surface structure of TCP induced osteoclastogenesis and ectopic bone formation in a process that is blocked by monocyte/macrophage depletion.     

SiO2 and ZnO dopants in three-dimensionally printed tricalcium phosphate bone tissue engineering scaffolds enhance osteogenesis and angiogenesis in vivo

Calcium phosphate (CaP) scaffolds with three-dimensionally-interconnected pores play an important role in mechanical interlocking and biological fixation in bone implant applications. CaPs alone, however, are only osteoconductive (able to guide bone growth). Much attention has been given to the incorporation of biologics and pharmacologics to add osteoinductive (able to cause new bone growth) properties to CaP materials. Because biologics and pharmacologics are generally delicate compounds and also subject to increased regulatory scrutiny, there is a need to investigate alternative methods to introduce osteoinductivity to CaP materials. In this study silica (SiO₂) and zinc oxide (ZnO) have been incorporated into three-dimensional printed β-tricalcium phosphate (β-TCP) scaffolds to investigate their potential to trigger osteoinduction in vivo. Silicon and zinc are trace elements that are common in bone and have also been shown to have many beneficial properties, from increased bone regeneration to angiogenesis. Implants were placed in bicortical femur defects introduced to a murine model for up to 16 weeks. The addition of dopants into TCP increased the capacity for new early bone formation by modulating collagen I production and osteocalcin production. Neovascularization was found to be up to three times more than the pure TCP control group. The findings from this study indicate that the combination of SiO₂ and ZnO dopants in TCP may be a viable alternative to introducing osteoinductive properties to CaPs.     

Osteoinduction of hydroxyapatite/β-tricalcium phosphate bioceramics in mice with a fractured fibula

Many studies have shown that calcium phosphate ceramics can induce bone formation in non-osseous sites without the application of any osteoinductive biomolecules, but the mechanisms of this phenomenon (intrinsic osteoinduction of bioceramics) remain unclear. In this study, we compared the intrinsic osteoinduction of porous hydroxyapatite/β-tricalcium phosphate (HA/β-TCP) implanted in mice at different sites. In 30 mice the left fibula was fractured and the right fibula was kept intact. A porous HA/β-TCP cylinder was implanted into both the left (group 1) and right (group 2) leg muscles of each animal. In addition, two HA/β-TCP cylinders were bilaterally implanted into leg subcutaneous pockets (group 3) in each of the remaining 15 mice. New bone formation was studied in the three groups by histology, histomorphometry and immunostaining. In group 1 new bone was observed at week 6 and bone marrow appeared at week 12. In group 2 new bone was observed at week 8 and bone marrow appeared at week 12. The new bone area percentage in group 1 was significantly higher than in group 2 at both weeks 8 and 12. In contrast, group 3 did not show any new bone within the period studied. These differences were explained based on the location of the implants and thus their proximity to the osteogenic environment of fracture healing. The results support the hypothesis that intrinsic osteoinduction by calcium phosphate ceramics is the result of adsorption of osteoinductive substances on the surface.     

Review paper: behavior of ceramic biomaterials derived from tricalcium phosphate in physiological condition

Various calcium phosphates are used for bone repair. Although hydroxyapatite (HA) sintered ceramics are widely used due to their osteoconductivity, its bioresorbability is so low that HA remains in the body for a long time after implantation. In contrast, tricalcium phosphate (TCP) ceramics show resorbable characters during bone regeneration, and can be completely substituted for the bone tissue after stimulation of bone formation. Therefore, much attention is paid to TCP ceramics for scaffold materials for supporting bone regeneration. This paper reviews bioresorbable properties of calcium phosphate ceramics derived from beta-TCP and alpha-TCP.     

Investigation as to the osteoinductivity of macroporous calcium phosphate cement in goats

For bone formation in critical-sized or poor healing defects, osteoinductive behavior of synthetic bone grafts is crucial. Although the osteoconductive behavior of calcium phosphate (CaP) cement is generally accepted, its osteoinductive potential is less reported. In this study, osteoinduction of porous CaP cement was investigated. Four goats received each six subcutaneous placed prehardened porous CaP cement implants. Implantation time was 3 and 6 months. After explantation, histological evaluation and scoring with a histological grading scale for soft-tissue implants were performed. The histological sections revealed that the implants degraded for more than 50% over time. The implants had lost their macroporous structure from 3 months on. A medium-thick fibrous capsule with a few inflammatory cells surrounded the implants after 3 months. This capsule significantly decreased in thickness after 6 months. Throughout the implant ingrowth of fibrous tissue was seen with scattered foci of inflammatory cells. Cement particles were surrounded by a layer of inflammatory cells. The massive inflammatory response in the interstice was seen after 3 months, which disappeared after 6 months implantation. No bone formation was detected in any of the specimens. The fast degradation and thereby collapsing of the porous structure of our CaP cement implant might have prevented osteoinduction.     

beta-TCP microporosity decreases the viability and osteoblast differentiation of human bone marrow stromal cells

Association of osteoprogenitor cells to calcium phosphate ceramics is currently under intense investigation, for its considered ability to induce bone formation and therefore to allow the successful repair of large bone defects. However, if the first experimental and clinical studies provided promising results, lack of new bone formation has been reported in a large number of animal experiments. In this context and since it has been reported that in some conditions, calcium phosphate ceramic microstructure induces ectopic bone formation, we investigated the effects of ceramic microporosity on the behavior of osteoprogenitor cells for the development of hybrid materials. Human bone marrow stromal cells (BMSCs) were seeded on beta-tricalcium phosphate (TCP) ceramics with 0, 25, or 45% microporosity and cell adhesion, viability, and osteoblastic differentiation have been studied for three weeks. Cell counts, measurement of MTS conversion, and LDH activity indicated that microporosity decreased the viability of BMSCs in a time and rate-dependent manner. In addition, microporosity inhibited osteoblastic differentiation as compared with nonmicroporous ceramics, as revealed by decreased alkaline phosphatase activity and osteocalcin secretion. Results of this in vitro study therefore highlight a negative role for beta-TCP microporosity in the behavior of human osteoprogenitor cells.     

Fibrin glue as an osteoinductive protein in a mouse model

Fibrin sealant or fibrin glue (FG) has been found to be effective as a wound-healing substance in surgery. However, its role in bone fracture healing and osseous tissue response is not fully understood. This ambiguity questions the potential of FG as an inductive protein. The present study was undertaken to evaluate the osteoinductive property of FG when coated with calcium phosphate and glass ceramics and implanted in the extraskeletal site of male Swiss albino mice. Implant materials used for this study were hydroxyapatite (HA) porous granules (300-350 microm), bioactive glass system (BGS)-AW type and calcium phosphate calcium silicate system (HABGS) non-porous granules (300-350 microm). Uncoated granules (control) and coated granules with 2.5 mg FG and 5 mg FG were implanted in the quadriceps muscle of mice and sacrificed after 28 days. Histologically, HA, BGS and HABGS implanted animal groups showed good healing response. However, neo-osteogenesis was observed only in the BGS and HABGS granules impregnated with FG. Furthermore, bone formation was observed to be more conspicuous in 5 mg FG coated BGS and HABGS granules when compared with 2.5 mg FG coated BGS and HABGS granules. Fluorochrome labeling proved that mineralization had already started by day 15 with FG preadsorbed BGS and HABGS granules. On the contrary, the uncoated granules did not show any de novo bone formation. This experimental study provides an evidence of the positive role of FG as a potential osteoinductive biologic tissue adhesive.     

Ceramic drug delivery: a perspective

Different ceramic substances are offered in the market as bone substitute materials. These include monophasic calcium phosphate ceramics of tricalciumphosphate (TCP) or hydroxyapatite (HA), biphasic calcium phosphate ceramics and multiphasic bio-glasses synthetic calcium phosphate cements. Ceramics with appropriate three-dimensional geometry are able to bind and concentrate bone morphogenetic proteins in circulation and may become osteoinductive (capable of osteogenesis) and can be effective carriers of bioactive peptide or bone cell seeds and are therefore potentially useful in tissue engineering and drug delivery. An attempt has been made to review various drug delivery applications of ceramics.     

Material-dependent bone induction by calcium phosphate ceramics: a 2.5-year study in dog

Bone induction by different calcium phosphate biomaterials has been reported previously. With regard to (1) whether the induced bone would disappear with time due to the absence of mechanical stresses and (2) whether this heterotopically formed bone would give rise to uncontrolled growth, a long-time investigation of porous hydroxyapatite ceramic (HA), porous biphasic calcium phosphate ceramic (TCP/HA, BCP), porous alpha-tricalcium phosphate ceramic (alpha-TCP) and porous beta-tricalcium phosphate ceramic (beta-TCP) was performed in dorsal muscles of dog, for 2.5 years. Histological observation, backscattered scanning electron microscopy observation and histomorphometric analysis were made on thin un-decalcified sections of retrieved samples. Normal compact bone with bone marrow was found in all HA implants (n = 4) and in all BCP implants (n = 4), 48 +/- 4% pore area was filled with bone in HA implants and 41 +/- 2% in BCP implants. Bone-like tissue, which was a mineralised bone matrix with osteocytes but lacked osteoblasts and bone marrow, was found in all beta-TCP implants (n = 4) and in one of the four alpha-TCP implants. Both normal bone and bone-like tissues were confined inside the pores of the implants. The results show that calcium phosphate ceramics are osteoinductive in muscles of dogs. Although the quality and quantity varied among different ceramics, the induced bone in both HA and BCP ceramics did neither disappear nor grow uncontrollably during the period as long as 2.5 years.     

Bone in-growth induced by biphasic calcium phosphate ceramic in femoral defect of dogs

Biphasic calcium phosphate (BCP) ceramics consisting of hydroxyapatite (HA) and tricalcium phosphate (TCP) has been used as a bone graft material during the last decade. In this paper, we report the bone in-growth induced by BCP ceramic in the experimentally created circular defects in the femur of dogs. This BCP ceramic consists of 55% hydroxyapatite (HA) and 45% b-tricalcium phosphate (TCP) prepared in situ by the microwave method. The defects were created as 4-mm holes on the lateral aspect of the femur of dogs and the holes were packed with the implant material. The defective sites were radiographed at a period of 4, 8, and 12 weeks postoperatively. The radiographical results showed that the process of ossification started after 4 weeks and the defect was completely filled with new woven bone after 12 weeks. Histological examination of the tissue showed the formation of osteoblast inducing the osteogenesis in the defect. The collageneous fibrous matrix and the complete Haversian system were observed after 12 weeks. The blood serum was collected postoperatively and biochemical assays for alkaline phosphatase activity were carried out. The measurement of alkaline phosphatase activity levels also correlated with the formation of osteoblast-like cells. This microwave-prepared BCP ceramic has proved to be a good biocompatible implant as well as osteoconductive and osteoinductive materials to fill bone defects.     

成骨诱导脂肪基质细胞在骨组织工程体内成骨中的作用

背景：以往研究认为,经过成骨诱导后的脂肪基质细胞通过转化为成骨细胞分泌骨基质进而修复骨缺损,然而并没有明确结论证实。 目的：将经过体外成骨诱导的脂肪基质细胞复合支架材料分别植入骨缺损区和非骨区,根据是否成骨,验证经过成骨诱导后的脂肪基质细胞是否转化为成骨细胞。 方法：取12月龄犬背部皮下脂肪,经胶原酶消化法获得单个核细胞,将培养的第3代细胞与双相磷酸钙陶瓷形成复合物。在犬下颌骨两侧制备长20 mm、高10 mm的箱状缺损,拔除术区牙齿,将细胞支架复合物植入一侧术区,空白侧留作对照;另外在犬背部皮下肌肉区植入细胞支架复合物及骨诱导性磷酸钙陶瓷材料,术后6周及12周经组织学检测骨缺损修复情况。 结果与结论：脂肪基质细胞复合双相磷酸钙陶瓷在骨缺损区成骨,在肌肉区未形成新骨;骨诱导性磷酸钙陶瓷在肌肉区形成新骨。提示成骨诱导并不能将脂肪基质细胞转化为成骨细胞,其确切机制有待进一步研究。     

Analysis of the roles of microporosity and BMP-2 on multiple measures of bone regeneration and healing in calcium phosphate scaffolds

Osteoinductive agents, such as BMP-2, are known to improve bone formation when combined with scaffolds. Microporosity (<20 μm) has also been shown to influence bone regeneration in calcium phosphate (CaP) scaffolds. However, many studies use only the term "osteoconductive" to describe the effects of BMP-2 and microporosity on bone formation, and do not assess the degree of healing that occurred. The objective of this study was to quantify the influence of BMP-2 and microporosity on bone regeneration and healing in biphasic calcium phosphate scaffolds using multiple measures including bone volume fraction, radial distribution, and specific surface area. These measures were quantitatively compared by analyzing microcomputed tomography data and used to formally define and assess healing. A custom image segmentation program was used to segment >100 samples, with 900 images each, that were implanted in porcine mandibular defects for 3, 6, 12 and 24 weeks. The assessment of healing presented in this work demonstrates the level of detail possible in evaluating scaffold-guided bone regeneration. The analysis shows that BMP-2 and microporosity accelerate healing up to 4-fold. BMP-2 and microporosity were shown to have different and complementary roles in bone formation that effect the time needed for a defect to heal.     

磷酸钙生物材料固有骨诱导性的研究现状与展望

磷酸钙陶瓷因与骨的无机组成相似,具有良好生物相容性和骨引导性,无抗原性,被广泛应用于骨缺损修复,但一直认为磷酸钙陶瓷是一类仅具有骨引导性,而无骨诱导性的生物活性材料。大量研究表明通过材料自身组成和结构优化,可赋予磷酸钙陶瓷骨诱导性。本文对磷酸钙骨诱导现象发现和确证、骨诱导过程和机制、影响骨诱导性的材料学因素和骨诱导与动物种属关系、骨诱导相关的间充质细胞来源、以及骨诱导性材料的应用研究现状进行综述,并对磷酸钙生物材料骨诱导性及相关的研究方向予以展望。     

A comparative study of biphasic calcium phosphate ceramics for human mesenchymal stem-cell-induced bone formation

For the repair of bone defects, a tissue engineering approach would be to combine cells capable of osteogenic (i.e. bone-forming) activity with an appropriate scaffolding material to stimulate bone regeneration and repair. Human mesenchymal stem cells (hMSCs), when combined with hydroxyapatite/beta-tricalcium phosphate (HA/TCP) ceramic scaffolds of the composition 60% HA/40% TCP (in weight %), have been shown to induce bone formation in large, long bone defects. However, full repair or function of the long bone could be limited due to the poor remodeling of the HA/TCP material. We conducted a study designed to determine the optimum ratio of HA to TCP that promoted hMSC induced bone formation yet be fully degradable. In a mouse ectopic model, by altering the composition of HA/TCP to 20% HA/80% TCP, hMSC bone induction occurred at the fastest rate in vivo over the other formulations of the more stable 100% HA, HA/TCP (76/24, 63/37, 56/44), and the fully degradable, 100% TCP. In vitro studies also demonstrated that 20/80 HA/TCP stimulated the osteogenic differentiation of hMSCs as determined by the expression of osteocalcin.     

Transplantation of cultured bone cells using combinations of scaffolds and culture techniques

The transplantation of cultured bone cells is expected to become a candidate for bone regeneration therapy. For the clinical application of this therapy, there remain several problems to be overcome, for example, the improvements of scaffolds and culture techniques. In this review article, two kinds of porous ceramics, a novel sintered porous hydroxyapatite and a porous beta-tricalcium phosphate (TCP), as well as a collagen-phosphosphoryn sponge are introduced as new scaffolds for bone regeneration. The former two ceramic scaffolds proved to be applicable for bone regeneration therapy. The collagen-phosphophoryn sponge proved to have bone formation ability in vivo. Moreover, for the application of this therapy to the regeneration of large bone defects, we improved the culture method by applying a low-pressure system and a perfusion system. Both culture systems accelerated the formation of bone in vivo in this transplantation model. Combinations of the scaffolds and culture techniques might be considered when designing therapeutic strategies.     

Efficacité et performance des substituts osseux pour remplacer les allogreffes et autogreffes

Although bone tissue possesses the capacity for regenerative growth, the bone repair process is impaired in many clinical and pathological situations. For example massive bone loss caused by trauma and tumor resection as well as deformities require reconstructive surgery. In this context, there was a critical need to develop implant technologies to promote bone healing. Cortical and cancellous bone grafts are the materials of choice for bone filling or reconstruction, but their clinical use involves some difficulties. Septic complications, viral transmission and unavailability of native bone have therefore led to the development of synthetic bone substitutes. Allograft bone, or tissue harvested from a cadaver, while more readily available, may carry with it the risk of disease transmission and is also difficult to shape [1–3]. A significant additional limitation of allograft bone is the delayed remodeling by the host. In the case of very large defects, the allograft may remain in the implant site throughout the patient's life, creating an area more prone to fracture or infection. The development of calcium phosphate ceramics and other related biomaterials for bone graft involved a better control of the process of biomaterials resorption and bone substitution. Synthetic bone graft materials available as alternatives to autogeneous bone for repair, substitution or augmentation, in particular synthetic biomaterials include, special glass ceramics described as bioactive glasses; calcium phosphates (calcium hydroxyapatite, HA; tricalcium phosphate, TCP; and biphasic calcium phosphate, BCP). These materials differ in composition and physical properties from each other and from bone; and must be take in consideration for more efficient bone ingrowth at the expense of the biomaterials and to adapt to new development of dedicated biomaterials. In the last decade synthetic calcium phosphate materials, principally calcium hydroxyapatite (HA) ceramics, was commercially used. However the concept of bioactivity (release of ions of biological interest) well described for glass ceramic was not particularly take in account for HA and other related biomaterials(ACP Amorphous Calcium Phosphate, CdA Calcium Phosphate deficient Apatite). HA until recently was considered to be non able to be resorbed. Calcium phosphate biomaterials differ in their solubility or extent of dissolution: ACP > > α-TCP > > β-TCP > CdA > > ACP. These ceramics are osteoconductive (act as a support for new bone formation requiring the presence of porosity) and able to be resorbed (degradable through chemical and cellular processes). They are also biocompatible (do not induce adverse local tissue reaction, immunogenicity or systematically toxicity); and more recently, some papers report osteoinductive properties associated to the chemical nature (biphasic Ca P) and the microstruture. Past decade, these bioceramics have been marketed and approved for use in humans as bone substitutes. Various presentations are currently used in orthopaedic and maxillo-facial surgery such as wedges, blocks or granules. Owing to their bone substitution properties, CaP ceramics have naturally been considered as a potential matrix for tissue engineering and the development of a bioactive drug delivery system (DDS) in bone sites. The paper presents the current knowledge on Calcium phosphate bioceramics, Bone tissue engineering and Calcium Phosphate Drug Delivery.     

骨诱导性磷酸钙陶瓷材料修复牙槽突裂

BACKGROUND: In our previous studies, we had prepared calcium phosphate ceramics with better ectopic osteogenesis. OBJECTIVE: To explore the effect of novel osteoinductive calcium phosphate ceramics in the repairing of alveolar cleft. METHODS: Bilaterl alveolar defects were created in nine immature beagles. Three months later, osteoinductive calcium phosphate ceramics with high modular surface (experimental group) and smooth surface (control group) were randomly implanted in each side of the defect. Meanwhile, the corresponding material was implanted into the thigh muscle. New bone formation in the implanted region, osteogenesis in the implanted region and muscle, and respair results were respectively observed by fluorescence microscope, light microscope and CT at 4, 8 and 12 weeks after implantation. RESULTS AND CONCLUSION: (1) Fluorescence microscope observation: A circular permutation of red, yellow and green fluorescent strip could be observed in both two groups. (2) Light microscope observation: At 12 weeks after implantation, in the experimental group, the bone reconstruction was obvious, the implant material was decomposed gradually, the gap was filled with a large number of mature bone that combined with the rest material closely, and numerous Haversian canals appeared; the control group was similar but slightly inferior to the experimental group in the quality of new bone. The experimental group material successfully induced heterotopic osteogenesis in muscle, while the control did not. (3) CT examination: The two group materials restored the appearance and continuity of the alveolar ridge, and made no effect on the eruption of permanent teeth in both sides of the defect. To conclude, our findings suggest that the novel osteoinductive calcium phosphate ceramic exhibits advantages in alveolar cleft repair with earlier osteogenesis activation, faster osteogenesis rate and more bone formation than those traditional materials.     

Discontinuous release of bone morphogenetic protein-2 loaded within interconnected pores of honeycomb-like polycaprolactone scaffold promotes bone healing in a large bone defect of rabbit ulna

The choice of an appropriate carrier and its microarchitectural design is integral in directing bone ingrowth into the defect site and determining its subsequent rate of bone formation and remodeling. We have selected a three-dimensional polycaprolactone (PCL) scaffold with an interconnected honeycomb-like porous structure to provide a conduit for vasculature ingrowth as well as an osteoconductive pathway to guide recruited cells responding to a unique triphasic release of osteoinductive bone morphogenetic proteins (BMP) from these PCL scaffolds. We hypothesize that the use of recombinant human bone morphogenetic protein 2 (rhBMP2)-PCL constructs promotes rapid union and bone regeneration of a large defect. Results of our pilot study on a unilateral 15 mm mid-diaphyseal segmental rabbit ulna defect demonstrated enhanced bone healing with greater amount of bone formation and bridging under plain radiography and microcomputed tomography imaging when compared with an empty PCL and untreated group after 8 weeks postimplantation. Quantitative measurements showed significantly higher bone volume fraction and trabecular thickness, with lower trabecular separation in the rhBMP2-treated groups. Histology evaluation also revealed greater mature bone formation spanning across the entire scaffold region compared with other groups, which showed no bone regeneration within the central defect zone. We highlight that it is the uniqueness of the scaffold having a highly porous network of channels that promoted vascular integration and allowed for cellular infiltration, leading to a discontinuous triphasic BMP2 release profile that mimicked the release profile during natural repair mechanisms in vivo. This study serves as preclinical evidence demonstrating the potential of combining osteoinductive rhBMP2 with our PCL constructs for the repair of large defects in a large animal model.