Mechanical properties of bioactive glass (13-93) scaffolds fabricated by robotic deposition for structural bone repair

There is a need to develop synthetic scaffolds to repair large defects in load-bearing bones. Bioactive glasses have attractive properties as a scaffold material for bone repair, but data on their mechanical properties are limited. The objective of the present study was to comprehensively evaluate the mechanical properties of strong porous scaffolds of silicate 13-93 bioactive glass fabricated by robocasting. As-fabricated scaffolds with a grid-like microstructure (porosity 47%, filament diameter 330 μm, pore width 300 μm) were tested in compressive and flexural loading to determine their strength, elastic modulus, Weibull modulus, fatigue resistance, and fracture toughness. Scaffolds were also tested in compression after they were immersed in simulated body fluid (SBF) in vitro or implanted in a rat subcutaneous model in vivo. As fabricated, the scaffolds had a strength of 86 ± 9 MPa, elastic modulus of 13 ± 2 GPa, and a Weibull modulus of 12 when tested in compression. In flexural loading the strength, elastic modulus, and Weibull modulus were 11 ± 3 MPa, 13 ± 2 GPa, and 6, respectively. In compression, the as-fabricated scaffolds had a mean fatigue life of ～10⁶ cycles when tested in air at room temperature or in phosphate-buffered saline at 37 °C under cyclic stresses of 1–10 or 2–20 MPa. The compressive strength of the scaffolds decreased markedly during the first 2 weeks of immersion in SBF or implantation in vivo, but more slowly thereafter. The brittle mechanical response of the scaffolds in vitro changed to an elasto-plastic response after implantation for longer than 2–4 weeks in vivo. In addition to providing critically needed data for designing bioactive glass scaffolds, the results are promising for the application of these strong porous scaffolds in loaded bone repair.     

Mechanical and microstructural properties of polycaprolactone scaffolds with one-dimensional,two-dimensional,and three-dimensional orthogonally oriented porous architectures produced by selective laser sintering

This article reports on the experimental determination and finite element modeling of tensile and compressive mechanical properties of solid polycaprolactone (PCL) and of porous PCL scaffolds with one-dimensional, two-dimensional and three-dimensional orthogonal, periodic porous architectures produced by selective laser sintering (SLS). PCL scaffolds were built using optimum processing parameters, ensuring scaffolds with nearly full density (>95%) in the designed solid regions and with excellent geometric and dimensional control (within 3–8% of design). The tensile strength of bulk PCL ranged from 10.5 to 16.1 MPa, its modulus ranged from 343.9 to 364.3 MPa, and the tensile yield strength ranged from 8.2 to 10.1 MPa. These values are consistent with reported literature values for PCL processed through various manufacturing methods. Across porosity ranged from 56.87% to 83.3%, the tensile strength ranged from 4.5 to 1.1 MPa, the tensile modulus ranged from 140.5 to 35.5 MPa, and the yield strength ranged from 3.2 to 0.76 MPa. The compressive strength of bulk PCL was 38.7 MPa, the compressive modulus ranged from 297.8 to 317.1 MPa, and the compressive yield strength ranged from 10.3 to 12.5 MPa. Across porosity ranged from 51.1% to 80.9%, the compressive strength ranged from 10.0 to 0.6 MPa, the compressive modulus ranged from 14.9 to 12.1 MPa, and the compressive yield strength ranged from 4.25 to 0.42 MPa. These values, while being in the lower range of reported values for trabecular bone, are the highest reported for PCL scaffolds produced by SLS and are among the highest reported for similar PCL scaffolds produced through other layered manufacturing techniques. Finite element analysis showed good agreement between experimental and computed effective tensile and compressive moduli. Thus, the construction of bone tissue engineering scaffolds endowed with oriented porous architectures and with predictable mechanical properties through SLS is demonstrated.     

Preconditioned 70S30C bioactive glass foams promote osteogenesis in vivo

Bioactive glass scaffolds (70S30C; 70% SiO₂ and 30% CaO) produced by a sol–gel foaming process are thought to be suitable matrices for bone tissue regeneration. Previous in vitro data showed bone matrix production and active remodelling in the presence of osteogenic cells. Here we report their ability to act as scaffolds for in vivo bone regeneration in a rat tibial defect model, but only when preconditioned. Pretreatment methods (dry, pre-wetted or preconditioned without blood) for the 70S30C scaffolds were compared against commercial synthetic bone grafts (NovaBone® and Actifuse®). Poor bone ingrowth was found for both dry and wetted sol–gel foams, associated with rapid increase in pH within the scaffolds. Bone ingrowth was quantified through histology and novel micro-CT image analysis. The percentage bone ingrowth into dry, wetted and preconditioned 70S30C scaffolds at 11 weeks were 10 ± 1%, 21 ± 2% and 39 ± 4%, respectively. Only the preconditioned sample showed above 60% material–bone contact, which was similar to that in NovaBone and Actifuse. Unlike the commercial products, preconditioned 70S30C scaffolds degraded and were replaced with new bone. The results suggest that bioactive glass compositions should be redesigned if sol–gel scaffolds are to be used without preconditioning to avoid excess calcium release.     

Repairing a critical-sized bone defect with highly porous modified and unmodified baghdadite scaffolds

This is the first reported study to prepare highly porous baghdadite (Ca₃ZrSi₂O₉) scaffolds with and without surface modification and investigate their ability to repair critical-sized bone defects in a rabbit radius under normal load. The modification was carried out to improve the mechanical properties of the baghdadite scaffolds (particularly to address their brittleness) by coating their surfaces with a thin layer (～400 nm) of polycaprolactone (PCL)/bioactive glass nanoparticles (nBGs). The β-tricalcium phosphate/hydroxyapatite (TCP/HA) scaffolds with and without modification were used as the control groups. All of the tested scaffolds had an open and interconnected porous structure with a porosity of ～85% and average pore size of 500 μm. The scaffolds (six per scaffold type and size of 4 mm × 4 mm × 15 mm) were implanted (press-fit) into the rabbit radial segmental defects for 12 weeks. Micro-computed tomography and histological evaluations were used to determine bone ingrowth, bone quality, and implant integration after 12 weeks of healing. Extensive new bone formation with complete bridging of the radial defect was evident with the baghdadite scaffolds (modified/unmodified) at the periphery and in close proximity to the ceramics within the pores, in contrast to TCP/HA scaffolds (modified/unmodified), where bone tended to grow between the ulna adjacent to the implant edge. Although the modification of the baghdadite scaffolds significantly improved their mechanical properties, it did not show any significant effect on in vivo bone formation. Our findings suggest that baghdadite scaffolds with and without modification can serve as a potential material to repair critical sized bone defects.     

Fabrication of HA/TCP scaffolds with a graded and porous structure using a camphene-based freeze-casting method

A room temperature camphene-based freeze-casting method was used to fabricate hydroxyapatite/tricalcium phosphate (HA/TCP) ceramic scaffolds. By varying the solid loading of the mixture and the freezing temperature, a range of structures with different pore sizes and strength characteristics were achieved. The macropore size of the HA/TCP bioceramics was in the range of 100–200 μm, 40–80 μm and less than 40 μm at solid loadings of 10, 20 and 30 vol.%, respectively. The initial level of solid loading played a primary role in the resulting porosity of the scaffolds. The porosity decreased from 72.5 to 31.4 vol.% when the solid loading was increased from 10 to 30 vol.%. This resulted in an increase in the compressive strength from 2.3 to 36.4 MPa. The temperature gradient, rather than the percentage porosity, influenced the pore size distribution. The compressive strength increased from 1.95 to 2.98 MPa when samples were prepared at 4 °C as opposed to 30 °C. The results indicated that it was possible to manufacture porous HA/TCP bioceramics, with compressive strengths comparable to cancellous bone, using the freeze-casting manufacturing technique, which could be of significant clinical interest.     

Incorporation of bioactive polyvinylpyrrolidone–iodine within bilayered collagen scaffolds enhances the differentiation and subchondral osteogenesis of mesenchymal stem cells

Polyvinylpyrrolidone–iodine (Povidone-iodine, PVP-I) is widely used as an antiseptic agent for lavation during joint surgery; however, the biological effects of PVP–I on cells from joint tissue are unknown. This study examined the biocompatibility and biological effects of PVP–I on cells from joint tissue, with the aim of optimizing cell-scaffold based joint repair. Cells from joint tissue, including cartilage derived progenitor cells (CPC), subchondral bone derived osteoblast and bone marrow derived mesenchymal stem cells (BM-MSC) were isolated. The concentration-dependent effects of PVP–I on cell proliferation, migration and differentiation were evaluated. Additionally, the efficacy and mechanism of a PVP–I loaded bilayer collagen scaffold for osteochondral defect repair was investigated in a rabbit model. A micromolar concentration of PVP–I was found not to affect cell proliferation, CPC migration or extracellular matrix production. Interestingly, micromolar concentrations of PVP–I promote osteogenic differentiation of BM-MSC, as evidenced by up-regulation of RUNX2 and Osteocalcin gene expression, as well as increased mineralization on the three-dimensional scaffold. PVP–I treatment of collagen scaffolds significantly increased fibronectin binding onto the scaffold surface and collagen type I protein synthesis of cultured BM-MSC. Implantation of PVP–I treated collagen scaffolds into rabbit osteochondral defect significantly enhanced subchondral bone regeneration at 6 weeks post-surgery compared with the scaffold alone (subchondral bone histological score of 8.80 ± 1.64 vs. 3.8 ± 2.19, p < 0.05). The biocompatibility and pro-osteogenic activity of PVP–I on the cells from joint tissue and the enhanced subchondral bone formation in PVP–I treated scaffolds would thus indicate the potential of PVP–I for osteochondral defect repair.     

Density–property relationships in collagen–glycosaminoglycan scaffolds

Collagen–glycosaminoglycan scaffolds for the regeneration of skin have previously been fabricated by freeze-drying a slurry containing a co-precipitate of collagen and glycosaminoglycan. The mechanical properties of the scaffold are low (e.g. the dry compressive Young’s modulus is roughly 30 kPa and the dry compressive strength is roughly 5 kPa). There is interest in using these scaffolds for tendon and ligament regeneration where there is a need for improved mechanical properties. Previous attempts to increase the mechanical properties of the scaffold by increasing the solid volume fraction of the scaffolds were limited by the increasing viscosity of the slurry, making it more difficult to mix and giving inhomogeneous scaffolds. Our recent work on mineralized collagen–glycosaminoglycan scaffolds used a vacuum filtration technique to increase the volume fraction of solids in the slurry, thereby increasing the density and mechanical properties of the scaffolds. In this work, we used this technique to fabricate collagen–glycosaminoglycan scaffolds with dry densities between 0.0076 and 0.0311 g cm^?3 and pore sizes between 250 and 350 μm, values appropriate for soft tissue growth. The compressive Young’s modulus and strength in the dry state increased from 32 to 127 kPa and from 5 to 19 kPa, respectively, with increasing density. The tensile Young’s modulus in the dry state increased from 295 to 3.1 MPa with increasing density. Finally, we showed that the attachment of cells onto the scaffold was directly proportional to the specific surface area of the scaffold, which defines the total internal surface area per volume of scaffold.     

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.     

Bone regeneration in strong porous bioactive glass (13-93) scaffolds with an oriented microstructure implanted in rat calvarial defects

There is a need for synthetic bone graft substitutes to repair large bone defects resulting from trauma, malignancy and congenital diseases. Bioactive glass has attractive properties as a scaffold material but factors that influence its ability to regenerate bone in vivo are not well understood. In the present work, the ability of strong porous scaffolds of 13-93 bioactive glass with an oriented microstructure to regenerate bone was evaluated in vivo using a rat calvarial defect model. Scaffolds with an oriented microstructure of columnar pores (porosity = 50%; pore diameter = 50?150 μm) showed mostly osteoconductive bone regeneration, and new bone formation, normalized to the available pore area (volume) of the scaffolds, increased from 37% at 12 weeks to 55% at 24 weeks. Scaffolds of the same glass with a trabecular microstructure (porosity = 80%; pore width = 100?500 μm), used as the positive control, showed bone regeneration in the pores of 25% and 46% at 12 and 24 weeks, respectively. The brittle mechanical response of the as-fabricated scaffolds changed markedly to an elastoplastic response in vivo at both implantation times. These results indicate that both groups of 13-93 bioactive glass scaffolds could potentially be used to repair large bone defects, but scaffolds with the oriented microstructure could also be considered for the repair of loaded bone.     

Evaluation of bone regeneration in implants composed of hollow HA microspheres loaded with transforming growth factor β1 in a rat calvarial defect model

Implants that serve simultaneously as an osteoconductive matrix and as a device for local growth factor delivery may be required for optimal bone regeneration in some applications. In the present study, hollow hydroxyapatite (HA) microspheres (106–150 μm) in the form of three-dimensional (3-D) scaffolds or individual (loose) microspheres were created using a glass conversion process. The capacity of the implants, with or without transforming growth factor β1 (TGF-β1), to regenerate bone in a rat calvarial defect model was compared. The 3-D scaffolds supported the proliferation and alkaline phosphatase activity of osteogenic MLO-A5 cells in vitro, showing their cytocompatibility. Release of TGF-β1 from the 3-D scaffolds into phosphate-buffered saline ceased after 2–3 days when ～30% of the growth factor was released. Bone regeneration in the 3-D scaffolds and the individual microspheres increased with time from 6 to 12 weeks, but it was significantly higher (23%) in the individual microspheres than in the 3-D scaffolds (15%) after 12 weeks. Loading with TGF-β1 (5 μg per defect) enhanced bone regeneration in the 3-D scaffolds and individual microspheres after 6 weeks, but had little effect after 12 weeks. 3-D scaffolds and individual microspheres with larger HA diameter (150–250 μm) showed better ability to regenerate bone. Based on these results, implants composed of hollow HA microspheres show promising potential as an osteoconductive matrix for local growth factor delivery in bone regeneration.     

Modified PHBV scaffolds by in situ UV polymerization: Structural characteristic,mechanical properties and bone mesenchymal stem cell compatibility

An ideal scaffold provides an interface for cell adhesion and maintains enough biomechanical support during tissue regeneration. Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) scaffolds with pore sizes ranging from 100 to 500 μm and porosity ～90% were prepared by the particulate-leaching method, and then modified by the introduction of polyacrylamide (PAM) on the inner surface of scaffolds using in situ UV polymerization, with the aim of enhancing the biological and mechanical properties of the PHBV scaffolds. The modified PHBV scaffolds had interconnected pores with porosity of 75.4–78.6% and pore sizes at peak volume from 20 to 50 μm. The compressive load and modulus were up to 62.45 N and 1.06 MPa, respectively. The water swelling percentage (WSP) of the modified PHBV scaffolds increased notably compared with that of the PHBV scaffolds, with the maximum WSP at 537%. Sheep bone mesenchymal stem cells (BMSC) were cultured on the PHBV and modified PHBV. The hydrophilic PAM chains did not influence BMSC viability or proliferation index, but the initial cell adhesion at 1 h of culture was enhanced significantly. Framing PHBV scaffold along with gel-like PAM chains inside is a novel model of inner surface modification for PHBV scaffolds, which shows potential in tissue engineering applications.     

Solvent-dependent properties of electrospun fibrous composites for bone tissue regeneration

Biodegradable polymer–ceramic composite scaffolds have gained importance in recent years in the field of orthopedic biomaterials and tissue engineering scaffolds for improving the rate of degradation and limited mechanical properties of bioactive ceramics. This study sought to create composites using the electrospinning process to achieve fibrous scaffolds with uniform fiber morphologies and uniform ceramic dispersions. Composites consisting of 20% hydroxyapatite/80% β-tricalcium phosphate (20/80 HA/TCP) and poly (ε-caprolactone) (PCL) were fabricated. The 20/80 HA/TCP composition was chosen as the ceramic component because of previous reports of greater bone tissue formation in comparison with HA or TCP alone. For electrospinning, PCL was dissolved in either methylene chloride (Composite–MC) or a combination of methylene chloride (80%) and dimethylformamide (20%) (Composite–MC + DMF). Composite–MC mats contained a bimodal distribution of fiber diameters with nanofibers between larger, micron-sized fibers with an average pore size of 79.6 ± 67 μm, whereas Composite–MC + DMF fibers had uniform fiber diameters with an average pore size of 7.0 ± 4.2 μm. Elemental mapping determined that the ceramic was distributed throughout the mat and inside the fiber for both composites. However, physical characterization using differential scanning calorimetry (DSC) and mechanical testing revealed that the ceramic in the mats produced with MC + DMF were more uniformly dispersed than the ceramic in the mats produced with MC alone. Maximum tensile stress and strain were significantly higher for Composite–MC + DMF mats compared with Composite–MC mats and were comparable with the mechanical properties of mats of PCL alone. For both composites, there was molecular interaction between the PCL and the ceramic, as demonstrated by a maximum increase of ～10 °C in the glass transition values with the addition of the ceramic, as confirmed by Fourier transform infrared analysis. In addition, the crystallization behavior of the composites suggested that the ceramic was acting as a nucleating agent. Cell viability studies using human mesenchymal stem cells (MSC) showed that both composite scaffolds supported cell growth. However, cell numbers at early time points in culture were significantly higher on mats produced from MC + DMF compared with mats prepared with MC alone. Further examination revealed that cells were able to infiltrate the pores of the Composite–MC mats, but remained on the outer surface of the Composite–MC + DMF and unfilled PCL mats during the culture period. The results of this study demonstrate that the solvent or solvent combination used in preparing the electrospun composite mats plays a critical role in determining its properties, which may, in turn, affect cell behavior.     

The stimulation of healing within a rat calvarial defect by mPCL–TCP/collagen scaffolds loaded with rhBMP-2

Bone morphogenetic proteins (BMPs) have been widely investigated for their clinical use in bone repair and it is known that a suitable carrier matrix to deliver them is essential for optimal bone regeneration within a specific defect site. Fused deposited modeling (FDM) allows for the fabrication of medical grade poly ?-caprolactone/tricalcium phosphate (mPCL–TCP) scaffolds with high reproducibility and tailor designed dimensions. Here we loaded FDM fabricated mPCL–TCP/collagen scaffolds with 5 μg recombinant human (rh)BMP-2 and evaluated bone healing within a rat calvarial critical-sized defect. Using a comprehensive approach, this study assessed the newly regenerated bone employing micro-computed tomography (μCT), histology/histomorphometry, and mechanical assessments. By 15 weeks, mPCL–TCP/collagen/rhBMP-2 defects exhibited complete healing of the calvarium whereas the non-BMP-2-loaded scaffolds showed significant less bone ingrowth, as confirmed by μCT. Histomorphometry revealed significantly increased bone healing amongst the rhBMP-2 groups compared to non-treated scaffolds at 4 and 15 weeks, although the % BV/TV did not indicate complete mineralisation of the entire defect site. Hence, our study confirms that it is important to combine microCt and histomorphometry to be able to study bone regeneration comprehensively in 3D. A significant up-regulation of the osteogenic proteins, type I collagen and osteocalcin, was evident at both time points in rhBMP-2 groups. Although mineral apposition rates at 15 weeks were statistically equivalent amongst treatment groups, micro-compression and push-out strengths indicated superior bone quality at 15 weeks for defects treated with mPCL–TCP/collagen/rhBMP-2. Consistently over all modalities, the progression of healing was from empty defect < mPCL–TCP/collagen < mPCL–TCP/collagen/rhBMP-2, providing substantiating data to support the hypothesis that the release of rhBMP-2 from FDM-created mPCL–TCP/collagen scaffolds is a clinically relevant approach to repair and regenerate critically-sized craniofacial bone defects.     

Electrospun nanostructured scaffolds for bone tissue engineering

The current challenge in bone tissue engineering is to fabricate a bioartificial bone graft mimicking the extracellular matrix (ECM) with effective bone mineralization, resulting in the regeneration of fractured or diseased bones. Biocomposite polymeric nanofibers containing nanohydroxyapatite (HA) fabricated by electrospinning could be promising scaffolds for bone tissue engineering. Nanofibrous scaffolds of poly-l-lactide (PLLA, 860 ± 110 nm), PLLA/HA (845 ± 140 nm) and PLLA/collagen/HA (310 ± 125 nm) were fabricated, and the morphology, chemical and mechanical characterization of the nanofibers were evaluated using scanning electron microscopy, Fourier transform infrared spectroscopy and tensile testing, respectively. The in vitro biocompatibility of different nanofibrous scaffolds was also assessed by growing human fetal osteoblasts (hFOB), and investigating the proliferation, alkaline phosphatase activity (ALP) and mineralization of cells on different nanofibrous scaffolds. Osteoblasts were found to adhere and grow actively on PLLA/collagen/HA nanofibers with enhanced mineral deposition of 57% higher than the PLLA/HA nanofibers. The synergistic effect of the presence of an ECM protein, collagen and HA in PLLA/collagen/HA nanofibers provided cell recognition sites together with apatite for cell proliferation and osteoconduction necessary for mineralization and bone formation. The results of our study showed that the biocomposite PLLA/collagen/HA nanofibrous scaffold could be a potential substrate for the proliferation and mineralization of osteoblasts, enhancing bone regeneration.     

Mesenchymal stem cell proliferation and differentiation on load-bearing trabecular Nitinol scaffolds

Bone tissue regeneration in load-bearing regions of the body requires high-strength porous scaffolds capable of supporting angiogenesis and osteogenesis. 70% porous Nitinol (NiTi) scaffolds with a regular 3-D architecture resembling trabecular bone were produced from Ni foams using an original reactive vapor infiltration technique. The “trabecular Nitinol” scaffolds possessed a high compressive strength of 79 MPa and high permeability of 6.9 × 10^?6 cm². The scaffolds were further modified to produce a near Ni-free surface layer and evaluated in terms of Ni ion release and human mesenchymal stem cell (hMSC) proliferation (AlamarBlue), differentiation (alkaline phosphatase activity, ALP) and mineralization (Alizarin Red S staining). Scanning electron microscopy was employed to qualitatively corroborate the results. hMSCs were able to adhere and proliferate on both as-produced and surface-modified trabecular NiTi scaffolds, to acquire an osteoblastic phenotype and produce a mineralized extracellular matrix. Both ALP activity and mineralization were increased on porous scaffolds compared to control polystyrene plates. Experiments in a model coculture system of microvascular endothelial cells and hMSCs demonstrated the formation of prevascular structures in trabecular NiTi scaffolds. These data suggest that load-bearing trabecular Nitinol scaffolds could be effective in regenerating damaged or lost bone tissue.     

Enhanced bone regeneration in rat calvarial defects implanted with surface-modified and BMP-loaded bioactive glass (13-93) scaffolds

The repair of large bone defects, such as segmental defects in the long bones of the limbs, is a challenging clinical problem. Our recent work has shown the ability to create porous scaffolds of silicate 13-93 bioactive glass by robocasting which have compressive strengths comparable to human cortical bone. The objective of this study was to evaluate the capacity of those strong porous scaffolds with a grid-like microstructure (porosity = 50%; filament width = 330 μm; pore width = 300 μm) to regenerate bone in a rat calvarial defect model. Six weeks post-implantation, the amount of new bone formed within the implants was evaluated using histomorphometric analysis. The amount of new bone formed in implants composed of the as-fabricated scaffolds was 32% of the available pore space (area). Pretreating the as-fabricated scaffolds in an aqueous phosphate solution for 1, 3 and 6 days to convert a surface layer to hydroxyapatite prior to implantation enhanced new bone formation to 46%, 57% and 45%, respectively. New bone formation in scaffolds pretreated for 1, 3 and 6 days and loaded with bone morphogenetic protein-2 (BMP-2) (1 μg per defect) was 65%, 61% and 64%, respectively. The results show that converting a surface layer of the glass to hydroxyapatite or loading the surface-treated scaffolds with BMP-2 can significantly improve the capacity of 13-93 bioactive glass scaffolds to regenerate bone in an osseous defect. Based on their mechanical properties evaluated previously and their capacity to regenerate bone found in this study, these 13-93 bioactive glass scaffolds, pretreated or loaded with BMP-2, are promising in structural bone repair.     

Injectable and porous PLGA microspheres that form highly porous scaffolds at body temperature

Injectable scaffolds are of interest in the field of regenerative medicine because of their minimally invasive mode of delivery. For tissue repair applications, it is essential that such scaffolds have the mechanical properties, porosity and pore diameter to support the formation of new tissue. In the current study, porous poly(dl-lactic acid-co-glycolic acid) (PLGA) microspheres were fabricated with an average size of 84 ± 24 μm for use as injectable cell carriers. Treatment with ethanolic sodium hydroxide for 2 min was observed to increase surface porosity without causing the microsphere structure to disintegrate. This surface treatment also enabled the microspheres to fuse together at 37 °C to form scaffold structures. The average compressive strength of the scaffolds after 24 h at 37 °C was 0.9 ± 0.1 MPa, and the average Young’s modulus was 9.4 ± 1.2 MPa. Scaffold porosity levels were 81.6% on average, with a mean pore diameter of 54 ± 38 μm. This study demonstrates a method for fabricating porous PLGA microspheres that form solid porous scaffolds at body temperature, creating an injectable system capable of supporting NIH-3T3 cell attachment and proliferation in vitro.     

Microwave-processed nanocrystalline hydroxyapatite: Simultaneous enhancement of mechanical and biological properties

Despite the excellent bioactivity of hydroxyapatite (HA) ceramics, poor mechanical strength has limited the applications of these materials primarily to coatings and other non-load-bearing areas as bone grafts. Using synthesized HA nanopowder, dense compacts with grain sizes in the nanometer to micrometer range were processed via microwave sintering between 1000 and 1150 °C for 20 min. Here we demonstrate that the mechanical properties, such as compressive strength, hardness and indentation fracture toughness, of HA compacts increased with a decrease in grain size. HA with 168 ± 86 nm grain size showed the highest compressive strength of 395 ± 42 MPa, hardness of 8.4 ± 0.4 GPa and indentation fracture toughness of 1.9 ± 0.2 MPa m^1/2. To study the in vitro biological properties, HA compacts with grain size between 168 nm and 1.16 μm were assessed for in vitro bone cell–material interactions with human osteoblast cell line. Vinculin protein expression for cell attachment and bone cell proliferation using MTT assay showed that surfaces with finer grains provided better bone cell–material interactions than coarse-grained samples. Our results indicate simultaneous improvements in mechanical and biological properties in microwave sintered HA compacts with nanoscale grain size.     

Characterization of a biodegradable electrospun polyurethane nanofiber scaffold: Mechanical properties and cytotoxicity

The current study analyzes the biodegradation of a polycarbonate polyurethane scaffold intended for the growth of a tissue-engineered annulus fibrosus (AF) disc component. Electrospun scaffolds with random and aligned nanofiber configurations were fabricated using a biodegradable polycarbonate urethane with and without an anionic surface modifier (anionic dihydroxyl oligomer), and the mechanical behavior of the scaffolds was examined during a 4 week biodegradation study. Both the tensile strength and initial modulus of aligned scaffolds (σ = 14 ± 1 MPa, E = 46 ± 3 MPa) were found to be higher than those of random fiber scaffolds (σ = 1.9 ± 0.4 MPa, E = 2.1 ± 0.2 MPa) prior to degradation. Following initial wetting of the scaffold, the initial modulus of the aligned samples showed a significant decrease (dry: 46 ± 3 MPa; pre-wetted: 9 ± 1 MPa, p < 0.001). The modulus remained relatively constant during the remainder of the 4 week incubation period (aligned at 4 weeks: 8.0 ± 0.3 MPa). The tensile strength for aligned fiber scaffolds was affected in the same manner. Similar changes were not observed for the initial modulus of the random scaffold configuration. Biodegradation of the scaffold in the presence of cholesterol esterase (a monocyte derived enzyme) yielded a 0.5 mg week^–1 weight loss. The soluble and non-soluble degradation products were found to be non-toxic to bovine AF cells grown in vitro. The consistent rate of material degradation along with stable mechanical properties comparable to those of native AF tissue and the absence of cytotoxic effects make this polymer a suitable biomaterial candidate for further investigation into its use for tissue-engineering annulus fibrosus.