Macroporous Scaffolds Research Articles

Template-assisted freeze casting of macroporous Ti6Al4V scaffolds with long-range order lamellar structure

A modified water-based freeze casting to fabricate long-range order lamellar Ti6Al4V scaffolds with macropores and high porosity has been developed by using the pre-frozen WS2 substrate as the template. Pores morphologies of the scaffolds relied on the previous ice crystal which had formed in templates and thus could be adjusted by altering the template structure. The decrease of gelatin in WS2 slurry transformed the bottom template from cell pore structure into the lamellar structure followed by improvement of lamellar orientation of the top Ti6Al4V scaffold. The pore widths of the scaffolds were in the range of 84–217 μm which were difficult to realize without the assisted template. The scaffolds also exhibited mechanical compatibility with bone tissues, indicating that the template-assisted freeze casting is a promising approach to fabricate macroporous Ti6Al4V implants.

Preparation and Characterization of Surface Heat Sintered Nanohydroxyapatite and Nanowhitlockite Embedded Poly (Lactic-co-glycolic Acid) Microsphere Bone Graft Scaffolds: In Vitro and in Vivo Studies.

In the context of using bone graft materials to restore and improve the function of damaged bone tissues, macroporous biodegradable composite bone graft scaffolds have osteoinductive properties that allow them to provide a suitable environment for bone regeneration. Hydroxyapatite (HAP) and whitlockite (WLKT) are the two major components of hard tissues such as bone and teeth. Because of their biocompatibility and osteoinductivity, we synthesized HAP (nHAP) and WLKT nanoparticles (nWLKT) by using the chemical precipitation method. The nanoparticles were separately incorporated within poly (lactic-co-glycolic acid) (PLGA) microspheres. Following this, the composite microspheres were converted to macroporous bone grafts with sufficient mechanical strength in pin or screw shape through surface sintering. We characterized physico-chemical and mechanical properties of the nanoparticles and composites. The biocompatibility of the grafts was further tested through in vitro cell adhesion and proliferation studies using rabbit bone marrow stem cells. The ability to promote osteogenic differentiation was tested through alkaline phosphate activity and immunofluorescence staining of bone marker proteins. For in vivo study, the bone pins were implanted in tibia bone defects in rabbits to compare the bone regeneration ability though H&E, Masson’s trichrome and immunohistochemical staining. The results revealed similar physico-chemical characteristics and cellular response of PLGA/nHAP and PLGA/nWLKT scaffolds but the latter is associated with higher osteogenic potential towards BMSCs, pointing out the possibility to use this ceramic nanoparticle to prepare a sintered composite microsphere scaffold for potential bone grafts and tissue engineered implants.

Novel synthesis method combining a foaming agent with freeze-drying to obtain hybrid highly macroporous bone scaffolds

Three-dimensional macroporous scaffolds are commonly used in bone tissue engineering applications since they provide sufficient space for cell migration and proliferation, facilitating bone ingrowth and implant vascularisation. The aim of this work was to combine two simple methods, freeze-drying and gas-foaming, in order to fabricate highly macroporous bone scaffolds made of chitosan/agarose matrix reinforced with nanohydroxyapatite. The secondary goal of this research was to comprehensively assess biomedical potential of developed biomaterials. In this work, it was demonstrated that simultaneous application of freeze-drying and gas-foaming technique allows to obtain hybrid (as proven by ATR-FTIR) macroporous bone scaffolds (pore diameter > 50 μm) characterized by high open (70 %) and interconnected porosity. Novel scaffolds were non-toxic, favoured osteoblasts adhesion and growth and induced apatite formation on their surfaces, indicating their high bioactivity that is essential for good implant osseointegration. Biomaterials were also prone to enzymatic degradation, degradation in acidified microenvironment (e.g. osteoclast-mediated), and slow degradation under physiological pH of 7.4. Moreover, the scaffolds revealed microstructure (70 % open porosity, SSA approx. 30 m2/g, high share of macropores with diameter in the range 100−410 μm) and compressive strength (1–1.4 MPa) comparable to cancellous bone, indicating that they are promising implants for cancellous bone regeneration.

Nanostructured degradable macroporous hydrogel scaffolds with controllable internal morphologies via reactive electrospinning

Creating micro/nanostructured hydrogels with tunable morphologies under cell-friendly processing conditions would enable rational engineering of hydrogel scaffolds for targeted biomedical applications. Herein, an all-aqueous single-step reactive electrospinning method is applied to prepare hydrogel networks with controlled morphologies on both the nanoscale and the microscale. Hydrazide and aldehyde-functionalized poly(oligo ethylene glycol methacrylate) (POEGMA) are co-spun from a double barrel syringe together with poly(ethylene oxide) (PEO) as an electrospinning aid. By varying the concentrations and molecular weights of PEO and/or POEGMA, various morphologies from pure fibers to beaded fibers to bead network morphologies with tunable bead sizes can be fabricated, all of which remain monolithically stable in water due to the dynamic covalent crosslinks formed within the gel structure. The rates and magnitudes of swelling, degradation, and mechanics of the resulting scaffolds can be tuned by independently controlling gel morphologies on the nanoscale (i.e. crosslink density within the gel) and the microscale (i.e. the network structure formed), with an atypical independence of swelling relative to the mechanics and degradation rate observed. Furthermore, the internal morphology of the networks is demonstrated to systematically alter both the cell responses within the scaffolds and the rate of protein release from the scaffolds, with small fibers showing optimal cell proliferation, bead networks exhibiting the slowest protein release kinetics and very high maintained cell viabilities post-electrospinning, and beaded fibers showing intermediate properties. Statement of SignificanceControlling the internal structure of hydrogels is critical to successfully applying hydrogels in biomedical applications such as tissue engineering or cell/drug delivery. However, current techniques to fabricate hydrogel scaffolds typically require additives or gelation processes that are poorly compatible with cells and/or require multi-step processes. In this paper, we describe the fabrication of hydrogel scaffolds with tunable feature sizes (from nanometer to micrometer scale) and structures (from all fibers to bead/fiber mixtures to a new “bead network” morphology) using a reactive electrospinning strategy leveraging dynamic hydrazone crosslinking. We show single-step cell/protein loading and systematic control over cell proliferation and protein release kinetics by systematically manipulating the scaffold morphologies and feature sizes, allowing facile customization of scaffold properties for targeted applications.

Engineered macroporous hydrogel scaffolds via pickering emulsions stabilized by MgO nanoparticles promote bone regeneration.

Hydrogels are appealing biomaterials for regenerative medicine since biomimetic modifications of their polymeric network can provide unique physical properties and emulate the native extracellular matrix (ECM). Meanwhile, therapeutic metal ions, such as magnesium ions (Mg2+), not only regulate cellular behaviours but also stimulate local bone formation and healing. However, the absence of a meaningful macroporous structure and the uncompromising mechanical strength are still challenges. Herein, we designed a macroporous composite hydrogel based on mild and fast thiol-ene click reactions. The Pickering emulsion method was adopted to form a macroporous structure and introduce MgO nanoparticles (NPs). The results show that the composite hydrogel possesses good mechanical strength and an evenly distributed macroporous structure. MgO NPs stabilized at the oil/water interface not only function as effective emulsion stabilizers, but also enhance the mechanical properties of hydrogels and mediate the sustained release of Mg2+. In vitro cell experiments demonstrated that the composite hydrogel displays good biocompatibility. More importantly, the release of Mg2+ ions from hydrogels can effectively promote the osteogenic differentiation of BMSCs. Furthermore, an in vivo study showed that macroporous hydrogels can provide a good extracellular matrix microenvironment for in situ osteogenesis and accelerate bone tissue regeneration.

Microencapsulation improves chondrogenesis in vitro and cartilaginous matrix stability in vivo compared to bulk encapsulation.

The encapsulation of cells into microgels is attractive for applications in tissue regeneration. While cells are protected against shear stress during injection, the assembly of microgels after injection into a tissue defect also forms a macroporous scaffold that allows effective nutrient transport throughout the construct. However, in most of current strategies that form microgel-based macroporous scaffold or higher-order structures, cells are seeded during or post the assembly process and not microencapsulated in situ. The objective of this study is to investigate the chondrogenic phenotype of microencapsulated fetal chondrocytes in a biocompatible, assembled microgel system vs. bulk gels and to test the stability of the constructs in vivo. Here, we demonstrate that cell microencapsulation leads to increased expression of cartilage-specific genes in a TGF-β1-dependent manner. This correlates, as shown by histological staining, with the ability of microencapsulated cells to deposit cartilaginous matrix after migrating to the surface of the microgels, while keeping a macroscopic granular morphology. Implantation of precultured scaffolds in a subcutaneous mouse model results in vessel infiltration in bulk gels but not in assembled microgels, suggesting a higher stability of the matrix produced by the cells in the assembled microgel constructs. The cells are able to remodel the microgels as demonstrated by the gradual disappearance of the granular structure in vivo. The biocompatible microencapsulation and microgel assembly system presented in this article therefore hold great promise as an injectable system for cartilage repair.

Substituted hydroxyapatite coatings of bone implants.

Surface modification of orthopedic and dental implants has been demonstrated to be an effective strategy to accelerate bone healing at early implantation times. Among the different alternatives, coating implants with a layer of hydroxyapatite (HAp) is one of the most used techniques, due to its excellent biocompatibility and osteoconductive behavior. The composition and crystalline structure of HAp allow for numerous ionic substitutions that provide added value, such as antibiotic properties or osteoinduction. In this article, we will review and critically analyze the most important advances in the field of substituted hydroxyapatite coatings. In recent years substituted HAp coatings have been deposited not only on orthopedic prostheses and dental implants, but also on macroporous scaffolds, thus expanding their applications towards bone regeneration therapies. Besides, the capability of substituted HAps to immobilize proteins and growth factors by non-covalent interactions has opened new possibilities for preparing hybrid coatings that foster bone healing processes. Finally, the most important in vivo outcomes will be discussed to understand the prospects of substituted HAp coatings from a clinical point of view.

A mesoporous encapsulated nanozyme for decontaminating two kinds of wastewater and avoiding secondary pollution.

Water pollution is becoming a major and serious issue all over the world. However, considering conventional methods for wastewater purification is chemically, energetically and technologically intensive, thus finding a reliable and energy-saving method is still a considerable conundrum. Herein, a mesoporous encapsulated dual-function complex nanozyme consisting of gold, platinum, and cobalt tri-metal is reported, and it not only shows high activity for water purification but also has outstanding resistance to the harsh conditions of the wastewater. This complex nanozyme simultaneously exhibits excellent peroxidase mimicking activity and catalase mimicking activity, which could handle two kinds of wastewater effectively in one system, thus avoiding the use of fresh H2O2 resources and greatly saving resources. In comparison with conventional methods, this nanozyme integrated the "oxidation" process and the "deoxidation" process together in one system very well, successfully avoiding the problem of secondary pollution in the current methods. Furthermore, the treated water is compatible with cells and safe for mice. Therefore, this highly-active nanozyme provides a resource-saving and environment-friendly method for wastewater decontamination, which will be a potential way to directly convert polluted water into bio-safe resources in the future.

Fabrication and characterization of polylactic acid/polycaprolactone composite macroporous micro-nanofiber scaffolds by phase separation

By using incompatible polymers, the preparation of scaffolds with a macroporous structure has overcome the use of porogens and carcinogenic solvents.

One-step synthesis of a macroporous Cu-g/C3N4 nanofiber electrocatalyst for efficient oxygen reduction reaction.

We report a one-step synthesis of a macroporous Cu-g/C3N4 nanofiber catalyst, in which Cu-nanodots (<10 nm) are well coupled with g/C3N4 nanosheets to form Cu-Nx nanorods on the macroporous carbon nanofiber scaffold. The catalyst with a high specific surface area of 514.9 m2 g-1 exposes abundant electroactive sites that facilitate the adsorption of oxygen intermediates and thus exhibits high ORR activity, such as a high half wave potential of 0.83 V and long-term stability over 1000 cycles. The catalyst is a potential substitute for noble metal catalysts.

Static and dynamic 3D culture of neural precursor cells on macroporous cryogel microcarriers

Neural precursor cells have been much studied to further our understanding of the far-reaching and controversial question of adult neurogenesis. Currently, differentiation of primary neural precursor cells from the mouse dentate gyrus via 2-dimentional in vitro culture yields low numbers of neurons, a major hindrance to the field of study. 3-dimentional “neurosphere” culture allows better 3D cell-cell contact, but control over cell differentiation is poor because nutrition and oxygen restrictions at the core of the sphere causes spontaneous differentiation, predominantly to glial cells, not neurons. Our group has developed macroporous scaffolds, which overcome the above-mentioned problems, allowing long-term culture of neural stem cells, which can be differentiated into a much higher yield of neurons. Herein we describe a method for culturing neural precursor cells on RGD peptide functionalized-heparin containing cryogel scaffolds, either in standard non-adherent well-plates (static culture) or in spinner flasks (dynamic culture). This method includes:•The synthesis and characterization of heparin based microcarriers.•A “static” 3D culture method for that does not require spinner flask equipment.•“Dynamic” culture in which cell loaded microcarriers are transferred to a spinner flask.

In Situ functionalization of Poly(hydroxyethyl methacrylate) Cryogels with Oligopeptides via β-Cyclodextrin-Adamantane Complexation for Studying Cell-Instructive Peptide Environment.

Oligopeptides are versatile cell modulators resembling pleiotropic activities of ECM proteins and growth factors. Studying the role of cell-instructive peptide signals within 3D scaffolds, yet poorly known, requires effective approaches to introducing bioactive sequences into appropriate materials. We synthesized RGD and GHK motif based peptides 1 and 2 linked to the terminal adamantyl group (Ad) and their fluorescent derivatives 3 and 4. Poly(hydroxyethyl methacrylate) (pHEMA) cryogels with additional PEG/β-cyclodextrin (CD) units were prepared as an inert macroporous scaffold capable to bind the adamantylated peptides via affinity CD-Ad complexation. According to toluidine blue staining, the CD moieties were effectively and stably incorporated in the pHEMA cryogels at nanomolar amounts per milligram of material. The CD component gradually increased the thickness and swelling ability of the polymer walls of cryogels, resulting in a noticeable decrease in macropore size and modulation of viscoelastic properties. The labeled peptides exhibited fast kinetics of specific binding to the CD-modified cryogels and were simultaneously immobilized by coincubation. The peptide loading approached ca. 0.31 mg per cm2 of cryogel sheet. A well-defined mitogenic effect of the immobilized peptides (2 < 1≪ 1 + 2) was revealed toward 3T3 and PC-12 cells. The synergistic action of RGD and GHK peptides induced a profound change in cell behavior/morphology attributed to a growth-factor-like activity of the composition. Altogether, our results provide an effective procedure for the preparation of CD-modified pHEMA cryogels and their uniform in situ functionalization with bioactive peptide(s) of interest and an informative study of cellular responses in the functionalized scaffolds.

Increased pore size of scaffolds improves coating efficiency with sulfated hyaluronan and mineralization capacity of osteoblasts

BackgroundDelayed bone regeneration of fractures in osteoporosis patients or of critical-size bone defects after tumor resection are a major medical and socio-economic challenge. Therefore, the development of more effective and osteoinductive biomaterials is crucial.MethodsWe examined the osteogenic potential of macroporous scaffolds with varying pore sizes after biofunctionalization with a collagen/high-sulfated hyaluronan (sHA3) coating in vitro. The three-dimensional scaffolds were made up from a biodegradable three-armed lactic acid-based macromer (TriLA) by cross-polymerization. Templating with solid lipid particles that melt during fabrication generates a continuous pore network. Human mesenchymal stem cells (hMSC) cultivated on the functionalized scaffolds in vitro were investigated for cell viability, production of alkaline phosphatase (ALP) and bone matrix formation. Statistical analysis was performed using student’s t-test or two-way ANOVA.ResultsWe succeeded in generating scaffolds that feature a significantly higher average pore size and a broader distribution of individual pore sizes (HiPo) by modifying composition and relative amount of lipid particles, macromer concentration and temperature for cross-polymerization during scaffold fabrication. Overall porosity was retained, while the scaffolds showed a 25% decrease in compressive modulus compared to the initial TriLA scaffolds with a lower pore size (LoPo). These HiPo scaffolds were more readily coated as shown by higher amounts of immobilized collagen (+ 44%) and sHA3 (+ 25%) compared to LoPo scaffolds. In vitro, culture of hMSCs on collagen and/or sHA3-coated HiPo scaffolds demonstrated unaltered cell viability. Furthermore, the production of ALP, an early marker of osteogenesis (+ 3-fold), and formation of new bone matrix (+ 2.5-fold) was enhanced by the functionalization with sHA3 of both scaffold types. Nevertheless, effects were more pronounced on HiPo scaffolds about 112%.ConclusionIn summary, we showed that the improvement of scaffold pore sizes enhanced the coating efficiency with collagen and sHA3, which had a significant positive effect on bone formation markers, underlining the promise of using this material approach for in vivo studies.

Cellulose-Derived Nanographene Oxide Reinforced Macroporous Scaffolds of High Internal Phase Emulsion-Templated Cross-Linked Poly(ε-caprolactone).

Cellulose-derived nanographene oxide (nGO)-type carbon dot reinforced porous scaffolds of poly(ε-caprolactone) (PCL) were developed as templates from high internal phase emulsions (HIPE). The mechanical strength, structural integrity, and reusability of the scaffolds were enhanced via in situ cross-linking. An oil-in-oil (o/o) HIPE of ε-caprolactone monomer (CL) was made for this purpose, and the ring-opening polymerization of a continuous phase comprised of CL, catalyst (Sn(Oct)2), and cross-linker (bis(caprolactone-4-yl)) (BCY) was carried out. The functionalization of scaffolds with nGO was assessed along with its role as an effective Pickering stabilizer of the HIPEs. The pore size and porosity of the scaffolds were governed by HIPE morphology, which in turn was controlled by the amount of nGO and the volume fraction of the dispersed phase. The nGO-functionalized scaffolds of cross-linked PCL thus prepared were characterized for their morphological structure, mechanical strength, and oil sorption capacity. Enhanced oil adsorption of nGO-functionalized scaffolds proved them to be of higher potency compared to those made of neat PCL. Superior compressive strength and reusability of scaffolds for oil adsorption up to 40 times while maintaining the structural integrity for ≥25 sorption-desorption cycles added extra value to such scaffolds. The scaffolds also had excellent cell viability as evaluated by MG63 osteoblast-like cells and some bioactivity in the form of calcium phosphate mineralization on the surface of the scaffolds.

Gelatin-Based Microribbon Hydrogels Guided Mesenchymal Stem Cells to Undergo Endochondral Ossification In Vivo with Bone-Mimicking Mechanical Strength

Most stem cell–based bone tissue engineering strategies to date yield bone through direct bone formation, which mimics intramembranous ossification. However, bone injuries often affect long bones which are formed through endochondral ossification, involving an initial cartilage template formation followed by remodeling to form bones. There remains a critical need to develop scaffolds that enhance stem cell–based bone formation through endochondral ossification with bone-mimicking mechanical strength. Here we evaluated the potential of gelatin-based microribbons (μRBs) as macroporous scaffolds for enhancing human mesenchymal stem cell (MSC)–based bone formation through endochondral ossification. This material platform was compared with conventional gelatin hydrogels (HGs) as controls. MSCs were encapsulated in μRB or HG scaffolds, primed in chondrogenic medium in vitro for 2 weeks, and then implanted in a mouse subcutaneous model with no additional factors. μRB scaffolds supported fast cartilage deposition by MSCs, which was completely remodeled and replaced by mineralized bone. Impressively, the compressive moduli of MSC-seeded μRB scaffolds increased from 10 to 3224 kPa by week 11, a range that mimics native bone. In comparison, while HG supported endochondral ossification, the speed was much slower, with less matrix deposition and only a modest increase in compressive modulus to 269 kPa. These results validate gelatin μRBs as a promising scaffold for repairing long bone defects by guiding robust endochondral ossification. Natural bone development and healing occurs through two distinct pathways: intramembranous ossification and endochondral ossification. Most bone injuries affect long bones, which are formed through endochondral ossification, involving an initial cartilage template formation followed by remodeling to form bones. However, scaffolds that can guide stem cell–based bone formation through endochondral ossification with bone-mimicking mechanical strength remain lacking. Here we report that macroporous gelatin-based microribbons (μRBs) accelerate endochondral ossification by human mesenchymal stem cells (MSCs) in vivo using a mouse subcutaneous model. Impressively, the mechanical properties of MSC-seeded μRB scaffolds increased over 300-fold over 11 weeks to bone-mimicking range, whereas conventional gelatin hydrogel controls reached less than 10% of the bone modulus. These results validate gelatin μRBs as a promising novel scaffold for repairing long bone defects by guiding robust endochondral ossification.

Composite Cryogel with Polyelectrolyte Complexes for Growth Factor Delivery.

Macroporous scaffolds composed of chitosan (CHI), hydroxyapatite (HA), heparin (Hep), and polyvinyl alcohol (PVA) were prepared with a glutaraldehyde (GA) cross-linker by cryogelation. Addition of PVA to the reaction mixture slowed down the formation of a polyelectrolyte complex (PEC) between CHI and Hep, which allowed more thorough mixing, and resulted in the development of the homogeneous matrix structure. Freezing of the CHI-HA-GA and PVA-Hep-GA mixture led to the formation of a non-stoichiometric PEC between oppositely charged groups of CHI and Hep, which caused further efficient immobilization of bone morphogenic protein 2 (BMP-2) possible due to electrostatic interactions. It was shown that the obtained cryogel matrix released BMP-2 and supported the differentiation of rat bone marrow mesenchymal stem cells (rat BMSCs) into the osteogenic lineage. Rat BMSCs attached to cryogel loaded with BMP-2 and expressed osteocalcin in vitro. Obtained composite cryogel with PEC may have high potential for bone regeneration and tissue engineering applications.

3D Bioprinting of Novel Biocompatible Scaffolds for Endothelial Cell Repair.

The aim of this study was to develop and evaluate an optimized 3D bioprinting technology in order to fabricate novel scaffolds for the application of endothelial cell repair. Various biocompatible and biodegradable macroporous scaffolds (D = 10 mm) with interconnected pores (D = ~500 µm) were fabricated using a commercially available 3D bioprinter (r3bEL mini, SE3D, USA). The resolution of the printing layers was set at ~100 µm for all scaffolds. Various compositions of polylactic acid (PLA), polyethylene glycol (PEG) and pluronic F127 (F127) formulations were prepared and optimized to develop semi-solid viscous bioinks. Either dimethyloxalylglycine (DMOG) or erythroprotein (EPO) was used as a model drug and loaded in the viscous biocompatible ink formulations with a final concentration of 30% (w/w). The surface analysis of the bioinks via a spectroscopic analysis revealed a homogenous distribution of the forming materials throughout the surface, whereas SEM imaging of the scaffolds showed a smooth surface with homogenous macro-porous texture and precise pore size. The rheological and mechanical analyses showed optimum rheological and mechanical properties of each scaffold. As the drug, DMOG, is a HIF-1 inducer, its release from the scaffolds into PBS solution was measured indirectly using a bioassay for HIF-1α. This showed that the release of DMOG was sustained over 48 h. The release of DMOG was enough to cause a significant increase in HIF-1α levels in the bioassay, and when incubated with rat aortic endothelial cells (RAECs) for 2 h resulted in transcriptional activation of a HIF-1α target gene (VEGF). The optimum time for the increased expression of VEGF gene was approximately 30 min and was a 3-4-fold increase above baseline. This study provides a proof of concept, that a novel bioprinting platform can be exploited to develop biodegradable composite scaffolds for potential clinical applications in endothelial cell repair in cardiovascular disease (CVD), or in other conditions in which endothelial damage occurs.

In vitro and in vivo biocompatibility of calcium-phosphate scaffolds three-dimensional printed by stereolithography for bone regeneration.

Stereolithography (SLA) is an interesting manufacturing technology to overcome limitations of commercially available particulated biomaterials dedicated to intra-oral bone regeneration applications. The purpose of this study was to evaluate the in vitro and in vivo biocompatibility and osteoinductive properties of two calcium-phosphate (CaP)-based scaffolds manufactured by SLA three-dimensional (3D) printing. Pellets and macro-porous scaffolds were manufactured in pure hydroxyapatite (HA) and in biphasic CaP (HA:60-TCP:40). Physico-chemical characterization was performed using micro X-ray fluorescence, scanning electron microscopy (SEM), optical interferometry, and microtomography (μCT) analyses. Osteoblast-like MG-63 cells were used to evaluate the biocompatibility of the pellets in vitro with MTS assay and the cell morphology and growth characterized by SEM and DAPI-actin staining showed similar early behavior. For in vivo biocompatibility, newly formed bone and biodegradability of the experimental scaffolds were evaluated in a subperiosteal cranial rat model using μCT and descriptive histology. The histological analysis has not indicated evidences of inflammation but highlighted close contacts between newly formed bone and the experimental biomaterials revealing an excellent scaffold osseointegration. This study emphasizes the relevance of SLA 3D printing of CaP-based biomaterials for intra-oral bone regeneration even if manufacturing accuracy has to be improved and further experiments using biomimetic scaffolds should be conducted.

Effect of conductive substrate on the photoelectrochemical properties of Cu2O film electrodes for methyl viologen reduction

Cuprous oxide (Cu2O) is a candidate for low-cost p-type semiconductor oxides for photoelectrochemical solar energy conversion. The Cu2O crystalline films electrodeposited on conductive substrates have been extensively studied as photocathodes because of their moderate photoelectrochemical performance, but their incident photon-to-current conversion efficiency (IPCE) was still limited because of the incompatibility between minority carrier diffusion length and light absorption depth in planar structures. One of the strategies to solve this issue is to decrease the thickness of the Cu2O films with keeping the loading amount. Herein, we used a sintered titanium microfiber felt as a three-dimensional macroporous scaffold for Cu2O crystalline thin films because it exhibits a large surface area compared with the conventional flat substrate. The effect of the Cu2O film thickness on the photocathode properties was investigated using methyl viologen as an acceptor of the photoexcited electrons. The Cu2O thin films deposited on Ti microfibers were superior to that of the Cu2O films deposited on dense planar substrates such as fluorine-doped tin oxide (FTO)-coated glass and titanium sheets. It was revealed that the large Cu2O–electrolyte and Cu2O–substrate interfaces resulted in the enhancement of IPCE for the photoelectrochemical cathodic reaction.

Pulp ECM-derived macroporous scaffolds for stimulation of dental-pulp regeneration process

ObjectiveRecent studies suggest xenogeneic extracellular matrices as potential regenerative tools in dental pulp regeneration. This study aimed to fabricate and characterize a novel three-dimensional macroporous pulp-derived scaffold that enables the attachment, penetration, proliferation and differentiation of mesenchymal stem cells. MethodBovine pulp was decellularized and characterized with histological and DNA content methods. This scaffold was prepared using finely milled lyophilized decellularized pulp extracellular matrix (ECM) digested with pepsin. Three different concentrations of ECM (1.50, 2.25 and 3.00mg/ml) were freeze-dried and were tested with/without chemical crosslinking. The specimens were subjected to physicochemical characterization, cell viability and quantitative real time polymerase chain reaction assessments with human bone marrow mesenchymal stem cells (hBMMSCs). All scaffolds were subcutaneously implanted in rats for two weeks and histological and immunostaining analyses were performed. ResultsHistological and DNA analysis confirmed complete decellularization. All samples demonstrated more than 97% porosity and 1.50mg/ml scaffold demonstrated highest water absorption. The highest cell viability and proliferation of hBMMSCs was observed on the 3.00mg/ml crosslinked scaffolds. The gene expression analysis showed a significant increase of dmp-1 and collagen-I on 3.00mg/ml crosslinked scaffolds compared to the other scaffolds. Histological examination of subcutaneous implanted scaffolds revealed low immunological response, and enhanced angiogenesis in cross-linked samples compared to non-crosslinked samples. SignificanceThe three-dimensional macroporous pulp-derived injectable scaffold developed and characterized in this study displayed potential for regenerative therapy. While the scaffold biodegradability was decreased by crosslinking, the biocompatibility of post-crosslinked scaffold was significantly improved.