Facilitate Cell Infiltration Research Articles

Reliable Fabrication of Mineral-Graded Scaffolds by Spin-Coating and Laser Machining for Use in Tendon-to-Bone Insertion Repair.

A reliable method for fabricating biomimetic scaffolds with a controllable mineral gradient to facilitate the surgical repair of tendon-to-bone injuries and the regeneration of the enthesis is reported. The gradient in mineral content is created by sequentially spin-coating with hydroxyapatite/poly(ε-caprolactone) suspensions containing hydroxyapatite nanoparticles in decreasing concentrations. To produce pores and facilitate cell infiltration, the spin-coated film is released and patterned with an array of funnel-shaped microchannels by laser machining. The unique design provided both mechanical (i.e., substrate stiffness) and biochemical (e.g., hydroxyapatite content) cues to spatially control the graded differentiation of mesenchymal stem cells. Immunocytochemical analysis of human mesenchymal stem cell-seeded scaffolds after 14 days of culture demonstrated the formation of a spatial phenotypic cell gradient from osteoblasts to mineralized chondrocytes based on the level of mineralization in the scaffold. By successfully recreating compositional and cellular features of the native tendon enthesis, the biomimetic scaffolds offer a promising avenue for improved tendon-to-bone repair.

Fabricating oxygen self-supplying 3D printed bioactive hydrogel scaffold for augmented vascularized bone regeneration

Limited cells and factors, inadequate mechanical properties, and necrosis of defects center have hindered the wide clinical application of bone-tissue engineering scaffolds. Herein, we construct a self-oxygenated 3D printed bioactive hydrogel scaffold by integrating oxygen-generating nanoparticles and hybrid double network hydrogel structure. The hydrogel scaffold possesses the characteristics of extracellular matrix; Meanwhile, the fabricated hybrid double network structure by polyacrylamide and CaCl2-crosslinked sodium carboxymethylcellulose endows the hydrogel favorable compressive strength and 3D printability. Furthermore, the O2 generated by CaO2 nanoparticles encapsulated in ZIF-8 releases steadily and sustainably because of the well-developed microporous structure of ZIF-8, which can significantly promote cell viability and proliferation in vitro, as well as angiogenesis and osteogenic differentiation with the assistance of Zn2+. More significantly, the synergy of O2 and 3D printed pore structure can prevent necrosis of defects center and facilitate cell infiltration by providing cells the nutrients and space they need, which can further induce vascular network ingrowth and accelerate bone regeneration in all areas of the defect in vivo. Overall, this work provides a new avenue for preparing cell/factor-free bone-tissue engineered scaffolds that possess great potential for tissue regeneration and clinical alternative.

Intelligent In situ Printing of Multimaterial Bioinks for First‐Aid Wound Care Guided by Eye‐In‐Hand Robot Technology

AbstractINSIGHT (INtelligent in situ printing Guided by Eye‐in‐Hand robot Technology), an innovative computer vision‐enabled system that combines a depth camera with a 6‐degree of freedom robot arm, empowering it to identify arbitrary areas at various angles through real time adjustments and to enable volumetric printing performed by dynamic image recognition based on color and contour differences is presented. Continuous targeting of multiple wounds at different locations is achieved. The optimized pneumatic valve synchronized with the INSIGHT can print multiple inks with diverse rheological properties to fabricate scaffolds and bandages with the capacity to treat various types of wounds. The design of dual printed modes, such as extrusion and spray, can significantly decrease printing time for large‐scale wounds on an ex vivo porcine model. INSIGHT demonstrates its ability to treat diabetic wounds, using a microgel‐based ink possessing an inherent porous microstructure to facilitate cell infiltration. In vivo verification highlights its adaptability to enable customized care for rapid emergency treatment of trauma patients.

Controlled pore anisotropy in chitosan-gelatin cryogels for use in bone tissue engineering.

In tissue engineering, the development of an appropriate scaffold is crucial to provide a framework for new tissue growth. The use of cryogels as scaffolds shows promise due to their macroporous structure, but the pore size, distribution, and interconnectivity is highly variable depending on the fabrication process. The objective of the current research is to provide a technique for controlled anisotropy in chitosan-gelatin cryogels to develop scaffolds for bone tissue engineering application. A mold was developed using additive manufacturing to be used during the freezing process in order to fabricate cryogels with a more interconnected pore structure. The scaffolds were tested to evaluate their porosity, mechanical strength, and to observe cell infiltration through the cryogel. It was found that the use of the mold allowed for the creation of designated pores within the cryogel structure which facilitated cell infiltration to the center of the scaffold without sacrificing mechanical integrity of the structure.

A novel porous hydrogel based on hybrid gelation for injectable and tough scaffold implantation and tissue engineering applications

Although there have been many advances in injectable hydrogels as scaffolds for tissue engineering or as payload-containing vehicles, the lack of adequate microporosity for the desired cell behavior, tissue integration, and successful tissue generation remains an important drawback. Herein, we describe an effective porous injectable system that allows in vivo formation of pores through conventional syringe injection at room temperature. This system is based on the differential melting profiles of photocrosslinkable salmon gelatin and physically crosslinked porogens of porcine gelatin (PG), in which PG porogens are solid beads, while salmon methacrylamide gelatin remains liquid during the injection procedure. After injection and photocrosslinking, the porogens were degraded in response to the physiological temperature, enabling the generation of a homogeneous porous structure within the hydrogel. The resultant porogen-containing formulations exhibited controlled gelation kinetics within a broad temperature window (18.5 ± 0.5–28.8 ± 0.8 °C), low viscosity (133 ± 1.4–188 ± 16 cP), low force requirements for injectability (17 ± 0.3–39 ± 1 N), robust mechanical properties after photo-crosslinking (100.9 ± 3.4–332 ± 13.2 kPa), and favorable cytocompatibility (>70% cell viability). Remarkably, in vivo subcutaneous injection demonstrated the suitability of the system with appropriate viscosity and swift crosslinking to generate porous hydrogels. The resulting injected porous constructs showed favorable biocompatibility and facilitated cell infiltration for desirable potential tissue remodeling. Finally, the porogen-containing formulations exhibited favorable handling, easy deposition, and good shape fidelity when used as bioinks in 3D bioprinting technology. This injectable porous system serves as a platform for various biomedical applications, thereby inspiring future advances in cell therapy and tissue engineering.

Warp-knitted fabric structures for a novel biomimetic artificial intervertebral disc for the cervical spine

As an attempt to better replicate the complex kinematics of a natural disc, a novel biomimetic artificial intervertebral disc replacement (bioAID) has been developed containing a swelling hydrogel core as nucleus pulposus, a fiber jacket as annulus fibrosus and metal endplates to connect the device to the adjacent vertebrae. The first prototype consisted of a weft-knitted fiber jacket, in which only a single fiber was used to create the jacket structure. This can endanger the structural integrity of the complete device upon yarn damage. Therefore, in this study, several warp-knitted textile structures were assessed to (1) ensure structural integrity, (2) while allowing for swelling constraint of the hydrogel and (3) behaving as one integrated unit similar to the natural IVD. Moreover, the fiber jacket should (4) act as a scaffold that allows bone ingrowth to ensure long-term stability and (5) have a good durability, (6) be wear resistant and (7) have good manufacturing feasibility with good quality control. In this study, 4 different stitch patterns, including 2 × 1 and 1 × 1 lapping with and without a pillar stitch, were produced. The effect of the stitch pattern and stitch density on the fabric mechanical properties and device swelling and compressive strength was assessed. As a next step, the effect of using multiple layers of fabrics, mimicking the layered structure of annulus fibrosus, on the functional capacity of the bioAID was characterized. All textile structures were capable of limiting the swelling of the hydrogel while withstanding its internal pressure and showing sufficient wear resistance. However, only the 2 × 1 and 2 × 1 with pillar stitch had a pore size range that was suitable for cell infiltration to facilitate osseointegration as well as having the highest strength of the complete device to ensure safety under compression loading. Incorporating different number of jacket layers of these two stitch patterns did not show any significant effect. When also taking the structural parameters into consideration, the 2 × 1 lapping design with 4 layers was able to constrain hydrogel swelling, provide a high compressive strength, could facilitate cell infiltration and had dimensions within the range of a natural intervertebral disc.

Multileveled Hierarchical Hydrogel with Continuous Biophysical and Biochemical Gradients for Enhanced Repair of Full-Thickness Osteochondral Defect.

The repair of hierarchical osteochondral defect requires sophisticated gradient reestablishment; however, few strategies for continuous gradient casting consider the relevance to clinical practice regarding cell adaptability, multiple gradient elements, and precise gradient mirroring native tissue. Here, a hydrogel with continuous gradients in nano-hydroxyapatite (HA) content, mechanical, and magnetism is developed using synthesized superparamagnetic HA nanorods (MagHA) that easily respond to a brief magnetic field. To precisely reconstruct osteochondral tissue, the optimized gradient mode is calculated according to magnetic resonance imaging (MRI) of healthy rabbit knees. Then, MagHA are patterned to form continuous biophysical and biochemical gradients, consequently generating incremental HA, mechanical, and electromagnetic cues under an external magnetic stimulus. To make such depth-dependent biocues work, an adaptable hydrogel is developed to facilitate cell infiltration. Furthermore, this approach is applied in rabbit full-thickness osteochondral defects equipped with a local magnetic field. Surprisingly, this multileveled gradient composite hydrogel repairs osteochondral unit in a perfect heterogeneous feature, which mimics the gradual cartilage-to-subchondral transition. Collectively, this is the first study that combines an adaptable hydrogel with magneto-driven MagHA gradients to achieve promising outcomes in osteochondral regeneration.

A Balance between Pro-Inflammatory and Pro-Reparative Macrophages is Observed in Regenerative D-MAPS.

Microporous annealed particle scaffolds (MAPS) are a new class of granular materials generated through the interlinking of tunable microgels, which produce an interconnected network of void space. These microgel building blocks can be designed with different mechanical or bio-active parameters to facilitate cell infiltration and modulate host response. Previously, changing the chirality of the microgel crosslinking peptides from L- to D-amino acids led to significant tissue regeneration and functional recovery in D-MAPS-treated cutaneous wounds. In this study, the immunomodulatory effect of D-MAPS in a subcutaneous implantation model is investigated. How macrophages are the key antigen-presenting cells to uptake and present these biomaterials to the adaptive immune systemisuncovered. A robust linker-specific IgG2b/IgG1 response to D-MAPS is detected as early as 14 days post-implantation. The fine balance between pro-regenerative and pro-inflammatory macrophage phenotypes is observed in D-MAPS as an indicator for regenerative scaffolds. The work offers valuable insights into the temporal cellular response to synthetic porous scaffolds and establishes a foundation for further optimization of immunomodulatory pro-regenerative outcomes.

Three-dimensional nanofibrous sponges with aligned architecture and controlled hierarchy regulate neural stem cell fate for spinal cord regeneration.

Background: Spinal cord injury (SCI) induces neuronal death and disrupts the nerve fiber bundles, which leads to severe neurological dysfunction and even permanent paralysis. A strategy combining biomimetic nanomaterial scaffolds with neural stem cell (NSC) transplantation holds promise for SCI treatment. Methods: Innovative three-dimensional (3D) nanofibrous sponges (NSs) were designed and developed by a combination of directional electrospinning and subsequent gas-foaming treatment. Immunofluorescence, mRNA sequencing, magnetic resonance imaging, electrophysiological analysis, and behavioral tests were used to investigate the in vitro and in vivo regenerative effects of the 3D NSs. Results: The generated 3D NSs exhibited uniaxially aligned nano-architecture and highly controllable hierarchical structure with super-high porosity (99%), outstanding hydrophilicity, and reasonable mechanical performance. They facilitated cell infiltration, induced cell alignment, promoted neuronal differentiation of NSCs, and enhanced their maturation mediated through cellular adhesion molecule pathways. In vivo, the NSC-seeded 3D NSs efficiently promoted axon reinnervation and remyelination in a rat SCI model, with new "neural relays" developing across the lesion gap. These histological changes were associated with regain of function, including increasing the neurological motor scores of SCI rats, from approximately 2 to 16 (out of 21), and decreasing the sensing time in the tape test from 140 s to 36 s. Additionally, the scaffolds led to restoration of ascending and descending electrophysiological signalling. Conclusion: The as-fabricated 3D NSs effectively regulate NSC fates, and an advanced combination of 3D NS design and transplanted NSCs enables their use as an ideal tissue-engineered scaffold for SCI repair.

3D Electrospun Nanofiber-Based Scaffolds: From Preparations and Properties to Tissue Regeneration Applications.

Electrospun nanofibers have been frequently used for tissue engineering due to their morphological similarities with the extracellular matrix (ECM) and tunable chemical and physical properties for regulating cell behaviors and functions. However, most of the existing electrospun nanofibers have a closely packed two-dimensional (2D) membrane with the intrinsic shortcomings of limited cellular infiltration, restricted nutrition diffusion, and unsatisfied thickness. Three-dimensional (3D) electrospun nanofiber-based scaffolds can provide stem cells with 3D microenvironments and biomimetic fibrous structures. Thus, they have been demonstrated to be good candidates for in vivo repair of different tissues. This review summarizes the recent developments in 3D electrospun nanofiber-based scaffolds (ENF-S) for tissue engineering. Three types of 3D ENF-S fabricated using different approaches classified into electrospun nanofiber 3D scaffolds, electrospun nanofiber/hydrogel composite 3D scaffolds, and electrospun nanofiber/porous matrix composite 3D scaffolds are discussed. New functions for these 3D ENF-S and properties, such as facilitated cell infiltration, 3D fibrous architecture, enhanced mechanical properties, and tunable degradability, meeting the requirements of tissue engineering scaffolds were discovered. The applications of 3D ENF-S in cartilage, bone, tendon, ligament, skeletal muscle, nerve, and cardiac tissue regeneration are then presented with a discussion of current challenges and future directions. Finally, we give summaries and future perspectives of 3D ENF-S in tissue engineering and clinical transformation.

An effective strategy for preparing macroporous and self-healing bioactive hydrogels for cell delivery and wound healing

Macroporous hydrogels have been attracting increasingly attention in cell delivery and tissue regeneration. Assembling microgels is an effective method for preparing macroporous hydrogels. However, current methods are limited to specific requirements to devices and hydrogel inherent properties. In addition, most of macroporous hydrogels lack self-healing abilities due to the permanent covalent crosslink of microgels. In this study, we proposed an effective strategy for preparing a bioactive macroporous self-healable hydrogel by producing microgels through squeezing methacrylated hyaluronic acid (MeHA) and 3-aminophenylboronic acid modified sodium alginate (SABA) nanoporous hydrogels through steel meshes and assembling the MeHA-SABA microgels under alkaline condition created by bioglass (BG) ionic products. Results demonstrated that the obtained macroporous hydrogels have significant larger pore size than nanoporous hydrogels, which facilitated cell infiltration, viability and proliferation. Meanwhile, the macroporous hydrogels had good self-healing abilities with the formation of dynamic B-O bonds. Furthermore, with the bioactivity of BG, the hydrogels could induce cell migration and ingrowth of cells and blood vessels. Thus, bioactivity and macroporous structure of the hydrogels can work synergistically to stimulate tissue regeneration. Taken together, this strategy is effective, simple and can be widely applied in fabricating binary components self-healable macroporous hydrogels as long as the two components are crosslinkable, indicating a great application potential in fabricating macroporous hydrogels for cell delivery and tissue regeneration.

Granular Cellulose Nanofibril Hydrogel Scaffolds for 3D Cell Cultivation.

The replacement of diseased and damaged organs remains an challenge in modern medicine. However, through the use of tissue engineering techniques, it may soon be possible to (re)generate tissues and organs using artificial scaffolds. For example, hydrogel networks made from hydrophilic precursor solutions can replicate many properties found in the natural extracellular matrix (ECM) but often lack the dynamic nature of the ECM, as many covalently crosslinked hydrogels possess elastic and static networks with nanoscale pores hindering cell migration without being degradable. To overcome this, macroporous colloidal hydrogels can be prepared to facilitate cell infiltration. Here, an easy method is presented to fabricate granular cellulose nanofibril hydrogel (CNF) scaffolds as porous networks for 3D cell cultivation. CNF is an abundant natural and highly biocompatible material that supports cell adhesion. Granular CNF scaffolds are generated by pre-crosslinking CNF using calcium and subsequently pressing the gel through micrometer-sized nylon meshes. The granular solution is mixed with fibroblasts and crosslinked with cell culture medium. The obtained granular CNF scaffold is significantly softer and enables well-distributed fibroblast growth. This cost-effective material combined with this efficient and facile fabrication technique allows for 3D cell cultivation in an upscalable manner.

Simultaneous nano- and microscale structural control of injectable hydrogels via the assembly of nanofibrous protein microparticles for tissue regeneration

Injectable hydrogels are advantageous as tissue regeneration scaffolds, as they can be delivered through a minimally invasive injection and seamlessly integrate with the target tissues. However, an important shortcoming of current injectable hydrogels is the lack of simultaneous control over their micro- and nanoscale structures. In this article, the authors report a strategy for developing injectable hydrogels that integrate a fibrous nanostructure and porous microstructure. The hydrogels are prepared by using novel nanofibrous microparticles as the building blocks. The protein based nanofibrous microparticles, fabricated by a spray freezing technology, can be injected through a syringe-needle system. A cell-compatible photocuring process can be deployed to connect the microparticles and form a mechanically robust hydrogel scaffold. The inter-particle voids combined to form the interconnected micropores and the diameter of the nanofibers (100–300 nm) closely mimics that of the native extracellular matrix. Compared to the non-porous hydrogels and non-fibrous hydrogels, the microparticle annealed nanofibrous (MANF) hydrogels potently enhance the osteogenic-marker expression (ALP, Runx2, OCT and BSP) and mineralization of human mesenchymal stem cells in vitro. MANF hydrogels also facilitate cell infiltration and enhance neovasculization in a subcutaneous implantation model in vivo. The capacity of MANF hydrogels to promote bone regeneration is investigated in a calvarial bone repair model. MANF hydrogels demonstrate significant higher bone regeneration after 8 weeks, indicating the significant role of microporosity and nanofibrous architecture in bone regeneration.

Novel biomimetic fiber incorporated scaffolds for tissue engineering.

From a structural perspective, an ideal scaffold ought to possess interconnected macro pores with sizes from 10 to 100 μm to facilitate cell infiltration and tissue formation, as well as similar topography to the natural extracellular matrix (ECM) which would have an impact on cell behavior such as cell adhesion and gene expression. For that purpose, here we developed a fabrication process to incorporate electrospun short fibers within freeze-dried scaffolds for tissue engineering applications. Briefly, PCL short fibers were first produced from electrospun fibers with ultrasonication method. They were then evenly dispersed in gelatin solution and freeze-dried to obtain fiber incorporated scaffolds. The resulting scaffolds exhibited hierarchical structure including major pores with sizes ranging from 50 to 150 μm and the short fibers dispersed in the thin walls of major pores, mimicking the fibrous feature of natural ECM. The short fibers were proven to modify the mechanical properties of scaffold and to facilitate cell adhesion and proliferation on the scaffold. As a promising scaffold for tissue engineering and regenerative medicine, the fiber incorporated scaffold may have further biomedical applications, in which the short fibers can act as drug release vehicles for growth factors or other biomolecules to promote vascularization.

ScCO2-foamed silk fibroin aerogel/poly(ε-caprolactone) scaffolds containing dexamethasone for bone regeneration

Bone scaffolds prepared with porogens and bioactive agents can accelerate bone tissue formation by providing a suitable 3D-porous structure that promotes cell colonization and differentiation towards the osteogenic lineage. In this work, scaffolds containing poly(ε-caprolactone) (PCL) as biopolymeric matrix, silk fibroin as cell adhesion promoter, and dexamethasone as osteogenic differentiation agent, were prepared by supercritical foaming (37 °C, 140 bar, 1 h), a solvent-free method providing a straightforward and effective route for the processing of bioactive grafts. Silk fibroin aerogels in the form of submicron-sized particles were herein developed for the first time and evaluated as porosity inducers. These aerogels incorporated in the scaffolds refined the porous structure and facilitated cell infiltration and the biological fluid transport. Dexamethasone was used in two different forms (base, DX and phosphate salt, DS) to unveil their role in bone regeneration. The morphology of the scaffolds was evaluated using mercury intrusion porosimetry, helium pycnometry and in silico structure modelling. Different release profiles were recorded when using DX or DS. The biological performance was assessed in in vivo tests in calvarial defects using a rat model. Results unveiled the interesting morphological properties of the scaffolds in terms of porosity, pore size distribution and interconnectivity, which are compatible with their application in bone repair. In vivo tests showed the importance of the dexamethasone form and release profile in promoting the bone tissue regeneration with a significant increase in the number of ossification foci and in bone repair extent 14 weeks post-implantation for certain formulations.

A nanofibrous bilayered scaffold for tissue engineering of small‐diameter blood vessels

The main challenge encountered in clinical efficacy of tissue engineered vascular grafts is development of a biomimetic scaffold. To establish a scaffold resembling the architecture of the native blood vessels, a bilayered small‐diameter nanofibrous tubular scaffold was fabricated by sequential electrospinning process. The inner layer of the scaffold was electrospun from a blend of fast degrading poly(glycerol sebacate) (PGS), a hydrophilic and elastomeric polymer, and slowly degrading polycaprolactone (PCL), with a weight ratio of 2:1, while the outer layer was electrospun using PCL. Our findings elucidated that nanofibrous PCL outer layer can improve the mechanical integrity and at the same time the presence of PGS within the nanofibers of the inner layer provides a nonthrombogenic interface. Additionally, electrospun PGS/PCL nanofibers with an appropriate balance between hydrophilic and hydrophobic characteristics served a suitable substrate for adhesion and proliferation of mesenchymal stem cells (MSCs); meanwhile, sufficient pore size provided by the inner layer facilitated cell infiltration into the interior of the scaffold.

Novel engineered tendon-fibrocartilage-bone composite with cyclic tension for rotator cuff repair.

Surgical repair of rotator cuff tears presents a significant clinical challenge with high failure rates and inferior functional outcomes. Graft augmentation improves repair outcomes; however, currently available grafting materials have limitations. Although cell-seeded decellularized tendon slices may facilitate cell infiltration, promote tendon incorporation, and preserve original mechanical strength, the unique fibrocartilage zone is yet to be successfully reestablished. In this study, we investigated the biological and mechanical properties of an engineered tendon-fibrocartilage-bone composite (TFBC) with cyclic tension (3% strain; 0.2Hz). Decellularized TFBCs seeded with bone marrow-derived mesenchymal stem cell (BMSCs) sheets and subjected to mechanical stimulation for up to 7days were characterised by histology, immunohistochemistry, scanning electron microscopy, mechanical testing, and transcriptional regulation. The decellularized TFBC maintained native enthesis structure and properties. Mechanically stimulated TFBC-BMSC constructs displayed increased cell migration after 7days of culture compared with static groups. The seeded cell sheet not only integrated well with tendon scaffold but also distributed homogeneously and aligned to the direction of stretch under dynamic culture. Developmental genes were regulated including scleraxis, which was significantly upregulated with mechanical stimulation. The Young's modulus of the cell-seeded constructs was significantly higher compared with the noncell-seeded controls. In conclusion, the results of this study reveal that the TFBC-BMSC composite provides an ideal multilayer construct for cell seeding and growth, with mechanical preconditioning further enhances cell penetration and differentiation. The BMSC cell sheet revitalised TFBC in conjunction with mechanical stimulation could serve as a novel and primed biological patch to improve rotator cuff repair.

Biomaterial Scaffolds in Regenerative Therapy of the Central Nervous System.

The central nervous system (CNS) is the most important section of the nervous system as it regulates the function of various organs. Injury to the CNS causes impairment of neurological functions in corresponding sites and further leads to long-term patient disability. CNS regeneration is difficult because of its poor response to treatment and, to date, no effective therapies have been found to rectify CNS injuries. Biomaterial scaffolds have been applied with promising results in regeneration medicine. They also show great potential in CNS regeneration for tissue repair and functional recovery. Biomaterial scaffolds are applied in CNS regeneration predominantly as hydrogels and biodegradable scaffolds. They can act as cellular supportive scaffolds to facilitate cell infiltration and proliferation. They can also be combined with cell therapy to repair CNS injury. This review discusses the categories and progression of the biomaterial scaffolds that are applied in CNS regeneration.

Regeneration of osteochondral defects in vivo by a cell-free cylindrical poly(lactide-co-glycolide) scaffold with a radially oriented microstructure.

A scaffold with an oriented porous architecture to facilitate cell infiltration and bioactive interflow between neo-host tissues is of great importance for in situ inductive osteochondral regeneration. In this study, a poly(lactide-co-glycolide) (PLGA) scaffold with oriented pores in its radial direction was fabricated via unidirectional cooling of the PLGA solution in the radial direction, following with lyophilization. Micro-computed tomography evaluation and scanning electron microscopy observation confirmed the radially oriented microtubular pores in the scaffold. The scaffold had porosity larger than 90% and a compressive modulus of 4MPa in a dry state. Culture of bone marrow stem cells in vitro revealed faster migration and regular distribution of cells in the poly(lactide-co-glycolide) scaffold with oriented pores compared with the random PLGA scaffold. The cell-free oriented macroporous PLGA scaffold was implanted into rabbit articular osteochondral defect in vivo for 12weeks to evaluate its inductive tissue regeneration function. Histological analysis confirmed obvious tide mark formation and abundant chondrocytes distributed regularly with obvious lacunae in the cartilage layer. Safranin O-fast green staining showed an obvious boundary between the two layers with distinct staining results, indicating the simultaneous regeneration of the cartilage and subchondral bone layers, which is not the case for the random poly(lactide-co-glycolide) scaffold after the same implantation in vivo. The oriented macroporous PLGA scaffold is a promising material for the in situ inductive osteochondral regeneration without the necessity of preseeding cells.

Development of hybrid scaffolds with natural extracellular matrix deposited within synthetic polymeric fibers.

A major challenge of tissue engineering is to generate materials that combine bioactivity with stability in a form that captures the robust nature of native tissues. Here we describe a procedure to fabricate a novel hybrid extracellular matrix (ECM)-synthetic scaffold biomaterial by cell-mediated deposition of ECM within an electrospun fiber mat. Synthetic polymer fiber mats were fabricated using poly(desamino tyrosyl-tyrosine carbonate) (PDTEC) co-spun with poly(ethylene glycol) (PEG) used as a sacrificial polymer. PEG removal increased the overall mat porosity and produced a mat with a layered structure that could be peeled into separate sheets of about 50 μm in thickness. Individual layers had pore sizes and wettability that facilitated cell infiltration over the depth of the scaffold. Confocal microscopy showed the formation of a highly interpenetrated network of cells, fibronectin fibrils, and synthetic fibers mimicking a complex ECM as observed within tissues. Decellularization did not perturb the structure of the matrix or the fiber mat. The resulting hybrid ECM-scaffold promoted cell adhesion and spreading and stimulated new ECM assembly by stem cells and tumor cells. These results identify a new technique for fabricating highly porous synthetic fibrous scaffolds and an approach to supplement them with natural biomimetic cues. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2162-2170, 2017.