Scaffold Layer Research Articles

Understanding the Significance of Layer Bonding in Melt Electrowriting.

Melt electrowriting (MEW) is a high-resolution additive manufacturing technology capable of depositing micrometric fibers onto a moving collector to form 3D scaffolds of controlled mechanical properties. While the critical role of layer bonding to achieve mechanical integrity in fused deposition modeling has been widely reported, it remains largely unknown in MEW, in part due to a lack of methods to assess it. Here, a systematic framework is developed to unravel the significance of layer bonding in MEW scaffolds and its ultimate effect on their mechanical properties. Results show that printing parameters, scaffold design, and print path have a strong impact on layer bonding strength of poly(ɛ-caprolactone) MEW scaffolds. This study demonstrates that a small increase of 5µm in fiber diameter can enhance the layer bonding strength by as much as 70%, greatly impacting the overall scaffold properties. A method is also established to control MEW scaffold layer bonding using a heated collector. Importantly, this study reveals that scaffold architecture alone is not responsible for the overall mechanical properties. Finally, a method to obtain tailored layer bond strengths within a given scaffold is established. This has significant implications as provides new possibilities to control mechanical properties of MEW scaffolds through layer bonding.

Phytotherapeutic Hierarchical PCL-Based Scaffolds as a Multifunctional Wound Dressing: Combining 3D Printing and Electrospinning.

This study focuses on developing hybrid scaffolds incorporating phytotherapeutic agents via a combination of three-dimensional (3D) printing and electrospinning to enhance mechanical properties and provide antibacterial activity, in order to address the limitations of traditional antibiotics. In this regard, 3D-printed polycaprolactone (PCL) struts are first fabricated using fused deposition modeling (FDM). Then, alkaline surface treatment is applied to improve the adhesion of electrospun nanofibers. Finally, peppermint oil (PEP) or clove oil (CLV)-incorporated PCL-gelatin (GEL) electrospun nanofibers are collected on top of the 3D-printed PCL scaffolds by electrospinning. Incorporating PEP or CLV into PCL-GEL electrospun nanofibers enhances the scaffold's layer detachment and adhesion force. In addition, the DPPH free radical scavenging activity assay indicates that incorporating PEP or CLV improves the antioxidant properties of the scaffolds. Further, antibacterial activity results reveal that PEP or CLV incorporated scaffolds exhibit inhibition against Staphylococcus aureus and Escherichia coli bacteria. Moreover, anti-inflammatory assays show that scaffolds reduce the concentration of nitric oxide (NO) released from Raw 264.7 macrophage-like cells. On the other hand, the phytotherapeutic hierarchical scaffolds have no toxic effect on normal human dermal fibroblast (NHDF) cells, and PEP or CLV enhance cell attachment and proliferation. Overall, incorporating natural phytotherapeutic agents into hierarchical scaffolds shows promise for advancing wound healing applications.

Multifunctional hernia repair biopatch: Development, characterization, in vitro and in vivo evaluation

Incisional hernia, a prevalent postoperative complication, is characterized by the protrusion of organs or tissues through damaged abdominal wall. Predisposing factors for hernias include obesity, wound infections, immunosuppression, and comorbidities. However, hernia patches currently in use, including the commercial polypropylene (PP) patch, still have limitations in providing the mechanical and biological properties necessary for abdominal wall regeneration. In this study, three dimensional (3D) printing and coaxial electrospinning methods were combined to create a multifunctional double layered hernia repair biopatch to overcome these limitations. The double-layer design of the biopatch serves a multifunctional role in addressing incisional hernia models, with 3D printed ciprofloxacin (CIP) loaded polycaprolactone (PCL)/gelatin (Ge) scaffold (3DCIP) layer and coaxially electrospun PCL/Ge/κ-carrageenan (κ-C) nanofiber (NF) layers. While 3DCIP layers provides temporary mechanical reinforcement to the damaged abdominal wall, prevention of adhesions to internal organs, and reduction of surgical site infections, NF layer serves as tissue regeneration and fast wound healing. The developed multifunctional hernia biopatches underwent comprehensive physical and chemical characterization, followed by in vitro and in vivo evaluations. These evaluations included comparisons with a commercial PP patch, which was used as the control in the experiments. The study successfully fabricated multifunctional hernia biopatches with excellent antibacterial properties, high mechanical robustness, and strong biocompatibility.

Improving biological and mechanical properties of bioprinted PCL-alginate-chondrocyte scaffolds for patellofemoral cartilage tissue regeneration

In this study, polycaprolactone (PCL) scaffolds have been employed as structural framework scaffolds for patellofemoral cartilage tissue regeneration. The biomechanical and biological properties of different scaffolds were investigated by varying alginate concentrations and the number of scaffold layers. Patellofemoral cartilage defects result in knee pain and reduced mobility, and they are usually treated with conventional methods, often with limited success. Generally, tissue-engineered PCL-alginate scaffolds fabricated by bioprinting technology show promise for enhanced cartilage regeneration due to the biocompatibility and mechanical stability of PCL. In addition, alginate is known for its cell encapsulation capabilities and for promoting cell viability. Biological and morphological assessments, utilizing water contact angle, cell adhesion tests, MTT assays, and scanning electron microscopy (SEM), informed the selection of the optimized scaffold. Comparative analyses between the initial optimal scaffolds with the same chemical composition also included flexural and compression tests and fracture surface observations using SEM. The controlled integration of PCL and alginate offers a hybrid approach, that assembles the mechanical strength of PCL and the bioactive properties of alginate for tissue reconstruction potential. This study aims to identify the most effective scaffold composition for patellofemoral articular cartilage tissue engineering, emphasizing cell viability, structural morphology, and mechanical integrity. The results showed that the optimum biomechanical and biological properties of scaffolds were obtained with a 10% alginate concentration in the monolayer of PCL structure. The findings contribute to regenerative medicine by advancing the understanding of functional tissue constructs, bringing us closer to addressing articular cartilage defects and related clinical challenges.

Fabrication of 3D engineered intestinal tissue producing abundant mucus by air–liquid interface culture using paper-based dual-layer scaffold

Fabrication of engineered intestinal tissues with the structures and functions as humans is crucial and promising as the tools for developing drugs and functional foods. The aim of this study is to fabricate an engineered intestinal tissue from Caco-2 cells by air–liquid interface culture using a paper-based dual-layer scaffold and analyze its structure and functions. Just by simply placing on a folded paper soaked in the medium, the electrospun gelatin microfiber mesh as the upper cell adhesion layer of the dual-layer scaffold was exposed to the air, while the lower paper layer worked to preserve and supply the cell culture medium to achieve stable culture over several weeks. Unlike the flat tissue produced using the conventional commercial cultureware, Transwell, the engineered intestinal tissue fabricated in this study formed three-dimensional villous architectures. Microvilli and tight junction structures characteristic of epithelial tissue were also formed at the apical side. Furthermore, compared to the tissue prepared by Transwell, mucus production was significantly larger, and the enzymatic activities of drug metabolism and digestion were almost equivalent. In conclusion, the air–liquid interface culture using the paper-based dual-layer scaffold developed in this study was simple but effective in fabricating the engineered intestinal tissue with superior structures and functions.

3D bioartificial stretchable scaffolds mimicking the mechanical hallmarks of human cardiac fibrotic tissue.

Human cardiac fibrotic tissues are characterized by a higher stiffness relative to healthy cardiac tissues. These tissues are unable to spontaneously contract and are subjected to passive mechanical stimulation during heart functionality. Moreover, scaffolds that can recapitulate the in vivo mechanical properties of the cardiac fibrotic tissues are lacking. Herein, this study aimed to design and fabricate mechanically stretchable bioartificial scaffolds with biomimetic composition and stiffness comparable to human cardiac fibrotic tissues. Poly(ε-caprolactone) (PCL) scaffolds with a stretchable mesh architecture were initially designed through structural and finite element method (FEM) analyses and subsequently fabricated by melt extrusion additive manufacturing (MEX). Scaffolds were surface functionalized by 3,4-dihydroxy-DL-phenylalanine (DOPA) polymerization (polyDOPA) to improve their interaction with natural polymers. Scaffold pores were then filled with different concentrations (5%, 7%, and 10% w/v) of gelatin methacryloyl (GelMA) hydrogels to support in vitro human cardiac fibroblasts (HCFs) 3D culture, thereby producing bioartificial PCL/GelMA scaffolds. Uniaxial tensile mechanical tests in static and dynamic conditions (1 Hz; 22% maximum strain) demonstrated that the bioartificial scaffolds had in vivo-like stretchability and similar stiffness to that of pathological cardiac tissue (tailored as a function of the number of PCL scaffold layers and GelMA hydrogel concentration). In vitro cell tests on bioartificial scaffolds using HCF-embedded GelMA hydrogels under static conditions displayed increasing cell viability, spreading, and cytoskeleton organization with decreasing GelMA hydrogel concentration. Moreover, α-smooth muscle actin (α-SMA)-positive cells were detected after 7 days of culture in static conditions followed by another 7 days of culture in dynamic conditions and not in HCF-loaded scaffolds cultured in static conditions for 14 days. These results suggested that in vitro culture under cyclic mechanical stimulations could induce an HCF phenotypic switch into myofibroblasts.

Scaffold providing spatial guidance enhances the healing of osteochondral defects and reduce adverse bone-cartilage crosstalk

Despite previous attempts using multiphasic scaffolds to organize cells and bioactive elements, osteochondral repair still faces challenges, such as thin cartilage layers and uneven bone-cartilage interfaces. These issues are likely exacerbated by detrimental biochemical interactions and the formation of new blood vessels within the osteochondral unit, impacting tissue repair quality. In this study, we devised a novel freeze-welding technique to create a unified porous scaffold. Chitosan was chosen as the primary material due to its structural and compositional similarity to glycosaminoglycans in the ECM. In particular, by meticulously adjusting freezing and solute conditions, we engineered the scaffold to have an axially aligned porous structure in the cartilage layer and a radially aligned porous structure in the subchondral layer (scaffold A/R), each designed to meet the unique repair needs of the respective tissues. This scaffold showed enhanced mechanical compressibility, shape retention, and interfacial bonding capabilities. The results demonstrated that this unique spatial scaffold structure offers significant advantages over purely axial scaffolds in guiding osteochondral regeneration. These advantages were evident in its ability to independently promote cartilage and bone growth, reduce undesirable bone-cartilage crosstalk, particularly in the context of vessel invasion in the cartilage layer, and optimize the remodeling of the osteochondral interface. This study offers valuable insights into the structural design patterns for osteochondral regeneration, highlighting the potential importance of structural scaffold design in guiding endogenous tissue remodeling during heterogeneous tissue regeneration.

Regulating ionic transport and interface chemistry via high-dielectric BaTiO3 porous scaffolds for aqueous Zn-ion batteries

Rechargeable aqueous Zn-ion batteries (AZIBs) are gaining considerable attention as alternatives to Li-ion batteries owing to their excellent safety, high energy density, low cost, and environmental friendliness. However, their Zn anodes suffer from severe dendritic growth, the hydrogen evolution reaction (HER), and corrosion, which limit the commercialization of AZIBs. To address this challenge, we designed a solution involving the development of a high-dielectric hydrophobic BaTiO3 (BTO) porous scaffold layer that is strategically applied to the Zn surface. This BTO layer effectively impedes the dendritic growth of Zn. The distinct polarizability of the BTO layer actively diminishes the electric-field gradient, thereby minimizing the interaction between the H2O molecules and the Zn surface. This phenomenon helps suppress dendritic growth, thus ensuring the uniform deposition of Zn and preventing the HER. The electrochemical performance of a symmetric cell containing the Zn@BTO electrode exhibits low-voltage polarization and stable charge–discharge performance for up to 500 h. In addition, a full-cell AZIB containing a high-capacity α-MnO2 cathode and Zn@BTO anode exhibits good rate capability and a long life of up to 500 cycles. This study demonstrates the development of a cost-effective dendrite-free Zn anode integrated with a highly dielectric hydrophobic BTO porous scaffold layer as a stable Zn–electrolyte interface layer for high-performance AZIBs.

Stimulation of Bone ingrowth using YSZ/ β-TCP/PCL composite bilayer coating on 316 LSS scaffold for orthopedic application

The demand towards ideal tissue construct using polymer and ceramics made the researchers to do the search of polymer composites with desirable properties. Material selection for the composite play a major role to get long life. The selection of materials can be based on the nature, location and tissue type, which should be regenerated. Normally, a bone construct has its own requirements like osteo conduction properties, Tendency to mineralize, Rate of degradation and osteogenic differentiation properties. The best approach is to create regenerative medicine using suitable devices. This work focuses on developing the scaffold using 316L stainless steel fabricated with dual coating. The first layer of coating was made using Yttria Stabilized Zirconia by electrophoretic deposition. While the top second layer made as electro spun over 316L stainless steel . The electro spun made with β-Tricalcium phosphate (β-TCP) incorporated in PCL (Polycaprolactone). The process of electrospinning preferred to achieve porous surface. The bilayer scaffold performance was evaluated by surface morphology, bioactivity, bioresorbable and biodegradability with its cell adhesion capacity were studied. The results of X-ray diffraction reveal the presence of nanoparticle on the top layer of 316L stainless steel scaffold. The surface topography studies using Field Emission Scanning Electron Microscopy (FESEM) confirms more than 70 % porosity. The functional group present in YSZ/β-TCP/PCL composite bilayer on 316L SS were determined using FTIR. The developed bilayer coated 316L Stainless steel scaffolds further evaluated by invitro studies to confirm the cell adhesion capability. MG 63 cell line isolated from the bone and possess fibroblast morphology was used for the assay. The cells were cultured and exposed to the developed scaffold, the cultured cells were exposed to MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) and the reduction of MTT directly measures the cell viability and the color change read through spectrophotometer. The cell growth and proliferation confirm its cell adhesion property. This present study helps in drug delivery using the nano fibers incorporated with bioresorbable TCP. The presence of TCP on the top of electro spun layer substantially supports the osteo conductivity through mineralization and degradation property.

Multifunctional structural color Chinese herb hydrogel patches for wound management

Chinese herb hydrogel has outstanding values in wound treatment. Tremendous effort is dedicated to developing functional and intelligent Chinese herb hydrogels for accelerating healing process. Herein, a novel multifunctional bilayer Chinese herb hydrogel patch for wound healing is presented. The patches are composed of a Ginsenoside compound K (GCK)-integrated chitosan inverse opal scaffold layer and a Bletilla striata polysaccharide (BP) hydrogel filler layer. As the photothermal conversion capacity of BP can trigger faster degradation of its hydrogel layer under near-infrared (NIR) light, the encapsulated BP can be controllably released to wound area and exhibit photothermal antibacterial and antioxidant effects. Besides, the inverse opal scaffold layer shows great mechanical property and moisturizing performance, and its released GCK can promote angiogenesis for long term therapeutic effects. Attractively, due to responsive structural color, the patch is endowed with self-reporting performance, which can real-time visually monitor the drug release degree. Taking advantages of these properties, the patches have been demonstrated to achieve satisfying therapeutic effects for in vivo wound treatment. Therefore, we believe that the proposed Chinese herb hydrogel patches are valuable as multifunctional patches for clinical treatment.

Enhanced efficiency and stability of inorganic CsPbBr3 perovskite solar cells based on carbon counter electrode by applying PbTiO3-coated TiO2 nanorod films as scaffold layers

The CsPbBr3 perovskite solar cells (PSCs) based on carbon counter electrode (CCE) are promising because of the advantages including fabrication simplicity and excellent stability, but their power conversion efficiencies (PCE) are low due to interfacial carrier recombination as the direct contact of the CsPbBr3 with electron transport materials (ETM). In this paper, PbTiO3 shells were grown onto TiO2 nanorods to form PbTiO3-coated TiO2 scaffold layer via a two-step method. The presence of PbTiO3 could promote crystallinity of the CsPbBr3 perovskite film and inhibits interface recombination at the CsPbBr3/TiO2 interface. Consequently, the carbon-based CsPbBr3 solar cells with PbTiO3/TiO2 scaffold layer achieves a high PCE of 7.28%, demonstrating an increase by 19.9% compared with the devices based on TiO2 NRs. Moreover, the champion devices exhibited excellent stability in ambient air, with the PCE value remaining at 93% over 28 days.

Janus silk fibroin/polycaprolactone-based scaffold with directionally aligned fibers and porous structure for bone regeneration

To promote bone repair, it is desirable to develop three-dimensional multifunctional fiber scaffolds. The densely stacked and tightly arranged conventional two-dimensional electrospun fibers hinder cell penetration into the scaffold. Most of the existing three-dimensional structural materials are isotropic and monofunctional. In this research, a Janus nanofibrous scaffold based on silk fibroin/polycaprolactone (SF/PCL) was fabricated. SF-encapsulated SeNPs demonstrated stability and resistance to aggregation. The outside layer (SF/PCL/Se) of the Janus nanofiber scaffold displayed a structured arrangement of fibers, facilitating cell growth guidance and impeding cell invasion. The inside layer (SF/PCL/HA) featured a porous structure fostering cell adhesion. The Janus fiber scaffold containing SeNPs notably suppressed S. aureus and E. coli activities, correlating with SeNPs concentration. In vitro, findings indicated considerable enhancement in alkaline phosphatase (ALP) activity of MC3T3-E1 osteoblasts and upregulation of genes linked to osteogenic differentiation with exposure to the SF/PCL/HA/Se Janus nanofibrous scaffold. Moreover, in vivo, experiments demonstrated successful critical bone defect repair in mouse skulls using the SF/PCL/HA/Se Janus nanofiber scaffold. These findings highlight the potential of the SF/PCL-based Janus nanofibrous scaffold, integrating SeNPs and nHA, as a promising biomaterial in bone tissue engineering.

In vitro assessment of Momordica charantia/Hypericum perforatum oils loaded PCL/Collagen fibers: Novel scaffold for tissue engineering.

The research on tissue engineering applications has been progressing to manufacture ideal tissue scaffold biomaterials. In this study, a double-layered electrospun biofiber scaffold biomaterial including Polycaprolactone (PCL)/Collagen (COL) fibrous inner layer and PCL/ Momordica charantia (MC) and Hypericum perforatum (HP) oils fibrous outer layer was developed to manufacture a functional, novel tissue scaffold with the advantageous mechanical and biological properties. The main approach was to combine the natural perspective using medicinal oils with an engineering point of view to fabricate a potential functional scaffold for tissue engineering. Medicinal plants MC and HP are rich in functional oils and incorporation of them in a tissue scaffold will unveil their potential to augment both new tissue formation and wound healing. In this study, a novel double-layered scaffold prototype was fabricated using electrospinning technique with two PCL fiber layers, first is composed of collagen, and second is composed of oils extracted from medicinal plants. Initially, the composition of plant oils was analyzed. Thereafter the biofiber scaffold layers were fabricated and were evaluated in terms of morphology, physicochemistry, thermal and mechanical features, wettability, in vitro bio-degradability. Double-layered scaffold prototype was further analyzed in terms of in vitro biocompatibility and antibacterial effect. The medicinal oils blend provided antioxidant and antibacterial properties to the novel PCL/Oils layer. The results signify that inner PCL/COL layer exhibited advanced biodegradability of 8.5% compared to PCL and enhanced wettability with 11.7° contact angle. Strength of scaffold prototype was 5.98 N/mm2 thanks to the elastic PCL fibrous matrix. The double-layered functional biofiber scaffold enabled 92% viability after 72 h contact with fibroblast cells and furthermore provided feasible attachment sites for the cells. The functional scaffold prototype's noteworthy mechanical, chemical, and biological features enable it to be suggested as a different novel biomaterial with the potential to be utilized in tissue engineering applications.

Piezoelectric ceramics with hierarchical macro- and micro-pore channels for sensing applications

Piezoelectric sensors have attracted increasing attention due to their high sensitivity to forces, pressures and acceleration. The use of porous piezoelectric ceramics with aligned pores formed by freeze casting for sensing application also exhibit advantages due to their combination of a high piezoelectric charge coefficient and low permittivity, which leads to an enhancement in sensor sensitivity compared to dense materials. To fabricate porous materials, the process of direct ink writing (DIW) exhibits superiority in terms of structural design as a result of its ability to provide layer-by-layer and mold-free rapid prototyping. This work provides the first demonstration of the fabrication of lead zirconate titanate (PZT) piezoelectric ceramic scaffolds with sophisticated hierarchical porous structures by simultaneous DIW and freeze casting. The morphology of the porous cells and individual layers of the hierarchical porous PZT ceramic scaffolds were designed by adjusting the rheological properties and optimizing the printing path. The effect of cell structure and number of layers on sensing performance was investigated in detail. Manufactured scaffolds with a sandwich structure exhibited a high output voltage (191 V) under the application of 1 N external force, with a sensitivity of 8.98 V·kPa−1. This work provides a new approach to structural design of piezoelectric sensors to inform future efforts in the fabrication of high-performance piezoelectric ceramics with hierarchical porosity with complex and bespoke geometries.

Investigation and optimization of In-Vitro behaviour of Perovskite barium titanate as a scaffold and protective coatings

The piezoelectric effect is widely known to have a significant physiological function in bone development, remodeling, and fracture repair. As a well-known piezoelectric material, barium titanate is particularly appealing as a scaffold layer to improve bone tissue engineering applications. Currently, the chemical bath deposition method is used to prepare green synthesized barium titanate coatings to improve mechanical and biological characteristics. Molarity of the solutions, an essential parameter in chemical synthesis, is changed at room temperature (0.1–1.2 Molar) to prepare coatings. The XRD spectra for as deposited coatings indicate amorphous behavior, while polycrystalline nature of coatings is observed after annealing (300 °C). Coatings prepared with solutions of relatively low molarities, i.e. from 0.1 to 0.8 M, exhibit mixed tetragonal – cubic phases. However, the tetragonal phase of Perovskite barium titanate is observed using solution molarities of 1.0 M and 1.2 M. Relatively high value of transmission, i.e. ∼80%, is observed for the coatings prepared with high molarities. Band gap of annealed coatings varies between 3.47 and 3.70 eV. For 1.2 M sample, the maximum spontaneous polarization (Ps) is 0.327x10−3 (μC/cm2) and the residual polarization (Pr) is 0.072x10−3 (μC/cm2). For 1.2M solution, a high hardness value (1510 HV) is recorded, with a fracture toughness of 28.80 MPam−1/2. Low values of weight loss, after dipping the coatings in simulated body fluid, is observed. The antibacterial activity of BaTiO3 is tested against E. coli and Bacillus subtilis. Drug encapsulation capability is also tested for different time intervals. As a result, CBD-based coatings are a promising nominee for use as scaffold and protective coatings.

3D-printed biomimetic scaffolds with precisely controlled and tunable structures guide cell migration and promote regeneration of osteochondral defect

Untreated osteochondral defects will develop into osteoarthritis, affecting patients’ quality of life. Since articular cartilage and subchondral bone exhibit distinct biological characteristics, repairing osteochondral defects remains a major challenge. Previous studies have tried to fabricate multilayer scaffolds with traditional methods or 3D printing technology. However, the efficacy is unsatisfactory because of poor control over internal structures or a lack of integrity between adjacent layers, severely compromising repair outcomes. Therefore, there is a need for a biomimetic scaffold that can simultaneously boost osteochondral defect regeneration in both structure and function. Herein, an integrated bilayer scaffold with precisely controlled structures is successfully 3D-printed in one step via digital light processing (DLP) technology. The upper layer has both ‘lotus- and radial-’ distribution pores, and the bottom layer has ‘lotus-’ pores to guide and facilitate the migration of chondrocytes and bone marrow mesenchymal stem cells, respectively, to the defect area. Tuning pore sizes could modulate the mechanical properties of scaffolds easily. Results show that 3D-printed porous structures allow significantly more cells to infiltrate into the area of ‘lotus- and radial-’ distribution pores during cell migration assay, subcutaneous implantation, and in situ transplantation, which are essential for osteochondral repair. Transplantation of this 3D-printed bilayer scaffold exhibits a promising osteochondral repair effect in rabbits. Incorporation of Kartogenin into the upper layer of scaffolds further induces better cartilage formation. Combining small molecules/drugs and precisely size-controlled and layer-specific porous structure via DLP technology, this 3D-printed bilayer scaffold is expected to be a potential strategy for osteochondral regeneration.

Multifunctional hydrogel/platelet-rich fibrin/nanofibers scaffolds with cell barrier and osteogenesis for guided tissue regeneration/guided bone regeneration applications

Periodontal defect seriously affects people's life health and quality. Guided tissue regeneration (GTR) and guided bone regeneration (GBR) have made great progress in periodontal disease treatment, but some deficiencies existed in commercial materials of GTR and GBR. For obtaining better therapeutic effects, multifunctional composite scaffolds containing different biological macromolecules were developed in this study. Chitosan/poly (γ-glutamic acid)/nano-hydroxyapatite hydrogels (CP/nHA) made by electrostatic interactions and lyophilization were filled in the bone defects to achieve osteogenesis. Platelet-rich fibrin (PRF) extracted from blood could accelerate bone formation by releasing various bioactive substances as middle layer of composite scaffolds. Polycaprolactone/gelatin nanofibers (PG) prepared by electrospinning were attached to the junction of soft and hard tissue, which could prevent fibrous tissue from infiltrating into bone defects. The composite scaffolds showed good morphology, biocompatibility, cell barriers and osteogenic differentiation in vitro. The excellent ability of bone formation was verified by implantation of triple-layered composite scaffolds into alveolar bone defects in rabbit in vivo. The hierarchical structure was conducive to personalized customization to meet the needs of different defects. All in all, the multifunctional scaffolds could play important roles of GTR and GBR in alveolar bone regeneration and provide good application prospect for bone repair in clinic.

Layered PCL scaffolds modified with bioactive additives fabricated by electrospinning and 3D-printing for the nasal bone and cartilage defects

This paper presents a method for fabricating layered materials supporting the reconstruction of nasal tissues by combining the 3D/fused deposition modeling (FDM) technology with electrospinning (ES). Polycaprolactone (PCL) scaffolds modified with bioglass (BG) and zinc (Zn)-doped BG were printed from the connectable filaments sticks. Then, using the ES, a nanofibrous membrane with particles of the pharmaceutical drug Osteogenon (OST), was applied directly on a spatial scaffold. The layered scaffolds were investigated by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and micro-computed tomography (µCT). The bioactivity and biocompatibility of the scaffolds were evaluated by in vitro studies. The results indicated that the presence of the OST drug in the top layer of scaffolds promoted bioactivity. The presence of a membrane had a positive effect on the production and secretion of aggrecan (Agg), and incorporation of BG particles significantly improved the production and secretion of collagen type II (Col2). Each of the introduced PCL modifications leads to articular cartilage cell (NHAC-kn) viability improvement and results in a decrease in the cytotoxicity of the material. Moreover, in the long-term culture, cell proliferation was positively affected by the introduction of the OST-modified membrane and also by the addition of Zn-doped BG particles.

3D bioprinting of a gelatin-alginate hydrogel for tissue-engineered hair follicle regeneration.

Hair follicle (HF) regeneration remains challenging, principally due to the absence of a platform that can successfully generate the microenvironmental cues of hair neogenesis. Here, we demonstrate a 3D bioprinting technique based on a gelatin/alginate hydrogel (GAH) to construct a multilayer composite scaffold simulating the HF microenvironment in vivo. Fibroblasts (FBs), human umbilical vein endothelial cells (HUVECs), dermal papilla cells (DPCs), and epidermal cells (EPCs) were encapsulated in GAH (prepared from a mixture of gelatin and alginate) and respectively 3D-bioprinted into the different layers of a composite scaffold. The bioprinted scaffold with epidermis- and dermis-like structure was subsequently transplanted into full-thickness wounds in nude mice. The multilayer scaffold demonstrated suitable cytocompatibility and increased the proliferation ability of DPCs (1.2-fold; P<0.05). It also facilitated the formation of self-aggregating DPC spheroids and restored DPC genes associated with hair induction (ALP, β-catenin, and α-SMA). The dermal and epidermal cells self-assembled successfully into immature HFs in vitro. HFs were regenerated in the appropriate orientation in vivo, which can mainly be attributed to the hierarchical grid structure of the scaffold and the dot bioprinting of DPCs. Our 3D printed scaffolds provide a suitable microenvironment for DPCs to regenerate entire HFs and could make a significant contribution in the medical management of hair loss. This method may also have broader applications in skin tissue (and appendage) engineering. STATEMENT OF SIGNIFICANCE: Hair loss remains a challenging clinical problem that influences quality of life. Three-dimensional (3D) bioprinting has become a useful tool for the fabrication of tissue constructs for transplantation and other biomedical applications. In this study, we used a 3D bioprinting technique based on a gelatin/alginate hydrogel to construct a multi-layer composite scaffold with cuticular and corium layers to simulate the microenvironment of dermal papilla cells (DPCs) in the human body. This new approach permits the controllable formation of self-aggregating spheroids of DPCs in a physiologically relevant extracellular matrix and the initiation of epidermal-mesenchymal interactions, which results in HF formation in vivo. The ability to regenerate entire HFs should have a significant impact on the medical management of hair loss.

Crystal structure and induced stability of trimeric BxpB: implications for the assembly of BxpB-BclA complexes in the exosporium of Bacillus anthracis.

The outermost exosporium layer of Bacillus anthracis spores, the causative agents of anthrax, is comprised of a basal layer and an external hair-like nap. The nap includes filaments composed of trimers of the collagen-like glycoprotein BclA. Essentially all BclA trimers are attached to the spore in a process in which part of the 38-residue amino-terminal domain (NTD) of BclA forms an extremely stable interaction with the basal layer protein BxpB. Evidence indicates that the BclA-BxpB interaction is direct and requires trimeric BxpB. To further investigate the nature of the BclA-BxpB interaction, we determined the crystal structure of BxpB. The structure was trimeric with each monomer consisting of 11 β strands with connecting loops. The structure did not include apparently disordered amino acids 1-19, which contain the only two cysteine residues of the 167-residue BxpB. The orientation of the structure reveals regions of BxpB that could be involved in interacting with the BclA NTD and with adjacent cysteine-rich proteins in the basal layer. Furthermore, the BxpB structure closely resembles that of the 134-residue carboxyl-terminal domain of BclA, which forms trimers that are highly resistant to heat and detergent. We demonstrated that BxpB trimers do not share this resistance. However, when BxpB trimers are mixed with a peptide containing residues 20-38 of BclA, they form a complex that is as stable as BclA-BxpB complexes extracted from spores. Together, our results provide new insights into the mechanism of BclA-BxpB attachment and incorporation into the exosporium. IMPORTANCE The B. anthracis exosporium plays major roles in spore survival and infectivity, but the complex mechanism of its assembly is poorly understood. Key steps in this process are the stable attachment of collagen-like BclA filaments to the major basal layer structural protein BxpB and the insertion of BxpB into an underlying basal layer scaffold. The goal of this study is to further elucidate these interactions thereby advancing our understanding of exosporium assembly, a process shared by many spore-forming bacteria including important human pathogens.