Bioactive Nanofibrous Scaffolds Research Articles

Bioactive Nanofibrous Scaffolds Incorporating Decellularized Cell-Derived Extracellular Matrix for Periodontal Tissue Engineering

Bioactive Nanofibrous Scaffolds Incorporating Decellularized Cell-Derived Extracellular Matrix for Periodontal Tissue Engineering

Cell loaded hydrogel containing Ag-doped bioactive glass-ceramic nanoparticles as skin substitute: Antibacterial properties, immune response, and scarless cutaneous wound regeneration.

An ideal tissue‐engineered dermal substitute should possess angiogenesis potential to promote wound healing, antibacterial activity to relieve the bacterial burden on skin, as well as sufficient porosity for air and moisture exchange. In light of this, a glass–ceramic (GC) has been incorporated into chitosan and gelatin electrospun nanofibers (240–360 nm), which MEFs were loaded on it for healing acceleration. The GC was doped with silver to improve the antibacterial activity. The bioactive nanofibrous scaffolds demonstrated antibacterial and superior antibiofilm activities against Gram‐negative and Gram‐positive bacteria. The nanofibrous scaffolds were biocompatible, hemocompatible, and promoted cell attachment and proliferation. Nanofibrous skin substitutes with or without Ag‐doped GC nanoparticles did not induce an inflammatory response and attenuated LPS‐induced interleukin‐6 release by dendritic cells. The rate of biodegradation of the nanocomposite was similar to the rate of skin regeneration under in vivo conditions. Histopathological evaluation of full‐thickness excisional wounds in BALB/c mice treated with mouse embryonic fibroblasts‐loaded nanofibrous scaffolds showed enhanced angiogenesis, and collagen synthesis as well as regeneration of the sebaceous glands and hair follicles in vivo.

Co-Electrospun Silk Fibroin and Gelatin Methacryloyl Sheet Seeded with Mesenchymal Stem Cells for Tendon Regeneration.

Silk fibroin (SF) is a promising biomaterial for tendon repair, but its relatively rigid mechanical properties and low cell affinity have limited its application in regenerative medicine. Meanwhile, gelatin-based polymers have advantages in cell attachment and tissue remodeling but have insufficient mechanical strength to regenerate tough tissue such as tendons. Taking these aspects into account, in this study, gelatin methacryloyl (GelMA) is combined with SF to create a mechanically strong and bioactive nanofibrous scaffold (SG). The mechanical properties of SG nanofibers can be flexibly modulated by varying the ratio of SF and GelMA. Compared to SF nanofibers, mesenchymal stem cells (MSCs) seeded on SG fibers with optimal composition (SG7) exhibit enhanced growth, proliferation, vascular endothelial growth factor production, and tenogenic gene expression behavior. Conditioned media from MSCs cultured on SG7 scaffolds can greatly promote the migration and proliferation of tenocytes. Histological analysis and tenogenesis-related immunofluorescence staining indicate SG7 scaffolds demonstrate enhanced in vivo tendon tissue regeneration compared to other groups. Therefore, rational combinations of SF and GelMA hybrid nanofibers may help to improve therapeutic outcomes and address the challenges of tissue-engineered scaffolds for tendon regeneration.

Spindle-Like Zinc Silicate Nanoparticles Accelerating Innervated and Vascularized Skin Burn Wound Healing.

The treatment of severe burn injuries is a crucial challenge in skin tissue engineering. Severe burns are always accompanied with large-area neurovascular networks damage, leading to the lack of excitation functions and difficulty in self-healing. Therefore, it is of great importance to develop biomaterials which can not only promote wound healing but also simultaneously reconstruct cutaneous neurovascular networks. In this study, Zn2 SiO4 (ZS) nanoparticles-incorporated bioactive nanofibrous scaffolds are designed for innervated and vascularized skin burn wound healing. ZS nanoparticles with spindle-like morphology are synthesized via a facile hydrothermal method. The incorporation of ZS nanoparticles endows the scaffolds with excellent angiogenic and neurogenic activities in vitro. Additionally, in vivo results show that the ZS nanoparticles-incorporated scaffolds have favorable re-epithelialization, innervation, and vascularization abilities through local release of bioactive Zn and Si ions from ZS nanoparticles, leading to rapid wound healing featuring with newly formed blood vessels and nerve fibers. Taken together, this study suggests that the spindle-like ZS nanoparticles are useful bioactive agents for stimulating vascularization and innervation of functional skin repair. The bioactive inorganic nanoparticles may be used for multifunctional tissue regeneration.

Three-dimensional endothelial cell incorporation within bioactive nanofibrous scaffolds through concurrent emulsion electrospinning and coaxial cell electrospraying

Nanofibrous scaffolds hold great promise in tissue engineering owing to their extracellular matrix (ECM)-mimicking architectures. Electrospinning, with its ease for producing nanofibrous scaffolds, has therefore been widely employed for various tissue engineering applications. However, electrospun nanofibrous scaffolds have faced the inherent challenge of three-dimensional (3D) cell distribution due to the small sizes of interconnected pores in these scaffolds when conventional approach of scaffold fabrication with subsequent cell seeding is adopted, which severely limits their applications in repairing/regenerating human body tissues with thick and vascularized structures. In this study, we demonstrate a method to directly place living endothelial cells within bioactive nanofibrous scaffolds in 3D through concurrent emulsion electrospinning and coaxial cell electrospraying. Using this concurrent manufacturing method, endothelial cells are encapsulated in hydrogel microspheres and deposited along with vascular endothelial growth factor (VEGF)-containing nanofibers in the scaffold fabrication process, resulting in nanofibrous scaffolds with 3D embedded cell-encapsulated microspheres. After selective disruption of the hydrogel microspheres, the encapsulated endothelial cells are released, yielding bioactive nanofibrous scaffolds with tissue-like 3D cell-incorporated nanofibrous structures. It is shown that cell viability is well preserved (>98%) during the concurrent manufacturing process and that a deep cell distribution (~100 μm) through the scaffold thickness has been achieved. With combined structural and biochemical cues via the 3D cell-incorporated architectures, endothelial cells can freely stretch, display enhanced intercellular connections, and maintain the phenotype in the bioactive nanofibrous scaffolds. Our investigations offer a promising platform technology for creating bioactive nanofibrous scaffolds with 3D cell incorporation and for overcoming inherent problems of electrospun nanofibrous scaffolds, which should open new avenues for biomanufacturing tissue-mimicking constructs with vascularized structures and complex anatomy. Statement of significanceElectrospun nanofibrous scaffolds face challenges in three-dimensional (3D) cell incorporation and vascularization. Enhancing cell penetration via enlarged interconnected pores is a common strategy to address that. However, there are conflicts between cell penetration and structural integrity for scaffolds formed using such strategy, as deep cell penetration, if possible, can only achieve in highly loose architectures. In this investigation, we demonstrate a concurrent emulsion electrospinning and coaxial cell electrospraying technique, realizing 3D endothelial cell incorporation in electrospun nanofibrous scaffolds independent of cell penetration. Our technology appropriately addresses the conflict between deep 3D cell incorporation and structural integrity. In the scaffolds, the 3D incorporated endothelial cells show well-preserved viability, phenotype and functions, implying improved vascularization potential.

Biomimetic Elastomeric Bioactive Siloxane‐Based Hybrid Nanofibrous Scaffolds with miRNA Activation: A Joint Physico‐Chemical‐Biological Strategy for Promoting Bone Regeneration

AbstractRapid and efficient disease‐induced or critical‐size bone regeneration remains a challenge in tissue engineering due to the lack of highly bioactive biomaterial scaffolds. Physical structures such as nanostructures, chemical components such as silicon elements, and biological factors such as genes have shown positive effects on bone regeneration. Herein, a bioactive photoluminescent elastomeric silicate‐based nanofibrous scaffold with sustained miRNA release is reported for promoting bone regeneration based on a joint physico‐chemical‐biological strategy. Bioactive nanofibrous scaffolds are fabricated by cospinning poly (ε‐caprolactone) (PCL), elastomeric poly (citrates‐siloxane) (PCS), and bioactive osteogenic miRNA nanocomplexes (denoted PPM nanofibrous scaffolds). The PPM scaffolds possess uniform nanostructures, significantly enhanced tensile stress (≈15 MPa) and modulus (≈32 MPa), improved hydrophilicity (30–60°), controlled biodegradation, and strong blue fluorescence. Bioactive miRNA complexes are efficiently loaded into the nanofibrous matrix and exhibit long‐term release for up to 70 h. The PPM scaffolds significantly promote the adhesion, proliferation, and osteoblast differentiation of bone marrow stem cells in vitro and enhanced rat cranial defect restoration (12 weeks) in vivo. This work reports an attractive joint physico‐chemical‐biological strategy for the design of novel cell/protein‐free bioactive scaffolds for synergistic tissue regeneration.

Highly efficient local delivery of endothelial progenitor cells significantly potentiates angiogenesis and full-thickness wound healing

Wound therapy with a rapid healing performance remains a critical clinical challenge. Cellular delivery is considered to be a promising approach to improve the efficiency of healing, yet problems such as compromised cell viability and functionality arise due to the inefficient delivery. Here, we report the efficient delivery of endothelial progenitor cells (EPCs) with a bioactive nanofibrous scaffold (composed of collagen and polycaprolactone and bioactive glass nanoparticles, CPB) for enhancing wound healing. Under the stimulation of CPB nanofibrous system, the viability and angiogenic ability of EPCs were significantly enhanced through the activation of Hif-1α/VEGF/SDF-1α signaling. In vivo, CPB/EPC constructs significantly enhanced the formation of high-density blood vessels by greatly upregulating the expressions of Hif-1α, VEGF, and SDF-1α. Moreover, owing to the increased local delivery of cells and fast neovascularization within the wound site, cell proliferative activity, granulation tissue formation, and collagen synthesis and deposition were greatly promoted by CPB/EPC constructs resulting in rapid re-epithelialization and regeneration of skin appendages. As a result, the synergistic enhancement of wound healing was observed from CPB/EPC constructs, which suggests the highly efficient delivery of EPCs. CPB/EPC constructs may become highly competitive cell-based therapeutic products for efficient impaired wound healing application. This study may also provide a novel strategy to develop bioactive cell therapy constructs for angiogenesis-related regenerative medicine. Statement of SignificanceThis paper reported a highly efficient local delivery of EPCs using bioactive glass-based CPB nanofibrous scaffold for enhancing angiogenesis and wound regeneration. In vitro study showed that CPB can promote the proliferation, migration, and tube formation of EPCs through upregulation of the Hif-1α/VEGF/SDF-1α signaling pathway, indicating that the bioactivity and angiogenic ability of EPCs can be highly maintained and promoted by the CPB scaffold. Moreover, CPB/EPC constructs effectively stimulated the regeneration of diabetic wounds with satisfactory vascularization and better healing outcomes in a full-thickness wound model, suggesting that the highly efficient delivery of EPCs to wound site facilitates angiogenesis and further leads to wound healing. The high angiogenic capacity and excellent healing ability make CPB/EPC constructs highly competitive in cell-based therapeutic products for efficient wound repair application.

A Biomimetic Emu Oil-Blended Electrospun Nanofibrous Mat for Maintaining Stemness of Adipose Tissue-Derived Stem Cells.

Emu oil (EO) with anti-inflammatory, antioxidative, and wound healing properties can be blended for preparing bioactive nanofibrous scaffold. Adipose tissue-derived stem cells (ADSCs) are promising candidates for tissue engineering, and preserving their stemness potential is vital for further therapeutic applications. The aim of this study was to fabricate EO-blended nanofiber and investigate its effect on proliferation, survival, and stemness preservation of ADSCs. Pure EO composition was characterized using a gas chromatograph mass spectrometer. EO-PCL-polyethylene glycol (PEG) nanofibers were successfully fabricated using an electrospinning technique and characterized by field emission scanning electron microscopy (FE-SEM) and fourier-transform infrared spectroscopy (FTIR). Cell viability and adhesion were measured using the MTT assay and FE-SEM. Finally, quantitative PCR (qPCR) was used to quantify the expression level of cell cycle regulated genes and pluripotency-associated transcription factors. Findings showed that 20% (w/w) of EO is the optimum oil content in the electrospun solution to achieve good morphology and ultrafine fibers. The relatively high optical densities and FE-SEM images indicated that EO highly supported cell adhesion and proliferation on the matrices. In addition, EO-PCL-PEG electrospun nanofibrous mats significantly upregulated the expression levels of cell cycle regulated genes (Cyclin D1, pRb, and P53) and stemness markers (Nanog, OCT-4, Rex-1, and Sox-2) than PCL-PEG nanofiber and tissue culture polystyrene in 7 and 14 days of cell culture. These results demonstrate that the EO-blended nanofibrous mat can be used as a bioactive scaffold to support cell adhesion and proliferation while simultaneously maintaining the stemness of ADSCs.

Nanofibre Based Smart Pharmaceutical Scaffolds for Wound Repair and Regenerations

Chronic wounds and ulcers are posing a devastating manifestation on the socioeconomic status across the globe along with the patient compliance. It reinforces a need for the development of successful alternative treatments for the chronic wound care and ulcer management practices. This review explores the progressive developments being made in the fabrication of electrospun nanofibrous scaffolds towards elimination of microbial infection from chronic wounds to accelerate the wound healing process. Functional three dimensional nanofibrous scaffolds produced by electrospinning have great potential in a wide spectrum of biomedical practices, such as tissue engineering, drug/gene delivery and wound dressing. Moreover, this review also highlights the materials and post modification methods, such as the functionaliation of electrospun nanofibrous scaffolds using growth factors, so that such smart and bioactive nanofibrous scaffolds could be made suitable for wound healing applications.

Tissue-engineering-based Strategies for Regenerative Endodontics

Stemming from in vitro and in vivo pre-clinical and human models, tissue-engineering-based strategies continue to demonstrate great potential for the regeneration of the pulp-dentin complex, particularly in necrotic, immature permanent teeth. Nanofibrous scaffolds, which closely resemble the native extracellular matrix, have been successfully synthesized by various techniques, including but not limited to electrospinning. A common goal in scaffold synthesis has been the notion of promoting cell guidance through the careful design and use of a collection of biochemical and physical cues capable of governing and stimulating specific events at the cellular and tissue levels. The latest advances in processing technologies allow for the fabrication of scaffolds where selected bioactive molecules can be delivered locally, thus increasing the possibilities for clinical success. Though electrospun scaffolds have not yet been tested in vivo in either human or animal pulpless models in immature permanent teeth, recent studies have highlighted their regenerative potential both from an in vitro and in vivo (i.e., subcutaneous model) standpoint. Possible applications for these bioactive scaffolds continue to evolve, with significant prospects related to the regeneration of both dentin and pulp tissue and, more recently, to root canal disinfection. Nonetheless, no single implantable scaffold can consistently guide the coordinated growth and development of the multiple tissue types involved in the functional regeneration of the pulp-dentin complex. The purpose of this review is to provide a comprehensive perspective on the latest discoveries related to the use of scaffolds and/or stem cells in regenerative endodontics. The authors focused this review on bioactive nanofibrous scaffolds, injectable scaffolds and stem cells, and pre-clinical findings using stem-cell-based strategies. These topics are discussed in detail in an attempt to provide future direction and to shed light on their potential translation to clinical settings.

Bioactive Nanofibrous Scaffolds for Regenerative Endodontics

Here we report the synthesis, materials characterization, antimicrobial capacity, and cytocompatibility of novel antibiotic-containing scaffolds. Metronidazole (MET) or Ciprofloxacin/(CIP) was mixed with a polydioxanone (PDS)polymer solution at 5 and 25 wt% and processed into fibers. PDS fibers served as a control. Scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), tensile testing, and high-performance liquid chromatography (HPLC) were used to assess fiber morphology, chemical structure, mechanical properties, and drug release, respectively. Antimicrobial properties were evaluated against those of Porphyromonas gingivalis/Pg and Enterococcus faecalis/Ef. Cytotoxicity was assessed in human dental pulp stem cells (hDPSCs). Statistics were performed, and significance was set at the 5% level. SEM imaging revealed a submicron fiber diameter. FTIR confirmed antibiotic incorporation. The tensile values of hydrated 25 wt% CIP scaffold were significantly lower than those of all other groups. Analysis of HPLC data confirmed gradual, sustained drug release from the scaffolds over 48 hrs. CIP-containing scaffolds significantly (p < .00001) inhibited biofilm growth of both bacteria. Conversely, MET-containing scaffolds inhibited only Pg growth. Agar diffusion confirmed the antimicrobial properties against specific bacteria for the antibiotic-containing scaffolds. Only the 25 wt% CIP-containing scaffolds were cytotoxic. Collectively, this study suggests that polymer-based antibiotic-containing electrospun scaffolds could function as a biologically safe antimicrobial drug delivery system for regenerative endodontics.

Carbon nanofibers produced from modified electrospun PAN/hydroxyapatite precursors as scaffolds for bone tissue engineering

In the current study we have proposed a method to obtain a carbon/HAp bioactive nanofibrous scaffold. The modified carbon nanofibrous nonwoven' fabrics were obtained by the use of electrospinning and subsequent stabilization and carbonization processes. The modified with HAp powder nanofibrous PAN nonwovens were thermally stabilized using a multi-stage process in the temperature ranging from 100°C to 300°C in an oxidative environment and then carbonized at 1000°C in argon atmosphere. The changes of properties of composite precursor membranes taking place during stabilization and carbonization processes were investigated using the methods of: DSC, TGA, FTIR, SEM, EDX, WAXD and mechanical tests. Bioactivity was determined by assessing the formation of crystalline apatite on the surface of membranes upon immersion in Simulated Body Fluid (SBF). The FTIR, SEM and WAXD investigation clearly prove that hydroxyapatite added to the electrospinning solution was present also in composites nanofibrous nonwovens after stabilization and carbonization process. It was found that due to HAp addition: the significant decrease of fibers average diameter occurs and that the average pore size for modified membranes is smaller than for the unmodified one. On the other hand it was shown that the ceramic additive protects fibers from mass reduction during the stabilization treatment. Finally a drastic increase of mineralization activity of nCF/HAp scaffolds as compared to their nCF counterparts has been proved.

P.146 PEEK implants for cranioplasty of complex cranial defects

In the current study we have proposed a method to obtain a carbon/HAp bioactive nanofibrous scaffold. The modified carbon nanofibrous nonwoven' fabrics were obtained by the use of electrospinning and subsequent stabilization and carbonization processes. The modified with HAp powder nanofibrous PAN nonwovens were thermally stabilized using a multi-stage process in the temperature ranging from 100 °C to 300 °C in an oxidative environment and then carbonized at 1000 °C in argon atmosphere. The changes of properties of composite precursor membranes taking place during stabilization and carbonization processes were investigated using the methods of: DSC, TGA, FTIR, SEM, EDX, WAXD and mechanical tests. Bioactivity was determined by assessing the formation of crystalline apatite on the surface of membranes upon immersion in Simulated Body Fluid (SBF). The FTIR, SEM and WAXD investigation clearly prove that hydroxyapatite added to the electrospinning solution was present also in composites nanofibrous nonwovens after stabilization and carbonization process. It was found that due to HAp addition: the significant decrease of fibers average diameter occurs and that the average pore size for modified membranes is smaller than for the unmodified one. On the other hand it was shown that the ceramic additive protects fibers from mass reduction during the stabilization treatment. Finally a drastic increase of mineralization activity of nCF/HAp scaffolds as compared to their nCF counterparts has been proved.

Bioactive Nanofibers: Synergistic Effects of Nanotopography and Chemical Signaling on Cell Guidance

Biodegradable nanofibers have tremendous potential for tissue repair. However, the combined effects of nanofiber organization and immobilized bioactive factors on cell guidance are not well understood. In this study, we developed aligned and bioactive nanofibrous scaffolds by immobilizing extracellular matrix protein and growth factor onto nanofibers, which simulated the physical and biochemical properties of native matrix fibrils. The aligned nanofibers significantly induced neurite outgrowth and enhanced skin cell migration during wound healing compared to randomly oriented nanofibers. Furthermore, the immobilized biochemical factors (as efficient as soluble factors) synergized with aligned nanofibers to promote highly efficient neurite outgrowth but had less effect on skin cell migration. This study shed light on the relative importance of nanotopography and chemical signaling in the guidance of different cell behavior.