Scaffolds For Delivery Research Articles

3D bioprinting of gene delivery scaffolds with controlled release

Localized gene delivery via engineered scaffolds offers spatiotemporal control of the gene vector release. Here, we explored the capability of digital light processing based bioprinting to fabricate 3D scaffolds in hydrogels for controlled gene delivery. We demonstrated the compatibility of the method with three representative hydrogel biomaterials for gene delivery. We further investigated the highly tunable release profile with these scaffolds by creating and combining distinct release mechanisms of diffusion and ion exchange. The efficacy of gene delivery of these scaffolds was validated in vitro using 293T cells. Results from this work could potentially facilitate the development of synergistic and personalized gene therapies.

Enhancing the properties of industrial waste nanocellulose bioaerogels using turmeric nano particles

In the present investigation, modified approach to enhance the properties of corrugated paper wastes nanocellulose (NC). The isolated NC was used with seafood wastes chitosan (CS) in the preparation of nano turmeric-loaded bioaerogel scaffolds. The results showed that the isolated NC had fibers length and diameters of 81 ± 12 nm and 7 ± 1.9 nm, respectively and crystallinity index of 77.9%, which indicates a maximum removal of impurity from the NC. CS had 64.3% crystallinity index with observable nonporous structures. Pure NC aerogel had the highest porosity, surface area and water absorption, but it showed very weak mechanical properties. Nano turmeric loaded bioaerogels showed enhanced mechanical properties and reduced water absorption capacity. Loaded bioaerogels exhibited higher antibacterial activity against S. aureus than E. coli with an inhibition zone of 32 mm and 27 mm respectively. Biocompatibility assay was conducted on murine fibroblast cells; the addition of nano turmeric significantly enhanced the proliferation of the cells compared with the control. These findings confirm the possibility and potential of smart conversion of wastes materials into advanced functional materials. Such bioaerogels can be of great interest in several biomedical applications such as wound healing, drug delivery and tissue scaffolding.

Engineering of a core-shell polyvinyl alcohol/gelatin fibrous scaffold for dual delivery of Thymus daenensis essential oil and Glycyrrhiza glabra L. extract as an antibacterial and functional wound dressing

Engineering of a core-shell polyvinyl alcohol/gelatin fibrous scaffold for dual delivery of Thymus daenensis essential oil and Glycyrrhiza glabra L. extract as an antibacterial and functional wound dressing

Periadventitial β-aminopropionitrile-loaded nanofibers reduce fibrosis and improve arteriovenous fistula remodeling in rats.

Arteriovenous fistula (AVF) postoperative stenosis is a persistent healthcare problem for hemodialysis patients. We have previously demonstrated that fibrotic remodeling contributes to AVF non-maturation and lysyl oxidase (LOX) is upregulated in failed AVFs compared to matured. Herein, we developed a nanofiber scaffold for the periadventitial delivery of β-aminopropionitrile (BAPN) to determine whether unidirectional periadventitial LOX inhibition is a suitable strategy to promote adaptive AVF remodeling in a rat model of AVF remodeling. Bilayer poly (lactic acid) ([PLA)-]- poly (lactic-co-glycolic acid) ([PLGA)] scaffolds were fabricated with using a two-step electrospinning process to confer directionality. BAPN-loaded and vehicle control scaffolds were wrapped around the venous limb of a rat femoral-epigastric AVF during surgery. AVF patency and lumen diameter were followed monitored using Doppler ultrasound surveillance and flow was measured before euthanasia. AVFs were harvested after 21 days for histomorphometry and immunohistochemistry. AVF compliance was measured using pressure myography. RNA from AVF veins was sequenced to analyze changes in gene expression due to LOX inhibition. Bilayer periadventitial nanofiber scaffolds extended BAPN release compared to the monolayer design (p < 0.005) and only released BAPN in one direction. Periadventitial LOX inhibition led to significant increases in AVF dilation and flow after 21 days. Histologically, BAPN trended toward increased lumen and significantly reduced fibrosis compared to control scaffolds (p < 0.01). Periadventitial BAPN reduced downregulated markers associated with myofibroblast differentiation including SMA, FSP-1, LOX, and TGF-β while increasing the contractile marker MYH11. RNA sequencing revealed differential expression of matrisome genes. Periadventitial BAPN treatment reduces fibrosis and promotes AVF compliance. Interestingly, the inhibition of LOX leads to increased accumulation of contractile VSMC while reducing myofibroblast-like cells. Periadventitial LOX inhibition alters the matrisome to improve AVF vascular remodeling.

Three-dimensional printing of microfiber- reinforced hydrogel loaded with oxymatrine for treating spinal cord injury.

Spinal cord injury (SCI) causes severe neural tissue damage and motor/sensory dysfunction. Since the injured spinal cord tissue has limited self-regeneration ability, several strategies, including cell therapy, drug delivery, and tissue engineering scaffold implantation, have been employed to treat SCI. However, each of these strategies fails to obtain desirable outcomes due to their respective limitations. In comparison, advanced tissue engineering scaffolds with appropriate topographical features, favorable composition, and sustained drug delivery capability can be employed to recruit endogenous neural stem cells (NSCs), induce neuronal differentiation, and facilitate neuron maturation. This can lead to the regeneration of injured spinal cord tissue and the recovery of motor function. In this study, fiber bundle-reinforced spinal cord extracellular matrix hydrogel scaffolds loaded with oxymatrine (OMT) were produced through nearfield direct write electrospinning. The spinal cord extracellular matrix-based hydrogel was then coated with OMT. The physical/chemical properties and in vitro degradation behavior of the composite scaffolds were investigated. The in vitro cell culture results showed that composite scaffolds loaded with OMT promoted the differentiation of NSCs into neurons and inhibited differentiation into astrocytes. The in vivo results showed that the composite scaffolds loaded with OMT recruited NSCs from the host tissue, promoted neuronal differentiation and axon extension at the lesion site, inhibited glial scar formation at/around the lesion site, and improved the recovery of motor function in rats with SCI. To sum up, 3D-printed microfiber-reinforced spinal cord extracellular matrix hydrogel scaffolds loaded with OMT are promising biomaterials for the treatment of SCI.

Flowerbed-Inspired Biomimetic Scaffold with Rapid Internal Tissue Infiltration and Vascularization Capacity for Bone Repair.

The favorable microstructure and bioactivity of tissue-engineered bone scaffolds are closely associated with the regenerative efficacy of bone defects. For the treatment of large bone defects, however, most of them fail to meet requirements such as adequate mechanical strength, highly porous structure, and excellent angiogenic and osteogenic activities. Herein, inspired by the characteristics of a "flowerbed", we construct a short nanofiber aggregates-enriched dual-factor delivery scaffold via 3D printing and electrospinning techniques for guiding vascularized bone regeneration. By the assembly of short nanofibers containing dimethyloxalylglycine (DMOG)-loaded mesoporous silica nanoparticles with a 3D printed strontium-contained hydroxyapatite/polycaprolactone (SrHA@PCL) scaffold, an adjustable porous structure can be easily realized by changing the density of nanofibers, while strong compressive strength will be acquired due to the framework role of SrHA@PCL. Owing to the different degradation performance between electrospun nanofibers and 3D printed microfilaments, a sequential release behavior of DMOG and Sr ions is achieved. Both in vivo and in vitro results demonstrate that the dual-factor delivery scaffold has excellent biocompatibility, significantly promotes angiogenesis and osteogenesis by stimulating endothelial cells and osteoblasts, and effectively accelerates tissue ingrowth and vascularized bone regeneration through activating the hypoxia inducible factor-1α pathway and immunoregulatory effect. Overall, this study has provided a promising strategy for constructing a bone microenvironment-matched biomimetic scaffold for bone regeneration.

Review on Bioinspired Design of ECM-Mimicking Scaffolds by Computer-Aided Assembly of Cell-Free and Cell Laden Micro-Modules

Tissue engineering needs bioactive drug delivery scaffolds capable of guiding cell biosynthesis and tissue morphogenesis in three dimensions. Several strategies have been developed to design and fabricate ECM-mimicking scaffolds suitable for directing in vitro cell/scaffold interaction, and controlling tissue morphogenesis in vivo. Among these strategies, emerging computer aided design and manufacturing processes, such as modular tissue unit patterning, promise to provide unprecedented control over the generation of biologically and biomechanically competent tissue analogues. This review discusses recent studies and highlights the role of scaffold microstructural properties and their drug release capability in cell fate control and tissue morphogenesis. Furthermore, the work highlights recent advances in the bottom-up fabrication of porous scaffolds and hybrid constructs through the computer-aided assembly of cell-free and/or cell-laden micro-modules. The advantages, current limitations, and future challenges of these strategies are described and discussed.

Novel Silicone-Grafted Alginate as a Drug Delivery Scaffold: Pharmaceutical Characterization of Gliclazide-Loaded Silicone-Based Composite Microcapsules

A novel gliclazide-loaded elastomeric carbohydrate pharmaceutical vehicle was successfully developed. This new siliconized alginate platform showed pseudoplastic rheology with a zeta potential ranging from (-43.8 mV to -75.5 mV). A Buchi-B390 encapsulator was employed to formulate different types of silicone-grafted alginate microcapsules loaded with gliclazide relying on the vibrational ionic gelation technology. The use of tetraethyl orthosilicate (TEOS) to crosslink the silicone elastomer (hydroxy terminated polydimethylsiloxane) of this new platform had improved the gliclazide encapsulation (>92.13% ± 0.76) of the free-flowing composite microcapsules, which showed good mechanical durability (up to 12 h in PBS pH 6.8) and promising results to sustain the drug release.

Research progress of hydrogels as delivery systems and scaffolds in the treatment of secondary spinal cord injury.

Secondary spinal cord injury (SSCI) is the second stage of spinal cord injury (SCI) and involves vasculature derangement, immune response, inflammatory response, and glial scar formation. Bioactive additives, such as drugs and cells, have been widely used to inhibit the progression of secondary spinal cord injury. However, the delivery and long-term retention of these additives remain a problem to be solved. In recent years, hydrogels have attracted much attention as a popular delivery system for loading cells and drugs for secondary spinal cord injury therapy. After implantation into the site of spinal cord injury, hydrogels can deliver bioactive additives in situ and induce the unidirectional growth of nerve cells as scaffolds. In addition, physical and chemical methods can endow hydrogels with new functions. In this review, we summarize the current state of various hydrogel delivery systems for secondary spinal cord injury treatment. Moreover, functional modifications of these hydrogels for better therapeutic effects are also discussed to provide a comprehensive insight into the application of hydrogels in the treatment of secondary spinal cord injury.

PET-RAFT to expand the surface-modification chemistry of melt coextruded nanofibers

Polymeric nanofibers have been widely used as scaffolds for tissue engineering, drug delivery, and filtration applications, among many others. This work describes new methods to modify chemically-inert fibers using PET-RAFT.

Reassembly Design of Ferritin Cages.

The ferritin family is distributed in nearly all organisms and protects them from iron-induced oxidative damage. Besides, its highly symmetrical structure and biochemical features make it an appealing material for biotechnological applications, such as building blocks for multidimensional assembly, templates for nano-reactors, and scaffolds for encapsulation and delivery of nutrients and drugs. Moreover, it is of great significance to construct ferritin variants with different properties, size, and shape to further broaden its application. In this chapter, we present a routine process of the ferritin redesign and the characterization method of the protein structure to provide a feasible scheme.

Targeting connexin 43 expression via scaffold mediated delivery of antisense oligodeoxynucleotide preserves neurons, enhances axonal extension, reduces astrocyte and microglial activation after spinal cord injury

Injury to the central nervous system (CNS) provokes an inflammatory reaction and secondary damage that result in further tissue damage and destruction of neurons away from the injury site. Upon injury, expression of connexin 43 (Cx43), a gap junction protein, upregulates and is responsible for the spread and amplification of cell death signals through these gap junctions. In this study, we hypothesise that the downregulation of Cx43 by scaffold-mediated controlled delivery of antisense oligodeoxynucleotide (asODN), would minimise secondary injuries and cell death, and thereby support tissue regeneration after nerve injuries. Specifically, using spinal cord injury (SCI) as a proof-of-principle, we utilised a fibre-hydrogel scaffold for sustained delivery of Cx43asODN, while providing synergistic topographical cues to guide axonal ingrowth. Correspondingly, scaffolds loaded with Cx43asODN, in the presence of NT-3, suppressed Cx43 up-regulation after complete transection SCI in rats. These scaffolds facilitated the sustained release of Cx43asODN for up to 25 days. Importantly, asODN treatment preserved neurons around the injury site, promoted axonal extension, decreased glial scarring, and reduced microglial activation after SCI. Our results suggest that implantation of such scaffold-mediated asODN delivery platform could serve as an effective alternative SCI therapeutic approach.

Structural and mechanical properties of folded protein hydrogels with embedded microbubbles.

Globular folded proteins are powerful building blocks to create biomaterials with mechanical robustness and inherent biological functionality. Here we explore their potential as advanced drug delivery scaffolds, by embedding microbubbles (MBs) within a photo-activated, chemically cross-linked bovine serum albumin (BSA) protein network. Using a combination of circular dichroism (CD), rheology, small angle neutron scattering (SANS) and microscopy we determine the nanoscale and mesoscale structure and mechanics of this novel multi-composite system. Optical and confocal microscopy confirms the presence of MBs within the protein hydrogel, their reduced diffusion and their effective rupture using ultrasound, a requirement for burst drug release. CD confirms that the inclusion of MBs does not impact the proportion of folded proteins within the cross-linked protein network. Rheological characterisation demonstrates that the mechanics of the BSA hydrogels is reduced in the presence of MBs. Furthermore, SANS reveals that embedding MBs in the protein hydrogel network results in a smaller number of clusters that are larger in size (∼16.6% reduction in number of clusters, 17.4% increase in cluster size). Taken together, we show that MBs can be successfully embedded within a folded protein network and ruptured upon application of ultrasound. The fundamental insight into the impact of embedded MBs in protein scaffolds at the nanoscale and mesoscale is important in the development of future platforms for targeted and controlled drug delivery applications.

Sequential anti-inflammatory and osteogenic effects of a dual drug delivery scaffold loaded with parthenolide and naringin in periodontitis.

Our pilot study showed that a 3-dimensional dual drug delivery scaffold (DDDS) loaded with Chinese herbs significantly increased the regenerated bone volume fraction. This study aimed to confirm the synergistic anti-inflammatory and osteogenic preclinical effects of this system. The targets and pathways of parthenolide and naringin were predicted. Three cell models were used to assess the anti-inflammatory effects of parthenolide and the osteogenic effects of naringin. First, the distance between the cementoenamel junction and alveolar bone crest (CEJ-ABC) and the bone mineral density (BMD) of surgical defects were measured in a rat model of periodontitis with periodontal fenestration defects. Additionally, the mRNA expression levels of matrix metallopeptidase 9 (MMP9) and alkaline phosphatase (ALP) were measured. Furthermore, the number of inflammatory cells and osteoclasts, as well as the protein expression levels of tumor necrosis factor-alpha (TNF-α) and levels of ALP were determined. Target prediction suggested prostaglandin peroxidase synthase (PTGS2) as a potential target of parthenolide, while cytochrome P450 family 19 subfamily A1 (CYP19A1) and taste 2 receptor member 31 (TAS2R31) were potential targets of naringin. Parthenolide mainly targeted inflammation-related pathways, while naringin participated in steroid hormone synthesis and taste transduction. In vitro experiments revealed significant anti-inflammatory effects of parthenolide on RAW264.7 cells, and significant osteogenic effects of naringin on bone marrow mesenchymal stem cells and MC3T3-E1 cells. DDDS loaded with parthenolide and naringin decreased the CEJ-ABC distance and increased BMD and ALP levels in a time-dependent manner. Inflammation was significantly alleviated after 14 days of DDDS treatment. Additionally, after 56 days, the DDDS group exhibited the highest BMD and ALP levels. DDDS loaded with parthenolide and naringin in a rat model achieved significant synergistic anti-inflammatory and osteogenic effects, providing powerful preclinical evidence.

Nanocellulose-gellan cross-linked scaffolds for vaginal delivery of fluconazole

The objective of this research is to formulate lyophilized fluconazole-loaded nanocellulose-gellan scaffolds cross-linked using trisodium trimetaphosphate as a vaginal drug delivery system. The effect of polymers (nanocellulose and gellan gum) and cross-linking agents on drug release and mucoadhesive strength were determined by approaching a two-factor three-level central composite experimental design. The optimal formulation of the fluconazole-loaded cross-linked rice or wheat nanocellulose-gellan based scaffolds comprised of the concentration of polymers (4.91 % w/v or 4.99 % w/v) and trisodium trimetaphosphate (16.43 % w/v or 15.83 % w/v), respectively. The infrared spectra confirmed the cross-linking of nanocellulose and gellan gum while the thermal graph revealed the higher thermal stability of cross-linked scaffolds. The diffractogram of the scaffolds unveiled their amorphous nature while the electron micrographs depict the porous nature of the fluconazole-loaded nanocellulose-gellan scaffolds. The phosphorylated cross-linked nanocellulose-gellan scaffolds represent more swelling (8-fold higher), porosity (>83 %), tensile strength (>34 MPa), and mucoadhesive strength (>1940 mN), and less enzymatic degradation rate over the non cross-linked scaffolds. The optimal batch of cross-linked nanocellulose-gellan scaffolds provided a sustained release of 99 % of fluconazole over 24 h with 1.19-fold higher ex-vivo vaginal permeation over the native scaffolds. In addition, the phosphorylated nanocellulose-gellan based scaffolds exhibit improved antifungal activity and non-cytotoxicity.

Type I Collagen-Apatite Fibrillar Nanocomposites: Mineralization, Crosslinking and Anti-Inflammatory Cocrystal Impregnation for Bone Tissue Engineering

Self-assembly and mineralization of type I collagen (Col) with nanocrystalline apatite (nAp), by adding a solution of Ca(OH)2 to a stirred Col-H3PO4 solution by fast dripping, allowed the preparation of Col/nAp fibrils with good crystallographic control of the mineral phase. In this work, in addition, we have cross-linked the mineralized fibers by using different reagents, namely glutaraldehyde (GTA), tannic acid (TA), 1-Ethyl-3-(3-dimethyl aminopropyl)-carbodiimide combined with N-Hydroxysuccinimide (EDC/NHS), and genipin (GP), aimed at producing different types of biopolymeric Col/nAp-based drug delivery scaffolds. In parallel, we have investigated two different methods to impregnate the scaffolds with molecules of the cocrystal diclofenac-metformin (DF-MET). The result, when using TA as a crosslinking reagent, shows the sequence of mineralized fibrils impregnation followed by crosslinking leads to maximum cocrystal molecule loading. The impregnated material is expected to be useful in settings with excessive and prolonged inflammation, since they affect negatively the fracture healing/bone repair processes, especially during the early stages of healing.

Methods and Limitations of Augmenting Mesenchymal Stem Cells for Therapeutic Applications.

Significance: Given their capacity for self-renewal, multilineage differentiation, and immunomodulatory potential, mesenchymal stem cells (MSCs) represent a promising modality of clinical therapy for both regenerative medicine and immune diseases. In this study, we review the key approaches and popular methods utilized to boost potency and modify functions of MSCs for clinical purposes as well as their associated limitations. Recent Advances: Several major domains of cell modification strategies are currently employed by investigators to overcome these deficits and augment the therapeutic potential of MSCs. Priming MSCs with soluble factors or pharmacologic agents as well as manipulating oxygen availability in culture have been demonstrated to be effective biochemical methods to augment MSC potential. Distinct genetic and epigenetic methods have emerged in recent years to modify the genetic expression of target proteins and factors thereby modulating MSCs capacity for differentiation, migration, and proliferation. Physical methods utilizing three-dimensional culture methods and alternative cell delivery systems and scaffolds can be used to recapitulate the native MSC niche and augment their engraftment and viability for in vivo models. Critical Issues: Unmodified MSCs have demonstrated only modest benefits in many preclinical and clinical studies due to issues with cell engraftment, viability, heterogeneity, and immunocompatibility between donor and recipient. Furthermore, unmodified MSCs can have low inherent therapeutic potential for which intensive research over the past few decades has been dedicated to improving cell functionality and potency.

Engineering thixotropic supramolecular gelatin-based hydrogel as an injectable scaffold for cell transplantation

Despite many efforts focusing on regenerative medicine, there are few clinically-available cell-delivery carriers to improve the efficacy of cell transplantation due to the lack of adequate scaffolds. Herein, we report an injectable scaffold composed of functionalized gelatin for application in cell transplantation. Injectable functionalized gelatin-based hydrogels crosslinked with reversible hydrogen bonding based on supramolecular chemistry were designed. The hydrogel exhibited thixotropy, enabling single syringe injection of cell-encapsulating hydrogels. Highly biocompatible and cell-adhesive hydrogels provide cellular scaffolds that promote cellular adhesion, spreading, and migration. The in vivo degradation study revealed that the hydrogel gradually degraded for seven days, which may lead to prolonged retention of transplanted cells and efficient integration into host tissues. In volumetric muscle loss models of mice, cells were transplanted using hydrogels and proliferated in injured muscle tissues. Thixotropic and injectable hydrogels may serve as cell delivery scaffolds to improve graft survival in regenerative medicine.

Next-Generation 3D Scaffolds for Nano-Based Chemotherapeutics Delivery and Cancer Treatment.

Cancer is the leading cause of death after cardiovascular disease. Despite significant advances in cancer research over the past few decades, it is almost impossible to cure end-stage cancer patients and bring them to remission. Adverse effects of chemotherapy are mainly caused by the accumulation of chemotherapeutic agents in normal tissues, and drug resistance hinders the potential therapeutic effects and curing of this disease. New drug formulations need to be developed to overcome these problems and increase the therapeutic index of chemotherapeutics. As a chemotherapeutic delivery platform, three-dimensional (3D) scaffolds are an up-and-coming option because they can respond to biological factors, modify their properties accordingly, and promote site-specific chemotherapeutic deliveries in a sustainable and controlled release manner. This review paper focuses on the features and applications of the variety of 3D scaffold-based nano-delivery systems that could be used to improve local cancer therapy by selectively delivering chemotherapeutics to the target sites in future.

Cell Infiltrative Inner Connected Porous Hydrogel Improves Neural Stem Cell Migration and Differentiation for Functional Repair of Spinal Cord Injury.

The disadvantages of cell-adaptive microenvironments and cellular diffusion out of the lesion have limited hydrogel-based scaffold transplantation treatment for neural connectivity, leading to permanent neurological disability from spinal cord injury. Herein, porous GelMA scaffold was prepared, in which the inner porous structure was optimized. The average pore size was 168 ± 71 μm with a porosity of 77.1%. The modulus of porous hydrogel was 593 ± 4 Pa compared to 1535 ± 85 Pa of bulk GelMA. The inner connected porous structure provided a cell-infiltrative matrix for neural stem cell migration and differentiation in vitro and eventually enhanced neuron differentiation and hindlimb strength and movement of animals in in vivo experiments. Furthermore, inflammation response and apoptosis were also alleviated after implantation. This work demonstrated that the porous hydrogel with appropriately connected micropores exhibit favorable cellular responses compared with traditional non-porous GelMA hydrogel. Taken together, our findings suggest that porous hydrogel is a promising scaffold for future delivery of stem cells and has prospects in material design for the treatment of spinal cord injury.