Microporous Scaffolds Research Articles

Superporous soy protein isolate matrices as superabsorbent dressings for successful management of highly exuding wounds: In vitro and in vivo characterization

Soy protein isolate (SPI) has received widespread attention of the biomedical research community primarily due to its good biocompatibility, biodegradability, high availability and low cost. Herein, glutaraldehyde cross-linked microporous sponge-like SPI scaffolds were prepared using the cryogelation technique for tissue engineering applications. The prepared SPI scaffolds possess an interconnected porous structure with approximately 90% porosity and an average pore size in the range of 45–92 μm. The morphology, porosity, swelling capacity and degradation rate of the cryogels were found to be dependent on the concentration of polymer to crosslinking agent. All cryogels were found to be elastic and able to maintain physical integrity even after being compressed to one—fifth of their original length during cyclic compression analysis. These cryogels showed excellent mechanical properties, immediate water-triggered shape restoration and absorption speed. Furthermore, cryogels outperformed cotton and gauze in terms of blood clotting and blood cell adherence. The in vitro and in vivo studies demonstrated the potency of SPI scaffolds for skin tissue engineering applications. Our findings showed that crosslinking with glutaraldehyde had no detrimental effects on cell viability. In addition, an in vivo wound healing study in rats validated them as good potential wound dressing materials.

Molecular weight of hyaluronic acid crosslinked into biomaterial scaffolds affects angiogenic potential.

While hyaluronic acid (HA)-based hydrogels have been used clinically for decades, the mechanisms by which HA exerts molecular weight-dependent bioactivity and how chemical modification and crosslinking may affect molecular weight-dependent bioactivity remain poorly understood. This knowledge gap presents a significant barrier to designing HA hydrogels with predictable bioactivities. As HA has been widely reported to have molecular weight-dependent effects on endothelial cells (ECs), we investigated how the molecular weight of HA in either soluble or crosslinked forms affects angiogenesis and interrogated CD44 clustering on the surface of endothelial cells as a candidate mechanism for these affects. Using soluble HA, our results show high molecular weight (HMW) HA, but not low molecular weight (LMW) HA, increased viability and tube formation in cultured human cerebral microvascular ECs (HCMVECs). No size of HA affected proliferation. When HCMVECs were cultured with crosslinked HA of varying molecular weights in the form of HA-based microporous annealed particle scaffold (HMAPS), the cell response was comparable to when cultured with soluble HA. Similarly, when implanted subcutaneously, HMAPS with HMW HA were more vascularized than those with LMW HA. We also show that antibody-mediated CD44 clustering resulted in HCMVECs with increased viability and tube-like structure formation in a manner comparable to exposure to HMW HA, suggesting that HMW acts through CD44 clustering. STATEMENT OF SIGNIFICANCE: Biomaterials based on hyaluronic acid (HA), a bioactive extracellular matrix polysaccharide, have been used in clinical products for several years. Despite the knowledge that HA molecular weight heavily influences its bioactivity, molecular weight has been largely ignored in the development of HA-based biomaterials. Given the high viscosity of high molecular weight HA typically found in native tissues, lower molecular weight polysaccharides have been used most commonly for biomaterial fabrication. By comparing the ability of injectable, microporous annealed particle scaffolds (MAPS) fabricated from variably sized HA to promote angiogenesis, this study demonstrates that MAPS with high molecular weight HA better support vascularization, likely through an unique ability to induce clustering of CD44 receptors on endothelial cells.

Polysaccharide-Based Composite Hydrogel with Hierarchical Microstructure for Enhanced Vascularization and Skull Regeneration.

Critical-size skull defects caused by trauma, infection, and tumor resection raise great demands for efficient bone substitutes. Herein, a hybrid cross-linked hierarchical microporous hydrogel scaffold (PHCLS) was successfully assembled by a multistep procedure, which involved (i) the preparation of poly(lactic-co-glycolic)/nanohydroxyapatite (PLGA-HAP) porous microspheres, (ii) embedding the spheres in a solution of dopamine-modified hyaluronic acid and collagen I (Col I) and cross-linking via dopamine polyphenols binding to (i) Col I amino groups (via Michael addition) and (ii) PLGA-HAP (via calcium ion chelation). The introduction of PLGA-HAP not only improved the diversity of pore size and pore communication inside the matrix but also greatly enhanced the compressive strength (5.24-fold, 77.5 kPa) and degradation properties to construct a more stable mechanical structure. In particular, the PHCLS (200 mg, nHAP) promoted the proliferation, infiltration, and angiogenic differentiation of bone marrow mesenchymal stem cells in vitro, as well as significant ectopic angiogenesis and mineralization with a storage modulus enhancement of 2.5-fold after 30 days. Meanwhile, the appropriate matrix microenvironment initiated angiogenesis and early osteogenesis by accelerating endogenous stem cell recruitment in situ. Together, the PHCLS allowed substantial skull reconstruction in the rabbit cranial defect model, achieving 85.2% breaking load strength and 84.5% bone volume fractions in comparison to the natural cranium, 12 weeks after implantation. Overall, this study reveals that the hierarchical microporous hydrogel scaffold provides a promising strategy for skull defect treatment.

Injectable, pore-forming, self-healing, and adhesive hyaluronan hydrogels for soft tissue engineering applications

Most existing injectable hydrogels are non-porous, thereby lacking a microporous structure to promote cell ingrowth. Also, most hydrogels do not effectively adhere to the host tissue. The present study describes an injectable double network hydrogel formed by combining two hyaluronic acid (HA) derivatives, namely dopamine grafted HA (DAHA) and methacrylated HA (HAMA). These constituents instantly form a physically crosslinked network through Fe3+-dopamine coordination, and confer fast gelation, pore formation, and self-healing properties to the hydrogel. Photocroslinked upon UV exposure, HAMA forms a chemically crosslinked network, thereby improving mechanical and degradation properties. The adhesive properties of this hydrogel are attributed to the presence of dopamine groups, inspired by mussel creatures. Proper modification of HA chains was confirmed by NMR spectroscopy. The physical, mechanical, rheological, and biological properties of the new hydrogels were quantified in wet laboratory conditions. The results revealed that the DAHA/HAMA hydrogel rapidly forms a self-healing microporous adhesive scaffold with a 26.9 µm pore size, 29.4 kPa compressive modulus, and 12.8 kPa adhesion strength in under 6 s. These findings suggest that the new hydrogel is a promising candidate for in situ repair of soft tissues, particularly mechanically dynamic ones such as the vocal folds, cartilage, and dermis.

Drug-loading ZIF-8 for modification of microporous bone scaffold to promote vascularized bone regeneration

Surface modification of microporous bone scaffolds using nanoparticles has been broadly studied in bone tissue engineering. Aiming at improving vascularized bone regeneration (VBR), zeolitic imidazolate framework-8 (ZIF-8) was encapsulated with dimethyloxallyl glycine (DMOG) and the drug-carrying nanoparticles (D@Z) could be uniformly coated onto the surface of the bone scaffold. The osteogenic and angiogenic actions of D@Z are closely correlated with the amount of slowly released DMOG, and in general, exhibited a favorable association. Then, the D7.5@Z group, which showed the greatest capacity to induce in vitro osteogenesis–angiogenesis coupling, was utilized for surface modification of the bone scaffold. Biological processes including phosphate-containing compound metabolic process, cell differentiation, cell proliferation and cell motility might contribute to enhanced ability to induce VBR by the coated scaffold and signaling pathways such as Rap1, Ras, phosphatidylinositol 3-kinase/protein kinase B (PI3K-AKT) and vascular endothelial growth factor (VEGF) signaling pathways participated in these processes. Finally, as depicted by in vitro real time-polymerase chain reaction (RT-PCR), Western blot (WB) and in vivo cranial bone defect model, the microporous scaffold coated with nano-D7.5@Z greatly promoted VBR. To conclude, nano-D@Z has significant promise for practical application in modification of microporous bone scaffolds to enhance VBR, and DMOG loading quantity has a beneficial influence on D@Z to improve osteogenesis–angiogenesis coupling.

Sustained release of resveratrol from fused deposition modelling guided 3D porous scaffold for bone tissue engineering

To promote bone regeneration, bone tissue engineering necessitates development of a three-dimensional porous scaffold with osteoconductive, osteoinductive, and osteogenic features. In this study, a milliporous polylactic acid (PLA) scaffold was 3D printed, into which a microporous polycaprolactone scaffold releasing resveratrol was embedded. Herein, resveratrol-loaded albumin nanoparticles (RSV-Np) were synthesized, homogenously mixed with polycaprolactone (10% w/v & 12% w/v) at 2 different concentrations, and subsequently embedded into the milliporous PLA scaffold. Further, morphological analysis by field emission scanning electron microscopic (FESEM) analysis demonstrated interconnected pores measuring 100–300 µm diameter, and homogenous embedment of RSV-Np throughout the surface. In addition, XRD analysis inferred that resveratrol was present in an amorphous phase in the scaffold, which improves dissolution of the drug, thereby advantageous for drug delivery applications. Furthermore, successful incorporation of RSV-Np was validated by Raman spectroscopy and Fourier transform infrared spectroscopy. The drug release study implied that RSV-Np concentration modulated the resveratrol release from scaffolds. Moreover, in vitro testing of the scaffolds for 16 days showed that resveratrol sustained release enhanced the osteogenic potential of scaffolds. In summary, such scaffolds can be a potential matrix for bone regeneration due to their inherent ability to release resveratrol in a controlled fashion.

In Vivo Bone Regeneration Capacity of Multiscale Porous Polycaprolactone-Based High Internal Phase Emulsion (PolyHIPE) Scaffolds in a Rat Calvarial Defect Model.

Globally, one of the most common tissue transplantation procedures is bone grafting. Lately, we have reported the development of polymerized high internal phase emulsions (PolyHIPEs) made of photocurable polycaprolactone (4PCLMA) and shown their potential to be used as bone tissue engineering scaffolds in vitro. However, it is essential to evaluate the in vivo performance of these scaffolds to investigate their potential in a clinically more relevant manner. Therefore, in this study, we aimed to compare in vivo performances of macroporous (fabricated using stereolithography), microporous (fabricated using emulsion templating), and multiscale porous (fabricated using emulsion templating and perforation) scaffolds made of 4PCLMA. Also, 3D-printed macroporous scaffolds (fabricated using fused deposition modeling) made of thermoplastic polycaprolactone were used as a control. Scaffolds were implanted into a critical-sized calvarial defect, animals were sacrificed 4 or 8 weeks after implantation, and the new bone formation was assessed by micro-computed tomography, dental radiography, and histology. Multiscale porous scaffolds that include both micro- and macropores resulted in higher bone regeneration in the defect area compared to only macroporous or only microporous scaffolds. When one-grade porous scaffolds were compared, microporous scaffolds showed better performance than macroporous scaffolds in terms of mineralized bone volume and tissue regeneration. Micro-CT results revealed that while bone volume/tissue volume (Bv/Tv) values were 8 and 17% at weeks 4 and 8 for macroporous scaffolds, they were significantly higher for microporous scaffolds, with values of 26 and 33%, respectively. Taken together, the results reported in this study showed the potential application of multiscale PolyHIPE scaffolds, in particular, as a promising material for bone regeneration.

Ultra-short laser structuring of 3D-microfabricated non-resorbable polystyrene polymer to anchor cellular adhesion

Tissue engineering is emerging as a preferred choice for regeneration of irreversible bone tissue defects. It’s main “weapons” are the artificially created biocompatible three-dimensional cellular matrices that mimic the extracellular matrix’s structure and provide a mechanically stable inert substrate for the natural growth of cells. These porous scaffolds provide sufficient inner surface for cell adhesion, migration, proliferation and protrusion “inside” the temporal implant, facilitating in that way the natural bone ingrowth and vascularization. This leads, eventually, to overgrowing the outer and inner surface of the biomimetic implant with recipient self-formed bone tissue, thus avoiding a possible immune response.In the current study 3D Polystyrene (PS) non-resorbable synthetic cellular bone scaffolds, produced by 3D microfabrication technology, were additionally structured by applying femtosecond (fs) laser radiation with variable parameters thus achieving optimal laser processing conditions, for creation of 3D matrix with enhanced fiber surface properties. The main aim is the establishment of surface structured microporous biomimetic scaffolds, providing improved cell adhesion, infiltration and formation of extracellular matrix. The 3D laser functionalized PS samples were investigated by Scanning Electron Microscopy (SEM), Energy Dispersive X-Ray analysis (EDX), confocal (optical profilometer) and roughness analyses for evaluation of their morphological and chemical properties before and after fs structuring was performed. Water contact angle (WCA) evaluation was also conducted. Future experimental work will include in vitro cellular studies for determining the optimal structuring laser parameters, providing osteoconductive and osteoinductive qualities of the 3D fs-microstructured PS scaffolds.

Biocompatibility enhancement via post-processing of microporous scaffolds made by optical 3D printer.

Providing a 3D environment that mimics the native extracellular matrix is becoming increasingly important for various applications such as cell function studies, regenerative medicine, and drug discovery. Among the most critical parameters to consider are the scaffold's complicated micro-scale geometry and material properties. Therefore, stereolithography based on photopolymerization is an emerging technique because of its ability to selectively form volumetric structures from liquid resin through localized polymerization reactions. However, one of the most important parameters of the scaffold is biocompatibility, which depends not only on the material but also on the exposure conditions and post-processing, which is currently underestimated. To investigate this systematically, microporous scaffolds with pore sizes of 0.05mm3 corresponding to a porosity of 16,4% were fabricated using the stereolithography printer Asiga PICO2 39 UV from the widely used resins FormLabs Clear and Flexible. The use of various polymers is usually limited for cells because, after wet chemical development, the non-negligible amount of remaining monomers intertwined in the photopolymerized structures is significantly toxic to cells. Therefore, the aim of this research was to find the best method to remove monomers from the 3D scaffold by additional UV exposure. For this purpose, a Soxhlet extractor was used for the first time, and the monomers were immersed in different alcohols. A Raman microspectroscopy was also used to investigate whether different post-processing methods affect DC (cross-linking) to find out if this specifically affects the biocompatibility of the scaffolds. Finally, mesenchymal stem cells from rat dental pulp were examined to confirm the increased biocompatibility of the scaffolds and their ability to support cell differentiation into bone tissue cells.

LaNiO3-rGO perovskite interface for sustainable decontaminants of emerging concerns under visible light photocatalysis

The environment is extremely damaged by the disposal of contaminants of emerging concerns like pharmaceutical and personal care products from various sources. In the present study, lanthanum nickelate and reduced graphene oxide (LaNiO3-rGO) interfaced composite material were synthesized through simple sonication and solvothermal techniques and characterized. The morphological studies showed that LaNiO3 nanoparticles were well interfaced with rGO matrixes with microporous scaffolds that can yield positive impacts on the potential attraction of trace organic pollutants through adsorption processes. Perovskite carbonaceous interface promotes the trapping of photogenerated holes during photocatalytic activities and provides a continuous pathway for radical production. The perovskite carbonaceous interfaces showed high photocatalytic activities towards the antibacterial potential and demineralization of complex organic pollutants like dye and antibiotic compounds under visible light. About 83% and 64% of photocatalytic degradation efficiency were observed in the removal of brilliant green and ciprofloxacin compounds respectively in an aqueous media using LaNiO3-rGO (1:1) photocatalyst. LaNiO3-rGO interfaced composite material also exhibited a potential antimicrobial activity against the Escherichia coli and Staphylococcus aureus microorganisms. Further, the catalyst toxicity study confirmed that the treated water showed lesser toxicity towards the germination and root growth of Vigna radiata.

Fabrication of bFGF/polydopamine-loaded PEEK implants for improving soft tissue integration by upregulating Wnt/β-catenin signaling

The difficulties associated with polyetheretherketone (PEEK) implants and soft tissue integration for craniomaxillofacial bone repair have led to a series of complications that limit the clinical benefits. In this study, 3D printed multi-stage microporous PEEK implants coated with bFGF via polydopamine were fabricated to enhance PEEK implant-soft tissue integration. Multistage microporous PEEK scaffolds prepared by sulfonation of concentrated sulfuric acid were coated with polydopamine, and then used as templates for electrophoretic deposition of bFGF bioactive factors. Achieving polydopamine and bFGF sustained release, the composite PEEK scaffolds possessed good mechanical properties, hydrophilicity, protein adhesion properties. The in vitro results indicated that bFGF/polydopamine-loaded PEEK exhibited good biocompatibility to rabbit embryonic fibroblasts (REF) by promoting cell proliferation, adhesion, and migration. Ribonucleic acid sequencing (RNA-seq) revealed that bFGF/polydopamine-loaded PEEK implants significantly upregulated the expression of genes and proteins associated with soft tissue integration and activated Wnt/β-catenin signaling in biological processes, but related expression of genes and proteins was significantly downregulated when the Wnt/β-catenin signaling was inhibited. Furthermore, in vivo bFGF/polydopamine-loaded PEEK implants exhibited excellent performance in improving the growth and adhesion of the surrounding soft tissue. In summary, bFGF/polydopamine-loaded PEEK implants possess soft tissue integration properties by activating the Wnt/β-catenin signaling, which have a potential translational clinical application in the future.

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.

Integrated data-driven modeling and experimental optimization of granular hydrogel matrices

Integrated data-driven modeling and experimental optimization of granular hydrogel matrices

Introducing Controlled Microporosity in Melt Electrowriting

AbstractMelt electrowriting (MEW) enables the electric field‐assisted digital fabrication of precisely defined scaffold architectures of micron‐sized fibers. However, charge accumulation and consequent disruption of the precoded pattern by fiber bridging prevents controlled printing at small interfiber distances. This, together with the periodical layer stacking characteristic for additive manufacturing, typically results in scaffolds with channel‐like macroporosity, which need to be combined with other biofabrication techniques to achieve the desired microporosity for cellular infiltration. Therefore, a design strategy is devised to introduce controlled interconnected microporosity directly in MEW scaffolds by an algorithm that creates arrays of bridging‐free parallel fibers, angularly shifted from layer to layer and starting at a random point to avoid periodical fiber stacking, and hence channel‐like pores while defining micropores. This work hypothesizes that pore size can be controlled, decoupled from fiber diameter, and the mechanical properties, including anisotropy ratio, can be tuned. The authors demonstrate this while leveraging the platform for both flat and seamless tubular scaffolds and characterize them via micro‐computed tomography and tensile loading. Lastly, successful cell ingrowth into the micropores and extracellular matrix formation are shown. This platform enables microporous scaffolds entirely via MEW that can be tailored to the architectural and mechanical requirements of the target tissues.

Extrahepatic transplantation of 3D cultured stem cell-derived islet organoids on microporous scaffolds.

Stem cell differentiation methods have been developed to produce cells capable of insulin secretion which are showing promise in clinical trials for treatment of type-1 diabetes. Nevertheless, opportunities remain to improve cell maturation and function. Three-dimensional (3D) culture has demonstrated improved differentiation and metabolic function in organoid systems, with biomaterial scaffolds employed to direct cell assembly and facilitate cell-cell contacts. Herein, we investigate 3D culture of human stem cell-derived islet organoids, with 3D culture initiated at the pancreatic progenitor, endocrine progenitor, or immature β-cell stage. Clusters formed by reaggregation of immature β-cells could be readily seeded into the microporous poly(lactide-co-glycolide) scaffold, with control over cell number. Culture of islet organoids on scaffolds at the early to mid-stage beta cell progenitors had improved in vitro glucose stimulated insulin secretion relative to organoids formed at the pancreatic progenitor stage. Reaggregated islet organoids were transplanted into the peritoneal fat of streptozotocin-induced diabetic mice, which resulted in reduced blood glucose levels and the presence of systemic human C-peptide. In conclusion, 3D cell culture supports development of islet organoids as indicated by insulin secretion in vitro and supports transplantation to extrahepatic sites that leads to a reduction of hyperglycemia in vivo.

3D printing microporous scaffolds from modular bioinks containing sacrificial, cell-encapsulating microgels.

Microgel-based biomaterials have inherent porosity and are often extrudable, making them well-suited for 3D bioprinting applications. Cells are commonly introduced into these granular inks post-printing using cell infiltration. However, due to slow cell migration speeds, this strategy struggles to achieve depth-independent cell distributions within thick 3D printed geometries. To address this, we leverage granular ink modularity by combining two microgels with distinct functions: (1) structural, UV-crosslinkable microgels made from gelatin methacryloyl (GelMA) and (2) sacrificial, cell-laden microgels made from oxidized alginate (AlgOx). We hypothesize that encapsulating cells within sacrificial AlgOx microgels would enable the simultaneous introduction of void space and release of cells at depths unachievable through cell infiltration alone. Blending the microgels in different ratios produces a family of highly printable GelMA : AlgOx microgel inks with void fractions ranging from 0.03 to 0.35. As expected, void fraction influences the morphology of human umbilical vein endothelial cells (HUVEC) within GelMA : AlgOx inks. Crucially, void fraction does not alter the ideal HUVEC distribution seen throughout the depth of 3D printed samples. This work presents a strategy for fabricating constructs with tunable porosity and depth-independent cell distribution, highlighting the promise of microgel-based inks for 3D bioprinting.

Fas Ligand-Modified Scaffolds Protect Stem Cell Derived β-Cells by Modulating Immune Cell Numbers and Polarization.

Stem cell derived β-cells have demonstrated the potential to control blood glucose levels and represent a promising treatment for Type 1 diabetes (T1D). Early engraftment post-transplantation and subsequent maturation of these β-cells are hypothesized to be limited by the initial inflammatory response, which impacts the ability to sustain normoglycemia for long periods. We investigated the survival and development of immature hPSC-derived β-cells transplanted on poly(lactide-co-glycolide) (PLG) microporous scaffolds into the peritoneal fat, a site being considered for clinical translation. The scaffolds were modified with biotin for binding of a streptavidin-FasL (SA-FasL) chimeric protein to modulate the local immune cell responses. The presence of FasL impacted infiltration of monocytes and neutrophils and altered the immune cell polarization. Conditioned media generated from SA-FasL scaffolds explanted at day 4 post-transplant did not impact hPSC-derived β-cell survival and maturation in vitro, while these responses were reduced with conditioned media from control scaffolds. Following transplantation, β-cell viability and differentiation were improved with SA-FasL modification. A sustained increase in insulin positive cell ratio was observed with SA-FasL-modified scaffolds relative to control scaffolds. These results highlight that the initial immune response can significantly impact β-cell engraftment, and modulation of cell infiltration and polarization may be a consideration for supporting long-term function at an extrahepatic site.

Microporous metallic scaffolds supported liquid infused icephobic construction

Hypothesis: Ice accretion on component surfaces often causes severe impacts or accidents. Liquid-infused surfaces (LIS) have drawn much attention as icephobic materials for ice mitigation in recent years due to their outstanding icephobicity. However, the durability of LIS constructions remains a big challenge, including mechanical vulnerability and rapid depletion of lubricants. The practical applications of LIS materials are significantly restrained, and the full potential of LIS for ice prevention has yet to be demonstrated.Experiments: A universal approach was proposed to introduce microporous metallic scaffolds in the LIS construction to increase the applicability and durability, and to prompt the potential of LIS for ice mitigation. Microporous Ni scaffolds were chosen to integrate with polydimethylsiloxane modified by silicone oil addition.Findings: The new LIS construction demonstrated significantly improved durability in icing/de-icing cyclic test, and it also offered a solution for the rapid oil depletion by restraining the deformation of the matrix material. Low ice adhesion strength could be maintained via a micro-crack initiation mechanism. The results indicated that the multi-phase LIS construction consisting of microporous Ni scaffolds effectively addressed the shackles of the icephobicity deterioration of LIS materials, confirming a new design strategy for the R&D of icephobic surfaces.

Gelatin Methacryloyl Granular Hydrogel Scaffolds: High-throughput Microgel Fabrication, Lyophilization, Chemical Assembly, and 3D Bioprinting.

The emergence of granular hydrogel scaffolds (GHS), fabricated via assembling hydrogel microparticles (HMPs), has enabled microporous scaffold formation in situ. Unlike conventional bulk hydrogels, interconnected microscale pores in GHS facilitate degradation-independent cell infiltration as well as oxygen, nutrient, and cellular byproduct transfer. Methacryloyl-modified gelatin (GelMA), a (photo)chemically crosslinkable, protein-based biopolymer containing cell adhesive and biodegradable moieties, has widely been used as a cell-responsive/instructive biomaterial. Converting bulk GelMA to GHSmay open a plethora of opportunities for tissue engineering and regeneration. In this article, we demonstrate the procedures of high-throughput GelMA microgel fabrication, conversion to resuspendable dry microgels (micro-aerogels), GHS formation via the chemical assembly of microgels, and granular bioink fabrication for extrusion bioprinting. We show how a sequential physicochemical treatment via cooling and photocrosslinking enables the formation of mechanically robust GHS.When light is inaccessible (e.g., during deep tissue injection), individually crosslinked GelMA HMPs may be bioorthogonally assembled via enzymatic crosslinking using transglutaminases. Finally, three-dimensional (3D) bioprinting of microporous GHSat low HMP packing density is demonstrated via the interfacial self-assembly of heterogeneously charged nanoparticles.

The potential of eggshell hydroxyapatite, collagen, and EGCG (HAp-Col-EGCG) scaffold as a pulp regeneration material

BackgroundHydrogel scaffold is a biomaterial that can facilitate cells in forming a tissue structure. It can promote cell adhesion, migration, and proliferation. Further research to find a new scaffold from natural resources is challenging, so this study aimed to characterize a hydrogel composite scaffold, which has the potential to be used as a regenerative material. MethodsThe formulation of HAp-Col-EGCG was mixed with different ratios of 1%, 2%, and 4% hydroxyapatite. We analyzed its injectability, pH, and gelation time. Scanning electron microscopy (SEM), energy X-ray Spectroscopy (EDX), and Fourier-transform infrared spectroscopy (FTIR) were used to evaluate the surface morphologies, element composition, and chemical properties of HAp-Col-EGCG. ResultsThe results showed that the injectability test was almost 90 % in all groups. There was no significant difference in the median value of the pH at 0, 20, and 60 min in all groups, but there was a significant difference at 40 min. The average gelation times in all groups were not significant. SEM-EDX showed a microporous scaffold, with the HAp particles well distributed in the collagen pores at a ratio of 1.9, 2.29, and 1.89 Ca/P. The FTIR results showed intermolecular bonds in the HAp-Col-EGCG scaffold. The X-ray diffraction analysis showed that collagen and EGCG did not affect the crystal structure and size of HAp. Cytotoxicity test showed more dental pulp cell viability at the 4 % HAp concentration at 514.35 ± 15.45. ConclusionThis study indicates that hydrogel scaffold from eggshell hydroxyapatite, collagen, and EGCG has a high potential for pulp regenerative therapy.