Stimuli-responsive biomaterials: smart avenue toward 4D bioprinting.

3D bioprinting is an advanced technology combining cells and bioactive molecules within a single bioscaffold; however, this scaffold cannot change, modify or grow in response to a dynamic implemented environment. Lately, a new era of smart polymers and hydrogels has emerged, which can add another dimension, e.g., time to 3D bioprinting, to address some of the current approaches' limitations. This concept is indicated as 4D bioprinting. This approach may assist in fabricating tissue-like structures with a configuration and function that mimic the natural tissue. These scaffolds can change and reform as the tissue are transformed with the potential of specific drug or biomolecules released for various biomedical applications, such as biosensing, wound healing, soft robotics, drug delivery, and tissue engineering, though 4D bioprinting is still in its early stages and more works are required to advance it. In this review article, the critical challenge in the field of 4D bioprinting and transformations from 3D bioprinting to 4D phases is reviewed. Also, the mechanistic aspects from the chemistry and material science point of view are discussed too.

Viral and non-viral gene therapy using 3D (bio)printing.

The overall success in launching discovered drugs is tightly restricted to the high rate of late-stage failures, which ultimately inhibits the distribution of medicines in markets. As a result, it is imperative that methods reliably predict the effectiveness and, more critically, the toxicity of medicine early in the drug development process before clinical trials be continuously innovated. We must stay up to date with the fast appearance of new infections and diseases by rapidly developing the requisite vaccinations and medicines. Modern in vitro models of disease may be used as an alternative to traditional disease models, and advanced technology can be used for the creation of pharmaceuticals as well as cells, drugs, and gene delivery systems to expedite the drug discovery procedure. Furthermore, in vitro models that mimic the spatial and chemical characteristics of native tissues, such as a 3D bioprinting system or other technologies, have proven to be more effective for drug screening than traditional 2D models. Viral and non-viral gene delivery vectors are a hopeful tool for combinatorial gene therapy, suggesting a quick way of simultaneously deliver multiple genes. A 3D bioprinting system embraces an excellent potential for gene delivery into the different cells or tissues for different diseases, in tissue engineering and regeneration medicine, in which the precise nucleic acid is located in the 3D printed tissues and scaffolds. Non-viral nanocarriers, in combination with 3D printed scaffolds, are applied to their delivery of genes and controlled release properties. There remains, however, a big obstacle in reaching the full potential of 3D models because of a lack of in vitro manufacturing of live tissues. Bioprinting advancements have made it possible to create biomimetic constructions that may be used in various drug discovery research applications. 3D bioprinting also benefits vaccinations, medicines, and relevant delivery methods because of its flexibility and adaptability. This review discusses the potential of 3D bioprinting technologies for pharmaceutical studies.

Development of Cerium Oxide-Laden GelMA/PCL Scaffolds for Periodontal Tissue Engineering.

This study investigated gelatin methacryloyl (GelMA) and polycaprolactone (PCL) blend scaffolds incorporating cerium oxide (CeO) nanoparticles at concentrations of 0%, 5%, and 10% w/w via electrospinning for periodontal tissue engineering. The impact of photocrosslinking on these scaffolds was evaluated by comparing crosslinked (C) and non-crosslinked (NC) versions. Methods included Fourier transform infrared spectroscopy (FTIR) for chemical analysis, scanning electron microscopy (SEM) for fiber morphology/diameters, and assessments of swelling capacity, degradation profile, and biomechanical properties. Biological evaluations with alveolar bone-derived mesenchymal stem cells (aBMSCs) and human gingival fibroblasts (HGFs) encompassed tests for cell viability, mineralized nodule deposition (MND), and collagen production (CP). Statistical analysis was performed using Kruskal-Wallis or ANOVA/post-hoc tests (α = 5%). Results indicate that C scaffolds had larger fiber diameters (~250 nm) compared with NC scaffolds (~150 nm). NC scaffolds exhibited higher swelling capacities than C scaffolds, while both types demonstrated significant mass loss (~50%) after 60 days (p < 0.05). C scaffolds containing CeO showed increased Young's modulus and tensile strength than NC scaffolds. Cells cultured on C scaffolds with 10% CeO exhibited significantly higher metabolic activity (>400%, p < 0.05) after 7 days among all groups. Furthermore, CeO-containing scaffolds promoted enhanced MND by aBMSCs (>120%, p < 0.05) and increased CP in 5% CeO scaffolds for both variants (>180%, p < 0.05). These findings underscore the promising biomechanical properties, biodegradability, cytocompatibility, and enhanced tissue regenerative potential of CeO-loaded GelMA/PCL scaffolds for periodontal applications.

Biofabrication Strategies for Oral Soft Tissue Regeneration.

Gingival recession, a prevalent condition affecting the gum tissues, is characterized by the exposure of tooth root surfaces due to the displacement of the gingival margin. This review explores conventional treatments, highlighting their limitations and the quest for innovative alternatives. Importantly, it emphasizes the critical considerations in gingival tissue engineering leveraging on cells, biomaterials, and signaling factors. Successful tissue-engineered gingival constructs hinge on strategic choices such as cell sources, scaffold design, mechanical properties, and growth factor delivery. Unveiling advancements in recent biofabrication technologies like 3D bioprinting, electrospinning, and microfluidic organ-on-chip systems, this review elucidates their precise control over cell arrangement, biomaterials, and signaling cues. These technologies empower the recapitulation of microphysiological features, enabling the development of gingival constructs that closely emulate the anatomical, physiological, and functional characteristics of native gingival tissues. The review explores diverse engineering strategies aiming at the biofabrication of realistic tissue-engineered gingival grafts. Further, the parallels between the skin and gingival tissues are highlighted, exploring the potential transfer of biofabrication approaches from skin tissue regeneration to gingival tissue engineering. To conclude, the exploration of innovative biofabrication technologies for gingival tissues and inspiration drawn from skin tissue engineering look forward to a transformative era in regenerative dentistry with improved clinical outcomes.

GelMA/TCP nanocomposite scaffold for vital pulp therapy.

Pulp capping is a necessary procedure for preserving the vitality and health of the dental pulp, playing a crucial role in preventing the need for root canal treatment or tooth extraction. Here, we developed an electrospun gelatin methacryloyl (GelMA) fibrous scaffold incorporating beta-tricalcium phosphate (TCP) particles for pulp capping. A comprehensive morphological, physical-chemical, and mechanical characterization of the engineered fibrous scaffolds was performed. In vitro bioactivity, cell compatibility, and odontogenic differentiation potential of the scaffolds in dental pulp stem cells (DPSCs) were also evaluated. A pre-clinical in vivo model was used to determine the therapeutic role of the GelMA/TCP scaffolds in promoting hard tissue formation. Morphological, chemical, and thermal analyses confirmed effective TCP incorporation in the GelMA nanofibers. The GelMA+20%TCP nanofibrous scaffold exhibited bead-free morphology and suitable mechanical and degradation properties. In vitro, GelMA+20%TCP scaffolds supported apatite-like formation, improved cell spreading, and increased deposition of mineralization nodules. Gene expression analysis revealed upregulation of ALPL, RUNX2, COL1A1, and DMP1 in the presence of TCP-laden scaffolds. In vivo, analyses showed mild inflammatory reaction upon scaffolds' contact while supporting mineralized tissue formation. Although the levels of Nestin and DMP1 proteins did not exceed those associated with the clinical reference treatment (i.e., mineral trioxide aggregate), the GelMA+20%TCP scaffold exhibited comparable levels, thus suggesting the emergence of differentiated odontoblast-like cells capable of dentin matrix secretion. Our innovative GelMA/TCP scaffold represents a simplified and efficient alternative to conventional pulp-capping biomaterials. STATEMENT OF SIGNIFICANCE: Vital pulp therapy (VPT) aims to preserve dental pulp vitality and avoid root canal treatment. Biomaterials that bolster mineralized tissue regeneration with ease of use are still lacking. We successfully engineered gelatin methacryloyl (GelMA) electrospun scaffolds incorporated with beta-tricalcium phosphate (TCP) for VPT. Notably, electrospun GelMA-based scaffolds containing 20% (w/v) of TCP exhibited favorable mechanical properties and degradation, cytocompatibility, and mineralization potential indicated by apatite-like structures in vitro and mineralized tissue deposition in vivo, although not surpassing those associated with the standard of care. Collectively, our innovative GelMA/TCP scaffold represents a simplified alternative to conventional pulp capping materials such as MTA and Biodentine™ since it is a ready-to-use biomaterial, requires no setting time, and is therapeutically effective.

Integration of Melt Electrowritten Polymeric Scaffolds and Bioprinting for Epithelial Healing via Localized Periostin Delivery.

Management of skin injuries imposes a substantial financial burden on patients and hospitals, leading to diminished quality of life. Periostin (rhOSF), an extracellular matrix component, regulates cell function, including a proliferative healing phase, representing a key protein to promote wound healing. Despite its proven efficacy in vitro, there is a lack of scaffolds that facilitate the in situ delivery of rhOSF. In addition, there is a need for a scaffold to not only support cell growth, but also to resist the mechanical forces involved in wound healing. In this work, we synthesized rhOSF-loaded mesoporous nanoparticles (MSNs) and incorporated them into a cell-laden gelatin methacryloyl (GelMA) ink that was bioprinted into melt electrowritten poly(ε-caprolactone) (PCL) microfibrous (MF-PCL) meshes to develop mechanically competent constructs. Diffraction light scattering (DLS) analysis showed a narrow nanoparticle size distribution with an average size of 82.7 ± 13.2 nm. The rhOSF-loaded hydrogels showed a steady and controlled release of rhOSF over 16 days at a daily dose of ∼40 ng/mL. Compared with blank MSNs, the incorporation of rhOSF markedly augmented cell proliferation, underscoring its contribution to cellular performance. Our findings suggest a promising approach to address challenges such as prolonged healing, offering a potential solution for developing robust, biocompatible, and cell-laden grafts for burn wound healing applications.

Biodegradable electrospun poly(L-lactide-co-ε-caprolactone)/polyethylene glycol/bioactive glass composite scaffold for bone tissue engineering.

The field of tissue engineering has witnessed significant advancements in recent years, driven by the pursuit of innovative solutions to address the challenges of bone regeneration. In this study, we developed an electrospun composite scaffold for bone tissue engineering. The composite scaffold is made of a blend of poly(L-lactide-co-ε-caprolactone) (PLCL) and polyethylene glycol (PEG), with the incorporation of calcined and lyophilized silicate-chlorinated bioactive glass (BG) particles. Our investigation involved a comprehensive characterization of the scaffold's physical, chemical, and mechanical properties, alongside an evaluation of its biological efficacy employing alveolar bone-derived mesenchymal stem cells. The incorporation of PEG and BG resulted in elevated swelling ratios, consequently enhancing hydrophilicity. Thermal gravimetric analysis confirmed the efficient incorporation of BG, with the scaffolds demonstrating thermal stability up to 250°C. Mechanical testing revealed enhanced tensile strength and Young's modulus in the presence of BG; however, the elongation at break decreased. Cell viability assays demonstrated improved cytocompatibility, especially in the PLCL/PEG+BG group. Alizarin red staining indicated enhanced osteoinductive potential, and fluorescence analysis confirmed increased cell adhesion in the PLCL/PEG+BG group. Our findings suggest that the PLCL/PEG/BG composite scaffold holds promise as an advanced biomaterial for bone tissue engineering.

Strontium-Doped Bioglass-Laden Gelatin Methacryloyl Hydrogels for Vital Pulp Therapy.

This study aimed to develop gelatin methacryloyl (GelMA)-injectable hydrogels incorporated with 58S bioactive glass/BG-doped with strontium for vital pulp therapy applications. GelMA hydrogels containing 0% (control), 5%, 10%, and 20% BG (w/v) were prepared. Their morphological and chemical properties were evaluated by scanning electron microscopy/SEM, energy dispersive spectroscopy/EDS, and Fourier transform infrared spectroscopy/FTIR (n = 3). Their swelling capacity and degradation ratio were also measured (n = 4). Cell viability (n = 8), mineralized matrix formation, cell adhesion, and spreading (n = 6) on DPSCs were evaluated. Data were analyzed using ANOVA/post hoc tests (α = 5%). SEM and EDS characterization confirmed the incorporation of BG particles into the hydrogel matrix, showing GelMA's (C, O) and BG's (Si, Cl, Na, Sr) chemical elements. FTIR revealed the main chemical groups of GelMA and BG, as ~1000 cm-1 corresponds to Si-O and ~1440 cm-1 to C-H. All the formulations were degraded by day 12, with a lower degradation ratio observed for GelMA+BG20%. Increasing the concentration of BG resulted in a lower mass swelling ratio. Biologically, all the groups were compatible with cells (p > 0.6196), and cell adhesion increased over time, irrespective of BG concentration, indicating great biocompatibility. GelMA+BG5% demonstrated a higher deposition of mineral nodules over 21 days (p < 0.0001), evidencing the osteogenic potential of hydrogels. GelMA hydrogels incorporated with BG present great cytocompatibility, support cell adhesion, and have a clinically relevant degradation profile and suitable mineralization potential, supporting their therapeutic potential as promising biomaterials for pulp capping.

Injectable cell-laden hybrid bioactive scaffold containing bioactive glass microspheres.

The rising incidence of bone disorders has resulted in the need for minimally invasive therapies to meet this demand. Injectable bioactive filler, alone or with cells, could be applied in a minimally invasive manner to fulfill irregular cavities in non-load bearing sites, which do not require high mechanical properties. Thermosensitive chitosan hydrogels that transition from a liquid to a mechanically stable solid at body temperature provide interesting features as in-situ injectable cytocompatible biomaterials, but they are not osteoconductive. Osteoconductivity can be applied in combination with bioactive ceramics e.g., 45S5-Bioglass® (BG). However, BG addition in chitosan hydrogels results in pH elevation, due to rapid ions release, which adversely affects gel formation, mechanical properties, and cytocompatibility. To address this, we created hybrid hydrogels, where BG is concentrated in chitosan-based microbeads, incorporated in in-situ gelling chitosan hydrogels. We then compared the hybrid hydrogels' properties to chitosan hydrogels with homogenously distributed BG. By varying the stirred emulsification process, BG percentage, and CH formulation, we could tune the microbeads' properties. Incorporation of BG microbeads drastically improved the hydrogel's compressive modulus in comparison to homogeneously distributed BG. It also strongly increased the survival and metabolic activities of encapsulated cells. Calcium/phosphate increase on BG microbeads suggests hydroxyapatite formation. The small diameter of microbeads allows minimally invasive injection through small needles. The feasibility of freezing and thawing microbeads provides the possibility of long-term storage for potential clinical applications. These data indicate that this hybrid hydrogel forms a promising injectable cell-laden bioactive biomaterial for the treatment of unloaded bone defects.

Recent advances in smart wearable sensors as electronic skin.

Flexible and multifunctional electronic devices and soft robots inspired by human organs, such as skin, have many applications. However, the emergence of electronic skins (e-skins) or textiles in biomedical engineering has made a great revolution in a myriad of people's lives who suffer from different types of diseases and problems in which their skin and muscles lose their appropriate functions. In this review, recent advances in the sensory function of the e-skins are described. Furthermore, we have categorized them from the sensory function perspective and highlighted their advantages and limitations. The categories are tactile sensors (including capacitive, piezoresistive, piezoelectric, triboelectric, and optical), temperature, and multi-sensors. In addition, we summarized the most recent advancements in sensors and their particular features. The role of material selection and structure in sensory function and other features of the e-skins are also discussed. Finally, current challenges and future prospects of these systems towards advanced biomedical applications are elaborated.

Engineered Biomimetic Membranes for Organ-on-a-Chip.

Organ-on-a-chip (OOC) systems are engineered nanobiosystems to mimic the physiochemical environment of a specific organ in the body. Among various components of OOC systems, biomimetic membranes have been regarded as one of the most important key components to develop controllable biomimetic bioanalysis systems. Here, we review the preparation and characterization of biomimetic membranes in comparison with the features of the extracellular matrix. After that, we review and discuss the latest applications of engineered biomimetic membranes to fabricate various organs on a chip, such as liver, kidney, intestine, lung, skin, heart, vasculature and blood vessels, brain, and multiorgans with perspectives for further biomedical applications.

Biomaterials in bone and mineralized tissue engineering using 3D printing and bioprinting technologies.

This review focuses on recently developed printable biomaterials for bone and mineralized tissue engineering. 3D printing or bioprinting is an advanced technology to design and fabricate complex functional 3D scaffolds, mimicking native tissue forin vivoapplications. We categorized the biomaterials into two main classes: 3D printing and bioprinting. Various biomaterials, including natural, synthetic biopolymers and their composites, have been studied. Biomaterial inks or bioinks used for bone and mineralized tissue regeneration include hydrogels loaded with minerals or bioceramics, cells, and growth factors. In 3D printing, the scaffold is created by acellular biomaterials (biomaterial inks), while in 3D bioprinting, cell-laden hydrogels (bioinks) are used. Two main classes of bioceramics, including bioactive and bioinert ceramics, are reviewed. Bioceramics incorporation provides osteoconductive properties and induces bone formation. Each biopolymer and mineral have its advantages and limitations. Each component of these composite biomaterials provides specific properties, and their combination can ameliorate the mechanical properties, bioactivity, or biological integration of the 3D printed scaffold. Present challenges and future approaches to address them are also discussed.

Prevascularized Micro-/Nano-Sized Spheroid/Bead Aggregates for Vascular Tissue Engineering.

Efficient strategies to promote microvascularization in vascular tissue engineering, a central priority in regenerative medicine, are still scarce; nano- and micro-sized aggregates and spheres or beads harboring primitive microvascular beds are promising methods in vascular tissue engineering. Capillaries are the smallest type and in numerous blood vessels, which are distributed densely in cardiovascular system. To mimic this microvascular network, specific cell components and proangiogenic factors are required. Herein, advanced biofabrication methods in microvascular engineering, including extrusion-based and droplet-based bioprinting, Kenzan, and biogripper approaches, are deliberated with emphasis on the newest works in prevascular nano- and micro-sized aggregates and microspheres/microbeads.

Emerging Phospholipid Nanobiomaterials for Biomedical Applications to Lab-on-a-Chip, Drug Delivery, and Cellular Engineering.

The design of advanced nanobiomaterials to improve analytical accuracy and therapeutic efficacy has become an important prerequisite for the development of innovative nanomedicines. Recently, phospholipid nanobiomaterials including 2-methacryloyloxyethyl phosphorylcholine (MPC) have attracted great attention with remarkable characteristics such as resistance to nonspecific protein adsorption and cell adhesion for various biomedical applications. Despite many recent reports, there is a lack of comprehensive review on the phospholipid nanobiomaterials from synthesis to diagnostic and therapeutic applications. Here, we review the synthesis and characterization of phospholipid nanobiomaterials focusing on MPC polymers and highlight their attractive potentials for applications in micro/nanofabricated fluidic devices, biosensors, lab-on-a-chip, drug delivery systems (DDSs), COVID-19 potential usages for early diagnosis and even treatment, and artificial extracellular matrix scaffolds for cellular engineering.

A rheological approach to assess the printability of thermosensitive chitosan-based biomaterial inks.

For extrusion-based bioprinting, the inks must be printable and rapidly present sufficient mechanical properties to support additional layers and provide a cohesive, manipulable structure. Thermosensitive hydrogels may be interesting candidates. However, the use of these materials is particularly challenging, since their rheological properties evolve with time and temperature. In this work, a rheological approach to characterize the printability of chitosan-based thermosensitive inks was developed. The method consists of evaluating: (1) the gelation kinetic at room temperature and at 37 °C; (2) shear-thinning behavior to estimate the shear rate applied during printing as a function of printing parameters; and (3) the viscosity after shear removal (recovery test) to simulate behaviour after biomaterial deposition. Hydrogels containing 2 and 3% w v-1 chitosan, combined with different gelling agents (sodium hydrogen carbonate (SHC), phosphate buffer, beta-glycerophosphate (BGP)) were tested, and compared with alginate/gelatin bioink as controls. To correlate the rheological studies with real printing conditions, a 3D-Discovery bioprinter was used to print hydrogels and the visual aspect of the printed structure was observed. Unconfined compressive tests were carried out to study the impact of applied shear rate during printing on the mechanical properties of printed structures. All pre-hydrogel solutions presented shear-thinning properties. The recovery of viscosity was found to depend on the hydrogel formulation, as well as the level of shear rate and the state of gelation at the time of printing. Formulations made with SHC and phosphate buffer presented too rapid gelation and phase separation, leading to poor printing results. One particularly promising formulation composed of SHC and BGP, when printed at a shear rate of 140 s-1, before its gelation time (t g ⩽ 15 min), resulted in good printability and 3D structures with rigidity comparable with the alginate/gelatin bioink. The methodology introduced in this paper could be used to evaluate the printability of other time- and temperature-dependent biomaterial inks in the future.

Biomaterials and tissue engineering for scar management in wound care.

Scars are a natural and unavoidable result from most wound repair procedures and the body's physiological healing response. However, they scars can cause considerable functional impairment and emotional and social distress. There are different forms of treatments that have been adopted to manage or eliminate scar formation. This review covers the latest research in the past decade on using either natural agents or synthetic biomaterials in treatments for scar reduction.