Platform For Tissue Engineering Research Articles

Bioprinting of gelatin-based materials for orthopedic application.

Bio-printed hydrogels have evolved as one of the best regenerative medicine and tissue engineering platforms due to their outstanding cell-friendly microenvironment. A correct hydrogel ink formulation is critical for creating desired scaffolds that have better fidelity after printing. Gelatin and its derivatives have sparked intense interest in various biomedical sectors because of their biocompatibility, biodegradability, ease of functionalization, and rapid gelling tendency. As a result, this report emphasizes the relevance of gelatin-based hydrogel in fabricating bio-printed scaffolds for orthopedic applications. Starting with what hydrogels and bio-printing are all about. We further summarized the different gelatin-based bio-printing techniques explored for orthopedic applications, including a few recent studies. We also discussed the suitability of gelatin as a biopolymer for both 3D and 4D printing materials. As extrusion is one of the most widely used techniques for bio-printing gelatin-based, we summarize the rheological features of gelatin-based bio-ink. Lastly, we also elaborate on the recent bio-printed gelatin-based studies for orthopedics applications, the potential clinical translation issues, and research possibilities.

Multifunctional MXene-Based Platforms for Soft and Bone Tissue Regeneration and Engineering.

MXenes and their composites hold great promise in the field of soft and bone tissue regeneration and engineering (TRE). However, there are challenges that need to be overcome, such as ensuring biocompatibility and controlling the morphologies of MXene-based scaffolds. The future prospects of MXenes in TRE include enhancing biocompatibility through surface modifications, developing multifunctional constructs, and conducting in vivo studies for clinical translation. The purpose of this perspective about MXenes and their composites in soft and bone TRE is to critically evaluate their potential applications and contributions in this field. This perspective aims to provide a comprehensive analysis of the challenges, advantages, limitations, and future prospects associated with the use of MXenes and their composites for soft and bone TRE. By examining the existing literature and research, the review seeks to consolidate the current knowledge and highlight the key findings and advancements in MXene-based TRE. It aims to contribute to the understanding of MXenes' role in promoting soft and bone TRE, addressing the challenges faced in terms of biocompatibility, morphology control, and tissue interactions.

Ion‐Engineered Microcryogels via Osteogenesis‐Angiogenesis Coupling and Inflammation Reversing Augment Vascularized Bone Regeneration

AbstractNative bone inherently requires a balanced ionic microenvironment to maintain bone homeostasis. Hence, scaffolds designed for the sustained release of therapeutic ions into bone defects hold great promise for bone regeneration. Magnesium (Mg) and silicon (Si) are essential elements, which play crucial roles in the process of bone regeneration, impacting immunomodulation, angiogenesis, and osteogenesis. Herein, porous cryogel‐type organic–inorganic composite microspheres are developed as injectable microscaffolds (denoted as GMN). GMN enables sustained release of Mg/Si ions at an optimized ratio, achieving the most significant synergistic effect on vascularized bone regeneration. Various conditioned media are obtained to explore angiogenesis‐osteogenesis coupling, as well as the crosstalk between bone marrow mesenchymal stromal cells (BMSCs) and macrophages. Meanwhile, autocrine and paracrine effects simultaneously play synergistic modulating functions in determining cell fates under the guidance of Mg/Si ions and biofactors secreted by cells. Overall, the Mg/Si ion‐engineering microscaffolds create a conducive microenvironment to efficiently augment the regeneration of vascularized bone tissue in vivo, offering a versatile platform for tissue engineering.

A systematic review on green and natural polymeric nanofibers for biomedical applications

Electrospinning is the simplest technique to produce ultrathin nanofibers, which enables the use of nanotechnology in various applications. Nanofibrous materials produced through electrospinning have garnered significant attention in biomedical applications due to their unique properties and versatile potential. In recent years, there has been a growing emphasis on incorporating sustainability principles into material design and production. However, electrospun nanofibers, owing to their reliance on solvents associated with significant drawbacks like toxicity, flammability, and disposal challenges, frequently fall short of meeting environmentally friendly standards. Due to the limited solvent choices and heightened concerns for safety and hygiene in modern living, it becomes imperative to carefully assess the implications of employing electrospun nanofibers in diverse applications and consumer products. This systematic review aims to comprehensively assess the current state of research and development in the field of “green and natural” electrospun polymer nanofibers as well as more fascinating and eco-friendly commercial techniques, solvent preferences, and other green routes that respect social and legal restrictions tailored for biomedical applications. We explore the utilization of biocompatible and biodegradable polymers sourced from renewable feedstocks, eco-friendly processing techniques, and the evaluation of environmental impacts. Our review highlights the potential of green and natural electrospun nanofibers to address sustainability concerns while meeting the demanding requirements of various biomedical applications, including tissue engineering, drug delivery, wound healing, and diagnostic platforms. We analyze the advantages, challenges, and future prospects of these materials, offering insights into the evolving landscape of environmentally responsible nanofiber technology in the biomedical field.

Control of Neuronal Survival and Development Using Conductive Diamond.

This study demonstrates the control of neuronal survival and development using nitrogen-doped ultrananocrystalline diamond (N-UNCD). We highlight the role of N-UNCD in regulating neuronal activity via near-infrared illumination, demonstrating the generation of stable photocurrents that enhance neuronal survival and neurite outgrowth and foster a more active, synchronized neuronal network. Whole transcriptome RNA sequencing reveals that diamond substrates improve cellular-substrate interaction by upregulating extracellular matrix and gap junction-related genes. Our findings underscore the potential of conductive diamond as a robust and biocompatible platform for noninvasive and effective neural tissue engineering.

Conductive GelMA/alginate/polypyrrole/graphene hydrogel as a potential scaffold for cardiac tissue engineering; Physiochemical, mechanical, and biological evaluations

Cardiac failure can be a life-threatening condition that, if left untreated, can have significant consequences. Functional hydrogel has emerged as a promising platform for cardiac tissue engineering. In the proposed study, gelatin methacrylate (GelMA) and alginate, as a primary matrix to maintain cell viability and proliferation, and polypyrrole and carboxyl-graphene, to improve mechanical and electrical properties, are thoroughly evaluated. Initially, a polymer blend of GelMA/Alginate (1:1) was prepared, and then the addition of 2–5 wt% of polypyrrole was evaluated. Next, the effect of incorporating graphene-carboxyl nanosheets (1, 2, and 3 wt%) within the optimized scaffold with 2 wt% polypyrrole was thoroughly studied. The electrical conductivity of the hydrogels was significantly enhanced from 0.0615 ± 0.007 S/cm in GelMA/alginate to 0.124 ± 0.04 S/cm with the addition of 5 wt% polypyrrole. Also, 3 wt% carboxyl graphene improved the electrical conductivity to 0.27 ± 0.09 S/cm. The compressive strength of carboxyl-graphene-containing hydrogel was in the range of 175 to 520 kPa, and tensile strength was 61 and 133 kPa. The simplicity and low-cost fabrication, tunable mechanical properties, optimal electrical conductivity, blood compatibility, and non-cytotoxicity of GelMA/alginate/polypyrrole/graphene biocomposite hydrogel is a promising construct for cardiac tissue engineering.

Mesenchymal stem cells aligned and stretched in self-assembling peptide hydrogels

The presented research highlights a novel approach using fmoc-protected peptide hydrogels for the encapsulation and stretching of mesenchymal stem cells (MSCs). This study utilized a custom mechanical stretching device with a PDMS chamber to stretch human MSCs encapsulated in Fmoc hydrogels. The study assessed the influence of various solvents on the self-assembly and mechanical properties of the hydrogels, and MSC viability and alignment. Particularly we focused on fluorenylmethoxycarbonyl-diphenylalanine (Fmoc-FF) prepared in dimethyl sulfoxide (DMSO), hexafluoro-2-propanol (HFP), and deionized water (DiH2O).Through molecular self-assembly of the peptide sequence into β-sheets connected by π-π aromatic stacking of F–F groups, the peptide hydrogel was found to form a stiff, hydrated gel with nanofiber morphology and a compressive modulus ranging from 174 to 277 Pa. Therefore, this hydrogel can mimic certain critical features of the extracellular matrix and collagen. Evaluations of MSCs cultured on the peptide hydrogels, including viability, morphology, and alignment assessments using various staining techniques, demonstrated that 3D-cultured MSCs in Fmoc-FF/HFP and Fmoc-FF/DMSO, followed by mechanical stretching, exhibited elongated morphology with distinct microfilament fibers compared to the control cells, which maintained a round and spherical F-actin shape. Notably, peptide gels with a concentration of 5 mM maintained 100 % MSC viability.The findings indicate the potential and specific conditions for successful cell encapsulation and alignment within peptide hydrogels, highlighting a promising tissue engineering platform through the encapsulation of MSCs in peptide nanofibers followed by a stretching process. By enhancing our understanding of MSC-peptide hydrogel interactions, this research contributes to the development of biomaterials tailored for regenerative medicine.

In Vitro and In Vivo Evaluation of 3D Printed Poly(Ethylene Glycol) Dimethacrylate-Based Photocurable Hydrogel Platform for Bone Tissue Engineering.

This study focuses to develop a unique hybrid hydrogel bioink formulation that incorporates poly(ethylene glycol) dimethacrylate (PEGDMA), gelatin (Gel), and methylcellulose (MC). This formulation achieves the necessary viscosity for extrusion-based three-dimensional (3D) printing of scaffolds intended for bone regeneration. After thorough optimization of the hybrid bioink system with Gel, three distinct scaffold groups are investigated in vitro: 0%, 3%, and 6% (w/v) Gel. These scaffold groups are examined for their morphology, mechanical strength, biodegradation, in vitro cell proliferation and differentiation, and in vivo bone formation using a rat cranial defect model. Among these scaffold compositions, the 3% Gel scaffold exhibits the most favorable characteristics, prompting further evaluation as a rat mesenchymal stem cell (rMSC) carrier in a critical-size cranial defect within a Lewis rat model. The compressive strength of all three scaffold groups range between 1 and 2MPa. Notably, the inclusion of Gel in the scaffolds leads to enhanced bioactivity and cell adhesion. The Gel-containing scaffolds notably amplify osteogenic differentiation, as evidenced by alkaline phosphatase (ALP) and Western blot analyses. The in vivo results, as depicted by microcomputed tomography, showcase augmented osteogenesis within cell-seeded scaffolds, thus validating this innovative PEGDMA-based scaffold system as a promising candidate for cranial bone defect healing.

Elecrospun nanofibers: The versatile platform as a drug delivery system in healthcare

Nanofibers are fabricated using several polymers, carbon, and semiconductors having cross-sectional diameter less than 100 nm. The exceptional state-of-art 3-D architecture and remarkable physico-chemical characteristics of nanofibers have found versatile applications in various fields. This review provides insights into preparation methods and applications of nanofibers. Nanofibers may be prepared by several methods like electrospinning, self-assembly, phase separation, anti-solvent induced polymer precipitation, etc.; however, electrospinning is the mostly employed technique for nanofibers fabrication. Electrospinning is known to be an electrohydrodynamic process in which a liquid droplet is electrically charged, resulting in the formation of a jet, which is then stretched and elongated to produce fiber(s). The nanofibers have an enormous surface area to volume ratio, resulting in a high percentage of porosity of nanofibrous mats. These characteristics allowed higher drug loading, especially biological macromolecules albeit of their large molecular size. Hence, the nanofibers fetched attention for their application in the healthcare and biomedical engineering sector for the treatment and management of diabetic foot ulcers, cancer, microbiological disorders, wound healing, and cardiovascular diseases. Additionally, nanofiber mats possess a 3-D architecture as well as nanofibers can either be oriented randomly or in an ordered fashion for tissue engineering-related drug delivery applications. The above-mentioned peculiarities enables nanofiber scaffolds to mimics human tissues and hence are being explored for their applications in tissue engineering, artificial organ, filtering aids for microbes, cosmeceutical industry, etc. Conclusively, the nanofibers can serve as an excellent drug delivery and tissue-engineering platform for the treatment in healthcare.

Noninvasive and Microinvasive Nanoscale Drug Delivery Platforms for Hard Tissue Engineering.

Bone tissue plays a crucial role in protecting internal organs and providing structural support and locomotion of the body. Treatment of hard tissue defects and medical conditions due to physical injuries, genetic disorders, aging, metabolic syndromes, and infections is more often a complex and drawn out process. Presently, dealing with hard-tissue-based clinical problems is still mostly conducted via surgical interventions. However, advances in nanotechnology over the last decades have led to shifting trends in clinical practice toward noninvasive and microinvasive methods. In this review article, recent advances in the development of nanoscale platforms for bone tissue engineering have been reviewed and critically discussed to provide a comprehensive understanding of the advantages and disadvantages of noninvasive and microinvasive methods for treating medical conditions related to hard tissue regeneration and repair.

Stimuli-responsive hydrogels: smart state of-the-art platforms for cardiac tissue engineering.

Biomedicine and tissue regeneration have made significant advancements recently, positively affecting the whole healthcare spectrum. This opened the way for them to develop their applications for revitalizing damaged tissues. Thus, their functionality will be restored. Cardiac tissue engineering (CTE) using curative procedures that combine biomolecules, biomimetic scaffolds, and cells plays a critical part in this path. Stimuli-responsive hydrogels (SRHs) are excellent three-dimensional (3D) biomaterials for tissue engineering (TE) and various biomedical applications. They can mimic the intrinsic tissues' physicochemical, mechanical, and biological characteristics in a variety of ways. They also provide for 3D setup, adequate aqueous conditions, and the mechanical consistency required for cell development. Furthermore, they function as competent delivery platforms for various biomolecules. Many natural and synthetic polymers were used to fabricate these intelligent platforms with innovative enhanced features and specialized capabilities that are appropriate for CTE applications. In the present review, different strategies employed for CTE were outlined. The light was shed on the limitations of the use of conventional hydrogels in CTE. Moreover, diverse types of SRHs, their characteristics, assembly and exploitation for CTE were discussed. To summarize, recent development in the construction of SRHs increases their potential to operate as intelligent, sophisticated systems in the reconstruction of degenerated cardiac tissues.

Hydrocolloids of Egg White and Gelatin as a Platform for Hydrogel-Based Tissue Engineering.

Innovative materials are needed to produce scaffolds for various tissue engineering and regenerative medicine (TERM) applications, including tissue models. Materials derived from natural sources that offer low production costs, easy availability, and high bioactivity are highly preferred. Chicken egg white (EW) is an overlooked protein-based material. Whilst its combination with the biopolymer gelatin has been investigated in the food technology industry, mixed hydrocolloids of EW and gelatin have not been reported in TERM. This paper investigates these hydrocolloids as a suitable platform for hydrogel-based tissue engineering, including 2D coating films, miniaturized 3D hydrogels in microfluidic devices, and 3D hydrogel scaffolds. Rheological assessment of the hydrocolloid solutions suggested that temperature and EW concentration can be used to fine-tune the viscosity of the ensuing gels. Fabricated thin 2D hydrocolloid films presented globular nano-topography and in vitro cell work showed that the mixed hydrocolloids had increased cell growth compared with EW films. Results showed that hydrocolloids of EW and gelatin can be used for creating a 3D hydrogel environment for cell studies inside microfluidic devices. Finally, 3D hydrogel scaffolds were fabricated by sequential temperature-dependent gelation followed by chemical cross-linking of the polymeric network of the hydrogel for added mechanical strength and stability. These 3D hydrogel scaffolds displayed pores, lamellae, globular nano-topography, tunable mechanical properties, high affinity for water, and cell proliferation and penetration properties. In conclusion, the large range of properties and characteristics of these materials provide a strong potential for a large variety of TERM applications, including cancer models, organoid growth, compatibility with bioprinting, or implantable devices.

Multimodal therapy strategy based on a bioactive hydrogel for repair of spinal cord injury

Traumatic spinal cord injury results in permanent and serious neurological impairment, but there is no effective treatment yet. Tissue engineering approaches offer great potential for the treatment of SCI, but spinal cord complexity poses great challenges. In this study, the composite scaffold consists of a hyaluronic acid-based hydrogel, decellularized brain matrix (DBM), and bioactive compounds such as polydeoxyribonucleotide (PDRN), tumor necrosis factor-α/interferon-γ primed mesenchymal stem cell-derived extracellular vesicles (TI-EVs), and human embryonic stem cell-derived neural progenitor cells (NPC). The composite scaffold showed significant effects on regenerative prosses including angiogenesis, anti-inflammation, anti-apoptosis, and neural differentiation. In addition, the composite scaffold (DBM/PDRN/TI-EV/NPC@Gel) induced an effective spinal cord regeneration in a rat spinal cord transection model. Therefore, this multimodal approach using an integrated bioactive scaffold coupled with biochemical cues from PDRN and TI-EVs could be used as an advanced tissue engineering platform for spinal cord regeneration.

Osteogenic differentiation of human adipose-derived mesenchymal stem cells in a bisphosphonate-functionalized polycaprolactone/gelatin scaffold

Recent trends in bone tissue engineering have focused on the development of biomimetic constructs with appropriate mechanical and physiochemical properties. Here, we report the fabrication of an innovative biomaterial scaffold based on a new bisphosphonate-containing synthetic polymer combined with gelatin. To this end, zoledronate (ZA)-functionalized polycaprolactone (PCL-ZA) was synthesized by a chemical grafting reaction. After adding gelatin to the PCL-ZA polymer solution, the porous PCL-ZA/gelatin scaffold was fabricated by the freeze-casting method. A scaffold with aligned pores and a porosity of 82.04 % was obtained. During in vitro biodegradability test, 49 % of its initial weight lost after 5 weeks. The elastic modulus of the PCL-ZA/gelatin scaffold was 31.4 MPa, and its tensile strength was 4.2 MPa. Based on the results of MTT assay, the scaffold had good cytocompatibility with human Adipose-Derived Mesenchymal Stem Cells (hADMSCs). Furthermore, cells grown in PCL-ZA/gelatin scaffold showed the highest mineralization and ALP activity compared to other test groups. Results of the RT-PCR test revealed that RUNX2, COL 1A1, and OCN genes were expressed in PCL-ZA/gelatin scaffold at the highest level, suggesting its good osteoinductive capacity. These results revealed that PCL-ZA/gelatin scaffold could be considered a proper biomimetic platform for bone tissue engineering.

Retracted: A Photoclick Thiol‐Ene Collagen‐Based Hydrogel Platform for Skeletal Muscle Tissue Engineering

AbstractUV‐cured collagen‐based hydrogels hold promise in skeletal muscle regeneration due to their soft elastic properties and porous architecture. However, the complex triple helix conformation of collagen and environmental conditions, i.e., molecular oxygen, pose risks to reaction controllability, wet‐state integrity, and reproducibility. To address this challenge, a photoclick hydrogel platform is presented through an oxygen‐insensitive thiol‐ene reaction between 2‐iminothiolane (2IT)‐functionalized type I collagen and multiarm, nonhomopolymerizable norbornene‐terminated polyethylene glycol (PEG). UV‐induced network formation is demonstrated by oscillatory time sweeps on the reacting thiol‐ene mixture, so that significantly increased storage moduli are measured and adjusted depending on the photoinitiator concentration. Variations in PEG functionality (4‐arm and 8‐arm) and PEG content generate hydrogels with skeletal muscle native stiffness (Ec = 1.3 ± 0.2‒11.5 ± 0.9 kPa), diffusion‐controlled swelling behavior and erosion‐driven degradability. In vitro, no cytotoxic effect is detected on C2C12 murine myoblasts, while myogenic differentiation is successfully accomplished on hydrogel‐seeded cells in then low serum culture medium. In vivo, 7 d subcutaneous implantation of selected thiol‐ene hydrogel in rats reveal higher cell infiltration, blood vessel formation, and denser tissue interface compared to a clinical gold standard collagen matrix (Mucograft, a trademark of Geistlich Biomaterials). These results, therefore, support the applicability and further development of this hydrogel platform for skeletal muscle regeneration.

Preparation of Cell-Loaded Microbeads as Stable and Injectable Delivery Platforms for Tissue Engineering

Cell transplants in therapeutic studies do not preserve their long-term function inside the donor body. In mesenchymal stem cell (MSC) transplants, transplanted cells disperse through the body and are prone to degradation by immune cells after the transplant process. Various strategies, such as usage of the immunosuppressive drugs to eliminate allograft rejection, are designed to increase the efficiency of cell therapy. Another strategy is the construction of biomimetic encapsulates using polymeric materials, which isolate stem cells and protect them from environmental effects. In this study, fibroblasts (L929) and MSCs were investigated for their improved viability and functionality once encapsulated inside the alginate microbeads under in vitro conditions for up to 12 days of incubation. Thus, uniform and injectable (<200 µm) cell-loaded microbeads were constructed by the electrostatically assisted spraying technique. Results showed that both L929 and MSCs cells continue their metabolic activity inside the microbeads during the incubation periods. Glucose consumption and lactic acid production levels of both cell lines were consistently observed. The released cell number on day 12 was found to be increased compared to day 0. Protein expression levels of both groups increased every day with the expected doubling rate. Hence, this strategy with a simple yet clever design to encapsulate either MSCs or L929 cells might outstand as a potential cell delivery platform for cell therapy-based tissue engineering.

Acoustic and Magnetic Stimuli-Based Three-Dimensional Cell Culture Platform for Tissue Engineering.

In a conventional two-dimensional (2D) culture method, cells are attached to the bottom of the culture dish and grow into a monolayer. These 2D culture methods are easy to handle, cost-effective, reproducible, and adaptable to growing many different types of cells. However, monolayer 2D cell culture conditions are far from those of natural tissue, indicating the need for a three-dimensional (3D) culture system. Various methods, such as hanging drop, scaffolds, hydrogels, microfluid systems, and bioreactor systems, have been utilized for 3D cell culture. Recently, external physical stimulation-based 3D cell culture platforms, such as acoustic and magnetic forces, were introduced. Acoustic waves can establish acoustic radiation force, which can induce suspended objects to gather in the pressure node region and aggregate to form clusters. Magnetic targeting consists of two components, a magnetically responsive carrier and a magnetic field gradient source. In a magnetic-based 3D cell culture platform, cells are aggregated by changing the magnetic force. Magnetic fields can manipulate cells through two different methods: positive magnetophoresis and negative magnetophoresis. Positive magnetophoresis is a way of imparting magnetic properties to cells by labeling them with magnetic nanoparticles. Negative magnetophoresis is a label-free principle-based method. 3D cell structures, such as spheroids, 3D network structures, and cell sheets, have been successfully fabricated using this acoustic and magnetic stimuli-based 3D cell culture platform. Additionally, fabricated 3D cell structures showed enhanced cell behavior, such as differentiation potential and tissue regeneration. Therefore, physical stimuli-based 3D cell culture platforms could be promising tools for tissue engineering.

Tissue-Engineered Injectable Gelatin-Methacryloyl Hydrogel-Based Adjunctive Therapy for Intervertebral Disc Degeneration.

Gelatin-methacryloyl (GelMA) hydrogels are photosensitive with good biocompatibility and adjustable mechanical properties. The GelMA hydrogel composite system is a prospective therapeutic material based on a tissue engineering platform for treating intervertebral disc (IVD) degeneration (IVDD). The potential application value of the GelMA hydrogel composite system in the treatment of IVDD mainly includes three aspects: first, optimization of the current clinical treatment methods, including conservative treatment and surgical treatment; second, regeneration of IVD cells to reverse or repair IVDD; and finally, IVDD instead of injury plays a biomechanical role. In this paper, we summarized and analyzed the preparation of GelMA hydrogels and their excellent biological characteristics as carriers and comprehensively demonstrated the research status and prospects of GelMA hydrogel composite systems in IVDD treatment. In addition, the challenges facing the application of GelMA hydrogel composite systems and the progress of research on new hydrogels modified by GelMA hydrogels are presented. Hopefully, this study will provide theoretical guidance for the future application of GelMA hydrogel composite systems in IVDD.

Evaluation of in vitro coculture of keratinocytes derived from foreskin and adipose-derived mesenchymal stem cells (AMSCs) on a multilayer oxygen-releasing electrospun scaffold based on PU/PCL.Sodium percarbonate (SPC)-gelatine/PU.

Despite significant advancements in tissue engineering and regenerative medicine during the last two decades, the fabrication of proper scaffolds with appropriate cells can still be considered a critical achievement in this field. Hypoxia is a major stumbling block to chronic wound healing, which restrains tissue engineering plans because a lack of oxygen may cause cell death. This study evaluated the cocultured human keratinocytes and human adipose-derived mesenchymal stem cells (AMSCs) on a multilayer oxygen-releasing electrospun scaffold based on PU/PCL.Sodium percarbonate (SPC)-gelatin/PU. The scaffold was characterized using Fourier transform infrared (FTIR) and scanning electron microscopy (SEM) methods. Flow cytometry confirmed mesenchymal stem cells, and then the 3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyltetrazolium bromide (MTT) assay and DAPI staining were used to assess the in vitro biocompatibility of the scaffold. The experimental results showed that the multilayer electrospun scaffold containing 2.5% SPC could efficiently produce oxygen. Furthermore, according to cell viability results, this structure makes a suitable substrate for the coculture of keratinocytes and AMSCs. Gene expression analysis of various markers such as Involucrin, Cytokeratin 10, and Cytokeratin 14 after 14 days confirmed that keratinocytes and AMSCs coculture on PU/PCL.SPC-gelatin/PUelectrospun scaffold promotes dermal differentiation and epithelial proliferation compared to keratinocytes single-cell culture. Therefore, our study supports using oxygen-releasing scaffolds as a potential strategy to hasten skin tissue regeneration. Based on the results, this structure is suggested as a promising candidate for cell-based skin tissue engineering. Given that the developed oxygen-generating polymeric electrospun scaffolds could be used as part of a future strategy for skin tissue engineering, the PU/PCL.SPC-gelatin/PUhybrid electrospun multilayer scaffold in combination with keratinocyte/AMSC coculture is proposed as an effective substrate for skin tissue engineering and regenerative medicine platforms.

IPSC-derived three-dimensional brain organoid models and neurotropic viral infections.

Progress in stem cell research has revolutionized the medical field for more than two decades. More recently, the discovery of induced pluripotent stem cells (iPSCs) has allowed for the development of advanced disease modeling and tissue engineering platforms. iPSCs are generated from adult somatic cells by reprogramming them into an embryonic-like state via the expression of transcription factors required for establishing pluripotency. In the context of the central nervous system (CNS), iPSCs have the potential to differentiate into a wide variety of brain cell types including neurons, astrocytes, microglial cells, endothelial cells, and oligodendrocytes. iPSCs can be used to generate brain organoids by using a constructive approachin three-dimensional (3D) culture in vitro. Recent advances in 3D brain organoid modeling have provided access to a better understanding of cell-to-cell interactions in disease progression, particularly with neurotropic viral infections. Neurotropic viral infections have been difficult to study in two-dimensional culture systems in vitro due to the lack of a multicellular composition of CNS cell networks. In recent years, 3D brain organoids have been preferred for modeling neurotropic viral diseases and have provided invaluable information for better understanding the molecular regulation of viral infection and cellular responses. Here we provide a comprehensive review of the literature on recent advances in iPSC-derived 3D brain organoid culturing and their utilization in modeling major neurotropic viral infections including HIV-1, HSV-1, JCV, ZIKV, CMV, and SARS-CoV2.