Characterization Of Scaffolds Research Articles

Endogenous Tissue Engineering for Chondral and Osteochondral Regeneration: Strategies and Mechanisms.

Increasing attention has been paid to the development of effective strategies for articular cartilage (AC) and osteochondral (OC) regeneration due to their limited self-reparative capacities and the shortage of timely and appropriate clinical treatments. Traditional cell-dependent tissue engineering faces various challenges such as restricted cell sources, phenotypic alterations, and immune rejection. In contrast, endogenous tissue engineering represents a promising alternative, leveraging acellular biomaterials to guide endogenous cells to the injury site and stimulate their intrinsic regenerative potential. This review provides a comprehensive overview of recent advancements in endogenous tissue engineering strategies for AC and OC regeneration, with a focus on the tissue engineering triad comprising endogenous stem/progenitor cells (ESPCs), scaffolds, and biomolecules. Multiple types of ESPCs present within the AC and OC microenvironment, including bone marrow-derived mesenchymal stem cells (BMSCs), adipose-derived mesenchymal stem cells (AD-MSCs), synovial membrane-derived mesenchymal stem cells (SM-MSCs), and AC-derived stem/progenitor cells (CSPCs), exhibit the ability to migrate toward injury sites and demonstrate pro-regenerative properties. The fabrication and characteristics of scaffolds in various formats including hydrogels, porous sponges, electrospun fibers, particles, films, multilayer scaffolds, bioceramics, and bioglass, highlighting their suitability for AC and OC repair, are systemically summarized. Furthermore, the review emphasizes the pivotal role of biomolecules in facilitating ESPCs migration, adhesion, chondrogenesis, osteogenesis, as well as regulating inflammation, aging, and hypertrophy-critical processes for endogenous AC and OC regeneration. Insights into the applications of endogenous tissue engineering strategies for in vivo AC and OC regeneration are provided along with a discussion on future perspectives to enhance regenerative outcomes.

A study on the synergistic chondro-inductive effects of collagen II and its high-resolution 3D-printed scaffolds

Collagen II is the most essential component of the cartilage-specific extracellular matrix (ECM). However, research on collagen II-based scaffold fabrication, structural optimization, and correlation to the chondrogenic activities of stem cells is currently limited, presumably due to challenges related to its hydrogel processability. Existing collagen II-containing scaffolds are mainly produced by freeze-drying and exhibit limited properties, such as pore interconnectivity and tortuosity. Additionally, the chondro-inductive capability of collagen II composition and its underlying mechanism remains unclear, warranting further research. In this study, we addressed the aforementioned issues by investigating and enhancing the rheological properties of collagen II-based hydrogel, resulting in a high printing resolution exceeding 150 μm. The collagen II composition reportedly facilitated the condensation and chondrogenic activities of mesenchymal stem cells (MSCs) compared to gelatin. Moreover, high-resolution collagen II-based scaffolds promoted cell proliferation and chondrogenic differentiation to a higher degree. Therefore, we optimized the compositional and structural characteristics of collagen II-based scaffolds for enhancing chondrogenic activities. We anticipate that this study will broaden our understanding of collagen II-based scaffold designs and condition optimizations for cartilage tissue engineering.

Tailored edible 3D porous scaffolds constructed by Pickering emulgel templating for cell-cultured fish meat

Cell-cultured fish meat have emerged as an innovative approach to resolving the issues in response to consumer demand. Food-grade scaffolds, which not only serve as the “comfortable home” for stem cells but also endow the final product with meat-like texture, play a vital role in the construction of cell-cultured fish meat. In this study, pea protein, an abundant and inexpensive plant protein, is used to construct edible three-dimensional (3D) porous scaffolds, which exhibit desirable stem cell proliferation and differentiation performance. Our findings demonstrate that it is feasible to apply the Pickering emulgel templating combined with double-crosslinking to the preparation of cell-cultured fish meat scaffolds. The pore structure of 3D porous scaffolds can be altered by varying the emulsification parameters on demand. The Pickering emulgels with large droplet size (around 40 μm) are conducive to the formation of 3D porous scaffolds with large enough pore size for the growth of stem cells. The now available result help to explore the plant protein-derived scaffolds, provide insights into the development of Pickering emulgel-based 3D porous scaffolds and illuminate the relationship between the structural characteristics of scaffolds and the biological functions of stem cells.

Engineering of gelatin scaffold by extracellular matrix of Sertoli cells for embryonic stem cell proliferation

Mimicking the microenvironment of seminiferous tubules plays an indispensable role in directing differentiation of stem cells toward germ cells in vitro. In this work, we fabricated electrospun gelatin (EG) mats (i.e., with diameter <500 nm) conditioned with Sertoli cells' extracellular matrix (ECM) to simulate both 3D structures and composition of normal testis tissue. Sertoli cells were isolated from mice testis and represented through immunocytochemistry (ICC) staining for expression of vimentin, a specific marker of Sertoli cells. The morphological characteristics of ECM-coated scaffold were investigated under scanning electron microscope (SEM). The efficient elimination of cells was confirmed by MTT assay. Furthermore, the cyto/biocompatibility of ECM-conditioned EG scaffold was determined for Sertoli cells and embryonic stem cells (ESCs), alone and as in co-culture. According to the results, the designed scaffold provided a mat for cell proliferation with negligible toxicity (almost 100% cell viability). SEM micrographs displayed cells with elongated shape and complete stretching morphology when compared with those cultured on scaffold without ECM. Moreover, an enhanced differentiation of ESCs toward sperm-generating cells was obtained through co-culturing of Sertoli cells and ESCs, where cell viability was found almost 100%. Our findings introduce the ECM-conditioned EG scaffold as a potentially influential engineered substrate for in vitro guidance of stem cells differentiation by mimicking the native microenvironment.

Localized delivery of metformin via 3D printed GelMA-Nanoclay hydrogel scaffold for enhanced treatment of diabetic bone defects

BackgroundDiabetic bone defects present significant challenges for individuals with diabetes. While metformin has been explored for bone regeneration via local delivery, its application in treating diabetic bone defects remains under-explored. In this study, we aim to leverage 3D printing technology to fabricate a GelMA-Nanoclay hydrogel scaffold loaded with metformin specifically for this purpose. The objective is to assess whether the in situ release of metformin can effectively enhance osteogenesis, angiogenesis, and immunomodulation in the context of diabetic bone defects. Materials and methodsUtilizing 3D printing technology, we constructed a GelMA-Nanoclay-Metformin hydrogel scaffold with optimal physical properties and biocompatibility. The osteogenic, angiogenic, and immunomodulatory characteristics of the hydrogel scaffold were thoroughly investigated through both in vitro and in vivo experiments. ResultsGelMA10%-Nanoclay8%-Metformin5mg/mL was selected as the bioink for 3D printing due to its favorable swelling rate, degradation rate, mechanical strength, and drug release rate. Through in vitro investigations, the hydrogel scaffold extract, enriched with metformin, demonstrated a substantial enhancement in the proliferation and migration of BMSCs within a high-glucose microenvironment. It effectively enhances osteogenesis, angiogenesis, and immunomodulation. In vivo experimental outcomes further underscored the efficacy of the metformin-loaded GelMA-Nanoclay hydrogel scaffold in promoting superior bone regeneration within diabetic bone defects. ConclusionsIn conclusion, while previous studies have explored local delivery of metformin for bone regeneration, our research is pioneering in its application to diabetic bone defects using a 3D printed GelMA-Nanoclay hydrogel scaffold. This localized delivery approach demonstrates significant potential for enhancing bone regeneration in diabetic patients, offering a novel approach for treating diabetic bone defects. The translational potential of this articleOur study demonstrates, for the first time, the successful loading of the systemic antidiabetic drug metformin onto a hydrogel scaffold for localized delivery. This approach exhibits significant efficacy in mending diabetic bone defects, presenting a promising new avenue for the treatment of such conditions.

Diffusion‐induced phase separation 3D printed scaffolds for dynamic tissue repair

AbstractMany hydrogen‐bonded cross‐linked hydrogels possess unique properties, but their limited processability hinders their potential applications. By incorporating a hydrogen bond dissociator (HBD) into these hydrogels, we developed injectable 3D printing inks termed diffusion‐induced phase separation (DIPS) 3D printing inks. Upon extrusion into water and subsequent diffusion of HBD, these ink cure rapidly. The DIPS‐printed scaffold retained most of the original hydrogel properties due to the regeneration of hydrogen bonds. Additionally, the reversible nature of hydrogen bonds provides DIPS 3D‐printed scaffolds with exceptional recycling and reprinting capabilities, resulting in a reduction in the waste of valuable raw ink materials or additives. Postprocessing introduces new crosslinking methods that modulate the mechanical properties and degradation characteristics of DIPS scaffolds over a broad range. Based on its suitable mechanical properties and bioactivity, we successfully repaired and functionally reconstructed a complex defect in penile erectile tissue using the DIPS scaffold in a rabbit model. In summary, this approach is relevant for various hydrogen‐bonded cross‐linked hydrogels that offer mild printing conditions and enable the incorporation of bioactive agents. They can be used as scaffolds for dynamic tissue reconstruction, wearable devices, or soft robots.

A Chitosan Scaffold Supports the Enhanced and Prolonged Differentiation of HiPSCs into Nucleus Pulposus-like Cells.

Intervertebral disc degeneration (IDD) is a progressive condition and stands as one of the primary causes of low back pain. Cell therapy that uses nucleus pulposus (NP)-like cells derived from human induced pluripotent stem cells (hiPSCs) holds great promise as a treatment for IDD. However, the conventional two-dimensional (2D) monolayer cultures oversimplify cell-cell interactions, leading to suboptimal differentiation efficiency and potential loss of phenotype. While three-dimensional (3D) culture systems like Matrigel improve hiPSC differentiation efficiency, they are limited by animal-derived materials for translation, poorly defined composition, short-term degradation, and high cost. In this study, we introduce a new 3D scaffold fabricated using medical-grade chitosan with a high degree of deacetylation. The scaffold features a highly interconnected porous structure, near-neutral surface charge, and exceptional degradation stability, benefiting iPSC adhesion and proliferation. This scaffold remarkably enhances the differentiation efficiency and allows uninterrupted differentiation for up to 25 days without subculturing. Notably, cells differentiated on the chitosan scaffold exhibited increased cell survival rates and upregulated gene expression associated with extracellular matrix secretion under a chemically defined condition mimicking the challenging microenvironment of intervertebral discs. These characteristics qualify the chitosan scaffold-cell construct for direct implantation, serving as both a structural support and a cellular source for enhanced stem cell therapy for IDD.

Vanadium and strontium co-doped hydroxyapatite enriched polycaprolactone matrices for effective bone tissue engineering: A synergistic approach

Scientific research targeted at enhancing scaffold qualities has increased significantly during the last few decades. This emphasis frequently centres on adding different functions to scaffolds in order to increase their usefulness as instruments in the field of regenerative medicine. This study aims to investigate the efficacy of a multifunctional sustainable polymer scaffold, specifically Polycaprolactone (PCL) embedded with hydroxyapatite co-doped with vanadium and strontium (HVS), for bone tissue engineering applications. Polycaprolactone was used to fabricate the scaffold, while hydroxyapatite co-doped with vanadium and strontium (HVS) served as the nanofiller. A thorough investigation of the physicochemical and biological characteristics of the HVS nanofiller was carried out using cutting-edge techniques including Dynamic Light Scattering (DLS), and X-ray Photoelectron Spectroscopy (XPS) and in vitro cell studies. A cell viability rate of more than 70 % demonstrated that the synthesised nanofiller was cytotoxic, but in an acceptable range. The mechanical, biological, and physicochemical properties of the scaffold were extensively evaluated after the nanofiller was integrated. The water absorption characteristics of scaffold were enhanced by the addition of HVS nanofillers, leading to increased swelling, porosity, and hydrophilicity. These improvements speed up the flow of nutrients and the infiltration of cells into the scaffold. The scaffold has been shown to have important properties that stimulate bone cell activity, including better biodegradability and improved mechanical strength, which increased from 5.30 ± 0.37 to 10.58 ± 0.42 MPa. Further, its considerable antimicrobial qualities, blood-compatible nature, and capacity to promote biomineralization strengthen its appropriateness for usage in biomedical applications. Mainly, enhanced Alkaline phosphatase (ALP) activity, Alizarin Red Staining (ARS) activity, and excellent cell adhesive properties, indicating the outstanding osteogenic potential observed in rat bone marrow-derived stromal cells (rBMSC). These combined attributes highlight the pivotal role of these nanocomposite scaffolds in the field of bone tissue engineering.

Enhancing 3D poly[ɛ]caprolactone scaffold properties through alkaline hydrolysis for improved chondrocyte culture: Morphological, physicochemical, and biocompatibility evaluations

This study investigates the impact of alkaline hydrolysis on the morphology, physicochemical properties, and mechanical strength of poly[ε]caprolactone (PCL) scaffolds fabricated using a melt-based electrohydrodynamic process. We systematically analyzed how a brief, 5-min alkaline hydrolysis treatment influenced scaffold characteristics compared to longer durations (15 and 30 min). Results demonstrated that the short-duration treatment significantly increased fiber diameter and reduced pore size while modifying the scaffold's surface chemistry. Importantly, the 5-min treatment optimized the water contact angle to 55–65°, enhancing hydrophilicity which facilitated superior cell attachment and proliferation of the C28/I2 immortalized human chondrocyte cell line. Mechanical testing revealed that while alkaline hydrolysis decreased the scaffold's Young's modulus, a 5-min treatment maintained sufficient mechanical integrity for potential tissue engineering applications, in contrast to the more substantial degradation observed with longer treatments.Biocompatibility evaluations showed that the scaffolds, particularly those treated for 5 min, supported cell viability and proliferation without inducing cytotoxic effects. The molecular analysis indicated a favorable upregulation of chondrogenic genes such as SOX9 and COL2A1, and a downregulation of COL1A1, suggesting potential benefits for cartilage tissue engineering. These findings highlight that precise control over the duration of alkaline hydrolysis can significantly enhance PCL scaffold properties, suggesting a scalable and cost-effective method for scaffold optimization in regenerative medicine applications. The results pave the way for further in vivo studies to validate these promising in vitro findings and explore the clinical applicability of the optimized scaffolds.

3D-printed porous tantalum artificial bone scaffolds: fabrication, properties, and applications.

Porous tantalum scaffolds offer a high degree of biocompatibility and have a low friction coefficient. In addition, their biomimetic porous structure and mechanical properties, which closely resemble human bone tissue, make them a popular area of research in the field of bone defect repair. With the rapid advancement of additive manufacturing, 3D-printed porous tantalum scaffolds have increasingly emerged in recent years, offering exceptional design flexibility, as well as facilitating the fabrication of intricate geometries and complex pore structures that similar to human anatomy. This review provides a comprehensive description of the techniques, procedures, and specific parameters involved in the 3D printing of porous tantalum scaffolds. Concurrently, the review provides a summary of the mechanical properties, osteogenesis and antibacterial properties of porous tantalum scaffolds. The use of surface modification techniques and the drug carriers can enhance the characteristics of porous tantalum scaffolds. Accordingly, the review discusses the application of these porous tantalum materials in clinical settings. Multiple studies have demonstrated that 3D-printed porous tantalum scaffolds exhibit exceptional corrosion resistance, biocompatibility, and osteogenic properties. As a result, they are considered highly suitable biomaterials for repairing bone defects. Despite the rapid development of 3D-printed porous tantalum scaffolds, they still encounter challenges and issues when used as bone defect implants in clinical applications. Ultimately, a concise overview of the primary challenges faced by 3D-printed porous tantalum scaffolds is offered, and corresponding insights to promote further exploration and advancement in this domain are presented.

Quantitative Evaluation of the Pore and Window Sizes of Tissue Engineering Scaffolds on Scanning Electron Microscope Images Using Deep Learning.

The morphological characteristics of tissue engineering scaffolds, such as pore and window diameters, are crucial, as they directly impact cell-material interactions, attachment, spreading, infiltration of the cells, degradation rate and the mechanical properties of the scaffolds. Scanning electron microscopy (SEM) is one of the most commonly used techniques for characterizing the microarchitecture of tissue engineering scaffolds due to its advantages, such as being easily accessible and having a short examination time. However, SEM images provide qualitative data that need to be manually measured using software such as ImageJ to quantify the morphological features of the scaffolds. As it is not practical to measure each pore/window in the SEM images as it requires extensive time and effort, only the number of pores/windows is measured and assumed to represent the whole sample, which may cause user bias. Additionally, depending on the number of samples and groups, a study may require measuring thousands of samples and the human error rate may increase. To overcome such problems, in this study, a deep learning model (Pore D2) was developed to quantify the morphological features (such as the pore size and window size) of the open-porous scaffolds automatically for the first time. The developed algorithm was tested on emulsion-templated scaffolds fabricated under different fabrication conditions, such as changing mixing speed, temperature, and surfactant concentration, which resulted in scaffolds with various morphologies. Along with the developed model, blind manual measurements were taken, and the results showed that the developed tool is capable of quantifying pore and window sizes with a high accuracy. Quantifying the morphological features of scaffolds fabricated under different circumstances and controlling these features enable us to engineer tissue engineering scaffolds precisely for specific applications. Pore D2, an open-source software, is available for everyone at the following link: https://github.com/ilaydakaraca/PoreD2.

Mechanistic study on the role of 3D-Printed biomimetic coral bone scaffolds in bone defect repair

With the continuous innovation and development of 3D printing technology, 3D-printed biomimetic coral bone scaffolds have demonstrated significant potential in the field of bone defect repair. This paper aims to explore in-depth the mechanistic study of 3D-printed biomimetic coral bone scaffolds in bone defect repair, by systematically reviewing relevant literature and analyzing their potential mechanisms in promoting bone growth and improving the success rate of bone defect repair. Firstly, this paper introduces the fabrication process and material characteristics of 3D-printed biomimetic coral bone scaffolds. Secondly, the paper discusses the mechanisms of 3D-printed biomimetic coral bone scaffolds in terms of biocompatibility, biomechanical performance, as well as their roles in vascularization and bone formation. Finally, the paper outlines future research directions for 3D-printed biomimetic coral bone scaffolds, including further optimization of material properties, improvement of printing precision, and expansion of clinical applications.

Electrospun drug-loaded scaffolds for nervous system repair.

Nervous system injuries, encompassing peripheral nerve injury (PNI), spinal cord injury (SCI), and traumatic brain injury (TBI), present significant challenges to patients' wellbeing. Traditional treatment approaches have limitations in addressing the complexity of neural tissue regeneration and require innovative solutions. Among emerging strategies, implantable materials, particularly electrospun drug-loaded scaffolds, have gained attention for their potential to simultaneously provide structural support and controlled release of therapeutic agents. This review provides a thorough exploration of recent developments in the design and application of electrospun drug-loaded scaffolds for nervous system repair. The electrospinning process offers precise control over scaffold characteristics, including mechanical properties, biocompatibility, and topography, crucial for creating a conducive environment for neural tissue regeneration. The large surface area of the resulting fibrous networks enhances biomolecule attachment, influencing cellular behaviors such as adhesion, proliferation, and migration. Polymeric electrospun materials demonstrate versatility in accommodating a spectrum of therapeutics, from small molecules to proteins. This enables tailored interventions to accelerate neuroregeneration and mitigate inflammation at the injury site. A critical aspect of this review is the examination of the interplay between structural properties and pharmacological effects, emphasizing the importance of optimizing both aspects for enhanced therapeutic outcomes. Drawing upon the latest advancements in the field, we discuss the promising outcomes of preclinical studies using electrospun drug-loaded scaffolds for nervous system repair, as well as future perspectives and considerations for their design and implementation. This article is categorized under: Implantable Materials and Surgical Technologies > Nanomaterials and Implants Implantable Materials and Surgical Technologies > Nanotechnology in Tissue Repair and Replacement Therapeutic Approaches and Drug Discovery > Emerging Technologies.

Developing Porous Fibrin Scaffolds with Tunable Anisotropic Features to Direct Myoblast Orientation.

Functional regeneration of anisotropically aligned tissues such as ligaments, microvascular networks, myocardium, or skeletal muscle requires a temporal and spatial series of biochemical and biophysical cues to direct cell functions that promote native tissue regeneration. When these cues are lost during traumatic injuries such as volumetric muscle loss (VML), scar formation occurs, limiting the regenerative capacity of the tissue. Currently, autologous tissue transfer is the gold standard for treating injuries such as VML but can result in adverse outcomes including graft failure, donor site morbidity, and excessive scarring. Tissue-engineered scaffolds composed of biomaterials, cells, or both have been investigated to promote functional tissue regeneration but are still limited by inadequate tissue ingrowth. These scaffolds should provide precisely tuned topographies and stiffnesses using proregenerative materials to encourage tissue-specific functions such as myoblast orientation, followed by aligned myotube formation and recovery of functional contraction. In this study, we describe the design and characterization of novel porous fibrin scaffolds with anisotropic microarchitectural features that recapitulate the native tissue microenvironment and offer a promising approach for regeneration of aligned tissues. We used directional freeze-casting with varied fibrin concentrations and freezing temperatures to produce scaffolds with tunable degrees of anisotropy and strut widths. Nanoindentation analyses showed that the moduli of our fibrin scaffolds varied as a function of fibrin concentration and were consistent with native skeletal muscle tissue. Quantitative morphometric analyses of myoblast cytoskeletons on scaffold microarchitectures demonstrated enhanced cell alignment as a function of microarchitectural morphology. The ability to precisely control the anisotropic features of fibrin scaffolds promises to provide a powerful tool for directing aligned tissue ingrowth and enhance functional regeneration of tissues such as skeletal muscle.

Optimization of Polycaprolactone and Type I Collagen Scaffold for Tendon Tissue Regeneration.

Introduction Collagen synthesis is vital for restoring musculoskeletal tissues, particularly in tendon and ligamentous structures. Tissue engineering utilizes scaffolds for cell adhesion and differentiation. Although synthetic scaffolds offer initial strength, their long-term stability is surpassed by biological scaffolds. Combining polycaprolactone (PCL) toughness with collagen in scaffold design, this study refines fabrication via electrospinning, aiming to deliver enduring biomimetic matrices for widespread applications in musculoskeletal repair. Methods Electrospinning employed four solutions with varied collagen and PCL concentrations, dissolved in chloroform, methanol, and hexafluoro-2-propanol. Solutions were combined to yield 60 mg/mL concentrations with different collagen/PCL ratios. Electrospinning at 12-14kV voltage produced scaffolds, followed by vacuum-drying. Collagen coating was applied to PCL and 15% collagen/PCL scaffolds using a 0.1% collagen solution. SEM characterized fiber morphology, tensile testing was conducted to determine the mechanical properties of the scaffold, and Fourier-transform infrared (FTIR) spectroscopy analyzed scaffold composition. Atomic force microscopy (AFM) analyzed the stiffness properties of individual fibers, and a finite element model was developed to predict the mechanical properties. Cell culture involved seeding human bone marrow mesenchymal stem cells onto scaffolds, which were assessed through Alamar Blue assay and confocal imaging. Results Various scaffolds (100% PCL, PCL-15% collagen, PCL-25% collagen, PCL-35% collagen) were fabricated to emulate the extracellular matrix, revealing collagen's impact on fiber diameter reduction with increasing concentration. Tensile testing highlighted collagen's initial enhancement of mechanical strength, followed by a decline beyond PCL-15% collagen. FTIR spectroscopy detected potential hydrogen bonding between collagen and PCL. A finite element model predicted scaffold response to external forces which was validated by the tensile test data. Cell viability and proliferation assays demonstrated successful plating on all scaffolds, with optimal proliferation observed in PCL-25% collagen. Confocal imaging confirmed stem cell integration into the three-dimensional material. Collagen coating preserved nanofiber morphology, with no significant changes in diameter. Coating of collagen significantly altered the tensile strength of the scaffolds at the macro scale. AFM highlighted stiffness differences between PCL and collagen-coated PCL mats at the single fiber scale. The coating process did not significantly enhance initial cell attachment but promoted increased proliferation on collagen-coated PCL scaffolds. Conclusion The study reveals collagen-induced mechanical and morphological alterations, influencing fiber alignment, diameter, and chemical composition while emphasizing scaffolds' vital role in providing a controlled niche for stem cell proliferation and differentiation. The optimization of each of these scaffold characteristics and subsequent finite element modeling can lead to highly repeatable and ideal scaffold properties for stem cell integration and proliferation.

Fabrication of Silk Fibroin‐Derived Fibrous Scaffold for Biomedical Frontiers

AbstractSilk fibroin (SF), a natural protein derived from silkworms, has emerged as a promising biomaterial due to its biocompatibility, biodegradability, degradation rate, and tunable mechanical properties. This review delves into the intrinsic attributes of SF that make it an attractive candidate for scaffold development in tissue engineering and regenerative medicine. The distinctiveness of this comprehensive review resides in its detailed exploration of recent advancements in the fabrication techniques of SF‐based fibrous scaffolds, namely electrospinning, freeze‐drying, and 3D printing. An in‐depth analysis of these fabrication techniques is conducted to illustrate their versatility in customizing essential scaffold characteristics, such as porosity, fiber diameter, and mechanical strength. The article meticulously discusses process parameters, advantages, and challenges of each fabrication technique, highlighting the innovative advancements made in the respective field. Furthermore, the review goes beyond fabrication techniques to provide an overview of the latest biomedical applications and research endeavors utilizing SF‐derived scaffolds. From nerve regeneration and wound healing to drug delivery, bone regeneration, and vascular tissue engineering, the diverse applications underscore the versatility of SF in adopting various biomedical challenges. Finally, the article emphasizes the need for standardized characterization techniques, scalable manufacturing processes, and long‐term in vivo studies.

An overview of nasal cartilage bioprinting: from bench to bedside.

Nasal cartilage diseases and injuries are known as significant challenges in reconstructive medicine, affecting a substantial number of individuals worldwide. In recent years, the advent of three-dimensional (3D) bioprinting has emerged as a promising approach for nasal cartilage reconstruction, offering potential breakthroughs in the field of regenerative medicine. This paper provides an overview of the methods and challenges associated with 3D bioprinting technologies in the procedure of reconstructing nasal cartilage tissue. The process of 3D bioprinting entails generating a digital 3D model using biomedical imaging techniques and computer-aided design to integrate both internal and external scaffold features. Then, bioinks which consist of biomaterials, cell types, and bioactive chemicals, are applied to facilitate the precise layer-by-layer bioprinting of tissue-engineered scaffolds. After undergoing in vitro and in vivo experiments, this process results in the development of the physiologically functional integrity of the tissue. The advantages of 3D bioprinting encompass the ability to customize scaffold design, enabling the precise incorporation of pore shape, size, and porosity, as well as the utilization of patient-specific cells to enhance compatibility. However, various challenges should be considered, including the optimization of biomaterials, ensuring adequate cell viability and differentiation, achieving seamless integration with the host tissue, and navigating regulatory attention. Although numerous studies have demonstrated the potential of 3D bioprinting in the rebuilding of such soft tissues, this paper covers various aspects of the bioprinted tissues to provide insights for the future development of repair techniques appropriate for clinical use.

Multidimensional 3D-Printed Scaffolds and Regeneration of Intrabony Periodontal Defects: A Systematic Review.

The utilization of regenerative techniques in periodontology involves tailoring tissue engineering principles to suit the oral cavity's unique environment. Advancements in computer-assisted technology, specifically utilizing cone beam computed tomography (CBCT), enabled the fabrication of 3D-printed scaffolds. The current review aims to explore whether 3D-printed scaffolds are effective in promoting osteogenesis in patients with periodontal defects. A thorough exploration was undertaken across seven electronic databases (PubMed, Scopus, ScienceDirect, Google Scholar, Cochrane, Web of Science, Ovid) to detect pertinent research in accordance with specified eligibility criteria, aligning with the PRISMA guidelines. Two independent reviewers undertook the screening and selection of manuscripts, executed data extraction, and evaluated the bias risk using the Newcastle-Ottawa Scale for non-randomized clinical trials and SYRCLE's risk of bias tool for animal studies. Initially, 799 articles were identified, refined by removing duplicates. After evaluating 471 articles based on title and abstract, 18 studies remained for full-text assessment. Eventually, merely two manuscripts fulfilled all the eligibility criteria concerning human trials. Both studies were prospective non-randomized clinical trials. Moreover, 11 animal studies were also included. The use of multidimensional, 3D-printed, customized scaffolds appears to stimulate periodontal regeneration. While the reported results are encouraging, additional studies are required to identify the ideal characteristics of the 3D scaffold to be used in the regeneration of periodontal tissue.

Directing ligament-mimetic bi-directional cell organization in scaffolds through zone-specific microarchitecture for ligament tissue engineering

Ligament tissues exhibit zone-specific anisotropic cell organization. The cells in ligament-proper are longitudinally oriented, whereas, the cells in epiligament are circumferentially oriented. Therefore, scaffolds developed to regenerate ligament tissues should possess adequate architectural features to govern ligament-mimetic bi-directional cell organization. The scaffold architectural features along with ligament-mimetic cell organization may ultimately yield neo-tissues with ligament-like extracellular matrix (ECM) structure and biomechanical properties. Towards this goal, we fabricated a silk/gelatin-based core–shell scaffold (csSG) with zone-specific anisotropic architectural features, wherein, the core of the scaffold possessed longitudinally aligned pores while the shell of the scaffold possessed parallel microgrooves that are aligned circumferentially around the surface of the scaffold. The ligament-mimetic architectural features significantly improved the mechanical properties of the scaffold. Moreover, architectural features of the csSG scaffold governed zone-specific anisotropic organization of cells. The cells in the core were longitudinally oriented as observed in the ligament-proper and the cells on the shell were circumferentially oriented as observed in epiligament. This bi-directional cell orientation partially mimicked the complex cellular network in native ligament tissue. Additionally, both the core and the shell individually supported fibrogenic differentiation of stem cells which further improved their potential for ligament tissue engineering. Further, the aligned pores of the core could govern unidirectional organization of ECM deposited by cells which is crucial for regenerating anisotropic tissues like ligaments. Finally, when implanted subcutaneously in mice, the scaffolds retained their anisotropic architecture for at least 2 weeks, were biocompatible, supported cell infiltration and governed anisotropic organization of cells and ECM. Taken together, the fabricated biomimetic csSG scaffold, through its zone-specific architectural features, could govern ligament-mimetic cellular and ECM organization which is ultimately expected to achieve regeneration of ligament tissues with native-like hierarchical structure and biomechanical properties. Consequently, this study introduces bi-directional structural parameters as design criteria for developing scaffolds for ligament tissue engineering.

Microstructure of the silk fibroin-based hydrogel scaffolds derived from the orb-web spider Trichonephila clavata

Due to the unique properties of the silk fibroin (SF) made from silkworm, SF-based hydrogels have recently received significant attention for various biomedical applications. However, research on the SF-based hydrogels isolated from spider silks has been rtricted due to the limited collection and preparation of naïve silk materials. Therefore, this study focused on the microstructural characteristics of hydrogel scaffolds derived from two types of woven silk glands: the major ampullate gland (MAG) and the tubuliform gland (TG), in the orb-web spider Trichonephila clavate. We compared these spider glands with those of the silk fibroin (SF) hydrogel scaffold extracted from the cocoon of the insect silkworm Bombyx mori. Our FESEM analysis revealed that the SF hydrogel has high porosity, translucency, and a loose upper structure, with attached SF fibers providing stability. The MAG hydrogel displayed even higher porosity, as well as elongated fibrous structures, and improved mechanical properties: while the TG hydrogel showed increased porosity, ridge-like or wall-like structures, and stable biocapacity formed by physical crosslinking. Due to their powerful and versatile microstructural characteristics, the MAG and TG hydrogels can become tailored substrates, very effective for tissue engineering and regenerative medicine applications.