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Microscale carriers have emerged as promising materials for nurturing cell growth and as delivery vehicles for regenerative therapies. Carriers based on granular hydrogels have proved advantageous, where "microgels" can be formulated to have a broad range of properties to guide the behavior of adherent cells. Herein, the fabrication of osteogenic microgel matrices through the incorporation of laponite nanoclays is demonstrated. Forming a jammed suspension provides a scaffolding where cells can adhere to the surface of the microgels, with pathways for migration and proliferation fostered by the interstitial volume. By varying the content and type of laponite—RD and XLG—the degree of osteogenesis can be tuned in embedded populations of adipose‐derived stem cells. The nano‐ and microstructured composite materials enhance osteogenesis at the transcript and protein level, leading to increased deposition of bone minerals and an increase in the compressive modulus of the assembled scaffold. Together, these microgel suspensions are promising materials for encouraging osteogenesis with scope for delivery via injection and stabilization to bone‐mimetic mechanical properties after matrix deposition. [ABSTRACT FROM AUTHOR]

Hematopoietic stem cell (HSC) transplantation is used to treat blood and immunodeficient diseases. HSC expansion techniques must be developed to prevent complications and ensure reliable therapeutic efficacy. Hence, several studies have attempted in vitro expansion of HSCs using scaffolds but failed to mimic the diverse and complex nature of HSC environments. Herein, an artificial HSC microenvironment, bone marrow (BM) niches is created, through in vivo engineering using carbonate apatite honeycomb scaffolds and the potential of these scaffolds in restoring lost hematopoietic function and immunity is investigated. BM niches are generated in every honeycomb channel, wherein HSCs are gradually aggregated. Compared to the actual BM, the scaffolds exhibit a 9.9‐ and 78‐fold increase in the number of stored CD45− CD34+ side scatterlow cells that are mainly considered HSCs at 8 and 12 weeks, respectively. The transplantation of the honeycomb scaffold containing HSCs and BM niches into immunocompromised mice increases peripheral blood chimerism and restores hematopoietic function and the number of immunocytes (monocytes and lymphocytes) to normal levels. This study contributes to the development of efficient HSC transplantation techniques. Additionally, in vivo‐engineered integrated tissues using honeycomb scaffolds can be used to elucidate the interplay between the BM niches and resident cells. [ABSTRACT FROM AUTHOR]

Introduction: Digital dermatitis (DD) in cattle appears with high prevalence; nevertheless, the knowledge on its pathogenesis is still limited. In this context, in vitro skin models represent a valuable tool to facilitate the study of DD. Methods: Two in vitro skin models were established using bovine distal limb skin: a skin explant model and an organotypic skin model. For the skin explant model, skin samples were cultured with an air-liquid interface for up to 7 days. Besides routine histopathological examination, readout parameters were Ki-67 and cleaved Caspase-3 stainings. For the organotypic model, primary keratinocytes were layered on top of a dermal equivalent containing mainly mitotically inactive fibroblasts and maintained for up to 21 days. At regular intervals (days 7, 14, and 21), cultured skin samples were taken for (immuno)histological analysis. Results: Both cultures could be maintained for the entire duration of the intended culture period. In the histopathological assessment, explant skin cultures showed ballooning degeneration of keratinocytes and segmental necrosis starting at day 5 of culturing. Initially, basal keratinocytes in the organotypic model differentiated as demonstrated by positive Keratin 14, Desmoglein-1, Loricrin, and Involucrin immunofluorescent stainings. Ki-67 was observed occasionally and suprabasally still after 21 days of culture. Conclusion: Both in vitro models proved dependable and constitute a viable option for replacing experiments on live animals, each with its own benefits. Whereas skin explants include all cell types available in vivo and can therefore reflect realistic cell-cell interactions and signaling pathways, the organotypic model offers a higher standardization and reproducibility. Depending on the focus of future studies, both models can be used for specific experimental purposes of bovine dermatological research in general or specialized questions concerning (infectious) claw diseases as, e.g., DD. [ABSTRACT FROM AUTHOR]

Tissue engineering is a multidisciplinary field that uses biomaterials to restore tissue function and assist with drug development. Over the last decade, the fabrication of three-dimensional (3D) multifunctional scaffolds has become commonplace in tissue engineering and regenerative medicine. Thanks to the development of 3D bioprinting technologies, these scaffolds more accurately recapitulate in vivo conditions and provide the support structure necessary for microenvironments conducive to cell growth and function. The purpose of this review is to provide a background on the leading 3D bioprinting methods and bioink selections for tissue engineering applications, with a specific focus on the growing field of developing multifunctional bioinks and possible future applications. [ABSTRACT FROM AUTHOR]

• Disciplines such as biomaterials, biofabrication, and tissue engineering are converging to create more advanced tissue-like constructs that accurately reproduce detailed physical and biological properties. • Researchers now have access to a range of tools that enable them to replicate microenvironmental features such as heterogeneity, anisotropy, hierarchical structures, compositional gradients and interfaces, and spatiotemporal cues. • Advanced models are strengthening our understanding of the mechanisms that govern tissue function and disease. • While not fully resembling the complexity of human tissue, engineered models enable researchers to investigate and adapt subsets of parameters, addressing targeted research questions. Advances in tissue engineering for both system modeling and organ regeneration depend on embracing and recapitulating the target tissue's functional and structural complexity. Microenvironmental features such as anisotropy, heterogeneity, and other biochemical and mechanical spatiotemporal cues are essential in regulating tissue development and function. Novel biofabrication strategies and innovative biomaterial design have emerged as promising tools to better reproduce such features. These facilitate a transition towards high-fidelity biomimetic structures, offering opportunities for a deeper understanding of tissue function and the development of superior therapies. In this review, we explore some of the key structural and compositional aspects of tissues, lay out how to achieve similar outcomes with current fabrication strategies, and identify the main challenges and promising avenues for future research. [ABSTRACT FROM AUTHOR]

Recent years have seen a surge of interest in the potential use of 3D bioprinting technologies for the development of tissues and organs with the aim of restoring, maintaining, or improving tissue function. Bioprinting studies frequently claim to be close to clinical translation, yet only report research at very early stages of the translational pipeline. Technology readiness levels (TRLs) are a systematic metric used to objectively assess the maturity of a technology, allowing researchers and stakeholders to understand how far a particular project has progressed from the conceptual stage towards clinical application. Bioprinting is an annex of additive manufacturing, as defined by the American Society for Testing and Materials (ASTM) and International Organization for Standardization (ISO) standards, characterized by the automated deposition of living cells and biomaterials. The tissue engineering and regenerative medicine (TE&RM) community has eagerly adopted bioprinting, while review articles regularly herald its imminent translation to the clinic as functional tissues and organs. Here we argue that such proclamations are premature and counterproductive; they place emphasis on technological progress while typically ignoring the critical stage-gates that must be passed through to bring a technology to market. We suggest the technology readiness level (TRL) scale as a valuable metric for gauging the relative maturity of a bioprinting technology in relation to how it has passed a series of key milestones. We suggest guidelines for a bioprinting-oriented scale and use this to discuss the state-of-the-art of bioprinting in regenerative medicine (BRM) today. Finally, we make corresponding recommendations for improvements to BRM research that would support its progression to clinical translation. [ABSTRACT FROM AUTHOR]

Physical-property-based patterning is an alternative biofabrication approach that relies on the application of a remote field, and the intrinsic properties of the objects or materials of interest, to simply create complex tissues. Both buoyant and gravitational forces can rapidly create tissue gradients without any hardware, highlighting the appeal of in situ density-based patterning. Recent advances now allow for the magnetic patterning of naturally diamagnetic cells using paramagnetic solutions, including crosslinkable hydrogels. New methods have been developed to change the compressibility of a cell, enabling differential cell separation and alignment within an acoustic field. The field of biofabrication is rapidly expanding with the advent of new technologies and material systems to engineer complex tissues. In this opinion article, we introduce an emerging tissue patterning method, physical-property-based patterning, that has strong translational potential given its simplicity and limited dependence on external hardware. Physical-property-based patterning relies solely on the intrinsic density, magnetic susceptibility, or compressibility of an object, its surrounding solution, and the noncontact application of a remote field. We discuss how physical properties can be exploited to pattern objects and design a variety of biologic tissues. Finally, we pose several open questions that, if addressed, could transform the status quo of biofabrication, pushing us one step closer to patterning tissues in situ. [ABSTRACT FROM AUTHOR]

Development of artificial dental pulp for the regeneration of pulpectomized root canal is of vital importance in maintaining the tooth's health and function. To achieve this objective, designing a pulp-like structure able to support tissue repair and vascularization is needed. In the current study, we developed a composite hydrogel based on collagen hydrogel and resveratrol-loaded chitosan nanoparticles. Various physicochemical and biological properties of the hydrogel system were investigated in vitro. The healing potential of the artificial pulp was studied in a rabbit model of dental pulp injury. Study revealed the composite hydrogel had significantly higher healing activity compared with simple hydrogel, as evidenced by the results of histopathological examinations. Real-time PCR assay showed that the composite hydrogel system could enhance the level of VEGF expression and modulated inflammation via decreasing NFKβ expression. [ABSTRACT FROM AUTHOR]

Background: The advent of tissue engineering and regenerative medicine has introduced innovative approaches to treating degenerative and traumatic injuries, particularly in cartilage, a tissue with limited self-repair capabilities. Among the various stem cell sources, umbilical cord-derived mesenchymal stromal cells (UC-MSCs) have garnered significant interest due to their non-invasive collection, minimal ethical concerns, and robust regenerative potential, particularly in cartilage regeneration. Methods: A comprehensive literature review was conducted using multiple databases, including PubMed, Scopus, Web of Science, and Google Scholar. Search terms focused on "umbilical cordderived mesenchymal stromal cells," "chondrogenesis," "cartilage regeneration," and related topics. Studies published in the past two decades were included, with selection criteria emphasizing methodological rigor and relevance to UC-MSC chondrogenesis. The review synthesizes findings from various sources to provide a thorough analysis of the potential of UC-MSCs in cartilage tissue engineering. Results: UC-MSCs exhibit significant chondrogenic potential, supported by their ability to differentiate into chondrocytes under specific conditions. Recent advancements include the development of biomaterial scaffolds and the application of genetic engineering techniques, such as CRISPR/Cas9, to enhance chondrogenic differentiation. Despite these advancements, challenges remain in standardizing cell isolation techniques, scaling up production for clinical use, and ensuring the long-term functionality of regenerated cartilage. Conclusion: UC-MSCs offer a promising solution for cartilage regeneration in the field of regenerative medicine. Ongoing research is focused on overcoming current challenges through the use of advanced technologies, including bioreactors and gene editing. Collaborative efforts among researchers, clinicians, and bioengineers are essential to translating the potential of UC-MSCs into effective clinical therapies, which could significantly advance tissue regeneration and therapeutic innovation. [ABSTRACT FROM AUTHOR]

Background Encapsulated pancreatic fluid collection (PFC) is a requisite for endoscopic drainage procedures. The 4-week threshold for defining walled-off necrosis does not capture the dynamic process of encapsulation. We aim to investigate the changes in the wall characteristics of PFC in acute necrotizing pancreatitis (ANP) by comparing baseline contrast-enhanced computed tomography (CECT) with follow-up CT scans. Methods This retrospective study comprised consecutive patients with ANP who underwent a baseline CECT within first 2 weeks and follow-up CECT in the third to fifth weeks of illness. Presence, extent, and encapsulation thickness (defined as enhancing wall around the collection) on baseline CECT were compared with follow-up CT (done in the third–fifth weeks of illness). Results Thirty patients (19 males and 11 females; mean age 41.5 ± 13.5 years) were included in the study. The mean time to first CECT was 10 ± 3.6 days. There were 58 collections. The most common site was the lesser sac (n = 29), followed by the left pararenal space (n = 15). At baseline CT, 52 (89.7%) collections had varying degree of encapsulation (15.3%, complete encapsulation). Complete encapsulation was seen in 52 and 82.6% collections in third and fourth week, respectively. All collections in fifth week and beyond were encapsulated. The wall was thicker on follow-up CECT scans (p < 0.01). The mean wall thickness was not significantly associated with the degree of encapsulation (p = 0.417). There was no significant association between the site and degree of encapsulation (p = 0.546). Conclusion Encapsulation is dynamic and collections may get "walled off" before 4 weeks. Walled-off collections should be defined based on imaging rather than a fixed 4-week revised Atlanta classification threshold. [ABSTRACT FROM AUTHOR]

Recently, micro/nanorobots (MNRs) with self-propulsion have emerged as a promising smart platform for diagnostic, therapeutic and theranostic applications. Especially, polymer-based MNRs have attracted huge attention due to their inherent biocompatibility and versatility, making them actively explored for various medical applications. As the translation of MNRs from laboratory to clinical settings is imperative, the use of appropriate polymers for MNRs is a key strategy, which can prompt the advancement of MNRs to the next phase. In this review, we describe the multifunctional versatile polymers in MNRs, and their biodegradability, motion control, cargo loading and release, adhesion, and other characteristics. After that, we review the theranostic applications of polymer-based MNRs to bioimaging, biosensing, drug delivery, and tissue engineering. Furthermore, we address the challenges that must be overcome to facilitate the translational development of polymeric MNRs with future perspectives. This review would provide valuable insights into the state-of-the-art technologies associated with polymeric MNRs and contribute to their progression for further clinical development. [Display omitted] • Micro/nanorobot (MNR) with self-propulsion is promising for biomedical applications. • Biocompatible and multifunctional MNRs can be designed using polymers. • Polymeric MNRs are used for imaging, sensing, drug delivery, and tissue engineering. • Challenges need to be overcome for using these MNRs in clinical settings. [ABSTRACT FROM AUTHOR]

In Brazil, around 80% of snakebites are caused by snakes of the genus Bothrops. A three-dimensional culture model was standardized and used to perform treatments with Bothrops erythromelas venom (BeV) and its antivenom (AV). The MRC-5 and L929 cell lines were cultured at increasing cell densities. Morphometric parameters were evaluated through images obtained from an inverted microscope: solidity, circularity, and Feret diameter. L929 microtissues (MT) showed better morphometric data, and thus they were used for further analysis. MT viability was assessed using the acridine orange and ethidium bromide staining method, which showed viable cells in the MT on days 5, 7, and 10 of cultivation. Histochemical and histological analyses were performed, including hematoxylin/eosin staining, which showed a good structure of the spheroids. Alcian blue staining revealed the presence of acid proteoglycans. Immunohistochemical analysis with ki-67 showed different patterns of cell proliferation. The MT were also subjected to pharmacological tests using the BeV, in the presence or absence of its AV. The results showed that the venom was not cytotoxic, but it caused morphological changes. The MT showed cell detachment, losing their structure. The antivenom was able to partially prevent the venom activities. [ABSTRACT FROM AUTHOR]

Bacteriophages have attracted great attention in the bioengineering field in diverse research areas from tissue engineering to therapeutic and clinical applications. Recombinant filamentous bacteriophage, carrying multiple copies of foreign peptides on protein capsid has been successfully used in the vaccine delivery setting, even if their plasma instability and degradation have limited their use on the pharmaceutical market. Encapsulation techniques in polymeric materials can be applied to preserve bacteriophage activity, extend its half-life, and finely regulate their release in the target environment. The main goal of this study was to provide tunable formulations of the bacteriophage encapsulated in polymeric microparticles (MPs). We used poly (lactic-co-glycolic-acid) as a biocompatible and biodegradable polymer with ammonium bicarbonate as a porogen to encapsulate bacteriophage expressing OVA (257–264) antigenic peptide. We demonstrate that nano-engineered fdOVA bacteriophages encapsulated in MPs preserve their structure and are immunologically active, inducing a strong immune response towards the delivered peptide. Moreover, MP encapsulation prolongs bacteriophage stability over time also at room temperature. Additionally, in this study, we show the ability of in silico-supported approach to predict and tune the release of bacteriophages. These results lay the framework for a versatile bacteriophage-based vaccine delivery system that could successfully generate robust immune responses in a sustained manner, to be used as a platform against cancer and new emerging diseases. Synopsis: administration of recombinant bacteriophage-loaded PLGA microparticles for antigen delivery. PLGA microparticles release the bacteriophages, inducing activation of dendritic cells and enhancing antigen presentation and specific T cell response. Bacteriophage-encapsulated microneedles potentially can be administered into human body and generate robust immune responses. [ABSTRACT FROM AUTHOR]

The temporomandibular joint (TMJ) disc is an essential protective but vulnerable fibrocartilage. Their high mechanical strength is vital in absorbing loads, reducing friction, and protecting the condylar surface. Many diseases can lead to the destruction or degeneration of the mechanical function of the TMJ disc. Unfortunately, conservative treatment is ineffective in restoring the defective mechanical properties of the discs. Tissue engineering has been investigated as a promising alternative treatment approach to approximate the properties of native tissue. However, it is difficult for tissue-engineered discs to obtain sufficient mechanical properties. Several approaches have been proposed to improve the mechanical properties of tissue-engineered constructs. In this review, we summarized the mechanical properties of native TMJ discs and discussed the current mechanical testing methods. We then summarized the current advances in improving the mechanical properties of TMJ disc tissue-engineered constructs. Moreover, existing challenges and outbreak directions are discussed. This review assists future research in better understanding the mechanical properties of both native and tissue-engineered TMJ discs. It provides new insights into future mechanical property enhancement for TMJ disc tissue engineering. [ABSTRACT FROM AUTHOR]

There is a deficit for bone tissue natural grafts that seek to be covered with synthetic substitutes. Scaffolds generated with 3D printing and electrospinning allow adequate mechanical properties maintaining a structure appropriate for cell growth. Here, a scaffold made up of three-dimensional (3D) printed PLA frameworks added with PCL/PLA/nHA nanofibers was manufactured. The framework showed mechanical properties similar to other reported bone substitutes, while the nanofibers showed diameters between 200 and 850 nm. Scaffolds were suitable for cell adhesion and proliferation when evaluated with fibroblasts, showing cell proliferation into the nanofiber network, a fundamental aspect in tissue engineering. [ABSTRACT FROM AUTHOR]

This review summarises the available results on the effects of cation substitution on the antibacterial and other bioproperties of different phosphates. Attention is given to bulk cation-substituted phosphates (alkali, alkaline earth, transition and lanthanide metals) and to thin films, glasses, and various composite materials. The cationic effects on the biological properties of phosphate bioceramics are also documented. These phosphate-based biomaterials, especially nanostructured ones, show high biocompatibility and enhanced antibacterial behaviour. Numerous articles have concluded that the cationically substituted phosphates are the best candidates for applications in many biomedical fields such as bone repair, tissue engineering, drug and gene delivery, magnetic targeting and hyperthermia treatment for cancer, bioimaging, and theranostics. [ABSTRACT FROM AUTHOR]

Acellular matrices have historically been applied as biologic scaffolds in surgery, wound care, and tissue engineering, albeit with inconsistent outcomes. One aspect that varies widely between products is the selection of decellularization protocol, yet few studies assess comparative effectiveness of these protocols in preserving mechanics, and protein content. This study characterizes bladder acellular matrix (BAM) using two different detergent and enzymatic protocols, evaluating effects on nuclei and DNA removal (≥90%), structure, tensile properties, and maintenance of extracellular matrix proteins. Porcine bladders were decellularized with 0.5% Sodium Dodecyl Sulfate (SDS) or 0.25% Trypsin-hypotonic-Triton X-100 hypertonic (TT)-based agitation protocols, followed by DNase/RNase agents. Characterization of BAM included decellularization efficacy (DAPI, DNA quantification), structure (histology and scanning electron microscopy), tensile testing (Instron 345C-1 mechanical tester), and protein presence and diversity (mass spectrometry). SDS and TT data was directly compared to the same native bladder using two-tailed paired t-tests. Native, TT, and SDS cohorts for tensile testing were compared using one-way ANOVA; Tukey's post-hoc tests for among group differences. Effective nuclei removal was achieved by SDS- and TT-based protocols. However, target DNA removal was achieved with SDS but not TT. SDS more effectively maintained qualitative tissue architecture compared to TT. The tensile modulus of the TT cohort increased, and stretchability decreased after decellularization in both SDS and TT. UTS was unaffected by either protocol. Higher overall diversity and quantity of core matrisome and matrisome-associated proteins was maintained in the SDS vs TT cohort post-decellularization. The results indicated that detergent selection affects multiple aspects of the resultant BAM biologic product. In the selected protocols, SDS was superior to TT efficacy, and maintenance of gross tissue architecture as well as maintenance of ECM proteins. Decellularization increased scaffold resistance to deformation in both cohorts. Future studies applying biologic scaffolds must consider the processing method and agents used to ensure that materials selected are optimized for characteristics that will facilitate effective translational use. [Display omitted] [ABSTRACT FROM AUTHOR]

Islet transplantation has been developed as an effective cell therapy strategy to treat the progressive life-threatening disease Type 1 diabetes (T1DM). To mimic the natural islets and achieve immune isolation, hydrogel encapsulation of multiple islet cell types is the current endeavor. Here, we present a microfiber loading with pancreatic α and β cells by microfluidic spinning for diabetes treatment. Benefiting from microfluidic technology, the cells could be controllably and continuously loaded in the alginate and methacrylated hyaluronic acid (Alg-HAMA) microfiber and maintained their high bioactivity. The resultant microfiber could then hold the capacity of dual-mode glucose responsiveness attributed to the glucagon and insulin secreted by the encapsulated pancreatic α and β cells. After transplantation into the brown adipose tissue (BAT), these cell-laden microfibers showed successful blood glucose control in rodents and avoided the occurrence of hypoglycemia. These results conceived that the multicellular microfibers are expected to provide new insight into artificial islet preparation, diabetes treatment, and regenerative medicine as well as tissue engineering. • The microfibers were generated with a double network of alginate and methacrylated hyaluronic acid. • The microfibers could simultaneously encapsulate pancreatic α and β cells and showed dual-mode glucose responsiveness. • The cell-laden microfibers maintained a long-term hormone-releasing function in vivo. • Cell-laden microfibers transplanted into diabetic mice could achieve glycemic control for 6 weeks. [Display omitted] [ABSTRACT FROM AUTHOR]

Currently, principles of tissue engineering and implantology are uniformly applied to all bone sites, disregarding inherent differences in collagen, mineral composition, and healing rates between craniofacial and long bones. These differences could potentially influence bone quality during the healing process. Evaluating bone quality during healing is crucial for understanding local mechanical properties in regeneration and implant osseointegration. However, site-specific changes in bone quality during healing remain poorly understood. In this study, we assessed newly formed bone quality in sub-critical defects in the maxilla and femur, while impairing collagen cross-linking using β-aminopropionitrile (BAPN). Our findings revealed that femoral healing bone exhibited a 73 % increase in bone volume but showed significantly greater viscoelastic and collagen changes compared to surrounding bone, leading to increased deformation during long-term loading and poorer bone quality in early healing. In contrast, the healing maxilla maintained equivalent hardness and viscoelastic constants compared to surrounding bone, with minimal new bone formation and consistent bone quality. However, BAPN-impaired collagen cross-linking induced viscoelastic changes in the healing maxilla, with no further changes observed in the femur. These results challenge the conventional belief that increased bone volume correlates with enhanced tissue-level bone quality, providing crucial insights for tissue engineering and site-specific implant strategies. The observed differences in bone quality between sites underscore the need for a nuanced approach in assessing the success of regeneration and implant designs and emphasize the importance of exploring site-specific tissue engineering interventions. Accurate measurement of bone quality is crucial for tissue engineering and implant therapies. Bone quality varies between craniofacial and long bones, yet it's often overlooked in the healing process. Our study is the first to comprehensively analyze bone quality during healing in both the maxilla and femur. Surprisingly, despite significant volume increase, femur healing bone had poorer quality compared to the surrounding bone. Conversely, maxilla healing bone maintained consistent quality despite minimal bone formation. Impaired collagen diminished maxillary healing bone quality, but had no further effect on femur bone quality. These findings challenge the notion that more bone volume equals better quality, offering insights for improving tissue engineering and implant strategies for different bone sites. [Display omitted] [ABSTRACT FROM AUTHOR]

Silk fibroin (SF) can be enzymatically crosslinked through tyrosine residues to fabricate hydrogels with good biocompatibility and tunable mechanical properties. Using tyramine substitution can increase the phenolic group content to facilitate the gelation kinetics and mechanical properties. In this study, a two-step chemical modification method is demonstrated to synthesize silk acid-tyramine (SA-TA) conjugates with a high phenolic group content (>7 mol%). The SA-TA shows rapid enzyme-catalyzed gelation property where the sol–gel transition takes less than 10 s at 37 °C, allowing cell encapsulation with uniform distribution while maintaining high cell viability (>90 %). Furthermore, the enzyme-catalyzed SA-TA hydrogels show enhanced storage modulus than enzyme-catalyzed SF hydrogels, long-term stability, and good cytocompatibility, indicating their great potential in 3D cell culture. The in vivo implantation study demonstrates that the SA-TA hydrogels are biodegradable with a mild immune response. This implies that SA-TA hydrogels can be applied in various medical applications, such as tissue engineering, cell delivery, and 3D bioprinting. In this study, a two-step chemical modification method is demonstrated to synthesize silk acid-tyramine (SA-TA) conjugates with a high phenolic group content (>7 mol%). Owing to the increased content of the phenolic group, the SA-TA shows rapid enzyme-catalyzed gelation property where the sol–gel transition takes less than 10 s at 37 °C, allowing cell encapsulation with uniform distribution while maintaining high cell viability (>90 %). Furthermore, the enzyme-catalyzed SA-TA hydrogels show enhanced storage modulus than enzyme-catalyzed SF hydrogels, long-term stability, and good cytocompatibility, indicating their great potential in 3D cell culture. The in vivo implantation study demonstrates that the SA-TA hydrogels are biodegradable with a mild immune response. This implies that SA-TA hydrogels can be applied in various medical applications, such as tissue engineering, cell delivery, and 3D bioprinting. [Display omitted] [ABSTRACT FROM AUTHOR]

Skin tissue engineering is undergoing tremendous expansion as a result from clinical needs, mandatory replacement of animal models and development of new technologies. Many approaches have been used to produce vascularized skin substitutes for grafting purposes showing the presence of capillary-like structures but with limited analysis of their in vitro maturation and plasticity. Such knowledge is however important for the development of tissue substitutes with improved implantation success as well as for validation of vascularization in vitro models, including as a readout in pharmacological analyses. For optimal interactions of cells with microenvironment and vasculature, we here used a cell sheet approach consisting in the sole production of matrix by the cells. In this context, we limited the density of endothelial cells seeded for self-assembly and rather relied on the stimulation of angiogenesis for the development of an extensive connected microvascular-like network. After detailed characterization of this network, we challenged its plasticity both during and after establishment of the skin substitute. We show that fine tuning of VEGF concentration and time of application differentially affects formation of capillary-like structures and their perivascular coverage. Furthermore, we performed a deep wound assay that displayed tissue repair and angiogenesis with unique characteristics of the physiological process. These studies demonstrate the importance of cell-derived microenvironment for the establishment of mature yet dynamic vascularized skin models allowing a wide range of pharmacological and basic investigations. The significant advancements in organ-on-chips and tissue engineering call for more relevant models including microvascularization with remodeling potential. While vascularized skin substitutes have been developed for years, focus has primarily been on the impact of microvascularization on implantation rather than on its in vitro characterization. We here developed a cell sheet-based vascularized skin substitute relying on angiogenesis, i.e. growth of vessel-like structures within the 3D model, rather than solely on endothelial cell self-assembly. We then characterized :1/ vascularization after modulation of angiogenic factor VEGF during the substitute construction; -2/ angiogenesis associated to tissue repair after deep mechanical wounding. These studies establish a solid physiologically relevant model for further investigation of skin cell interactions and in vitro wound healing. [Display omitted] [ABSTRACT FROM AUTHOR]

Tissue engineering presents a promising approach for the treatment of meniscal injuries, yet the development of meniscal scaffolds that exhibit both superior biomechanical properties and biocompatibility remains a considerable challenge. In this study, decellularized skin matrix (DSM) scaffolds were first prepared using porcine skin through decellularization and freeze-drying techniques. The DSM scaffold has favorable porosity, hydrophilicity, and biocompatibility. Importantly, the collagen content and tensile modulus of the scaffold are comparable to those of native meniscus (44.13 ± 2.396 mg/g vs. 42.41 ± 2.40 mg/g and 103.30 ± 2.98 MPa vs. 128.80 ± 9.115 MPa). Subsequently, the peptide PFSSTKT (PFS) with mesenchymal stem cells (MSCs) recruitment capability was used to modify DSM to construct DSM-PFS scaffolds. Compared to the DSM scaffold, the optimized DSM-PFS scaffold enhanced in vitro collagen and glycosaminoglycan (GAG) production and upregulated the expression of cartilage-specific genes. Furthermore, the DSM-PFS scaffold was more effective in recruiting MSCs in vitro. In vivo studies in rabbit models showed that the DSM-PFS scaffold successfully promoted meniscus tissue regeneration. Three months post-implantation, meniscus tissue formation can be observable, and after six months, the neo-meniscus exhibited tissue structure and tensile properties similar to the native meniscus. Notably, the DSM-PFS scaffold exhibited significant chondroprotective effects, slowing osteoarthritis (OA) progression. In conclusion, the DSM-PFS scaffold may represent a promising candidate for future applications in meniscus tissue engineering. We developed a decellularized skin matrix (DSM) meniscus scaffold using whole-layer porcine skin, demonstrating superior biomechanical strength and biocompatibility. Following modification with the stem cell-recruiting peptide PFS, the optimized DSM-PFS scaffold outperformed the DSM scaffold in cell attraction, collagen and glycosaminoglycan production, and cartilage-specific gene expression. Implanted into rabbit knee joints, the cell-free DSM-PFS scaffold induced meniscal tissue formation within three months, achieving the histological structure and tensile strength of the native meniscus by six months. Moreover, it significantly protected the cartilage. Our findings provide new insights into the fabrication of scaffolds for meniscal tissue engineering, with the DSM-PFS scaffold emerging as an ideal candidate for future applications. [Display omitted] [ABSTRACT FROM AUTHOR]

Bilateral communication between bones and muscles is essential for healing composite bone–muscle injuries from orthopedic surgeries and trauma. However, these injuries are often characterized by exaggerated inflammation, which can disrupt bone–muscle crosstalk, thereby seriously delaying the healing of either tissue. Existing approaches are largely effective at healing single tissues. However, simultaneous healing of multiple tissues remains challenging, with little research conducted to date. Here we introduce collagen patches that overcome this overlooked issue by harnessing the plasticity of macrophage phenotypes. Phosphatidylserine liposomes (PSLs) capable of shifting the macrophage phenotype from inflammatory M1 into anti-inflammatory/prohealing M2 were coated on collagen patches via a layer-by-layer method. Original collagen patches failed to improve tissue healing under inflammatory conditions coordinated by M1 macrophages. In contrast, PSL-coated collagen patches succeeded in accelerating bone and muscle healing by inducing a microenvironment dominated by M2 macrophages. In cell experiments, differentiation of preosteoblasts and myoblasts was completely inhibited by secretions of M1 macrophages but unaffected by those of M2 macrophages. RNA-seq analysis revealed that type I interferon and interleukin-6 signaling pathways were commonly upregulated in preosteoblasts and myoblasts upon stimulation with M1 macrophage secretions, thereby compromising their differentiation. This study demonstrates the benefit of PSL-mediated M1-to-M2 macrophage polarization for simultaneous bone and muscle healing, offering a potential strategy toward simultaneous regeneration of multiple tissues. Existing approaches for tissue regeneration, which primarily utilize growth factors, have been largely effective at healing single tissues. However, simultaneous healing of multiple tissues remains challenging and has been little studied. Here we demonstrate that collagen patches releasing phosphatidylserine liposomes (PSLs) promote M1-to-M2 macrophage polarization and are effective for simultaneous healing of bone and muscle. Transcriptome analysis using next-generation sequencing reveals that differentiation of preosteoblasts and myoblasts is inhibited by the secretions of M1 macrophages but promoted by those of M2 macrophages, highlighting the importance of timely regulation of M1-to-M2 polarization in tissue regeneration. These findings provide new insight to tissue healing of multiple tissues. [Display omitted] [ABSTRACT FROM AUTHOR]

Lymphedema microsurgery is an emerging treatment modality, with dissimilar long-term outcomes. One of the main technical challenges in lymphatic microsurgery is the identification and availability of suitable donor vessels for anastomosis. Tissue engineering using biomaterials has demonstrated promise in addressing vessel quality issues in other fields, but its application in microsurgery is still limited. Decellularized cellulose tubes were developed and bioengineered by decellularizing stems of Taraxacum-Ruderalia. The microscopic structure, mechanical properties, and residual DNA content of the cellulose tubes were evaluated. Human and murine skin fibroblasts and dermal lymphatic endothelial cells were isolated and cultured for recellularization studies. Biocompatibility, proliferative capacity, and ex-vivo endothelialization of the cellulose tubes were assessed as potential interposition grafts. Finally, the engineered cellulose tubes were assessed as interposing xenografts for lymphovenous anastomoses (LVA) in an ex-vivo swine limb model. The decellularized cellulose tubes exhibited a suitable microscopic structure, mechanical properties, and low residual DNA content. The tubes showed adequate biocompatibility, supported cell proliferation, and facilitated spontaneous ex-vivo endothelialization of lymphatic endothelial cells. In the swine limb model, LVA using the engineered cellulose tubes was successfully performed. This translational study presents the use of decellularized cellulose tubes as an adjunct for micro and supermicrosurgical reconstruction. The developed tubes demonstrated favorable structural, mechanical, and biocompatible properties, making them a potential candidate for improving long-term outcomes in lymphedema surgical treatment. The next translational step would be trialing the obtained tubes in a microsurgical in-vivo model. [ABSTRACT FROM AUTHOR]

The co-cultivation of mammalian cells with microalgae is an innovative concept to overcome one major limitation in tissue engineering and regenerative medicine: shortage of oxygen. Under illumination, photosynthetically active microalgae can produce oxygen, consequently increasing the potential construct dimension. In such co-cultures, mammalian cells are exposed to light, which does not reflect their natural conditions and can negatively influence their viability and functionality. In this study, 3D extrusion bioprinted pancreatic murine Islets of Langerhans should be co-cultivated with the bioprinted green microalgae species Scenedesmus sp. to improve the oxygen supply of the Islets in a potential insulin-producing implant. After defining the microalgae partner and a suitable co-culture medium that supported the functionality of both cell types, the focus was put on the effects of long-term red, blue and white light illumination on this system. First, the direct influence of light of different wavelengths on both microalgae and different mammalian cell types was investigated to find an ideal illumination regime, followed by an investigation into the influence of light on the culture medium. It was found that long-term exposure to blue or white light is detrimental to mammalian cells itself as well as the medium, while long-wave light has no effect on the function of mammalian cells. [ABSTRACT FROM AUTHOR]

Bone defects in animals can arise from various causes, including diseases, neoplasms, and most commonly, trauma. Comminuted fractures that exceed the critical size may heal poorly due to deficient or interrupted vascularization, resulting in an insufficient number of progenitor cells necessary for bone regeneration. In this context, 3D printing techniques using poly-L-lactic acid/graphene oxide (PLLA/GO) aim to address this issue by creating customized scaffolds combined with canine placenta hydrogel and mesenchymal stem cells for use in goat mandibles, compared to a control group using titanium plate fixation. Ten canine placentas were decellularized and characterized using histological techniques. A hydrogel derived from the canine placenta extracellular matrix (cpECM) was produced to improve cell attachment to the scaffolds. In vitro cytotoxicity and cell adhesion to the cpECM hydrogel were assessed by scanning electron microscopy (SEM). The resulting biomaterials, cpECM hydrogel and PLLA/GO scaffolds, maintained their functional structure and supported cell adhesion, maintenance, and proliferation in vitro. Thermography showed that PLLA/GO scaffolds with cpECM hydrogel performed effectively, similar to the control group. Computed tomography scans revealed bone calluses, suggesting an ongoing repair process. These findings demonstrate the innovative technological potential of these materials for use in surgical interventions. Future studies on PLLA/GO scaffolds will provide further insights into their effects on goat models. [ABSTRACT FROM AUTHOR]

We describe the development of a bioink to bioprint human induced pluripotent stem cells (hiPSCs) for possible cardiac tissue engineering using a gelatin methacrylate (GelMA)-based hydrogel. While previous studies have shown that GelMA at a low concentration (5% w/v) allows for the growth of diverse cells, its 3D printability has been found to be limited by its low viscosity. To overcome that drawback, making the hydrogel both compatible with hiPSCs and 3D-printable, we developed an extrudable GelMA-based bioink by adding xanthan gum (XG). The GelMA-XG composite hydrogel had an elastic modulus (~9 kPa) comparable to that of cardiac tissue, and enabled 3D printing with high values of printing accuracy (83%) and printability (0.98). Tests with hiPSCs showed the hydrogel's ability to promote their proliferation within both 2D and 3D cell cultures. The tests also showed that hiPSCs inside hemispheres of the hydrogel were able to differentiate into cardiomyocytes, capable of spontaneous contractions (average frequency of ~0.5 Hz and amplitude of ~2%). Furthermore, bioprinting tests proved the possibility of fabricating 3D constructs of the hiPSC-laden hydrogel, with well-defined line widths (~800 μm). [ABSTRACT FROM AUTHOR]

Cyanobacteria, commonly known as blue-green algae, are prevalent in freshwater systems and have gained interest for their potential in medical applications, particularly in skin regeneration. Among these, Synechococcus sp. strain PCC 7002 stands out because of its rapid proliferation and capacity to be genetically modified to produce growth factors. This study investigates the safety of Synechococcus sp. PCC 7002 when used in scaffolds for skin regeneration, focusing on systemic inflammatory responses in a murine model. We evaluated the following three groups: scaffolds colonized with genetically engineered bacteria producing hyaluronic acid, scaffolds with wild-type bacteria, and control scaffolds without bacteria. After seven days, we assessed systemic inflammation by measuring changes in cytokine profiles and lymphatic organ sizes. The results showed no significant differences in spleen, thymus, and lymph node weights, indicating a lack of overt systemic toxicity. Blood cytokine analysis revealed elevated levels of IL-6 and IL-1β in scaffolds with bacteria, suggesting a systemic inflammatory response, while TNF-α levels remained unaffected. Proteome profiling identified distinct cytokine patterns associated with bacterial colonization, including elevated inflammatory proteins and products, indicative of acute inflammation. Conversely, control scaffolds exhibited protein profiles suggestive of a rejection response, characterized by increased levels of cytokines involved in T and B cell activation. Our findings suggest that Synechococcus sp. PCC 7002 does not appear to cause significant systemic toxicity, supporting its potential use in biomedical applications. Further research is necessary to explore the long-term effects and clinical implications of these responses. [ABSTRACT FROM AUTHOR]

The study addresses the challenge of temperature sensitivity in pristine gelatin hydrogels, widely used in biomedical applications due to their biocompatibility, low cost, and cell adhesion properties. Traditional gelatin hydrogels dissolve at physiological temperatures, limiting their utility. Here, we introduce a novel method for creating stable hydrogels at 37 °C using pristine gelatin through photopolymerization without requiring chemical modifications. This approach enhances consistency and simplifies production and functionalization of the gelatin with bioactive molecules. The stabilization mechanism involves the partial retention of the triple-helix structure of gelatin below 25 °C, which provides specific crosslinking sites. Upon activation by visible light, ruthenium (Ru) acts as a photosensitizer that generates sulphate radicals from sodium persulphate (SPS), inducing covalent bonding between tyrosine residues and "locking" the triple-helix conformation. The primary focus of this work is the characterization of the mechanical properties, swelling ratio, and biocompatibility of the photopolymerized gelatin hydrogels. Notably, these hydrogels supported better cell viability and elongation in normal human dermal fibroblasts (NHDFs) compared to GelMA, and similar performance was observed for human pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). As a proof of concept for functionalization, gelatin was modified with selenous acid (GelSe), which demonstrated antioxidant and antimicrobial capacities, particularly against E. coli and S. aureus. These results suggest that pristine gelatin hydrogels, enhanced through this new photopolymerization method and functionalized with bioactive molecules, hold potential for advancing regenerative medicine and tissue engineering by providing robust, biocompatible scaffolds for cell culture and therapeutic applications. [ABSTRACT FROM AUTHOR]

This study investigates the application of a drop‐on‐demand (DOD) thermal inkjet (TIJ)‐based bioprinting system for the fabrication of cell‐laden hydrogel microparticles (HMPs) with tunable sizes. The TIJ bioprinting technique involves the formation of vapor bubbles within the print chamber through thermal energy, expelling small droplets of bio‐ink onto a substrate. The study employs a heat‐treated saponified gelatin‐based bio‐ink, HSP‐GelMA. This bio‐ink is modified through methacrylic anhydride functionalization and undergoes subsequent saponification and heat treatment processes. Various concentrations of SPAN 80 surfactant in mineral oil were evaluated to assess their influence on HMP size and stability. The results indicate a direct correlation, with higher SPAN 80 concentrations resulting in smaller and more stable HMPs. The study further investigates the influence of jetting volume on HMP size distribution, revealing that larger jetting volumes lead to increased HMP sizes, attributed to droplet coalescence. This is supported by our further study via a Monte Carlo simulation, which shows that the mean droplet diameter grows approximately linear with the number of dispensed droplets. In addition, the study demonstrates the capability of the TIJ bioprinting system to achieve multimaterial encapsulation within HMPs, exemplified by staining living cells with distinct cytoplasmic membrane dyes. The presented approach provides insights into the controlled fabrication of cell‐laden HMPs, highlighting the versatility of the TIJ bioprinting system for potential applications in tissue engineering and drug delivery. [ABSTRACT FROM AUTHOR]

Scaffold‐based tissue engineering provides an efficient approach for repairing uterine tissue defects and restoring fertility. In the current study, a novel trilayer tissue engineering scaffold with high similarity to the uterine tissue in structure was designed and fabricated via 4D printing, electrospinning and 3D bioprinting for uterine regeneration. Highly stretchable poly(l‐lactide‐co‐trimethylene carbonate) (PLLA‐co‐TMC, "PTMC" in short)/thermoplastic polyurethane (TPU) polymer blend scaffolds were firstly made via 4D printing. To improve the biocompatibility, porous poly(lactic acid‐co‐glycolic acid) (PLGA)/gelatin methacryloyl (GelMA) fibers incorporated with polydopamine (PDA) particles were produced on PTMC/TPU scaffolds via electrospinning. Importantly, estradiol (E2) was encapsulated in PDA particles. The bilayer scaffolds thus produced could provide controlled and sustained release of E2. Subsequently, bone marrow derived mesenchymal stem cells (BMSCs) were mixed with gelatin methacryloyl (GelMA)‐based inks and the formulated bioinks were used to fabricate a cell‐laden hydrogel layer on the bilayer scaffolds via 3D bioprinting, forming ultimately biomimicking trilayer scaffolds for uterine tissue regeneration. The trilayer tissue engineering scaffolds thus formed exhibited a shape morphing ability by transforming from the planar shape to tubular structures when immersed in the culture medium at 37°C. The trilayer tissue engineering scaffolds under development would provide new insights for uterine tissue regeneration. [ABSTRACT FROM AUTHOR]

Droplet‐based bioprinting has shown remarkable potential in tissue engineering and regenerative medicine. However, it requires bioinks with low viscosities, which makes it challenging to create complex 3D structures and spatially pattern them with different materials. This study introduces a novel approach to bioprinting sophisticated volumetric objects by merging droplet‐based bioprinting and cryobioprinting techniques. By leveraging the benefits of cryopreservation, we fabricated, for the first time, intricate, self‐supporting cell‐free or cell‐laden structures with single or multiple materials in a simple droplet‐based bioprinting process that is facilitated by depositing the droplets onto a cryoplate followed by crosslinking during revival. The feasibility of this approach is demonstrated by bioprinting several cell types, with cell viability increasing to 80%–90% after up to 2 or 3 weeks of culture. Furthermore, the applicational capabilities of this approach are showcased by bioprinting an endothelialized breast cancer model. The results indicate that merging droplet and cryogenic bioprinting complements current droplet‐based bioprinting techniques and opens new avenues for the fabrication of volumetric objects with enhanced complexity and functionality, presenting exciting potential for biomedical applications. [ABSTRACT FROM AUTHOR]

Bioprinting is a highly promising application area of additive manufacturing technology that has been widely used in various fields, including tissue engineering, drug screening, organ regeneration, and biosensing. Its primary goal is to produce biomedical products such as artificial implant scaffolds, tissues and organs, and medical assistive devices through software‐layered discrete and numerical control molding. Despite its immense potential, bioprinting technology still faces several challenges. It requires concerted efforts from researchers, engineers, regulatory bodies, and industry stakeholders are principal to overcome these challenges and unlock the full potential of bioprinting. This review systematically discusses bioprinting principles, applications, and future perspectives while also providing a topical overview of research progress in bioprinting over the past two decades. The most recent advancements in bioprinting are comprehensively reviewed here. First, printing techniques and methods are summarized along with advancements related to bioinks and supporting structures. Second, interesting and representative cases regarding the applications of bioprinting in tissue engineering, drug screening, organ regeneration, and biosensing are introduced in detail. Finally, the remaining challenges and suggestions for future directions of bioprinting technology are proposed and discussed. Bioprinting is one of the most promising application areas of additive manufacturing technology that has been widely used in various fields. It aims to produce biomedical products such as artificial implant scaffolds, tissues and organs, and medical assistive devices. This review systematically discusses bioprinting principles, applications, and future perspectives, which provides a topical description of the research progress of bioprinting. [ABSTRACT FROM AUTHOR]

Respiratory diseases, claim over eight million lives annually. However, the transition from preclinical to clinical phases in research studies is often hindered, partly due to inadequate representation of preclinical models in clinical trials. To address this, we conducted a proof‐of‐concept study using an ex vivo model to identify lung pathologies and to screen therapeutics in a humanized rodent model. We extracted and decellularized mouse heart‐lung tissues using a detergent‐based technique. The lungs were then seeded and cultured with human cell lines (BEAS‐2B, A549, and Calu3) for 6−10 days, representing healthy lungs, cancerous states, and congenital pathologies, respectively. By manipulating cultural conditions and leveraging the unique characteristics of the cell lines, we successfully modeled various pathologies, including advanced‐stage solid tumors and the primary phase of SARS‐CoV‐2 infection. Validation was conducted through histology, immunofluorescence staining, and pathology analysis. Additionally, our study involved pathological screening of the efficacy and impact of key anti‐neoplastic therapeutics (Cisplatin and Wogonin) in cancer models. The results highlight the versatility and strength of the ex vivo model in representing crucial lung pathologies and screening therapeutics during the preclinical phase. This approach holds promise for bridging the gap between preclinical and clinical research, aiding in the development of effective treatments for respiratory diseases, including lung cancer. [ABSTRACT FROM AUTHOR]

Due to the absence of blood vessels and nerves, the regenerative potential of articular cartilage is significantly constrained. This implies that the impact of a ruptured cartilage extends to the entire joint. Osteoarthritis, a health condition, may arise due to injury and the progressive breakdown of joint tissues. The progression of osteoarthritis can be accelerated by the extensive degradation of articular cartilage, which is ranked as the third most prevalent musculoskeletal disorder necessitating rehabilitation, following low back pain and fractures. The existing therapeutic interventions for cartilage repair exhibit limited efficacy and seldom achieve complete restoration of both functional capacity and tissue homeostasis. Emerging technological advancements in the field of tissue engineering hold significant promise for the development of viable substitutes for cartilage tissue, capable of exhibiting functional properties. The overarching strategy involves ensuring that the cell source is enriched with bioactive molecules that facilitate cellular differentiation and/or maturation. This review provides a comprehensive summary of recent advancements in the field of cartilage tissue engineering. Additionally, it offers an overview of recent clinical trials that have been conducted to examine the latest research developments and clinical applications pertaining to weakened articular cartilage and osteoarthritis. [ABSTRACT FROM AUTHOR]

Tissue engineering offers a new horizon for restoring the function of damaged tissues and organs. Here, bone regeneration potential of three-dimensional (3D) scaffold made of collagen/beta-tricalcium phosphate/ginger hydroalcoholic extract (COL-ß-TCP-GIN) loaded with stem cells was evaluated. The scaffolds with different component ratios were fabricated using a freeze dryer to obtain the optimum composition. The scaffolds' chemical, physical, and biological characteristics were evaluated using scanning electron microscope, fourier transform infrared spectroscopy, tensile testing machine, and cytotoxicity assay. The optimum scaffold's bone repairing potential was assessed with loaded synovial membrane mesenchymal stem cells (SM-MSCs) in mandibular bone defect of a rat animal model after two months. The ß-TCP component up to 30% could increase the tensile strength of the freeze-dried scaffold. In comparison, the GIN up to 5% was selected as a sufficient amount to be incorporated with the scaffolds. The morphology of scaffolds showed a suitable porosity for cells to proliferate and migrate. In vitro cytotoxicity results showed that GIN increased the cell viability up to 7 days. Regarding in vivo bone regeneration study, histopathology and stereology assessments showed the mandibular bone formation in COL/β-TCP/GIN scaffolds with SM-MSCs group significantly increased compared to COL/β-TCP/GIN without cells and sham groups. These results demonstrated the effectiveness of COL/β-TCP/GIN scaffold with SM-MSCs to induce bone formation, and this composite can be applied in dental and reconstructive surgery. [ABSTRACT FROM AUTHOR]

Purpose: Cardiac tissue engineering is suggested as a promising approach to overcome problems associated with impaired myocardium. This is the first study to investigate the use of BC and gelatin for cardiomyocyte adhesion and growth. Methods: Bacterial cellulose (BC) membranes were produced by Komagataeibacter xylinus and coated or mixed with gelatin to make gelatin-coated BC (BCG) or gelatin-mixed BC (mBCG) scaffolds, respectively. BC based-scaffolds were characterized via SEM, FTIR, XRD, and AFM. Neonatal rat-ventricular cardiomyocytes (nr-vCMCs) were cultured on the scaffolds to check the capability of the composites for cardiomyocyte attachment, growth and expansion. Results: The average nanofibrils diameter in all scaffolds was suitable (~ 30–65 nm) for nr-vCMCs culture. Pore diameter (≥ 10 µm), surface roughness (~ 182 nm), elastic modulus (0.075 ± 0.015 MPa) in mBCG were in accordance with cardiomyocyte requirements, so that mBCG could better support attachment of nr-vCMCs with high concentration of gelatin, and appropriate surface roughness. Also, it could better support growth and expansion of nr-vCMCs due to submicron scale of nanofibrils and proper elasticity (~ 0.075 MPa). The viability of nr-vCMCs on BC and BCG scaffolds was very low even at day 2 of culture (~ ≤ 40%), but, mBCG could promote a metabolic active state of nr-vCMCs until day 7 (~ ≥ 50%). Conclusion: According to our results, mBCG scaffold was the most suitable composite for cardiomyocyte culture, regarding its physicochemical and cell characteristics. It is suggested that improvement in mBCG stability and cell attachment features may provide a convenient scaffold for cardiac tissue engineering. [ABSTRACT FROM AUTHOR]

Degenerative spinal pathology is a widespread medical issue, and spine fusion surgeries are frequently performed. In this study, we fabricated an injectable bioactive click chemistry polymer cement for use in spinal fusion and bone regrowth. Taking advantages of the bioorthogonal click reaction, this cement can be crosslinked by itself eliminating the addition of a toxic initiator or catalyst, nor any external energy sources like UV light or heat. Furthermore, nano‐hydroxyapatite (nHA) and microspheres carrying recombinant human bone morphogenetic protein‐2 (rhBMP‐2) and recombinant human vascular endothelial growth factor (rhVEGF) were used to make the cement bioactive for vascular induction and osteointegration. After implantation into a rabbit posterolateral spinal fusion (PLF) model, the cement showed excellent induction of new bone formation and bridging bone, achieving results comparable to autograft control. This is largely due to the osteogenic properties of nano‐hydroxyapatite (nHA) and the released rhBMP‐2 and rhVEGF growth factors. Since the availability of autograft sources is limited in clinical settings, this injectable bioactive click chemistry cement may be a promising alternative for spine fusion applications in addressing various spinal conditions. [ABSTRACT FROM AUTHOR]

Replicating the architecture of extracellular matrices (ECM) is crucial in tissue engineering to support tissues’ natural structure and functionality. The ECM’s structure plays a significant role in directing cell alignment. Electrospinning is an effective technique for fabricating nanofibrous substrates that mimic the architecture of extracellular matrices (ECM). This study aims to evaluate the efficacy of resin 3D-printed scaffolds made from a low-conductivity material (i.e., a resin composed of methacrylated oligomers, monomers, and photoinitiators) in directing the alignment of electrospun polycaprolactone (PCL) nanofibers. Six 3D-printed scaffolds were fabricated using stereolithography (SLA) technology and strategically positioned on an aluminum foil collector plate during electrospinning. The structured geometry of the scaffolds, rather than the local electric field distribution, is hypothesized to guide nanofiber alignment. Images acquired through the scanning electron microscopy (SEM) were used to analyze and statistically quantify the nanofibrous scaffolds to evaluate the alignment of nanofibers over the scaffolds compared to a set of randomly deposited control nanofiber samples in the absence of the 3D printed scaffolds. SEM images also showed significant alignment of nanofibers within the pores of scaffolds, using histograms as a means for indicating the distribution of orientation angles. Statistical analysis revealed that this distribution deviates from normality due to the deviations in the tails and the existence of relatively smaller peaks at angles relative to 0°, particularly within a range of ± 50° and ± 40°. It is further found that the average peak orientation angle relative to 0° had a maximum probability of 0.014. Furthermore, the statistical analysis confirmed the distribution and significant differences in orientation between test samples with 3D-printed scaffolds and control samples. These findings demonstrate the effectiveness of resin 3D-printed scaffolds, particularly their geometric filtering effect, leading to controlled nanofiber alignment, which is proposed to be beneficial for enhancing cell adhesion, proliferation, and cell migration in tissue engineering applications. [ABSTRACT FROM AUTHOR]

Background: Cryopreservation is a crucial procedure for safeguarding cells or other biological constructs, showcasing considerable potential for applications in tissue engineering and regenerative medicine. Aims: This study aimed to evaluate the effectiveness of different cryopreservation conditions on human cells viability. Methods: A set of cryopreserved data from Department of Tissue Engineering and Regenerative Medicine (DTERM) cell bank were analyse for cells attachment after 24 h being revived. The revived cells were analysed based on different cryopreservation conditions which includes cell types (skin keratinocytes and fibroblasts, respiratory epithelial, bone marrow mesenchymal stem cell (MSC); cryo mediums (FBS + 10% DMSO; commercial medium); storage durations (0 to > 24 months) and locations (tank 1–2; box 1–5), and revival methods (direct; indirect methods). Human dermal fibroblasts (HDF) were then cultured, cryopreserved in different cryo mediums (HPL + 10% DMSO; FBS + 10% DMSO; Cryostor) and stored for 1 and 3 months. The HDFs were revived using either direct or indirect method and cell number, viability and protein expression analysis were compared. Results: In the analysis cell cryopreserved data; fibroblast cells; FBS + 10% DMSO cryo medium; storage duration of 0–6 months; direct cell revival; storage in vapor phase of cryo tank; had the highest number of vials with optimal cell attachment after 24 h revived. HDFs cryopreserved in FBS + 10% DMSO for 1 and 3 months with both revival methods, showed optimal live cell numbers and viability above 80%, higher than other cryo medium groups. Morphologically, the fibroblasts were able to retain their phenotype with positive expression of Ki67 and Col-1. HDFs cryopreserved in FBS + 10% DMSO at 3 months showed significantly higher expression of Ki67 (97.3% ± 4.62) with the indirect revival method, while Col-1 expression (100%) was significantly higher at both 1 and 3 months compared to other groups. Conclusion: In conclusion, fibroblasts were able to retain their characteristics after various cryopreservation conditions with a slight decrease in viability that may be due to the thermal-cycling effect. However, further investigation on the longer cryopreservation periods should be conducted for other types of cells and cryo mediums to achieve optimal cryopreservation outcomes. Highlights: Cryopreservation optimizes cell recovery, viability for tissue engineering. Fibroblasts maintain phenotype in FBS + 10% DMSO, but longer periods need scrutiny. [ABSTRACT FROM AUTHOR]

Congenital Heart Defects (CHDs) are the most common congenital anomalies, affecting between 4 and 75 per 1000 live births. Cardiovascular patches (CVPs) are frequently used as part of the surgical armamentarium to reconstruct cardiovascular structures to correct CHDs in pediatric patients. This review aims to evaluate the history of cardiovascular patches, currently available options, clinical applications, and important features of these patches. Performance and outcomes of different patch materials are assessed to provide reference points for clinicians. The target audience includes clinicians seeking data on clinical performance as they make choices between different patch products, as well as scientists and engineers working to develop patches or synthesize new patch materials. [ABSTRACT FROM AUTHOR]

Objective. Currently, the summaries of research on periodontal ligament stem cells (PDLSCs) are mainly reviews, and the systematic evaluation of all relevant studies is lacking. The aim of our study was to reveal the research status and developmental trends of PDLSCs using bibliometric analyses. Methods. Publications on PDLSC from 2004 to 2023 in the PubMed database were searched and then screened according to certain inclusion and exclusion criteria. Two researchers browsed the included papers and recorded information such as the research type and research model. The VOSviewer software was used to analyze the distribution of authors, journals, and institutions. The contents and directions of PDLSC research were summarized by analyzing high‐frequency keywords. The CiteSpace software was used to monitor burst words, determine hot factors, and indicate developmental trends. Results. During the past two decades, the number of studies on PDLSCs increased. China published the most related papers. The primary type of article was basic research. Among core journals, the Journal of Periodontal Research had the highest number of publications. The Fourth Military Medical University (China) was leading in the number of articles on PDLSCs. Research topics mainly included mechanism of periodontal diseases, tissue engineering and regeneration, biological characteristics of PDLSCs, and comparison with other stem cells. Infectious inflammation and mechanical stimulation were important pathological conditions and research topics. Conclusion. The research of PDLSCs is still in a rapid development stage. Our study provides new insights into the current research status and future trend in this field. [ABSTRACT FROM AUTHOR]

Cell therapy has emerged as a viable approach for treating damaged organs or tissues, particularly with advancements in stem cell research and regenerative medicine. The innovative technique of cell sheet engineering offers the potential to create a cell-dense lamellar structure that preserves the extracellular matrix (ECM) secreted by cells, along with the cell-matrix and intercellular junctions formed during in vitro cultivation. In recent years, significant progress has been made in developing cell sheet engineering technology. A variety of novel materials and methods were utilized for enzyme-free cell detachment during the cell sheet formation process. The complexity of cell sheet structures increased to meet advanced usage demands. This review aims to provide an overview of the preparation methods and types of cell sheets, thereby enhancing the understanding of this rapidly evolving technology and offering a fresh perspective on the development and future application of cell sheet engineering. [ABSTRACT FROM AUTHOR]

BACKGROUND: Due to the very limited ability of articular cartilage to repair itself, articular cartilage defects caused by natural degeneration or trauma often cannot be repaired on their own, which triggers or aggravates osteoarthritis and even leads to severe disability. Therefore, the repair treatment after articular cartilage injury has become an urgent clinical problem. OBJECTIVE: To summarize the hot research topics and development trends in the field of articular cartilage repair using bibliometric analysis. METHODS: Literature related to articular cartilage repair was searched from the Web of Science Core Collection from 2000 to 2023, and bibliometric and visualization analyses were carried out using VOSviewer, Citespeace and Bibliometrix R-package. RESULTS AND CONCLUSION: The annual publication volume in the field of articular cartilage repair shows an overall increasing trend, with the United States, China, and Germany being the top three countries in terms of publication volume, and the research institutions focus on universities and hospitals, with Harvard University, New York Hospital for Special Surgery, and Shanghai Jiao Tong University being the top three institutions in terms of publication volume. AMERICAN JOURNAL OF SPORTS MEDICINE is the journal that publishes the most research literature in the field, and BIOMATERIALS is the journal with the highest number of citations in the field. “Injectable hydrogels for cartilage and bone tissue engineering” is the most cited document published in the last decade, and the author with the most publications is Madry Henning, an active author in the field. The active authors in this field have formed a number of stable research teams with each other, and the cooperation between different teams needs to be further strengthened. Intra-articular injections, high tibial osteotomies, hydrogels, drug delivery, inflammation, cartilage regeneration, and scaffolds are the current hot topics of research in this field, and the application of 3D printing technology, bio-inks, silk proteins, injectable hydrogels, and exosomes in articular cartilage repair may be the frontier of research in this field. Integrating various innovative technologies and methods for more effective, durable and functional cartilage tissue regeneration and repair, and facilitating the clinical translation of the relevant technologies and methods by conducting more high-quality clinical studies may be the future research trend in this field. [ABSTRACT FROM AUTHOR]

One of the hallmarks of tissue repair is the production of reactive oxygen species (ROS), which modulate processes such as cell proliferation. Although several attempts have been made to manipulate ROS levels to increase tissue repair, the lack of techniques able to remotely manipulate the redox homeostasis with spatio–temporal fashion has hindered its progress. Herein, a new approach for tuning the ROS levels using magnetic nanoparticles (MNPs) that act as nanoheaters when exposed to an alternating magnetic field is presented. Two manganese–iron oxide (MnxFe3−xO4) MNPs (with a low and a high Mn2+ content) are designed and probed for the possibility of modulating the ROS balance by magneto–thermal stimulation in the invertebrate model organism Hydra vulgaris, able to fully regenerate. By evaluating the expression of selected genes involved in the maintenance of ROS homeostasis and proliferation pathways, a biphasic modulation of the ROS levels played by the MNPs is found. While MNPs with a lower Mn2+ content are able to positively modulate the regeneration potential under magnetostimulation, MNPs with a higher Mn2+ content cause a different redox imbalance, negatively affecting the regeneration dynamic. This innovative approach reveals a novel way of manipulating redox homeostasis that can advance in the field of tissue engineering. [ABSTRACT FROM AUTHOR]

Tumor extracellular matrices (ECM) exhibit aberrant changes in composition and mechanics compared to normal tissues. Proteoglycans (PG) are vital regulators of cellular signaling in the ECM with the ability to modulate receptor tyrosine kinase (RTK) activation via their sulfated glycosaminoglycan (sGAG) side chains. However, their role on tumor cell behavior is controversial. Here, it is demonstrated that PGs are heavily expressed in lung adenocarcinoma (LUAD) patients in correlation with invasive phenotype and poor prognosis. A bioengineered human lung tumor model that recapitulates the increase of sGAGs in tumors in an organotypic matrix with independent control of stiffness, viscoelasticity, ligand density, and porosity, is developed. This model reveals that increased sulfation stimulates extensive proliferation, epithelial‐mesenchymal transition (EMT), and stemness in cancer cells. The focal adhesion kinase (FAK)‐phosphatidylinositol 3‐kinase (PI3K) signaling axis is identified as a mediator of sulfation‐induced molecular changes in cells upon activation of a distinct set of RTKs within tumor‐mimetic hydrogels. The study shows that the transcriptomic landscape of tumor cells in response to increased sulfation resembles native PG‐rich patient tumors by employing integrative omics and network modeling approaches. [ABSTRACT FROM AUTHOR]

Urogenital reconstructive malformation surgery is sometimes hampered by lack of tissue for the repair. We have previously shown that autologous micrografting allows for single-staged scaffold cellularization after surgical implantation. Here, a collagen-based scaffold reinforced with biodegradable mesh and a stent was implanted as a bladder conduit in ten full-grown female minipigs. We aimed to assess short-term regenerative outcomes, safety, and feasibility of implanting tubular urinary micrografted scaffolds versus acellular controls. Five scaffolds were embedded with autologous urothelial micrografts harvested perioperatively. After six weeks, all animals were assessed by cystoscopy, CT-urography, and microanatomical assessment of the urinary conduits. The procedure proved technically feasible within the confines of a regular surgical theater, with duration-times comparable to corresponding conventional procedures. No animals experienced postoperative complications, and all implanted conduits were patent at follow-up. Improved tissue regeneration was observed in the micrografted conduits compared with the acellular controls, including increased luminal epithelialization, increased cell proliferation, decreased cell apoptosis, and increased conduit vascularization. We concluded that single-staged on-site construction and implantation of tissue engineered urinary conduits proved feasible and safe, with improved regenerative potentials in micrografted conduits. This study presents a new approach to urinary conduits, and merits further investigations for advancement towards clinical translation. [ABSTRACT FROM AUTHOR]

B. mori silk has been extensively utilized to create Regenerated Silk Fibroin solutions (RSF) for a wide range of technological applications, including tissue engineering, drug delivery, biomaterials, and adhesives. In this study, an RSF‐dopamine (DA) composite is introduced that yields easily deployable hydrogel fibers possessing adhesive properties that can be released on demand to capture and retrieve loads from a distance. The RSF‐DA serves as an artificial dope, combining the functional attributes of silk fibroin (i.e., hydrogel formation) and dopamine (i.e., adhesion) to instantly generate adjustable hydrogel fibers displaying a sticky behavior. The mechanical strength and adhesive characteristics of this material are assessed using tensile and lap‐shear tests. Furthermore, the possibility of tuning these properties is demonstrated by adding chitosan (Ch) and borate ions (BB), leading to remarkable mechanical and adhesive performances up to 107 MPa and 280 kPa, respectively, which allows the retrieval of objects from the ejected structure. This process can be finely tuned to achieve a controlled fabrication of instantaneously formed adhesive hydrogel fibers for manifold applications, mimicking living organisms’ ability to eject tunable adhesive functional threads. [ABSTRACT FROM AUTHOR]

Tympanic membrane (TM) perforation is a serious ear discomfort that can cause hearing loss and make the middle ear vulnerable to infections. In this study, a unique TM patch is designed to mimic the structure of the natural eardrum for tissue engineering of TM perforations. Gelatin methacryloyl (GelMA)‐based TM patches are equipped with microneedles (MNs) to better adhere to the perforation site and developed using the digital light processing (DLP) based 3D printing technique. To impart biofunctionality to the 3D‐printed patches, their surfaces are coated with gentamicin (GEN) loaded poly(vinyl alcohol) (PVA) nanoparticles (NPs) using the Electrohydrodynamic Atomization (EHDA) method. The physicochemical characteristics, drug release behaviour, antimicrobial properties and biocompatibility of GelMA, PVA NP‐coated GelMA, and GEN@PVA NP‐coated GelMA patches are investigated. Morphological analyses confirmed that 3D‐printed GelMA patches are fabricated in desired sizes and geometries and successfully coated with NPs. In vitro antibacterial tests revealed that GEN@PVA NP‐coated GelMA patches have antibacterial activities against Staphylococcus aureus and Escherichia coli. Moreover, in vitro cell culture studies indicated that all GelMA‐based patches have no cytotoxic effect on L929 mouse fibroblast cells. Considering all, these specially designed biofunctional 3D‐printed GelMA patches can be an effective therapeutic approach for repairing TM perforations. [ABSTRACT FROM AUTHOR]