Functional Tissue Engineering Research Articles

3D printed titanium carbide MXene-coated polycaprolactone scaffolds for guided neuronal growth and photothermal stimulation

The exploration of neural circuitry is paramount for comprehending the computational mechanisms and physiology of the brain. Despite significant advances in materials and fabrication techniques, controlling neuronal connectivity and response in 3D remains a formidable challenge. Here, we introduce a method for engineering the growth of 3D neural circuits with the capability for optical stimulation. We fabricate bioactive interfaces by melt electrospinning writing (MEW) 3D polycaprolactone (PCL) scaffolds followed by coating with titanium carbide (Ti3C2Tx MXene). Beyond enhancing hydrophilicity, cell adhesion, and electrical conductivity, the Ti3C2Tx MXene coating enables optocapacitance-based neuronal stimulation, induced by localized temperature increases upon illumination. This approach offers a pathway for additive manufacturing of neural tissues endowed with optical control, facilitating functional tissue engineering and neural circuit computation.

Biofabrication of nanocomposite-based scaffolds containing human bone extracellular matrix for the differentiation of skeletal stem and progenitor cells

Autograft or metal implants are routinely used in skeletal repair. However, they fail to provide long-term clinical resolution, necessitating a functional biomimetic tissue engineering alternative. The use of native human bone tissue for synthesizing a biomimetic material ink for three-dimensional (3D) bioprinting of skeletal tissue is an attractive strategy for tissue regeneration. Thus, human bone extracellular matrix (bone-ECM) offers an exciting potential for the development of an appropriate microenvironment for human bone marrow stromal cells (HBMSCs) to proliferate and differentiate along the osteogenic lineage. In this study, we engineered a novel material ink (LAB) by blending human bone-ECM (B) with nanoclay (L, Laponite®) and alginate (A) polymers using extrusion-based deposition. The inclusion of the nanofiller and polymeric material increased the rheology, printability, and drug retention properties and, critically, the preservation of HBMSCs viability upon printing. The composite of human bone-ECM-based 3D constructs containing vascular endothelial growth factor (VEGF) enhanced vascularization after implantation in an ex vivo chick chorioallantoic membrane (CAM) model. The inclusion of bone morphogenetic protein-2 (BMP-2) with the HBMSCs further enhanced vascularization and mineralization after only seven days. This study demonstrates the synergistic combination of nanoclay with biomimetic materials (alginate and bone-ECM) to support the formation of osteogenic tissue both in vitro and ex vivo and offers a promising novel 3D bioprinting approach to personalized skeletal tissue repair.Graphic abstract

Effect of different acids during collagen extraction the bone and fins from purple-spotted bigeye (Priacanthus tayenus Richardson, 1846) and their physicochemical properties

This study was carried out to determine the effectiveness of different acids (acetic, lactic and citric)-on the extraction of collagen from the bone and fins of purple-spotted bigeye (Priacanthus tayenus Richardson, 1846) particularly on the yield and physicochemical properties. The physicochemical properties were characterized by electrophoretic pattern, X-ray diffraction, FT-IR, colour and pH. The citric acid-extracted collagen (CAC) has a higher percentage of yield (1.93±0.57), followed by lactic acid-extracted collagen (LAC) (1.43±0.42) and acetic acid-extracted collagen (AAC) (0.83±0.18) although those collagens did not differ significantly (p>0.05), and those acid solubilized collagens (ASC) contained about 96.82 - 96.89 mg/g of hydroxyproline. The ASCs obtained showed similar electrophoretic patterns due to the presence of identical α chains (α1 and α2) and classified as type I collagen. UV absorption spectrum in all ASCs was approximately around 231.0 - 231.5 nm. For diffraction angle analysis, first diffraction was detected at 7.23 - 7.41° and second diffraction was at 19.41 - 20.29° in all ASCs. Major absorption peaks of FT-IR spectrum, viz. amide A, amide B and amide I-III were present in the collagens with different wavenumbers. The results indicated that the physicochemical properties of ASCs extracted in the present study were comparable with collagens reported from other fish species. The findings suggested that ASCs from purple-spotted bigeye bones and fins could serve as alternative source of collagen in functional food, pharmaceutics and tissue engineering production, particularly found in acetic acidextracted collagen because of its properties.

Tissue Engineering for Cardiovascular Regeneration: Brief Review on Scaffold Fabrication

Tissue engineering is the combination of engineering and fundamental sciences to develop an artificial organ that derived from the tissue human sources. It involves the construction of three main pillars include cell sources, scaffold materials and biological factors. These three important elements are necessary to be incorporated and integrated well, to construct a functional artificial tissue engineering product. Few applications can be associated with the expansion of tissue engineering where cardiovascular regeneration is one of the targets of tissue engineering. Among the fabrication techniques, electrospinning, three-dimensional printing, molding and decellularization, are four engineering methods that are commonly used in fabricating scaffolds for cardiovascular tissue engineering. One of the purposes of the emerging of cardiovascular tissue engineering is the limitation of current commercialized patches or membranes that often cause post-complications following the cardiovascular treatments. This review paper covering a brief introduction on the tissue engineering for cardiovascular regeneration, focusing on the scaffold fabrication.

Mechanical Stimulation of Adipose-DerivedStromal/Stem Cells for Functional Tissue Engineering of the Musculoskeletal System via Cyclic Hydrostatic Pressure, Simulated Microgravity, and Cyclic Tensile Strain.

It is critical that human adipose-derived stromal/stem cell (hASC) tissue engineering therapies possess appropriate mechanical properties in order to restore the function of the load-bearing tissues of the musculoskeletal system. In an effort to elucidate hASC response to mechanical stimulation and develop mechanically robust tissue-engineered constructs, recent research has utilized a variety of mechanical loading paradigms, including cyclic tensile strain, cyclic hydrostatic pressure, and mechanical unloading in simulated microgravity. This chapter will describe the methods for applying these mechanical stimuli to hASC to direct differentiation for functional tissue engineering of the musculoskeletal system.

Construction of ultrasonically treated collagen/silk fibroin composite scaffolds to induce cartilage regeneration

A novel tissue-specific functional tissue engineering scaffold for cartilage repair should have a three-dimensional structure, good biosafety and biological activity, and should be able to promote cartilage tissue regeneration. This study aimed to determine the effect of ultrasound-treated collagen/silk fibroin (Col/SF) composite scaffolds with good mechanical properties and high biological activity on cartilage repair. The characteristics of the scaffolds with different Col/SF ratios (7:3, 8:2, and 9:1) were determined by scanning electron microscopy, Fourier-transform infrared spectroscopy, and porosity, water absorption, and compression tests. In vitro evaluations revealed the biocompatibility of the Col/SF scaffolds. Results suggested that the optimal ratio of Col/SF composite scaffolds was 7:3. The Col/SF scaffolds induced adipose-derived stem cells to undergo chondrogenic differentiation under chondrogenic culture conditions. The efficiency of Col/SF scaffolds for cartilage regeneration applications was further evaluated using an in vivo model of full-thickness articular cartilage defects in New Zealand rabbits. The Col/SF scaffolds effectively promoted osteochondral regeneration as evidenced by macroscopic, histological, and immunohistochemical evaluation. The study demonstrates that ultrasound-treated Col/SF scaffolds show great potential for repairing cartilage defects.

Vascular reconstruction of the decellularized biomatrix for whole-organ engineering-a critical perspective and future strategies.

Whole-organ re-engineering is the most challenging goal yet to be achieved in tissue engineering and regenerative medicine. One essential factor in any transplantable and functional tissue engineering is fabricating a perfusable vascular network with macro- and micro-sized blood vessels. Whole-organ development has become more practical with the use of the decellularized organ biomatrix (DOB) as it provides a native biochemical and structural framework for a particular organ. However, reconstructing vasculature and re-endothelialization in the DOB is a highly challenging task and has not been achieved for constructing a clinically transplantable vascularized organ with an efficient perfusable capability. Here, we critically and articulately emphasized factors that have been studied for the vascular reconstruction in the DOB. Furthermore, we highlighted the factors used for vasculature development studies in general and their application in whole-organ vascular reconstruction. We also analyzed in detail the strategies explored so far for vascular reconstruction and angiogenesis in the DOB for functional and perfusable vasculature development. Finally, we discussed some of the crucial factors that have been largely ignored in the vascular reconstruction of the DOB and the future directions that should be addressed systematically.

Shape-adapted self-gelation hydrogel powder for high-performance hemostasis and wound healing

In the battlefield, accidents, first aid, and hospital environments, uncontrollable bleeding caused by severe wound infections pose a serious risk of death and high costs. However, performing ultrafast hemostasis without any external conditions remains challenging. In this paper, polyacrylic acid, polyacrylamide, tea polyphenols, and Yunnan Baiyao (PAA/PAAm/T/YNBY, PPTY) are used to prepare self-gelling powder with shape adaptability. When the mass fraction of PAA and PAAm is 10 wt% and 8 wt%, the hydrogel can form the most intermolecular hydrogen bonds, exhibiting the best adhesion ability and mechanical strength. When the amount of YNBY was 0.1 wt%, the hydrogel had the most suitable cellular compatibility and hemostatic ability. At this time, the hydrogel powder can achieve the fastest self-gelling (< 3 s), the strongest adhesion (interface adhesion > 499.4 N m − 1), and the fastest hemostatic speed (< 30 s). With the addition of a small amount of YNBY (0.1 wt%), the hemostatic efficiency of the hydrogel can be greatly improved. The combination of hydrogel and YNBY not only prolongs the duration of YNBY ingredient efficacy but also enhances its biocompatibility. This easy-to-use but effective hydrogel can stop bleeding, promote wound healing in emergencies, and has opportunities in functional materials, tissue engineering, biomedical engineering, and clinical medicine.

Self‐Adaptive MoO3−x Subnanometric Wires Incorporated Scaffolds for Osteosarcoma Therapy and Bone Regeneration

AbstractResidual tumor cells and bone tissue defects are two critical challenges in clinical osteosarcoma treatment. Herein, a subnanomedicine concept is proposed by developing a self‐adaptive functional tissue engineering scaffold constructed by integrating MoO3−x subnanometric wires onto 3D printing bioactive glass scaffolds. The MoO3−x subnanometric wires are synthesized by a one‐pot hydrothermal method, which aggregate in an acidic tumor microenvironment and react with hydrogen peroxide to produce reactive oxygen species for specific chemodynamic therapy. However, they can degrade rapidly under physiological conditions without causing toxicity. Moreover, self‐adaptively enhanced photothermal conversion enables tumor‐targeting photothermal therapy while enhancing chemodynamic therapy. Additionally, the Mo5+‐Mo6+ transition enables lipid peroxide accumulation and glutathione depletion, thereby resulting in the deactivation of glutathione peroxidase 4 protein and ferroptosis through Western blot analysis, confocal laser scanning microscopy observation, and mRNA transcriptome analysis. In addition to its robustness against osteosarcoma, the constructed scaffolds can stimulate rat bone mesenchymal stem cell differentiation and proliferation as well as promote osteogenesis in bone defects. Therefore, this multifunctional scaffold not only validates the subnanomedicine concept but also provides a promising clinical strategy for bone tissue engineering.

Biomimetic Approaches in Cardiac Tissue Engineering: Replicating the Native Heart Microenvironment.

Cardiovascular diseases, including heart failure, pose significant challenges in medical practice, necessitating innovative approaches for cardiac repair and regeneration. Cardiac tissue engineering has emerged as a promising solution, aiming to develop functional and physiologically relevant cardiac tissue constructs. Replicating the native heart microenvironment, with its complex and dynamic milieu necessary for cardiac tissue growth and function, is crucial in tissue engineering. Biomimetic strategies that closely mimic the natural heart microenvironment have gained significant interest due to their potential to enhance synthetic cardiac tissue functionality and therapeutic applicability. Biomimetic approaches focus on mimicking biochemical cues, mechanical stimuli, coordinated electrical signaling, and cell-cell/cell-matrix interactions of cardiac tissue. By combining bioactive ligands, controlled delivery systems, appropriate biomaterial characteristics, electrical signals, and strategies to enhance cell interactions, biomimetic approaches provide a more physiologically relevant environment for tissue growth. The replication of the native cardiac microenvironment enables precise regulation of cellular responses, tissue remodeling, and the development of functional cardiac tissue constructs. Challenges and future directions include refining complex biochemical signaling networks, paracrine signaling, synchronized electrical networks, and cell-cell/cell-matrix interactions. Advancements in biomimetic approaches hold great promise for cardiovascular regenerative medicine, offering potential therapeutic strategies and revolutionizing cardiac disease modeling. These approaches contribute to the development of more effective treatments, personalized medicine, and improved patient outcomes. Ongoing research and innovation in biomimetic approaches have the potential to revolutionize regenerative medicine and cardiac disease modeling by replicating the native heart microenvironment, advancing functional cardiac tissue engineering, and improving patient outcomes.

Rapid biomimetic mineralization of keratin scaffold via electrodeposition with potential for bone regeneration

For bone regeneration, rapid mineralization is of particular importance in the preparation of biomimetic bone scaffolds, as for avoiding the drawbacks of prolonged mineralization which will cause the loss of growth factors and degradation. In this research, we developed a relatively rapid mineralization approach for constructing biomimetic keratin scaffold that exhibited highly interconnected pore and natural bone mimetic calcium phosphate coating by applying an electrodeposition technique. Mineralized keratin scaffold was obtained by freeze drying followed by electrodeposition for rapid biomimetic mineralization. The mineral coating morphology, component and crystal structure could be controllably tailored by manipulating the deposition electrode, voltage and duration. A satisfying coating of apatite layer on keratin scaffold could be obtained within a couple of hours by electrodeposition. By increasing the voltage and duration, a more favored amount of apatite coating which dominated by HA crystal could be formed. In addition, cell regeneration assay showed that mineralized biomimetic keratin scaffold exhibited more suitable supporting platform for the proliferation and osteoblastic differentiation of MC3T3 cell over pure keratin scaffold. Thus, the rapid electrodeposition mineralization approach presented in this work could be highly desired for fabricating biomimetic scaffolds in which biological molecules were loaded for functional bone tissue engineering applications.

Bio-Enhanced Neoligaments Graft Bearing FE002 Primary Progenitor Tenocytes: Allogeneic Tissue Engineering & Surgical Proofs-of-Concept for Hand Ligament Regenerative Medicine.

Hand tendon/ligament structural ruptures (tears, lacerations) often require surgical reconstruction and grafting, for the restauration of finger mechanical functions. Clinical-grade human primary progenitor tenocytes (FE002 cryopreserved progenitor cell source) have been previously proposed for diversified therapeutic uses within allogeneic tissue engineering and regenerative medicine applications. The aim of this study was to establish bioengineering and surgical proofs-of-concept for an artificial graft (Neoligaments Infinity-Lock 3 device) bearing cultured and viable FE002 primary progenitor tenocytes. Technical optimization and in vitro validation work showed that the combined preparations could be rapidly obtained (dynamic cell seeding of 105 cells/cm of scaffold, 7 days of co-culture). The studied standardized transplants presented homogeneous cellular colonization in vitro (cellular alignment/coating along the scaffold fibers) and other critical functional attributes (tendon extracellular matrix component such as collagen I and aggrecan synthesis/deposition along the scaffold fibers). Notably, major safety- and functionality-related parameters/attributes of the FE002 cells/finished combination products were compiled and set forth (telomerase activity, adhesion and biological coating potentials). A two-part human cadaveric study enabled to establish clinical protocols for hand ligament cell-assisted surgery (ligamento-suspension plasty after trapeziectomy, thumb metacarpo-phalangeal ulnar collateral ligamentoplasty). Importantly, the aggregated experimental results clearly confirmed that functional and clinically usable allogeneic cell-scaffold combination products could be rapidly and robustly prepared for bio-enhanced hand ligament reconstruction. Major advantages of the considered bioengineered graft were discussed in light of existing clinical protocols based on autologous tenocyte transplantation. Overall, this study established proofs-of-concept for the translational development of a functional tissue engineering protocol in allogeneic musculoskeletal regenerative medicine, in view of a pilot clinical trial.

A Hierarchical 3D Graft Printed with Nanoink for Functional Craniofacial Bone Restoration

AbstractAn ideal craniofacial bone repair graft shall not only focus on the repair ability but also the regeneration of natural architecture with occlusal loads‐related function restoration. However, such functional bone tissue engineering scaffold has rarely been reported. Herein, a hierarchical 3D graft is proposed for rebuilding craniofacial bone with both natural structure and healthy biofunction reconstruction. Inspired by the bone healing process, an organic–inorganic nanoink with ultrasmall calcium phosphate oligomers and bone morphogenetic protein‐2 incorporated is developed for spatiotemporal guidance of new bone. Based on such homogeneous nanoink, a biomimetic graft, including a cortical layer containing Haversian system, and a cancellous layer featured with triply periodic minimum surface macrostructures, is fabricated via projection‐based 3D printing method, and the layers are loaded with distinct concentrations of bioactive factors for regenerating new bone with gradient density. The graft exhibits excellent osteogenic and angiogenic potential in vitro, and accelerates revascularization and reconstructs neo‐bone with original morphology in vivo. Benefiting from such natural architecture, loading force is widely transferred with reduced stress concentration around the inserted dental implant. Taken from native physiochemical and structural cues, this wstudy provides a novel strategy for functional tissue engineering through designing function‐oriented biomaterials.

Cell Electrospinning: A Mini-Review of the Critical Processing Parameters and Its Use in Biomedical Applications.

Functional tissue engineering is a widely studied area of research with increasing importance in regenerative medicine, as well as in the development of in vitro models used for drug discovery and mimicking diseased tissues, among other applications. Electrospinning (ES) is one of the most widely used methods in these fields. It has attracted considerable interest because it can produce materials resembling the extracellular matrix of native tissues. The micro/nanofibers produced by this method provide a cell-friendly environment that promotes cellular activities. Cell electrospinning (C-ES) is based on the fundamental ES process and enables the encapsulation of viable cells in a micro/nanofibrous mesh. In this review, the process of C-ES and the materials used in this process are discussed. This work also discusses the applications of C-ES in tissue engineering, focusing on recent advances in this field.

Biomaterializing the advances in uterine tissue engineering.

The uterus is considered to be a unique wound-healing model and distinguished by the repeated shedding of the endometrium and self-traceless regeneration. Common curettage, cesarean section, and other operations often cause endometrial and myometrial defects and obstetric and gynecological complications, leading to a high demand for uterine repair or partial replacement. However, the structure and function of the uterus are complicated. Functional uterine tissue engineering requires highly specialized biomaterials with a natural extracellular microenvironment. Currently, no biomaterial can fully simulate the structural and biomechanical properties of the uterus. Many efforts have been made to develop highly functional materials and tissue structures that may provide uterine tissue engineering constructs for reducing obstetric and gynecological complications. Continuous efforts will likely facilitate the development of scalable cells and biomaterial technologies for clinical use. This review summarizes the recent applications of biomaterials and tissue engineering in rebuilding a portion of or the entire uterus.

Evolution of functional tissue engineering for tendon and ligament repair.

This review paper is motivated by a Back-to-Basics presentation given by the author at the 2022 Orthopaedic Research Societymeeting in Tampa, Florida. I was tasked with providing a brief history of research leading up to the introduction of functional tissue engineering (FTE) for tendon and ligament repair. Beginning in the 1970s, this timeline focused on two common orthopedic soft tissue problems, anterior cruciate ligament ruptures in the knee and supraspinatus tendon injuries in the shoulder. Historic changes in the field over the next 5decades revealed a transformation from a focus more on mechanics (called "bioMECHANICS") on a larger (tissue) scale to a more recent focus on biology (called "mechanoBIOLOGY") on a smaller (cellular and molecular) scale. Early studies by surgeons and engineers revealed the importance of testing conditions for ligaments and tendons (e.g., high strain rates while avoiding subject disuse and immobility) and the need to measure in vivo forces in these tissues. But any true tissue engineering and regeneration in these early decades was limited more to the use of auto-, allo- and xenografts than actual generation of stimulated cell-scaffold constructs in culture. It was only after the discovery of tissue engineering in 1988 and the recognition of frequent rotator cuff injuries in the early 1990s, that biologists joined surgeons and engineers to discover mechanical and biological testing criteria for FTE. This review emphasizes the need for broader and more inclusive collaborations by surgeons, biologists and engineers in the short term with involvement of those in biomaterials, manufacturing, and regulation of new products in the longer term.

Aligned multi-walled carbon nanotube-embodied hydrogel via low magnetic field: A strategy for engineering aligned injectable scaffolds

Injectable scaffolds are a promising strategy to restore and regenerate damaged and diseased tissues. They require minimally invasive procedure and allow the formation of an in-situ structure of any shape. However, the formation of 3D in-situ structure with aligned morphologies using a method which could be easily transferred to clinical settings remains a challenge. Herein, the rational design of an aligned injectable hydrogel-based scaffold via remote-induced alignment is reported. Carboxylated multi-walled carbon nanotubes (cMWCNT) are aligned into hydrogel via low magnetic field. The uniform dispersion and alignment of cMWCNT into the hydrogel are clearly demonstrated by small angle neutron scattering. The obtained aligned cMWCNT-embodied hydrogel is stable over 7 days at room temperature and as well at body temperature (i.e. 37 °C). As unique approach, the formation of MWCNT-hydrogel composite is investigated combining rheology with molecular dynamic and quantum mechanical calculations. The increase of MWCNT concentration into the hydrogel decreases the total energy promoting structural stabilization and increase of stiffness. The remote aligning of injectable hydrogel-based scaffold opens up horizons in the engineering of functional tissues which requires specific cell orientation.

Enhancing cartilage repair with optimized supramolecular hydrogel-based scaffold and pulsed electromagnetic field.

Functional tissue engineering strategies provide innovative approach for the repair and regeneration of damaged cartilage. Hydrogel is widely used because it could provide rapid defect filling and proper structure support, and is biocompatible for cell aggregation and matrix deposition. Efforts have been made to seek suitable scaffolds for cartilage tissue engineering. Here Alg-DA/Ac-β-CD/gelatin hydrogel was designed with the features of physical and chemical multiple crosslinking and self-healing properties. Gelation time, swelling ratio, biodegradability and biocompatibility of the hydrogels were systematically characterized, and the injectable self-healing adhesive hydrogel were demonstrated to exhibit ideal properties for cartilage repair. Furthermore, the new hydrogel design introduces a pre-gel state before photo-crosslinking, where increased viscosity and decreased fluidity allow the gel to remain in a semi-solid condition. This granted multiple administration routes to the hydrogels, which brings hydrogels the ability to adapt to complex clinical situations. Pulsed electromagnetic fields (PEMF) have been recognized as a promising solution to various health problems owing to their noninvasive properties and therapeutic potentials. PEMF treatment offers a better clinical outcome with fewer, if any, side effects, and wildly used in musculoskeletal tissue repair. Thereby we propose PEMF as an effective biophysical stimulation to be 4th key element in cartilage tissue engineering. In this study, the as-prepared Alg-DA/Ac-β-CD/gelatin hydrogels were utilized in the rat osteochondral defect model, and the potential application of PEMF in cartilage tissue engineering were investigated. PEMF treatment were proven to enhance the quality of engineered chondrogenic constructs in vitro, and facilitate chondrogenesis and cartilage repair in vivo. All of the results suggested that with the injectable self-healing adhesive hydrogel and PEMF treatment, this newly proposed tissue engineering strategy revealed superior clinical potential for cartilage defect treatment.

Dynamic Deformation Calculation of Articular Cartilage and Cells Using Resonance-Driven Laser Scanning Microscopy.

The deformation of articular cartilage and its cells at the micro-scale during dynamic activities such as gait has high mechanoregulatory importance. Measuring the cellular geometries during such dynamics has been limited by the rate of microscopic image acquisition. The introduction of resonating mirrors for image rasterization (resonant scanning), rather than the conventional servo control (galvano scanning), has significantly improved the scanning rate by more than 100×. However, the high scanning rate comes at the cost of image quality, thereby posing challenges in image processing. Here, resonance-driven 3-D laser microscopy is used to observe the transient, micro-scale deformation of articular cartilage and its cells under osmotic challenge conditions. Custom image segmentation and deformable registration software were implemented for analysis of the resonance-scanned microscopy data. The software exhibited robust and accurate performance on the osmotic swelling measurements, as well as quantitative validation testing. The resonance-scanning protocol and developed analysis software allow for simultaneous strain calculation of both the local tissue and cells, and are thus a valuable tool for real-time probing of the cell-matrix interactions that are highly relevant in the fields of orthopedic biomechanics, cell mechanobiology, and functional tissue engineering.

Stretchable and Electroactive Crosslinked Gelatin for Biodevice and Cell Culture Applications.

Biomimetic substrates that incorporate functionality such as electroactivity and mechanical flexibility, find utility in a variety of biomedical applications. Toward these uses, nature-derived materials such as gelatin offer inherent biocompatibility and sustainable sourcing. However, issues such as high swelling, poor mechanical properties, and lack of stability at biological temperatures limit their use. The enzymatic crosslinking of gelatin via microbial transglutaminase (mTG) yields flexible and robust large area substrates that are stable under physiological conditions. Here, we demonstrate the fabrication and characterization of strong, stretchable, conductive mTG crosslinked gelatin thin films. Incorporation of the conductive polymer poly(3,4-ethylenedioxythiophene) polystyrene sulfonate in the gel matrix with a bioinspired polydopamine surface coating is used to enable conductivity with enhanced mechanical properties such as extensibility and flexibility, in comparison to plain gelatin or crosslinked gelatin films. The electroconductive substrates are conducive to cell growth, supporting myoblast cell adhesion, viability, and proliferation and could find use in creating active cell culture systems incorporating electrical stimulation. The substrates are responsive to motion such as stretching and bending while being extremely handleable and elastic, making them useful for applications such as electronic skin and flexible bioelectronics. Overall, this work presents facile, yet effective development of bioinspired conductive composites as substrates for bio-integrated devices and functional tissue engineering.