A review about the role of additives in nerve tissue engineering: growth factors, vitamins, and drugs.

Notably the integration of additives such as growth factors, vitamins, and drugs with scaffolds promoted nerve tissue engineering. This study tried to provide a concise review of all these additives that facilitates nerve regeneration. An attempt was first made to provide information on the main principle of nerve tissue engineering, and then to shed light on the effectiveness of these additives on nerve tissue engineering. Our research has shown that growth factors accelerate cell proliferation and survival, while vitamins play an effective role in cell signalling, differentiation, and tissue growth. They can also act as hormones, antioxidants, and mediators. Drugs also have an excellent and necessary effect on this process by reducing inflammation and immune responses. This review shows that growth factors were more effective than vitamins and drugs in nerve tissue engineering. Nevertheless, vitamins were the most commonly used additive in the production of nerve tissue.

Assembling Spheroids of Rat Primary Neurons Using a Stress-Free 3D Culture System.

Neural injuries disrupt the normal functions of the nervous system, whose complexities limit current treatment options. Because of their enhanced therapeutic effects, neurospheres have the potential to advance the field of regenerative medicine and neural tissue engineering. Methodological steps can pose challenges for implementing neurosphere assemblies; for example, conventional static cultures hinder yield and throughput, while the presence of the necrotic core, time-consuming methodology, and high variability can slow their progression to clinical application. Here we demonstrate the optimization of primary neural cell-derived neurospheres, developed using a high-throughput, stress-free, 3D bioreactor. This process provides a necessary baseline for future studies that could develop co-cultured assemblies of stem cells combined with endothelial cells, and/or biomaterials and nanomaterials for clinical therapeutic use. Neurosphere size and neurite spreading were evaluated under various conditions using Image J software. Primary neural cells obtained from the hippocampi of three-day-old rat pups, when incubated for 24 h in a reactor coated with 2% Pluronic and seeded on Poly-D-Lysine-coated plates establish neurospheres suitable for therapeutic use within five days. Most notably, neurospheres maintained high cell viability of ≥84% and expressed the neural marker MAP2, neural marker ß-Tubulin III, and glial marker GFAP at all time points when evaluated over seven days. Establishing these factors reduces the variability in developing neurospheres, while increasing the ease and output of the culture process and maintaining viable cellular constructs.

Promoting neural transdifferentiation of BMSCs via applying synergetic multiple factors for nerve regeneration.

The strategy of using multipotential stem cells like bone mesenchymal stromal cells (BMSCs) for nerve tissue engineering is proven feasible. The promotion effects on neural transdifferentiation of BMSCs from factors including nerve growth factor (NGF), laminin and electrical stimulation (ES) have been reported, while it is not known if these factors can achieve a strong synergetic impact when the cells are cultured on conductive substrates. In this study, it was identified that any single factor (NGF, laminin, or conductive substrate) combined with ES demonstrated the capacity to induce BMSCs transdifferentiating into neural cells, while the efficiency was found in the order of NGF > laminin > conductive substrate. The combination of any two of the factors would be more efficient in inducing the neural transdifferentiation than individual factor. As expected, the strongest promotion in inducing BMSCs to transdifferentiate into neural cells was identified when BMSCs were cultured on laminin-treated conductive nanofibrous mesh in the presence of NGF and under proper ES simultaneously, showing significant synergetic efficiency from these multiple factors. Studies on the Notch-1 signaling pathway, a main negative regulator of neurogenesis, revealed these factors sharing a similar molecular mechanism in regulating the neural transdifferentiation of BMSCs. The results suggested that satisfactory nerve regeneration might be achievable if these synergetic multiple factors could be involved in nerve guidance conduit design, especially, when BMSCs were applied as co-implanted cells.

Effect of nanocomposite coating and biomolecule functionalization on silk fibroin based conducting 3D braided scaffolds for peripheral nerve tissue engineering.

In this work, the effects of carbon nanofiber (CNF) dispersed poly-ε-caprolactone (PCL) nanocomposite coatings and biomolecules functionalization on silk fibroin based conducting braided nerve conduits were studied for enhancing Neuro 2a cellular activities. A unique combination of biomolecules (UCM) and varying concentrations of CNF (5, 7.5, 10% w/w) were dispersed in 10% (w/v) PCL solution for coating on degummed silk threads. The coated silk threads were braided to develop the scaffold structure. As the concentration of CNF increased in the coating, the electrical impedance decreased up to 400 Ω indicating better conductivity. The tensile and dynamic mechanical property analysis showed better mechanical properties in CNF coated samples. In vitro cytocompatibility analysis proved the non-toxicity of the developed braided conduits. Cell attachment, growth and proliferation were significantly enhanced on the biomolecule functionalized nanocomposite coated silk braided structure, exhibiting their potential for peripheral nerve regeneration and recovery.

Prospects of Natural Polymeric Scaffolds in Peripheral Nerve Tissue-Regeneration.

Tissue-engineering is emerging field and can be considered as a novel therapeutic intervention in nerve tissue-regeneration. The various pitfalls associated with the use of autografts in nerve-regeneration after injuries have inspired researchers to explore the possibilities using various natural polymers. In this context, the present chapter summarizes the advances of the various types of natural polymeric scaffolds such as fibrous scaffolds, porous scaffolds, and hydrogels in nerve-regeneration and repair process. The functionalization of the scaffolds with wide-range of biomolecules and their biocompatibility analysis by employing various cells (e.g., mesenchymal, neural progenitor stem cells) along with the in vivo regeneration outcomes achieved upon implantation are discussed here. Besides, the various avenues that have been explored so far in nerve tissue-engineering, the use of the extracellular matrix in enhancing the functional polymeric scaffolds and their corresponding outcomes of regeneration are mentioned. We conclude with the present challenges and prospects of efficient exploration of natural polymeric scaffolds in the future to overcome the problems of nerve-regeneration associated with various nerve injuries and neurodegenerative disorders.

Co-assembling bioactive short peptide nanofibers coated silk scaffolds induce neurite outgrowth of PC12 cells.

Controlling biomolecular-cell interactions is crucial for the design of scaffolds for tissue engineering (TE). Regenerated silk fibroin (RSF) has been extensively used as TE scaffolds, however, RSF showed poor attachment of neuronal cells, such as rat pheochromocytoma (PC12) cells. In this work, amphiphilic peptides containing a hydrophobic isoleucine tail (I3) and laminin or fibronectin derived peptides (IKVAV, PDSGR, YIGSR, RGDS and PHSRN) were designed for promoting scaffold-cell interaction. Three of them (I3KVAV, I3RGDS and I3YIGSR) can self-assemble into nanofibers, therefore, were used to enhance the application of RSF in neuron TE. Live / dead assays revealed that the peptides exhibited negligible cytotoxicity against PC12 cells. The specific interaction between PC12 cells and the peptides were investigate using atomic force microscopy (AFM). The results indicated a synergistic effect in the designed peptides, promoting cellular attachment, proliferation and morphology changes. In addition, AFM results showed that co-assembling peptides I3KVAV and I3YIGSR possesses the best regulation of proliferation and attachment of PC12 cells, consistent with immunofluorescence staining results. Moreover, cell culture with hydrogels revealed that a mixture of peptides I3KVAV and I3YIGSR can also promote 3D neurites outgrowth. The approach of combining two different self-assembling peptides shows great potential for nerve regeneration applications.

Interactions of Cells and Biomaterials for Nerve Tissue Engineering: Polymers and Fabrication.

Neural injuries affect millions globally, significantly impacting their quality of life. The inability of these injuries to heal, limited ability to regenerate, and the lack of available treatments make regenerative medicine and tissue engineering a promising field of research for developing methods for nerve repair. This review evaluates the use of natural and synthetic polymers, and the fabrication methods applied that influence a cell's behavior. Methods include cross-linking hydrogels, incorporation of nanoparticles, and 3D printing with and without live cells. The endogenous cells within the injured area and any exogenous cells seeded on the polymer construct play a vital role in regulating healthy neural activity. This review evaluates the body's local and systemic reactions to the implanted materials. Although numerous variables are involved, many of these materials and methods have exhibited the potential to provide a biomaterial environment that promotes biocompatibility and the regeneration of a physical and functional nerve. Future studies may evaluate advanced methods for modifying material properties and characterizing the tissue-biomaterial interface for clinical applications.

Mathematical modelling with Bayesian inference to quantitatively characterize therapeutic cell behaviour in nerve tissue engineering.

Cellular engineered neural tissues have significant potential to improve peripheral nerve repair strategies. Traditional approaches depend on quantifying tissue behaviours using experiments in isolation, presenting a challenge for an overarching framework for tissue design. By comparison, mathematical cell-solute models benchmarked against experimental data enable computational experiments to be performed to test the role of biological/biophysical mechanisms, as well as to explore the impact of different design scenarios and thus accelerate the development of new treatment strategies. Such models generally consist of a set of continuous, coupled, partial differential equations relying on a number of parameters and functional forms. They necessitate dedicated in vitro experiments to be informed, which are seldom available and often involve small datasets with limited spatio-temporal resolution, generating uncertainties. We address this issue and propose a pipeline based on Bayesian inference enabling the derivation of experimentally informed cell-solute models describing therapeutic cell behaviour in nerve tissue engineering. We apply our pipeline to three relevant cell types and obtain models that can readily be used to simulate nerve repair scenarios and quantitatively compare therapeutic cells. Beyond parameter estimation, the proposed pipeline enables model selection as well as experiment utility quantification, aimed at improving both model formulation and experimental design.

Applications of chitosan-based biomaterials: From preparation to spinal cord injury neuroprosthetic treatment.

Spinal cord injury (SCI)-related disabilities are a serious problem in the modern society. Further, the treatment of SCI is highly challenging and is urgently required in clinical practice. Research on nerve tissue engineering is an emerging approach for improving the treatment outcomes of SCI. Chitosan (CS) is a cationic polysaccharide derived from natural biomaterials. Chitosan has been found to exhibit excellent biological properties, such as nontoxicity, biocompatibility, biodegradation, and antibacterial activity. Recently, chitosan-based biomaterials have attracted significant attention for SCI repair in nerve tissue engineering applications. These studies revealed that chitosan-based biomaterials have various functions and mechanisms to promote SCI repair, such as promoting neural cell growth, guiding nerve tissue regeneration, delivering nerve growth factors, and as a vector for gene therapy. Chitosan-based biomaterials have proven to have excellent potential for the treatment of SCI. This review aims to introduce the recent advances in chitosan-based biomaterials for SCI treatment and to highlight the prospects for further application.

The effect of conductive aligned fibers in an injectable hydrogel on nerve tissue regeneration.

Injectable hydrogels are a promising treatment option for nervous system injuries due to the difficulty to replace lost cells and nervous factors but research on injectable conductive hydrogels is limited and these scaffolds have poor electromechanical properties. This study developed a chitosan/beta-glycerophosphate/salt hydrogel and added conductive aligned nanofibers (polycaprolactone/gelatin/single-wall carbon nanotube (SWCNT)) for the first time and inspired by natural nerve tissue to improve their biochemical and biophysical properties. The results showed that the degradation rate of hydrogels is proportional to the regrowth of axons and these hydrogels' mechanical (hydrogels without nanofibers or SWCNTs and hydrogels containing these additions have the same Young's modulus as the brain and spinal cord or peripheral nerves, respectively) and electrical properties, and the interconnective structure of the scaffolds have the ability to support cells.

Effect of Electrical Stimulation on PC12 Cells Cultured in Different Hydrogels: Basis for the Development of Biomaterials in Peripheral Nerve Tissue Engineering.

Extensive damage to peripheral nerves is a health problem with few therapeutic alternatives. In this context, the development of tissue engineering seeks to obtain materials that can help recreate environments conducive to cellular development and functional repair of peripheral nerves. Different hydrogels have been studied and presented as alternatives for future treatments to emulate the morphological characteristics of nerves. Along with this, other research proposes the need to incorporate electrical stimuli into treatments as agents that promote cell growth and differentiation; however, no precedent correlates the simultaneous effects of the types of hydrogel and electrical stimuli. This research evaluates the neural differentiation of PC12 cells, relating the effect of collagen, alginate, GelMA, and PEGDA hydrogels with electrical stimulation modulated in four different ways. Our results show significant correlations for different cultivation conditions. Electrical stimuli significantly increase neural differentiation for specific experimental conditions dependent on electrical frequency, not voltage. These backgrounds allow new material treatment schemes to be formulated through electrical stimulation in peripheral nerve tissue engineering.

Hypoxic pre-conditioned adipose-derived stem/progenitor cells embedded in fibrin conduits promote peripheral nerve regeneration in a sciatic nerve graft model.

Recent results emphasize the supportive effects of adipose-derived multipotent stem/progenitor cells (ADSPCs) in peripheral nerve recovery. Cultivation under hypoxia is considered to enhance the release of the regenerative potential of ADSPCs. This study aimed to examine whether peripheral nerve regeneration in a rat model of autologous sciatic nerve graft benefits from an additional custom-made fibrin conduit seeded with hypoxic pre-conditioned (2% oxygen for 72 hours) autologous ADSPCs (n = 9). This treatment mode was compared with three others: fibrin conduit seeded with ADSPCs cultivated under normoxic conditions (n = 9); non-cell-carrying conduit (n = 9); and nerve autograft only (n = 9). A 16-week follow-up included functional testing (sciatic functional index and static sciatic index) as well as postmortem muscle mass analyses and morphometric nerve evaluations (histology, g-ratio, axon density, and diameter). At 8 weeks, the hypoxic pre-conditioned group achieved significantly higher sciatic functional index/static sciatic index scores than the other three groups, indicating faster functional regeneration. Furthermore, histologic evaluation showed significantly increased axon outgrowth/branching, axon density, remyelination, and a reduced relative connective tissue area. Hypoxic pre-conditioned ADSPCs seeded in fibrin conduits are a promising adjunct to current nerve autografts. Further studies are needed to understand the underlying cellular mechanism and to investigate a potential application in clinical practice.

Proliferation and differentiation study of melatonin functionalized polycaprolactone/gelatin electrospun fibrous scaffolds for nerve tissue engineering.

Melatonin (MLT), a pineal neurohormone with multiple neuroprotective, is often used for peripheral nerve recovery and regenerated nerve proliferation. In this study, Polycaprolactone/Gelatin (PG) fibrous electrospun scaffolds with various percentages of MLT (0, 1, 2, and 4%wt) were fabricated for nerve cell growth, the effects of different concentrations of MLT within PG fibers (PG, PGMLT1, PGMLT2, and PGMLT4) on the proliferation and differentiation for PC12 cells were quantitatively evaluated. The microstructures and morphologies of these scaffolds were analyzed by FE-SEM and digital camera. Fourier transform infrared spectrometer (FTIR), X-ray photoelectron spectroscopy (XPS), and Water Contact Angle (WCA) were used to study the composition, ratio and properties of MLT functionalized PG scaffolds. MTT and CLSM analysis showed that appropriate amount of MLT was beneficial to the proliferation of PC12 cell. MLT can also promote cell differentiation, neurite germination, the expression levels of MAP2 mRNA and protein were dramatically increased on the composite scaffolds with the increase of MLT content, moderate addition of MLT (PGMLT2, 2%) had a prominent enhancement for neurite length. This work would provide a more comprehensive reference for further researches on MLT functionalized composite scaffolds and suggest that high-performance PGMLT fibrous scaffolds could be a promising alternative for nerve repair.

Bacterial cellulose-based composites for nerve tissue engineering.

Nerve injuries and neurodegenerative disorders are very serious and costly medical challenges. Damaged nerve tissue may not be able to heal and regain its function, and scar tissue may restrict nerve cell regeneration. In recent years, new electroactive biomaterials have attracted widespread attention in the neural tissue engineering field. Bacterial cellulose (BC) due to its unique properties such as good mechanical properties, high water retention, biocompatibility, high crystallinity, large surface area, high purity, very fine network, and inability to absorb in the human body due to cellulase deficiency, can be considered a promising treatment for neurological injuries and disorders that require long-term support. However, BC lacks electrical activity, but can significantly improve the nerve regeneration rate by combining with conductive structures. Electrical stimulation has been shown to be an effective means of increasing the rate and accuracy of nerve regeneration. Many factors, such as the intensity and pattern of electrical current, have positive effects on cellular activity, including cell adhesion, proliferation, migration and differentiation, and cell-cell/tissue/molecule/drug interaction. This study discusses the importance and essential role of BC-based biomaterials in neural tissue regeneration and the effects of electrical stimulation on cellular behaviors.

A narrative review of current therapies in unilateral recurrent laryngeal nerve injury caused by thyroid surgery.

OBJECTIVE: To summarize and compare current common treatments in recurrent laryngeal nerve injury (RLNI). In addition, we introduced nerve tissue engineering technology in RLNI animal models. This review is a comprehensive summary of current therapies for unilateral RLNI. BACKGROUND: RLNI is a common complication in thyroid surgery. Although preoperative imaging and intraoperative nerve monitoring are widely applied, some damage to nerves is still inevitable. Currently, advances in nerve repair and regeneration have been made, but relatively few studies have focused on RLNI. In this review, we summarized and compared current common treatments in RLNI. METHODS: We searched the literature on PubMed and Web of Science, and chose studies about RLNI in thyroid surgery. Keywords included the following terms: "recurrent laryngeal nerve regeneration", "injection laryngoplasty", "type I thyroplasty", "arytenoid adduction", and "nerve tissue engineering technology". Only English language studies were included. In the section on nerve tissue engineering technology, we described the application in detail in the table below. CONCLUSIONS: Currently, the majority of treatments could obtain different effects to some extent, but there are still shortcomings that need to be overcome further overcome. Therefore, potential exploration such as nerve tissue engineering technology is worthy of attention.

Hypocapnia Stimuli-Responsive Engineered Exosomes Delivering miR-218 Facilitate Sciatic Nerve Regeneration.

Therapeutic strategies of microRNAs (miRNAs) and exosomes have been systematically explored as an enhancing application by paracrine and modulating cellular activity after internalization of recipient cells in vitro, and progressively developed to meet the requirements of peripheral nerve regeneration in vivo. However, how to obtain exosomes with superior properties and effectively deliver miRNAs becomes a key challenge. Hypocapnia environment might play unexpected outcomes in strengthening exosome function when culturing adipose-derived stem cells (ASCs). Previously, we discovered the intensive regulation of miR-218 on the differentiation of ASCs. In the present study, we analyzed the functional differences of secreted exosomes in response to hypocapnia stimulation, and explored the application in combination with miR-218 to facilitate sciatic nerve regeneration. Our results indicated that the delivery system of engineered exosomes derived from ASCs remarkably loads upregulated miR-218 and promotes cellular activity in the recipient cells (PC12 cells), and hypocapnia stimuli-responsive exosomes exhibit strengthening properties. Furthermore, in a sciatic nerve injury model, exosomes delivering miR-218 combined with engineered scaffold facilitated the regeneration of injured sciatic nerves. In the hypocapnia-stimulated exosome group, more encouraging promotion was revealed on the regeneration of motor and nerve fibers. Hypoc-miR-218-ASC exosomes are suggested as a promising cell-free strategy for peripheral nerve repair.

Conductive polypyrrole-coated electrospun chitosan nanoparticles/poly(D,L-lactide) fibrous mat: influence of drug delivery and Schwann cells proliferation.

For nerve tissue engineering (NTE), scaffolds with the ability to release drugs under control and support the rapid proliferation of cells are very important for the repair of nerve defects. This study aimed to fabricate a conductive drug-loaded fiber mat by electrospinning and assess its potential as a scaffold for Schwann cells proliferation. The conductive polypyrrole (PPy) was coated on an electrospun poly (D, L-lactide) (PLA) fibrous mat, which was simultaneously embedded with protein-loaded chitosan nanoparticles and ibuprofen as a model small molecule drug. The fibrous mat shows suitable conductivity, mechanical properties, and hydrophilicity for NTE. For drug release and degradation studies, the fibrous mat can achieve sustained release of bovine serum albumin (BSA) and ibuprofen, and the PPy coating can increase the surface wettability and conductivity while slowing down the degradation of the fibrous mat. The application of electrical stimulation (ES) to the fibrous mat can accelerate the release of ibuprofen, but there was no significant effect on the release rate of the protein. The fibrous mat showed no cytotoxicityin vitro, and Schwann cells (SCs) can adhere, grow, and proliferate well on mats. At the 120th hour of culturein vitro, the relative growth rate of SCs on the conductive drug-loaded fibrous mat reached 198.22 ± 2.34%, which was an increase of 37.93% compared to the SCs on the drug-loaded fibrous mat with ES. The density and elongation of SCs on the conductive drug-loaded fibrous mat were greater than those on the PLA fibrous mat, indicating that the conductive polypyrrole-coated electrospun chitosan nanoparticles/PLA fibrous mat has good potential for application in nerve regeneration.

In vivo near-infrared fluorescent fibrin highlights growth of nerve during regeneration across a nerve gap.

Significance: Exogenous extracellular matrix (ECM) proteins, such as fibrinogen and the thrombin-polymerized scaffold fibrin, are used in surgical repair of severe nerve injuries to supplement ECM produced via the injury response. Monitoring the dynamic changes of fibrin during nerve regeneration may shed light on the frequent failure of grafts in the repair of long nerve gaps. Aim: We explored whether monitoring of fibrin dynamics can be carried out using nerve guidance conduits (NGCs) containing fibrin tagged with covalently bound fluorophores. Approach: Fibrinogen was conjugated to a near-infrared (NIR) fluorescent dye. NGCs consisting of silicone tubes filled with the fluorescent fibrin were used to repair a 5-mm gap injury in rat sciatic nerve ( n = 6 ). Results: Axonal regeneration in fluorescent fibrin-filled NGCs was confirmed at 14 days after implantation. Intraoperative fluorescence imaging after implantation showed that the exogenous fibrin was embedded in the early stage regenerative tissue. The fluorescent signal temporarily highlighted a cable-like structure within the conduit and gradually degraded over two weeks. Conclusions: This study, for the first time, visualized in vivo intraneural fibrin degradation, potentially a useful prospective indicator of regeneration success, and showed that fluorescent ECM, in this case fibrin, can facilitate imaging of regeneration in peripheral nerve conduits without significantly affecting the regeneration process.

Three-Dimensional Engineered Peripheral Nerve: Toward a New Era of Patient-Specific Nerve Repair Solutions.

Reconstruction of peripheral nerve injuries (PNIs) with substance loss remains challenging because of limited treatment solutions and unsatisfactory patient outcomes. Currently, nerve autografting is the first-line management choice for bridging critical-sized nerve defects. The procedure, however, is often complicated by donor site morbidity and paucity of nerve tissue, raising a quest for better alternatives. The application of other treatment surrogates, such as nerve guides, remains questionable, and it is inefficient in irreducible nerve gaps. More importantly, these strategies lack customization for personalized patient therapy, which is a significant drawback of these nerve repair options. This negatively impacts the fascicle-to-fascicle regeneration process, critical to restoring the physiological axonal pathway of the disrupted nerve. Recently, the use of additive manufacturing (AM) technologies has offered major advancements to the bioengineering solutions for PNI therapy. These techniques aim at reinstating the native nerve fascicle pathway using biomimetic approaches, thereby augmenting end-organ innervation. AM-based approaches, such as three-dimensional (3D) bioprinting, are capable of biofabricating 3D-engineered nerve graft scaffolds in a patient-specific manner with high precision. Moreover, realistic in vitro models of peripheral nerve tissues that represent the physiologically and functionally relevant environment of human organs could also be developed. However, the technology is still nascent and faces major translational hurdles. In this review, we spotlighted the clinical burden of PNIs and most up-to-date treatment to address nerve gaps. Next, a summarized illustration of the nerve ultrastructure that guides research solutions is discussed. This is followed by a contrast of the existing bioengineering strategies used to repair peripheral nerve discontinuities. In addition, we elaborated on the most recent advances in 3D printing and biofabrication applications in peripheral nerve modeling and engineering. Finally, the major challenges that limit the evolution of the field along with their possible solutions are also critically analyzed. Impact statement Complex nerve injuries, including critical-sized gaps (>3 cm loss of substance), gaps involving nerve bifurcations, and those associated with ischemic environments, are difficult to manage. A biomimetic, personalized peripheral nerve tissue surrogate to address these injuries is lacking. The peripheral nerve repair market currently represents a multi-billion-dollar industry that is projected to expand. Given the clinical and economical dilemmas posed by this medical condition, it is crucial to devise novel and effective nerve substitutes. In this review article, we discuss progress in three-dimensional printing technologies, including biofabrication and nerve computer-aided design modeling, toward achieving a patient-specific and biomimetic nerve repair solution.

Hair Follicle Stem Cells for Tissue Regeneration.

With the positive outcomes of various cell therapies currently under preclinical and clinical studies, there is a significant interest in novel stem cell sources with unique therapeutic properties. Studies over the past two decades or so demonstrated the feasibility to isolate multipotent/pluripotent stem cells from hair follicles. The easy accessibility, high proliferation, and differentiation ability as well as lack of ethical concerns associated with this stem cell source make hair follicle stem cells (HFSCs) attractive candidate for cell therapy and tissue engineering. This review discusses the various stem cell types identified in rodent and human hair follicles and ongoing studies on the potential use of HFSCs for skin, bone, cardiovascular, and nerve tissue engineering. Impact statement Hair follicle stem cells are an autologous stem cell source, and recent preclinical and clinical studies demonstrated its unique properties to support and accelerate tissue regeneration, making it an attractive candidate for cell therapy and tissue engineering.