A Nature-Derived, Hetero-Structured, Pro-Healing Bioadhesive Patch for High-Performance Sealing of Wet Tissues.

Tissue adhesives are promising alternatives to sutures and staples to achieve wound closure and hemostasis. However, they often do not work well on tissues that are soaked in blood or other biological fluids, and organs that are typically exposed to a variety of harsh environments such as different pH values, nonhomogeneous distortions, continuous expansions and contractions, or high pressures. In this study, a nature-derived multilayered hetero-bioadhesive patch (skin secretion of Andrias davidianus (SSAD)-Patch) based on hydrophilic/hydrophobic pro-healing bioadhesives derived from the SSAD is developed, which is designed to form pressure-triggered strong adhesion with wet tissues. The SSAD-Patch is successfully applied for the sealing and healing of tissue defects within 10 s in diverse extreme injury scenarios in vivo including rat stomach perforation, small intestine perforation, fetal membrane defect, porcine carotid artery incision, and lung lobe laceration. The findings reveal a promising new type of self-adhesive regenerative SSAD-Patch, which is potentially adaptable to broad applications (under different pH values and air or liquid pressures) in sutureless wound sealing and healing.

An Adhesive Bioink toward Biofabrication under Wet Conditions.

Three-dimensional (3D) bioprinting is driving significant innovations in biomedicine over recent years. Under certain scenarios such as in intraoperative bioprinting, the bioinks used should exhibit not only cyto/biocompatibility but also adhesiveness in wet conditions. Herein, an adhesive bioink composed of gelatin methacryloyl, gelatin, methacrylated hyaluronic acid, and skin secretion of Andrias davidianus is designed. The bioink exhibits favorable cohesion to allow faithful extrusion bioprinting in wet conditions, while simultaneously showing good adhesion to a variety of surfaces of different chemical properties, possibly achieved through the diverse bonds presented in the bioink formulation. As such, this bioink is able to fabricate sophisticated planar and volumetric constructs using extrusion bioprinting, where the dexterity is further enhanced using ergonomic handheld bioprinters to realize in situ bioprinting. In vitro experiments reveal that cells maintain high viability; further in vivo studies demonstrate good integration and immediate injury sealing. The characteristics of the bioink indicate its potential widespread utility in extrusion bioprinting and will likely broaden the applications of bioprinting toward situations such as in situ dressing and minimally invasive tissue regeneration.

Systemic antibiotics increase microbiota pathogenicity and oral bone loss.

Periodontitis is the most widespread oral disease and is closely related to the oral microbiota. The oral microbiota is adversely affected by some pharmacologic treatments. Systemic antibiotics are widely used for infectious diseases but can lead to gut dysbiosis, causing negative effects on the human body. Whether systemic antibiotic-induced gut dysbiosis can affect the oral microbiota or even periodontitis has not yet been addressed. In this research, mice were exposed to drinking water containing a cocktail of four antibiotics to explore how systemic antibiotics affect microbiota pathogenicity and oral bone loss. The results demonstrated, for the first time, that gut dysbiosis caused by long-term use of antibiotics can disturb the oral microbiota and aggravate periodontitis. Moreover, the expression of cytokines related to Th17 was increased while transcription factors and cytokines related to Treg were decreased in the periodontal tissue. Fecal microbiota transplantation with normal mice feces restored the gut microbiota and barrier, decreased the pathogenicity of the oral microbiota, reversed the Th17/Treg imbalance in periodontal tissue, and alleviated alveolar bone loss. This study highlights the potential adverse effects of long-term systemic antibiotics-induced gut dysbiosis on the oral microbiota and periodontitis. A Th17/Treg imbalance might be related to this relationship. Importantly, these results reveal that the periodontal condition of patients should be assessed regularly when using systemic antibiotics in clinical practice.

Injectable decellularized dental pulp matrix-functionalized hydrogel microspheres for endodontic regeneration.

The sufficient imitation of tissue structures and components represents an effective and promising approach for tissue engineering and regenerative medicine applications. Dental pulp disease is one of the most common oral diseases, although functional pulp regeneration remains challenging. Herein, we propose a strategy that employs hydrogel microspheres incorporated with decellularized dental pulp matrix-derived bioactive factors to simulate a pulp-specific three-dimensional (3D) microenvironment. The dental pulp microenvironment-specific microspheres constructed by this regenerative strategy exhibited favorable plasticity, biocompatibility, and biological performances. Human dental pulp stem cells (hDPSCs) cultured on the constructed microspheres exhibited enhanced pulp-formation ability in vitro. Furthermore, the hDPSCs-microcarriers achieved the regeneration of pulp-like tissue and new dentin in a semi-orthotopic model in vivo. Mechanistically, the decellularized pulp matrix-derived bioactive factors mediated the multi-directional differentiation of hDPSCs to regenerate the pulp tissue by eliciting the secretion of crucial bioactive cues. Our findings demonstrated that a 3D dental pulp-specific microenvironment facilitated by hydrogel microspheres and dental pulp-specific bioactive factors regenerated the pulp-dentin complex and could be served as a promising treatment option for dental pulp disease. STATEMENT OF SIGNIFICANCE: Injectable bioscaffolds are increasingly used for regenerative endodontic treatment. Despite their success related to their ability to load stem cells, bioactive factors, and injectability, conventional bulk bioscaffolds have drawbacks such as ischemic necrosis in the central region. Various studies have shown that ischemic necrosis in the central region can be corrected by injectable hydrogel microspheres. Unfortunately, pristine microspheres or microspheres without dental pulp-specific bioactive factor would oftentimes fail to regulate stem cells fates in dental pulp multi-directional differentiation. Our present study reported the biofabrication of dental pulp-derived decellularized matrix functionalized gelatin microspheres, which contained dental pulp-specific bioactive factors and have the potential application in endodontic regeneration.

Investigation of Cell Adhesion and Cell Viability of the Endothelial and Fibroblast Cells on Electrospun PCL, PLGA and Coaxial Scaffolds for Production of Tissue Engineered Blood Vessel.

Endothelialization of artificial scaffolds is considered an effective strategy for increasing the efficiency of vascular transplantation. This study aimed to compare the biophysical/biocompatible properties of three different biodegradable fibrous scaffolds: Poly (ɛ-caprolactone) (PCL) alone, Poly Lactic-co-Glycolic Acid (PLGA) alone (both processed using Spraybase® electrospinning machine), and Coaxial scaffold where the fiber core and sheath was made of PCL and PLGA, respectively. Scaffold structural morphology was assessed by scanning electron microscope and tensile testing was used to investigate the scaffold tension resistance over time. Biocompatibility studies were carried out with human umbilical vein endothelial cells (HUVEC) and human vascular fibroblasts (HVF) for which cell viability (and cell proliferation over a 4-day period) and cell adhesion to the scaffolds were assessed by cytotoxicity assays and confocal microscopy, respectively. Our results showed that all biodegradable polymeric scaffolds are a reliable host to adhere and promote proliferation in HUVEC and HVF cells. In particular, PLGA membranes performed much better adhesion and enhanced cell proliferation compared to control in the absence of polymers. In addition, we demonstrate here that these biodegradable membranes present improved mechanical properties to construct potential tissue-engineered vascular graft.

Combination wound healing using polymer entangled porous nanoadhesive hybrids with robust ROS scavenging and angiogenesis properties.

Nanoadhesives can achieve tight wound closure by connecting biomacromolecules from both sides. However, previously developed adhesive systems suffered from suboptimal wound healing efficiency due to the lack of interparticle cohesion, sufficient reactive oxygen species (ROS)-scavenging sites, and angiogenesis consideration. Herein, we developed a polymer entangled porous nanoadhesive system to address the above challenge by synergy of three functional components. Firstly, hybrid mesoporous silica nanoparticles with highly integrated polydopamine (MS-PDA) were prepared by templated synthesis. The entangling between PVA polymer and MS-PDA contributed to much stronger cohesion between nanoparticles, which led to 75% larger adhesion strength. As confirmed by in vitro and in vivo evaluations, the highly exposed catechol groups boosted the scavenging activity of ROS (1.8-4.1 fold enhancement as compared with nonporous counterpart). Consequently, more macrophages exhibited anti-inflammatory phenotype, leading to 2-2.6 fold lower pro-inflammatory cytokine levels. Moreover, the sustained release of bioactive SiO44- by the disintegration of nanoparticles contributed to ∼3-fold higher expression of VEGF and enhanced new blood vessel formation, as well as better wound repair. This platform can provide a new paradigm for developing multifunctional nanoadhesive systems in treating skin wounds. STATEMENT OF SIGNIFICANCE: PVA polymer entangled mesoporous nanoadhesives of polydopamine (PDA)/silica hybrids with the combination of excellent wound closure effect, boosted ROS-scavenging activity, and significant angiogenesis ability were developed for improving the suboptimal skin wound healing efficiency. This strategy not only greatly advances our ability to rationally integrate repairing elements in nanoadhesives for manipulating combined processes of interfacial events during wound healing, but also offers general implications toward application of polymers to reinforce the adhesion strength in nanoadhesive systems. In addition, our findings on the impacts of pore effects mediated ROS species conversion and polymer entanglement may also trigger great interests and facilitate the development/broad application of therapeutic adhesives.

Perceived impact of the COVID-19 pandemic on infection containment training and mental state of dental residents in China: A longitudinal study.

Background: COVID-19 has presented a challenge for dental settings and dental schools: how to continue providing dental care and maintain education during the pandemic while remaining healthy. We highlight the necessity of infection containment control training for dental residents and rethink the tasks of safeguarding trainees' health and cultivating their abilities to deal with public health crises in the future. This paper may also serve as a health policy reference for policy makers. Objective: The study aimed to compare the formats, frequency, contents, emphasis, and test scores of infection containment control training pre- and post-pandemic. Besides, after the COVID-19 outbreak, we assessed the increased anxiety level, communication difficulties, and confidence of dental residents impacted by the pandemic. Methods: A total of 251 dental residents in Stomatological Hospital of Chongqing Medical University were recruited to complete a questionnaire of their routine involvement in infection control training before and after the COVID-19 outbreak. A self-designed 10-point Likert scale was used to assess the increased anxiety level, communication difficulties, and confidence in facing with the future public health crisis impacted by the pandemic. Results: After the outbreak, although more trainees chose online assessment than offline assessment, most of them (74.90%) still preferred in-person training rather than online training. Contents that trainees had been focusing on were affected by the COVID-19 outbreak. Thereafter, they were more inclined to learn crisis management. Over half of the participants (56.17%) participated in training more frequently after the outbreak. However, postgraduate students participated in training less frequently than others after the outbreak (p < 0.01). First-year trainees accounted for the majority in the population who emphasized considerably on infection control training and whose test scores had increased after the outbreak. In addition, the percentage of women scoring increasingly in post-pandemic assessment was significantly higher than that of men. In this study, the average increased anxiety level caused by COVID-19 was 5.51 ± 2.984, which was positively related to communication difficulties with patients caused by the pandemic. The trainees whose homes were located in Hubei Province showed higher increased anxiety levels (8.29 ± 2.93) impacted by the pandemic than the trainees from other provinces (p < 0.05). However, the former's confidence in coping with future public health crises was not significantly different from that of others (p > 0.05). Conclusions: Owing to the impact of COVID-19, the contents that the trainees focused on, frequency, emphasis, and test scores of infection containment control training were changed. Some recommendations have been provided for policy makers to attach importance to crisis-based training to cultivate dental residents in the post-pandemic era.

A rational synthesis of ultrasmall palladium-based alloys with superhydrophilicity as biocompatible agents and recyclable catalysts.

As essential controlling parameters, the local surface area (size distribution) and polarity property of the surface molecules can determine the catalytic activity and biocompatibility directly. Here, ultrasmall palladium-based alloys (FePd, FePdCo, and FePdCu NCs) were developed to serve as artificial degradation catalysts with superhydrophilicity (SPh), biocompatibility, and reusability, which were referred to as "biocatalysts". As synthesized in aqueous solvent with negative surface potential while dispersing in water medium, because of the surface biological molecules effect. The obtained alloys illustrated a size distribution of about 3.5 nm. Additionally, owing to SPh property, these alloys could be stored in water up to 30 days without any precipitation and retained their monodisperse morphology in colloidal solutions. The cytotoxicity assessment of the alloys by exposing to L-929 cells over 3 days indicated that it maintained cell viability of >80% even up to 390 µg mL-1 (concentration of alloys). Furthermore, they exhibited an obvious enhancement in the catalytic performance for degrading Rhodamine B (RhB) and 4-nitrophenol (4-NP). The recyclable utilization of biocatalysts demonstrates durable stability even after 8 reduction cycles.

Bioinspired Andrias davidianus-Derived wound dressings for localized drug-elution.

Local drug delivery has received increasing attention in recent years. However, the therapeutic efficacy of local delivery of drugs is still limited under certain scenarios, such as in the oral cavity or in wound beds after resection of tumors. In this study, we introduce a bioinspired adhesive hydrogel derived from the skin secretions of Andrias davidianus (SSAD) as a wound dressing for localized drug elution. The hydrogel was loaded with aminoguanidine or doxorubicin, and its controlled drug release and healing-promoting properties were verified in a diabetic rat palatal mucosal defect model and a C57BL/6 mouse melanoma-bearing model, respectively. The results showed that SSAD hydrogels with different pore sizes could release drugs in a controllable manner and accelerate wound healing. Transcriptome analyses of the palatal mucosa suggested that SSAD could significantly upregulate pathways linked to cell adhesion and extracellular matrix deposition and had the ability to recruit keratinocyte stem cells to defect sites. Taken together, these findings indicate that property-controllable SSAD hydrogels could be a promising biofunctional wound dressing for local drug delivery and promotion of wound healing.

A Natural Hydrogel with Prohealing Properties Enhances Tendon Regeneration.

Tendon regeneration and reduction of peritendinous adhesion remain major clinical challenges. This study addresses these challenges by adopting a unique hydrogel derived from the skin secretion of Andrias davidianus (SSAD) and taking advantage of its biological effects, adhesiveness, and controllable microstructures. The SSAD-derived hydrogel contains many cytokines, which could promote tendon healing. In vitro, leach liquid of SSAD powder could promote tendon stem/progenitor cells migration. In vivo, the SSAD-derived hydrogel featuring double layers possesses strong adhesiveness and could reconnect ruptured Achilles tendons of Sprague-Dawley rats without suturing. The intimal SSAD-derived hydrogel, with a pore size of 241.7 ± 21.0 µm, forms the first layer of the hydrogel to promote tendon healing, and the outer layer SSAD-derived hydrogel, with a pore size of 3.3 ± 1.4 µm, reducing peritendinous adhesion by serving as a dense barrier. Additionally, the SSAD-derived hydrogel exhibits antioxidant and antibacterial characteristics, which further contribute to the reduction of peritendinous adhesion. In vivo studies suggest that the SSAD-derived hydrogel reduces peritendinous adhesion, increases collagen fiber deposition, promotes cell proliferation, and improves the biomechanical properties of the regenerated tendons, indicating better functional restoration. The SSAD-derived bilayer hydrogel may be a feasible biomaterial for tendon repair in the future.

Correction: Antimicrobial peptide modification enhances the gene delivery and bactericidal efficiency of gold nanoparticles for accelerating diabetic wound healing.

Correction for 'Antimicrobial peptide modification enhances the gene delivery and bactericidal efficiency of gold nanoparticles for accelerating diabetic wound healing' by Song Wang et al., Biomater. Sci., 2018, 6, 2757-2772, DOI: 10.1039/C8BM00807H.

A Bioinspired Hemostatic Powder Derived from the Skin Secretion of Andrias davidianus for Rapid Hemostasis and Intraoral Wound Healing.

High-performance hemostasis has become increasingly essential in treating various traumas. However, available topical hemostats still have various drawbacks and side-effects. Herein, hemostatic powders derived from the skin secretion of Andrias davidianus (SSAD) with controllable particle size are prepared using feasible frozen-ball milling following lyophilization for hemorrhage-control. Scanning electron microscopy, rheometry, and Brunauer-Emmett-Teller test are used to characterize the coagulation-promoting surface topography, rheological properties, and porous structure of the SSAD particles. The blood-coagulation assays showed that the SSAD powders can induce blood-absorption in a particle size-dependent manner. Particle sizes of the SSAD powders larger than 200 µm and smaller than 800 µm greatly affect the blood-clotting rate. Associated with the thromboelastography (TEG) and amino acid/protein composition analyses, the accessibility and diffusion of blood are mainly dependent on the wettability, adhesivity, and clotting factors of the SSAD particles. Rapid hemostasis in vivo further involves three hemorrhage models (liver, femoral artery, and tail) as well as an oral wound model, which suggest favorable hemostatic and simultaneous regenerative effects of the SSAD hemostatic powder. Considering its degradability and good biocompatibility, SSAD can be an optimal candidate for a new class of inexpensive, natural, and promising hemostatic and wound-dressing agent.

Fabrication and Characterization of PCL/PLGA Coaxial and Bilayer Fibrous Scaffolds for Tissue Engineering.

Electrospinning is an innovative new fibre technology that aims to design and fabricate membranes suitable for a wide range of tissue engineering (TE) applications including vascular grafts, which is the main objective of this research work. This study dealt with fabricating and characterising bilayer structures comprised of an electrospun sheet made of polycaprolactone (PCL, inner layer) and an outer layer made of poly lactic-co-glycolic acid (PLGA) and a coaxial porous scaffold with a micrometre fibre structure was successfully produced. The membranes' propriety for intended biomedical applications was assessed by evaluating their morphological structure/physical properties and structural integrity when they underwent the degradation process. A scanning electron microscope (SEM) was used to assess changes in the electrospun scaffolds' structural morphology such as in their fibre diameter, pore size (µm) and the porosity of the scaffold surface which was measured with Image J software. During the 12-week degradation process at room temperature, most of the scaffolds showed a similar trend in their degradation rate except the 60 min scaffolds. The coaxial scaffold had significantly less mass loss than the bilayer PCL/PLGA scaffold with 1.348% and 18.3%, respectively. The mechanical properties of the fibrous membranes were measured and the coaxial scaffolds showed greater tensile strength and elongation at break (%) compared to the bilayer scaffolds. According to the results obtained in this study, it can be concluded that a scaffold made with a coaxial needle is more suitable for tissue engineering applications due to the improved quality and functionality of the resulting polymeric membrane compared to the basic electrospinning process. However, whilst fabricating a vascular graft is the main aim of this research work, the biological data will not present in this paper.

Degradation and Characterisation of Electrospun Polycaprolactone (PCL) and Poly(lactic-co-glycolic acid) (PLGA) Scaffolds for Vascular Tissue Engineering.

The current study aimed to evaluate the characteristics and the effects of degradation on the structural properties of Poly(lactic-co-glycolic acid) (PLGA)- and polycaprolactone (PCL)-based nanofibrous scaffolds. Six scaffolds were prepared by electrospinning, three with PCL 15% (w/v) and three with PLGA 10% (w/v), with electrospinning processing times of 30, 60 and 90 min. Both types of scaffolds displayed more robust mechanical properties with increased spinning times. The tensile strength of both scaffolds with 90-min electrospun membranes did not show a significant difference in their strengths, as the PCL and PLGA scaffolds measured at 1.492 MPa ± 0.378 SD and 1.764 MPa ± 0.7982 SD, respectively. All membranes were shown to be hydrophobic under a wettability test. A degradation behaviour study was performed by immersing all scaffolds in phosphate-buffered saline (PBS) solution at room temperature for 12 weeks and for 4 weeks at 37 °C. The effects of degradation were monitored by taking each sample out of the PBS solution every week, and the structural changes were investigated under a scanning electron microscope (SEM). The PCL and PLGA scaffolds showed excellent fibre structure with adequate degradation, and the fibre diameter, measured over time, showed slight increase in size. Therefore, as an example of fibre water intake and progressive degradation, the scaffold's percentage weight loss increased each week, further supporting the porous membrane's degradability. The pore size and the porosity percentage of all scaffolds decreased substantially over the degradation period. The conclusion drawn from this experiment is that PCL and PLGA hold great promise for tissue engineering and regenerative medicine applications.

3D printing of robust and biocompatible poly(ethylene glycol)diacrylate/nano-hydroxyapatite composites via continuous liquid interface production.

Three-dimensional (3D) printing technology with satisfactory speed and accuracy has been a powerful force in biomaterial processing. Early studies on 3D printing of biomaterials mainly focused on their biocompatibility and cellular viability while rarely attempted to produce robust specimens. Nonetheless, the biomedical applications of polymers can be severely limited by their inherently weak mechanical properties particularly in bone tissue engineering. In this study, continuous liquid interface production (CLIP) is applied to construct 3D objects of nano-hydroxyapatite (n-HA) filled polymeric biomaterials with complex architectures. Notably, the bioactive and osteoconductive n-HA endows the 3D prints of poly(ethyleneglycol)diacrylate (PEGDA) composites with a high compression strength of 6.5 ± 1.4 MPa, about 342% improvement over neat PEGDA. This work demonstrates the first successful attempt on CLIP 3D printing of n-HA nanocomposites, providing a feasible, cost-effective and patient-specific solution to various fields in the biomedical industry.

Honeycomb-structured carbon aerogels from nanocellulose and skin secretion of Andrias davidianus for highly compressible binder-free supercapacitors.

Nanocellulose-derived carbon is a promising material in energy storage because of its sustainability, low environmental impact, and large specific surface area. Herein, the skin secretion of Andrias davidianus (SSAD) is applied for the first time as the bio-nitrogen source to dope carbon aerogels from nanocellulose. Cellulose nanocrystal (CNC) is discovered to be very effective to address the dispersion problem of SSAD in water. After being homogeneously mixed with cellulose nanofiber (CNF), honeycomb-structured nanofibrous carbon aerogels are obtained via unidirectional freeze-drying of the SSAD/CNC/CNF mixture followed by high-temperature carbonization. Impressively, unlike those fragile carbon aerogels in many early works, the present ones exhibit outstanding elasticity in repeated compression and release tests. Moreover, a symmetric binder-free supercapacitor is assembled from the carbon aerogels, which exhibits improved electrochemical capacitive properties and cycling stability. And even after 500 compression and release cycles, the supercapacitor can still maintain high capacitive performance, indicating its superiorities in durability and electrochemical stability.

Aligned electrospun cellulose scaffolds coated with rhBMP-2 for both in vitro and in vivo bone tissue engineering.

Physical properties of scaffolds such as nanofibers and aligned structures have been reported to exert profound effects on the growth and differentiation of stem cells due to their homing-effect features and contact guidance. However, the biological function of aligned nanofibers utilized as bone-scaffold has not been rigorously characterized. In the present study, aligned electrospun cellulose/CNCs nanocomposite nanofibers (ECCNNs) loaded with bone morphogenic protein-2 (BMP-2) were used for the first time to investigate (1) in vitro osteogenic differentiation of human mesenchymal stem cells (BMSCs) and (2) in vivo collagen assembly direction and cortical bone regeneration. Aligned ECCNNs scaffolds loaded with BMP-2 possess good biological compatibility. The growth orientation of BMSCs followed the underlying aligned nanofiber morphology, accompanied with increased alizarin red stain, alkaline phosphatase (ALP) activity and calcium content in vitro while, a rabbit calvaria bone defect model was used in an in vivo study with micro CT and histology analyses.

Surface-Adaptive and Initiator-Loaded Graphene as a Light-Induced Generator with Free Radicals for Drug-Resistant Bacteria Eradication.

Since generating toxic reactive oxygen species is largely dependent on oxygen, bacteria-infected wounds' hypoxia significantly inhibits photodynamic therapy's antibacterial efficiency. Therefore, a novel therapeutic method for eradicating multidrug-resistant bacteria is developed based on the light-activated alkyl free-radical generation (that is oxygen independent). According to the polydopamine-coated carboxyl graphene (PDA@CG), an initiator-loaded and pH-sensitive heat-producible hybrid of bactericides was synthesized. According to fluorescence/thermal imaging, under the low pH of the bacterial infection sites, this platform turned positively charged, which allows their accumulation in local infection site. The plasmonic heating effects of PDA@CG can make the initiator decomposed to generate alkyl radical (R•) under the followed near-infrared light irradiation. As a result, oxidative stress can be elevated, DNA damages in bacteria can be caused, and finally even multidrug-resistance death can be caused under different oxygen tensions. Moreover, our bactericidal could promote wound healing in vivo and negligible toxicity in vivo and in vitro and eliminate abscess. Accordingly, this study proves that combination of oxygen-independent free-radical-based therapy along with a stimulus-responsiveness moiety not only can be used as an effective treatment of multidrug-resistant bacteria infection, but also creates a use of a variety of free radicals for treatment of multidrug-resistant bacteria infection wounds.

Identification of Glycine Receptor α3 as a Colchicine-Binding Protein.

Colchicine (Col) is considered a kind of highly effective alkaloid for preventing and treating acute gout attacks (flares). However, little is known about the underlying mechanism of Col in pain treatment. We have previously developed a customized virtual target identification method, termed IFPTarget, for small-molecule target identification. In this study, by using IFPTarget and ligand similarity ensemble approach (SEA), we show that the glycine receptor alpha 3 (GlyRα3), which play a key role in the processing of inflammatory pain, is a potential target of Col. Moreover, Col binds directly to the GlyRα3 as determined by the immunoprecipitation and bio-layer interferometry assays using the synthesized Col-biotin conjugate (linked Col and biotin with polyethylene glycol). These results suggest that GlyRα3 may mediate Col-induced suppression of inflammatory pain. However, whether GlyRα3 is the functional target of Col and serves as potential therapeutic target in gouty arthritis requires further investigations.