Naringin acts as a TRPV1 antagonist to attenuate UVB-induced senescence and damage in HaCaT cells.

This study aimed to explore the mechanism of naringin (Nar) in alleviating ultraviolet B (UVB)-induced HaCaT cell senescence and damage. Human keratinocytes (HaCaT cells) were divided into control, UVB, UVB + Nar, UVB + Cap, and UVB + Nar + Cap groups. Analysis was performed using the MTT assay to assess cell viability, flow cytometry to measure the apoptosis level, SA-ß-Gal staining to observe cellular senescence, and Western blot to assess protein levels of TRPV1, p16, p53, p21, matrix metalloproteinase (MMP)-1, and MMP-9. Both UVB irradiation and capsaicin (Cap) treatment upregulated the expression of TRPV1 in HaCaT cells, inhibited cell proliferation, promoted apoptosis, and increased the expression of p16, p53, p21, MMP-1, and MMP-9. Nar treatment reversed the above effects via inhibition of TRPV1 expression, thereby relieving senescence and cell damage induced by UVB irradiation. Taken together, these findings suggest that Nar can reduce UVB-induced senescence and damage in HaCaT cells by acting as an antagonist of TRPV1.

Adsorption and Visible Photocatalytic Synergistic Removal of a Cationic Dye with the Composite Material BiVO4/MgAl-LDHs.

Adsorption and photocatalysis are effective in removing organic pollutants from wastewater. This study is based on the memory effects of MgAl-layered double hydroxides (MgAl-LDHs) after high-temperature calcination. By introducing bismuth vanadate (BiVO4) during the reformation of the layered structure via contact with water, a composite material BiVO4/MgAl-LDHs with enhanced adsorption and visible light catalytic performance was synthesized. The effects of the calcination temperature, ratio, initial methylene blue (MB) concentration, and catalyst dosage on the adsorption and photocatalytic performance were investigated. The BiVO4/MgAl-LDHs showed better photocatalytic performance than the pure BiVO4 and MgAl-LDHs. Under the optimal conditions, the proportion of MB adsorbed in 20 min was 66.1%, and the percentage of MB degraded during 100 min of photolysis was 92.4%. The composite photocatalyst showed good chemical stability and cyclability, and the adsorption-degradation rate was 86% after four cycles. Analyses of the adsorption and photocatalytic mechanisms for the composite material showed that synergistic adsorption and visible light photocatalysis contributed to the excellent catalytic performance of the BiVO4/MgAl-LDHs. A highly adsorbent photocatalytic composite material exhibiting outstanding performance was prepared via a simple, cost-effective, and environmentally friendly method, providing reference information for the removal of organic pollutants from liquids.

Evolutionary origin and establishment of a dioecious diploid-tetraploid complex.

Polyploids recurrently emerge in angiosperms, but most polyploids are likely to go extinct before establishment due to minority cytotype exclusion, which may be specifically a constraint for dioecious plants. Here we test the hypothesis that a stable sex-determination system and spatial/ecological isolation facilitate the establishment of dioecious polyploids. We determined the ploidy levels of 351 individuals from 28 populations of the dioecious species Salix polyclona, and resequenced 190 individuals of S. polyclona and related taxa for genomic diversity analyses. The ploidy survey revealed a frequency 52% of tetraploids in S. polyclona, and genomic k-mer spectra analyses suggested an autopolyploid origin for them. Comparisons of diploid male and female genomes identified a female heterogametic sex-determining factor on chromosome 15, which probably also acts in the dioecious tetraploids. Phylogenetic analyses revealed two diploid clades and a separate clade/grade of tetraploids with a distinct geographic distribution confined to western and central China, where complex mountain systems create higher levels of environmental heterogeneity. Fossil-calibrated phylogenies showed that the polyploids emerged during 7.6-2.3 million years ago, and population demographic histories largely matched the geological and climatic history of the region. Our results suggest that inheritance of the sex-determining system from the diploid progenitor as intrinsic factor and spatial isolation as extrinsic factor may have facilitated the preservation and establishment of polyploid dioecious populations.

The male-heterogametic sex determination system on chromosome 15 of Salix triandra and Salix arbutifolia reveals ancestral male heterogamety and subsequent turnover events in the genus Salix.

Dioecious Salix evolved more than 45 million years ago, but have homomorphic sex chromosomes, suggesting that turnover event(s) prevented major differentiation. Sex chromosome turnover events have been inferred in the sister genus Populus. The genus Salix includes two main clades, Salix and Vetrix, with several previously studied Vetrix clade species having female-heterogametic (ZW) or male-heterogametic (XY) sex-determining systems (SDSs) on chromosome 15, while three Salix clade species have XY SDSs on chromosome 7. We here studied two basal taxa of the Vetrix clade, S. arbutifolia and S. triandra using S. purpurea as the reference genome. Analyses of whole genome resequencing data for genome-wide associations (GWAS) with the sexes and genetic differentiation between the sexes (FST values) showed that both species have male heterogamety with a sex-determining locus on chromosome 15, suggesting an early turnover event within the Vetrix clade, perhaps promoted by sexually antagonistic or (and) sex-ratio selection. Changepoint analysis based on FST values identified small sex-linked regions of ~3.33 Mb and ~2.80 Mb in S. arbutifolia and S. triandra, respectively. The SDS of S. arbutifolia was consistent with recent results that used its own genome as reference. Ancestral state reconstruction of SDS suggests that at least two turnover events occurred in Salix.

Investigation on characterization of novel anti-bacterial chitosan/gelatin composite membranes loaded with quercetin via electrophoretic deposition.

Peri-implantitis is characterized by inflammation resulting from bacterial infections in peri-implant connective tissue. The purpose of this study was to prepare and characterize chitosan/gelatin (CSG)-based membranes with antibacterial agents to functionalize the surface of titanium (Ti) implants. CSG membranes were prepared on Ti substrates via electrophoretic deposition (EPD). Quercetin, an active flavonoid responsible for fulfilling various plant functions, was introduced as an antibacterial agent to be loaded into the membrane during preparation. The fabrication of quercetin-loaded CSG membranes via EPD was also investigated. Fluorescent microscope, Attenuated Total Reflection Fourier transform infrared spectroscopy, and X-ray diffraction results verified the entrapment of quercetin. The membranes swelled by 150% of mass after rehydration. The antibacterial effects of quercetin on Gram-positive bacteria, such as Staphylococcus aureus and methicillin-resistant Staphylococcus aureus, were verified by spread-plate, scanning electron microscopy, and live/dead staining. Cytological experiments showed that the biocompatibility of rat bone marrow mesenchymal stromal cells was promoted by quercetin-loaded membranes, exclusively in the group with the highest content of quercetin. The quercetin-loaded groups also enhanced the antineoplastic activity of MG-63 cells. These results suggested that quercetin-loaded CSG membranes were successfully fabricated via EPD. Thus, biocompatible and antibacterial membranes could be a potential strategy to functionalize Ti implants.

Cell-scaffold interactions in tissue engineering for oral and craniofacial reconstruction.

Tissue engineering (TE) is critical in oral and craniofacial reconstruction. One of the most popular topics on the biomaterial-based tissue regeneration process may be the interaction between cells and scaffolds. An increasing number of studies have identified the variables affecting cell-scaffold interaction. The creation and investigation of new scaffolds for TE and regenerative medicine based on specific interactions have become possible owing to these findings. This review discusses the effects of various types of scaffold materials on cells in TE. Because the intrinsic properties of scaffolds are essential, the influence of the physical, chemical, mechanical, and biological characteristics of scaffold materials on cell-scaffold interaction that has been discovered in recent research is elaborated in this review. The components carried by scaffolds, the degradation process, and the role of degraded products in cell-scaffold interactions are examined. Further, the roles of cells, including cell types, functions, and adhesion mechanisms, and extracellular matrix are discussed. Finally, the latest research progress on cell-scaffold interactions among various engineered tissues or organs in the oral and craniofacial region is summarized. A deeper understanding of cell-scaffold interactions is anticipated to benefit the development of TE and regenerative medicine.

Antimicrobial peptide GL13K immobilized onto SLA-treated titanium by silanization: antibacterial effect against methicillin-resistant Staphylococcus aureus (MRSA).

Infection is the main reason for implant failure, and the incidence of drug-resistant bacterial infection has increased in recent years. Further, methicillin-resistant Staphylococcus aureus (MRSA)-related implant infection has become a serious worldwide threat. New strategies, other than antibiotics, to tackle drug-resistance, are of high clinical significance. Antimicrobial peptides show clear superiority over conventional antibiotics in inhibiting drug-resistant bacteria. In the present study, we combined the antimicrobial peptide, GL13K, with sandblasting and acid-etching (SLA)-treated titanium using a silane coupling agent. Field emission scanning electron microscopy images showed the morphology of the coating. Attenuated total reflectance Fourier transform infrared spectroscopy and X-ray photoelectron spectroscopy results confirmed loading of GL13K, and the hydrophilicity of the SLA-GL13K coating was evaluated by water contact angle analysis. The releasing study of samples showed that the coating has a sustained releasing profile. SLA-GL13K coating exhibited strong contact- and release-killing abilities against MRSA, E. coli, and S. aureus. Meanwhile, Cell Counting Kit 8 analysis and examination of cell morphology demonstrated that the SLA-GL13K coating had good cytocompatibility at antibacterial concentrations. Overall, all these results suggest that SLA-GL13K coating can be successfully fabricated using silanization, and is a promising candidate for controlling MRSA-induced implant-related infection.

Fabrication and functionalization of biocompatible carboxymethyl chitosan/gelatin membranes via anodic electrophoretic deposition.

Peri-implant surgical site infection is a significant challenge in oral implant surgery. Numerous surface functionalization methods, including electrophoretic deposition, have been studied to functionalize implant surfaces to prevent peri-implantitis. However, it is still challenging to load anti-inflammatory agents having negative charges into electrophoretic deposition membranes. The present study aimed to use water-soluble chitosan derivatives to fabricate negatively charged carboxymethyl chitosan/gelatin (CMCG) composite membranes on titanium (Ti) substrates via anodic electrophoretic deposition (AED). Membranes incorporating different amounts of gelatin were labeled as CMC, CMCG4, CMCG6, and CMCG8. X-ray diffraction and Fourier transform infrared spectroscopy tests verified that CMCG could be deposited on Ti disks via AED. The result of the contact angle test showed that groups incorporating gelatin had a certain degree of hydrophobicity. After rehydration, the membranes swelled by approximately 200% in weight. Fluorescence microscopy and scanning electron microscopy images showed that bone marrow stromal cells (BMSCs) on membranes stretched well, showing a good cell adhesion ability. The CCK-8 test demonstrated that CMCG6 had the highest proliferation rate. Cell apoptosis studies showed that CMCG could inhibit apoptosis of BMSCs statistically. It suggests that the CMCG membrane fabricated by AED would be a potent candidate for surface functionalization of biomaterials with negative charges.

The phylogeny of Salix revealed by whole genome re-sequencing suggests different sex-determination systems in major groups of the genus.

BACKGROUND AND AIMS: The largest genus of Salicaceae sensu lato, Salix, has been shown to consist of two main clades: clade Salix, in which species have XY sex-determination systems (SDSs) on chromosome 7, and clade Vetrix including species with ZW SDSs on chromosome 15. Here, we test the utility of whole genome re-sequencing (WGR) for phylogenomic reconstructions of willows to infer changes between different SDSs. METHODS: We used more than 1 TB of WGR data from 70 Salix taxa to ascertain single nucleotide polymorphisms on the autosomes, the sex-linked regions (SLRs) and the chloroplast genomes, for phylogenetic and species tree analyses. To avoid bias, we chose reference genomes from both groups, Salix dunnii from clade Salix and S. purpurea from clade Vetrix. KEY RESULTS: Two main largely congruent groups were recovered: the paraphyletic Salix grade and the Vetrix clade. The autosome dataset trees resolved four subclades (C1-C4) in Vetrix. C1 and C2 comprise species from the Hengduan Mountains and adjacent areas and from Eurasia, respectively. Section Longifoliae (C3) grouped within the Vetrix clade but fell into the Salix clade in trees based on the chloroplast dataset analysis. Salix triandra from Eurasia (C4) was revealed as sister to the remaining species of clade Vetrix. In Salix, the polyploid group C5 is paraphyletic to clade Vetrix and subclade C6 is consistent with Argus's subgenus Protitea. Chloroplast datasets separated both Vetrix and Salix as monophyletic, and yielded C5 embedded within Salix. Using only diploid species, both the SLR and autosomal datasets yielded trees with Vetrix and Salix as well-supported clades. CONCLUSION: WGR data are useful for phylogenomic analyses of willows. The different SDSs may contribute to the isolation of the two major groups, but the reproductive barrier between them needs to be studied.

Chromosome-scale assembly of the genome of Salix dunnii reveals a male-heterogametic sex determination system on chromosome 7.

Sex determination systems in plants can involve either female or male heterogamety (ZW or XY, respectively). Here we used Illumina short reads, Oxford Nanopore Technologies (ONT) long reads and Hi-C reads to assemble the first chromosome-scale genome of a female willow tree (Salix dunnii), and to predict genes using transcriptome sequences and available databases. The final genome sequence of 328 Mb in total was assembled in 29 scaffolds, and includes 31,501 predicted genes. Analyses of short-read sequence data that included female and male plants suggested a male heterogametic sex-determining factor on chromosome 7, implying that, unlike the female heterogamety of most species in the genus Salix, male heterogamety evolved in the subgenus Salix. The S. dunnii sex-linked region occupies about 3.21 Mb of chromosome 7 in females (representing its position in the X chromosome), probably within a pericentromeric region. Our data suggest that this region is enriched for transposable element insertions, and about one-third of its 124 protein-coding genes were gained via duplications from other genome regions. We detect purifying selection on the genes that were ancestrally present in the region, though some have been lost. Transcriptome data from female and male individuals show more male- than female-biased genes in catkin and leaf tissues, and indicate enrichment for male-biased genes in the pseudo-autosomal regions. Our study provides valuable genomic resources for further studies of sex-determining regions in the family Salicaceae, and sex chromosome evolution.

Protective roles of FICZ and aryl hydrocarbon receptor axis on alveolar bone loss and inflammation in experimental periodontitis.

AIM: The aryl hydrocarbon receptor (AhR)-ligand axis has been shown to be involved in inflammatory diseases and bone homeostasis. However, the activation of AhR signalling pathway and the possible functions of AhR ligands in periodontitis are underexplored. This study investigated the expression of the AhR target gene cytochrome P450 subfamily B member 1 (CYP1B1) and the functions and mechanisms of the AhR ligand 6 formylindolo[3,2-b]carbazole (FICZ) in periodontitis. MATERIALS AND METHODS: CYP1B1 expression was detected in human periodontitis samples, mice with ligature-induced periodontitis and lipopolysaccharide (LPS)-induced inflammation in periodontal ligament cells (PDLCs) in vitro. FICZ was administered topically or systemically. The therapeutic functions of FICZ were detected via qPCR, micro-computed tomography and immunohistochemistry. Finally, the mechanisms of AhR signalling in periodontitis were investigated by cell assays. RESULTS: CYP1B1 expression was downregulated in periodontitis. FICZ rescued the alveolar bone loss and mitigated the inflammatory cytokines in periodontitis mice. In vitro, FICZ pre-treatment reduced the LPS-induced inflammation in PDLCs via the increased phosphorylation of STAT3. Additionally, FICZ prompted the mineralization of PDLCs via activation of the Wnt/ß-catenin signalling pathway. CONCLUSION: AhR signalling pathway is suppressed in periodontitis and the AhR ligand FICZ can prevent periodontitis.

A thermosensitive chitosan-based hydrogel for sealing and lubricating purposes in dental implant system.

BACKGROUND: Mechanical and biological complications associated with implant systems happen frequently in the clinic. PURPOSE: To develop a chitosan (CS)-based thermosensitive hydrogel for sealing and lubricating purposes in dental implant system. MATERIALS AND METHODS: In this study, a thermosensitive hydrogel made up of CS, ß-glycerophosphate pentahydrate (ß-GP), and povidone-iodine (PVP-I), called CS/ß-GP/PVP-I thermosensitive hydrogel, was fabricated. Three experimental groups with different volume ratios of CS to ß-GP were prepared, namely 16/4, 13/7, and 10/10 groups. The surface topography of the different groups and their physicochemical characteristics were examined by SEM, FTIR, and X-ray diffraction analysis. The cytotoxicity of the hydrogel was examined by CCK-8 test. In vitro antibacterial efficiency was analyzed by the spread plate method. Sealing ability was detected by incubating two-piece implants in Escherichia coli suspension. Lubricating ability of the hydrogel was evaluated by the removal torque test with a calibrated digital torque meter. RESULTS: The CS/ß-GP/PVP-I thermosensitive hydrogel was fabricated and showed a highly porous structure under SEM. An in vitro cytotoxicity test demonstrated that 13/7 group displayed no cytotoxicity. Furthermore, all three groups showed obviously antibacterial effects. In the sealing ability test, 16/4 group showed the best sealing ability. The removal torque of 16/4 group and 13/7 group was significantly greater than control group. CONCLUSIONS: Based on our findings, it could be concluded that the thermosensitive and antibacterial CS/ß-GP/PVP-I hydrogel with sealing and lubricating ability was successfully prepared. The hydrogel had better sealing and lubricating effects when the volume ratio of CS to ß-GP was 16/4.

Periodontal regeneration via chemoattractive constructs.

AIM: Chemoattractants, such as stromal cell-derived factor-1α (SDF-1α), can offer an advantage for periodontal regeneration by recruiting the patient's own stem cells to stimulate self-repair. We here developed a chemoattractive construct for periodontal regeneration using SDF-1α and evaluated its efficacy in vivo. MATERIALS AND METHODS: SDF-1α was loaded on gelatin sponge and tested in vitro for SDF-1α release. Subsequently, SDF-1α constructs were implanted into rat periodontal defects for 1 and 6 weeks, with unloaded materials and empty defects as controls. The regenerative efficacy was evaluated by micro-CT, histological and histomorphometrical analyses. RESULTS: In vitro results showed limited SDF-1α release up to 35 days. In contrast, SDF-1α constructs significantly improved periodontal defect regeneration in terms of alveolar bone height, new bone area and functional ligament length. Additionally, SDF-1α constructs decreased the inflammatory response at Week 6. CONCLUSION: Chemoattractive constructs significantly improved periodontal regeneration in terms of alveolar bone height, new bone area and functional ligament length.

Evaluation of a platelet lysate bilayered system for periodontal regeneration in a rat intrabony three-wall periodontal defect.

With currently available therapies, full regeneration of lost periodontal tissues after periodontitis cannot be achieved. In this study, a combined compartmentalized system was tested, composed of (a) a platelet lysate (PL)-based construct, which was placed along the root aiming to regenerate the root cementum and periodontal ligament, and (b) a calcium phosphate cement composite incorporated with hyaluronic acid microspheres loaded with PL, aiming to promote the regeneration of alveolar bone. This bilayered system was assessed in a 3-wall periodontal defect in Wistar rats. The periodontal healing and the inflammatory response of the materials were scored for a period up to 6 weeks after implantation. Furthermore, histomorphometrical measurements were performed to assess the epithelial downgrowth, the formation of alveolar bone, and the formation of new connective tissue attachment. Our data showed that the stabilization of platelet-origin proteins on the root surface increased the overall periodontal healing score and restricted the formation of long epithelial junctions. Nevertheless, the faster degradation of the cement component with incorporated hyaluronic acid microspheres compromised the stability of the system, which hampered the periodontal regeneration. Overall, in this work, we proved the positive therapeutic effect of the immobilization of a PL-based construct over the root surface in a combined compartmentalized system to assist predictable healing of functional periodontium. Therefore, after optimization of the hard tissue analogue, the system should be further elaborated in (pre)clinical validation studies.

Preparation and functionalization of acetylsalicylic acid loaded chitosan/gelatin membranes from ethanol-based suspensions via electrophoretic deposition.

Acetylsalicylic acid (Aspirin, ASP), a frequently used analgesic and antipyretic drug, has prevailed for decades due to its multiple functions. To increase its solubility and maintain the topical and low-dose application, ethanol with electrophoretic deposition (EPD) was introduced. It was initially investigated to fabricate acetylsalicylic acid loaded chitosan/gelatin (CS/G) membranes via simple electrophoretic deposition under mild conditions. The spectrophotometry, SEM, FTIR and XRD results confirmed the entrapment of acetylsalicylic acid. FTIR spectra also indicated that new bonds were formed between acetylsalicylic acid and the CS/G membranes. The contact angle study confirmed the good hydrophilicity of the samples' surfaces. The mechanical strengths of membranes were promoted due to the introduction of ethanol. The in vitro cellular study revealed the capacity of promoting osteogenic differentiation and little influence on the cell viability for BMSCs. All these results suggested that acetylsalicylic acid loaded CS/G membranes could be successfully fabricated via EPD and used for functionalizing the titanium substrate. These membranes loaded with other functional reagents, hydrosoluble or liposoluble, may also be promising for use in medical applications.

Resolution of inflammation in periodontitis: a review.

Inflammation is a physiological response to an injury or infection. It is supposed to be self-limiting, stopping when the situation recovers to normal to protect the tissue. This self-limiting action is called "resolution of inflammation". Currently, periodontitis is thought to be the result of failed resolution of inflammation; specifically, it is the result of excessive inflammation that leads to gingival recession and alveolar bone loss. In this review, we will focus on the processes of resolution of inflammation in periodontitis, which may be a therapeutic target of periodontitis.

The mutual effects between macrophages and cartilage templates in the process of subcutaneous endochondral bone formation.

The interplay between implants and the recipient immune environment is key to the long-term effectiveness of bone tissue engineering. In this study, we aimed to investigate the mutual effects between macrophages and cartilage templates in the process of subcutaneous osteogenesis. Primary mice bone marrow derived mesenchymal stem cells (BMSCs) were seeded into gelatin sponge and chondrogenically cultured for 4 weeks in vitro to form cartilage templates. The constructs were then implanted subcutaneously in monocyte-depleted mice or normal C57BL/6 mice. Implants harvested at two months showed inferior osteogenic quality in monocyte-depleted mice compared with that of normal mice. In normal mice, the cartilage templates recruited a high ratio of alternatively activated macrophages (CAM or M2) to classically activated macrophages (AAM or M1), compared with empty sponge. In vitro co-culture assay of macrophages with cartilage templates also showed that the cartilage templates polarized macrophages to the M2 phenotype and that these effects were even stronger than those of primary BMSCs. In turn, the co-culture of polarized macrophages with cartilage templates showed that compared to M0 or M2, M1 significantly increased the expressions of osteogenic and angiogenic markers of cartilage templates. These data suggested that macrophages seem to be indispensable in the osteogenesis of cartilage templates and that cartilage templates have a favorable immunomodulatory ability to polarize macrophages to the M2 phenotype. M1 was the contributing phenotype of macrophages that promoted the osteogenesis and angiogenesis of cartilage templates. Macrophages and cartilage templates cooperate to achieve endochondral bone formation.

Bone regeneration in critically sized rat mandible defects through the endochondral pathway using hydroxyapatite-coated 3D-printed Ti6Al4V scaffolds.

The endochondral approach has been proved to be a promising pathway in bone tissue engineering. However, whether it is suitable for repairing critically sized mandible defects is unknown. We designed Ti6Al4V scaffolds with a suitable shape and pore size by a 3D-printing selective-laser-melting technique to implement this approach. In order to improve the surface bioactivity of the scaffolds, hydroxyapatite (HA) coatings (HA/L group and HA/H group) of different crystallite size were prepared on the scaffolds via electrochemical deposition. Rat bone mesenchymal stem cells (BMSCs) were seeded onto the scaffolds and chondrogenically differentiated in vitro for 4 weeks and then the scaffolds were implanted into critically sized rat mandible defects for 8 weeks. The bare scaffold and HA coatings were characterized with field emission scanning electron microscopy, water contact angle measurements and X-ray diffractometry. Cell proliferation results showed that the bioactivity of the HA coatings could better improve the growth rate of BMSCs compared with the bare surface. Additionally, safranin O staining showed abundant cartilage matrix and chondrocytes in the HA coated scaffold. Analyses using qPCR detected higher expression of chondrogenic-related gene Col2α1 and vegfα in the HA coated groups, especially in the HA/H group. Together these data demonstrate that the HA coating could improve the chondrogenic differentiation of BMSCs. In vivo, methylene blue staining of histological sections and micro-computed tomography revealed that the HA-coated groups, especially the HA/H group, increased new bone formation via endochondral ossification compared with the control group. Therefore, this strategy provides an alternative method to improve bone formation in mandible defects via the endochondral pathway and the scaffold with larger HA crystals was superior to those with smaller HA crystals.

Evaluation of antibacterial, angiogenic, and osteogenic activities of green synthesized gap-bridging copper-doped nanocomposite coatings.

Titanium (Ti) and its alloys have been widely used in clinics for years. However, their bio-inert surface challenges application in patients with compromised surgical conditions. Numerous studies were conducted to modify the surface topography and chemical composition of Ti substrates, for the purpose of obtaining antibacterial, angiogenic, and osteogenic activities. In this study, using green electrophoretic deposition method, we fabricated gap-bridging chitosan-gelatin (CSG) nanocomposite coatings incorporated with different amounts of copper (Cu; 0.01, 0.1, 1, and 10 mM for Cu I, II, III, and IV groups, respectively) on the Ti substrates. Physicochemical characterization of these coatings confirmed that Cu ions were successfully deposited into the coatings in a metallic status. After rehydration, the coatings swelled by 850% in weight. Mechanical tests verified the excellent tensile bond strength between Ti substrates and deposited coatings. All Cu-containing CSG coatings showed antibacterial property against both Gram-negative Escherichia coli and Gram-positive Staphylococcus aureus. The antibacterial property was positively correlated with the Cu concentration. In vitro cytocompatibility evaluation demonstrated that activities of bone marrow stromal cells were not impaired on Cu-doped coatings except for the Cu IV group. Moreover, enhanced angiogenic and osteogenic activities were observed on Cu II and Cu III groups. Overall, our results suggested that Cu-doped CSG nanocomposite coating is a promising candidate to functionalize Ti materials with antibacterial, angiogenic, and osteogenic properties.