A Mussel-Inspired Persistent ROS-Scavenging, Electroactive, and Osteoinductive Scaffold Based on Electrochemical-Driven In Situ Nanoassembly.

Conductive polymers are promising for bone regeneration because they can regulate cell behavior through electrical stimulation; moreover, they are antioxidative agents that can be used to protect cells and tissues from damage originating from reactive oxygen species (ROS). However, conductive polymers lack affinity to cells and osteoinductivity, which limits their application in tissue engineering. Herein, an electroactive, cell affinitive, persistent ROS-scavenging, and osteoinductive porous Ti scaffold is prepared by the on-surface in situ assembly of a polypyrrole-polydopamine-hydroxyapatite (PPy-PDA-HA) film through a layer-by-layer pulse electrodeposition (LBL-PED) method. During LBL-PED, the PPy-PDA nanoparticles (NPs) and HA NPs are in situ synthesized and uniformly coated on a porous scaffold from inside to outside. PDA is entangled with and doped into PPy to enhance the ROS scavenging rate of the scaffold and realize repeatable, efficient ROS scavenging over a long period of time. HA and electrical stimulation synergistically promote osteogenic cell differentiation on PPy-PDA-HA films. Ultimately, the PPy-PDA-HA porous scaffold provides excellent bone regeneration through the synergistic effects of electroactivity, cell affinity, and antioxidative activity of the PPy-PDA NPs and the osteoinductivity of HA NPs. This study provides a new strategy for functionalizing porous scaffolds that show great promise as implants for tissue regeneration.

A Mussel-Inspired Conductive, Self-Adhesive, and Self-Healable Tough Hydrogel as Cell Stimulators and Implantable Bioelectronics.

A graphene oxide conductive hydrogel is reported that simultaneously possesses high toughness, self-healability, and self-adhesiveness. Inspired by the adhesion behaviors of mussels, our conductive hydrogel shows self-adhesiveness on various surfaces and soft tissues. The hydrogel can be used as self-adhesive bioelectronics, such as electrical stimulators to regulate cell activity and implantable electrodes for recording in vivo signals.

Microparticle entrapment for drug release from porous-surfaced bone implants.

Metallic bone implants face interfacial concerns, such as infection and insufficient bone formation. Combination of drug-loaded microparticles with the implant surface is a promising approach to reducing the concerns. The present study reports a simple method for this purpose. Drug-loaded chitosan and alginate microparticles were separately prepared by emulsion methods. Dry microparticles were introduced into porous titanium (Ti) coatings on Ti discs, and induced to agglomerate in pores by wetting with water. Agglomerates were stably entrapped in the pores: 77-82% retained in the coating after immersion in a water bath for 7 d. Discs carrying drug-loaded microparticles showed a rapid release within 6 h and a subsequent slow release up to 1 d. After coculture with Staphylococcus epidermidis for 24 h, the discs formed inhibition zones, confirming antibacterial properties. These suggest that the microparticle entrapment-based method is a promising method for reducing some of the bone-implant interfacial concerns.

Chitosan/bovine serum albumin co-micropatterns on functionalized titanium surfaces and their effects on osteoblasts.

Chitosan (CS)/bovine serum albumin (BSA) micropatterns were prepared on functionalized Ti surfaces by micro-transfer molding (µ-TM). µ-TM realized the spatially controlled immobilization of cells and offered a new way of studying the interaction between micropatterns and cells. Two kinds of micropatterns were produced: (1) microgrooves representing a discontinuously grooved co-micropattern, with the rectangular CS region separated by BSA walls; (2) microcylinders representing a continuously interconnected co-micropattern, with the net-like CS region separated by BSA cylinders. A comparison of cell behaviors on the two types of micropatterns indicated that the shape rather than the size had a dominant effect on cell proliferation. The micropattern size in the same range of cell diameters favored cell proliferation. However, cell differentiation was more sensitive to the size rather than to the shape of the micropatterns. In conclusion, cell behavior can be regulated by micropatterns integrating different materials.

The W9 peptide inhibits osteoclastogenesis and osteoclast activity by downregulating osteoclast autophagy and promoting osteoclast apoptosis.

The W9 peptide has been shown to act as a receptor activator for nuclear factor-κB ligand (RANKL) antagonist and tumor necrosis factor (TNF)-α antagonist, which can promote bone formation and inhibit bone resorption. Studies on the W9 peptide at the cellular level have mainly focused on osteoblasts, and little research on the mechanism by which the W9 peptide regulates osteoclasts has been reported, which was the aim of this work. In this study, a rat mandibular defect model was established in vivo and implanted with hydrogel containing the W9 peptide for 2 weeks and 4 weeks, and histochemical staining was used to evaluate the formation of new bone and the changes in osteoclasts. RAW264.7 cells were cultured in vitro for osteoclast induction, and different concentrations of W9 peptide were added. Tartrate resistant acid phosphatase staining, monodansylcadaverine staining, TdT-mediated dUTP Nick-End Labeling assay, real-time PCR and Western blot were used to detect osteoclast differentiation, autophagy and apoptosis. Our results showed that the W9 peptide could reduce osteoclastogenesis and osteoclast activity induced by RANKL, and these effects were partly due to the inhibition of osteoclast autophagy. On the other hand, the W9 peptide could promote mature osteoclast apoptosis, in which autophagy might play an antagonistic role. Taken together, these results suggest that the W9 peptide inhibits osteoclastogenesis and osteoclast activity by downregulating osteoclast autophagy and promoting osteoclast apoptosis. Our results will benefit the development and application of new small molecule peptides for the treatment of bone resorption diseases.

Sponge-Like Macroporous Hydrogel with Antibacterial and ROS Scavenging Capabilities for Diabetic Wound Regeneration.

Hydrogels with soft and wet properties have been intensively investigated for chronic disease tissue repair. Nevertheless, tissue engineering hydrogels containing high water content are often simultaneously suffered from low porous size and low water-resistant capacities, leading to undesirable surgery outcomes. Here, a novel sponge-like macro-porous hydrogel (SM-hydrogel) with stable macro-porous structures and anti-swelling performances is developed via a facile, fast yet robust approach induced by Ti3 C2 MXene additives. The MXene-induced SM-hydrogels (80% water content) with 200-300 µm open macropores, demonstrating ideal mass/nutrient infiltration capability at ≈20-fold higher water/blood-transport velocity over that of the nonporous hydrogels. Moreover, the highly strong interactions between MXene and polymer chains endow the SM-hydrogels with excellent anti-swelling capability, promising equilibrium SM-hydrogels with identical macro-porous structures and toughened mechanical performances. The SM-hydrogel with versatile functions such as facilitating mass transport, antibacterial (bacterial viability in (Acrylic acid-co-Methacrylamide dopamine) copolymer-Ti3 C2 MXene below 25%), and reactive oxygen species scavenging capacities (96% scavenging ratio at 120 min) synergistically promotes diabetic wound healing (compared with non-porous hydrogels the wound closure rate increased from 39% to 81% within 7 days). Therefore, the durable SM-hydrogels exhibit connective macro-porous structures and bears versatile functions induced by MXene, demonstrating its great potential for wound tissue engineering.

Preparation and characterization of a novel porous titanium scaffold with 3D hierarchical porous structures.

This study aims to prepare a novel porous titanium (Ti) scaffold in order to improve the biocompatibility of the metallic implants. Porous Ti was produced by a Liquid foaming method and subsequent chemical treatments. It was found that the scaffold had three-dimensionally hierarchical porous structures with pore size ranging from nanometer to micrometer scale, and it also had activated surface. Mechanical test results showed that the scaffold also has sufficient compressive strength to meet the requirements of implantation. Protein adsorption results indicated that the novel scaffolds significantly enhanced the protein adsorption.

Self-adhesive hydrogels for tissue engineering.

Hydrogels consisting of a three-dimensional hydrophilic network of biocompatible polymers have been widely used in tissue engineering. Owing to their tunable mechanical properties, hydrogels have been applied in both hard and soft tissues. However, most hydrogels lack self-adhesive properties that enable integration with surrounding tissues, which may result in suture or low repair efficacy. Self-adhesive hydrogels (SAHs), an emerging class of hydrogels based on a combination of three-dimensional hydrophilic networks and self-adhesive properties, continue to garner increased attention in recent years. SAHs exhibit reliable and suitable adherence to tissues, and easily integrate into tissues to promote repair efficiency. SAHs are designed either by mimicking the adhesion mechanism of natural organisms, such as mussels and sandcastle worms, or by using supramolecular strategies. This review summarizes the design and processing strategies of SAHs, clarifies underlying adhesive mechanisms, and discusses their applications in tissue engineering, as well as future challenges.

The Synergy of Topographical Micropatterning and Ta|TaCu Bilayered Thin Film on Titanium Implants Enables Dual-Functions of Enhanced Osteogenesis and Anti-Infection.

Poor osteogenesis and implant-associated infection are the two leading causes of failure for dental and orthopedic implants. Surface design with enhanced osteogenesis often fails in antibacterial activity, or vice versa. Herein, a surface design strategy, which overcomes this trade-off via the synergistic effects of topographical micropatterning and a bilayered nanostructured metallic thin film is presented. A specific microgrooved pattern is fabricated on the titanium surface, followed by sequential deposition of a nanostructured copper (Cu)-containing tantalum (Ta) (TaCu) layer and a pure Ta cap layer. The microgrooved patterns coupled with the nanorough Ta cap layer shows strong contact guidance to preosteoblasts and significantly enhances the osteogenic differentiation in vitro, while the controlled local sustained release of Cu ions is responsible for high antibacterial activity. Importantly, rat calvarial defect models in vivo further confirm that the synergy of microgrooved patterns and the Ta|TaCu bilayered thin film on titanium surface could effectively promote bone regeneration. The present effective and versatile surface design strategy provides significant insight into intelligent surface engineering that can control biological response at the site of healing in dental and orthopedic implants.

The characteristics of mussel-inspired nHA/OSA injectable hydrogel and repaired bone defect in rabbit.

With the rapid development of minimally invasive techniques in orthopedics, minimally invasive surgery combined with injection of bone repair materials has attracted increasing attention for the treatment of bone defects. Inspired by material in mussels, we decorated nanohydroxyapatite (nHA) with dopamine (DA) to form polydopamine (PDA)-decorated nHA (PHA). Then, we introduced PHA into a Schiff base reaction of oxidized sodium alginate (OSA) and gelatin (Gel) to prepare an injectable bone repair hydrogel under mild conditions. Subsequently, the injectability, morphology, mechanical, swelling, and degradation properties of the hydrogel were studied. Then, bone marrow mesenchymal stem cells (BMSCs) were cocultured with the hydrogels to investigate the cytotoxicity, proliferation, and osteogenic differentiation properties of the hydrogel. Finally, the bone repair ability of the OSA-Gel-PHA hydrogels was explored using a 12 weeks rabbit bone defect model. The results of tube inversion and rheological test showed that the hydrogel gelation time was 3-7 min, which will be suitable for clinical operation. The results of SEM, compression tests, rheological tests, swelling and degradation properties tests showed that the addition of PHA changed the microstructure of the hydrogels from porous structure to layered structure, not only improved the compressive strength, toughness, elastic modulus, storage modulus of the hydrogels, but also reduced the swelling properties and degradation rate of the hydrogels, making it more conducive to clinical operations. The results of laser confocal, MTT assay, alkaline phosphatase (ALP) assay and in vivo animal experiments showed that the addition of PHA not only nontoxic, but also promoted the adhesion, proliferation and osteogenic differentiation of BMSCs, as well as the repair and maturation of bone. The application of injectable bone repair materials not only solves the problem of bone defect filling, but also avoids a series of complications caused by open surgery, providing new method for the treatment of bone defects.

[Preparation of porous Ti metal composite scaffold with bioactivity].

In this work, the titanium powder was used for preparing highly interconnected porous scaffolds by impregnating polymer method. Subsequently, the electrochemical method and the biomimetic mineralization method were adopted to deposit calcium phosphate coatings on the sintered scaffold which was supposed to improve the scaffold bioactivity. The experimental results show, with the use of the two methods, the scaffolds are successfully covered by the deposition of the nano-net structure calcium phosphate coating, and they hold their three dimensional interconnected porous structures. Therefore, this kind of bioactive composite scaffold with such mechanical strength as that of woven bone should be a promising bone graft in clinical applications.

Mussel-inspired hybrid coating functionalized porous hydroxyapatite scaffolds for bone tissue regeneration.

The scaffold for bone tissue engineering should possess proper porosity, adequate mechanical properties, cell affinity for cell attachment, and the capability to bind bioactive agents to induce cell differentiation. In this study, we successfully prepared a porous hydroxyapatite (HA) scaffold that is functionalized by poly(L-lysine)/polydopamine (PLL/PDA) hybrid coating. The PLL/PDA coating takes advantages of the high protein and cell affinity of PDA, as well as the biodegradability of PLL. Therefore, the coating can anchor bone morphogenic protein-2 (BMP2) to the HA scaffold via catechol chemistry under a mild condition so as to protect the bioactivity of BMP2. Meanwhile, the coating can also release BMP2 in a tunable and sustainable manner as the PLL degrades in the physiological environment. The BMP2-entrapped PLL/PDA coating on the HA scaffold can more efficiently promote osteogenic differentiation of bone marrow stromal cells (BMSCs) in vitro and induce ectopic bone formation to a much greater level in vivo compared with a bare HA scaffold that delivers BMP2 in a burst manner. All of these results suggest that the PDA-mediated catechol modification of the HA scaffold can be an effective strategy to develop sustainable protein delivery system, and that the PLL/PDA-coated HA scaffold could be a promising candidate for bone tissue engineering applications.

A strong, tough, and osteoconductive hydroxyapatite mineralized polyacrylamide/dextran hydrogel for bone tissue regeneration.

The design of hydrogels with adequate mechanical properties and excellent bioactivity, osteoconductivity, and capacity for osseointegration is essential to bone repair and regeneration. However, it is challenging to integrate all these properties into one bone scaffold. Herein, we developed a strong, tough, osteoconductive hydrogel by a facile one-step micellar copolymerization of acrylamide and urethacrylate dextran (Dex-U), followed by the in situ mineralization of hydroxyapatite (HAp) nanocrystals. We show that the soft, flexible, and hydrophobically associated polyacrylamide (PAAm) network is strengthened by the stiff crosslinked Dex-U phase, and that the mineralized HAp simultaneously improves the mechanical properties and osteoconductivity. The obtained HAp mineralized PAAm/Dex-U hydrogel (HAp-PADH) has extremely high compressive strength (6.5 MPa) and enhanced fracture resistance (over 2300 J m-2), as compared with pure PAAm hydrogels. In vitro, we show that the mineralized HAp layer promotes the adhesion and proliferation of osteoblasts, and effectively stimulates osteogenic differentiation. Through the in vivo evaluation of hydrogels in a femoral condyle defect rabbit model, we show regeneration of a highly mineralized bone tissue and direct bonding to the HAp-PADH interface. These findings confirm the excellent osteoconductivity and osseointegration ability of fabricated HAp-PADH. The present HAp-PADH, with its superior mechanical properties and excellent osteoconductivity, should have great potential for bone repair and regeneration. STATEMENT OF SIGNIFICANCE: We developed a strong, tough, and osteoconductive hydrogel by a facile one-step micellar copolymerization of acrylamide and urethane methacrylate dextran (Dex-U), followed by the in situ mineralization of hydroxyapatite (HAp) nanocrystals. The hydrophobic micellar copolymerization and introduction of the stiff crosslinked Dex-U phase endowed the soft polyacrylamide (PAAm) network with enhanced strength and toughness. The in situ mineralized HAp nanocrystals on the hydrogels further improved the mechanical properties of the hydrogels and promoted osteogenic differentiation of cells. Mechanical tests together with in vitro and in vivo evaluations confirmed that the HAp mineralized PAAm/Dex-U hydrogel (HAp-PADH) achieved a combination of superior mechanical properties and excellent osseointegration, and thus may offer a promising candidate for bone repair and regeneration.

Engineering High-Resolution Micropatterns Directly onto Titanium with Optimized Contact Guidance to Promote Osteogenic Differentiation and Bone Regeneration.

Topographical cues play an important role in directing cell behavior, and thus, extensive research efforts have been devoted to fabrication of surface patterns and exploring the contact guidance effect. However, engineering high-resolution micropatterns directly onto metallic implants remains a grand challenge. Moreover, there still lacks evidence that allows translation of in vitro screening to in vivo tissue response. Herein, we demonstrate a fast, cost-effective, and feasible approach to the precise fabrication of shape- and size-controlled micropatterns on titanium substrates using a combination of photolithography and inductively coupled plasma-based dry etching. A titanium TopoChip containing 34 microgrooved patterns with varying geometry parameters and a flat surface as the control was designed for a high-throughput in vitro study of the contact guidance of osteoblasts. The correlation between the surface pattern dimensions, cell morphological characteristics, proliferation, and osteogenic marker expression was systematically investigated in vitro. Furthermore, the surface with the highest osteogenic potential in vitro along with representative controls was evaluated in rat cranial defect models. The results show that microgrooved pattern parameters have almost no effect on osteoblast proliferation but significantly regulate the cell morphology, orientation, focal adhesion (FA) formation, and osteogenic differentiation in vitro. In particular, a specific groove pattern with a ridge width of 3 µm, groove width of 7 µm, and depth of 2 µm can most effectively align the cells through regulating the distribution of FAs, resulting in an anisotropic actin cytoskeleton, and thereby promoting osteogenic differentiation. In vivo, microcomputed tomography and histological analyses show that the optimized pattern can apparently stimulate new bone formation. This study not only offers a microfabrication method that can be extended to fabricate various shape- and size-controlled micropatterns on titanium alloys but also provides insight into the surface structure design of orthopedic and dental implants for enhanced bone regeneration.

Mussel-inspired cryogels for promoting wound regeneration through photobiostimulation, modulating inflammatory responses and suppressing bacterial invasion.

Wound healing is a complex and dynamic process, and involves a series of events, which create a unique microenvironment at the wound sites. It is highly desirable to develop multi-functional skin substitutes which can play their roles in the whole healing processes to enhance the final healing efficiency. Herein, we fabricated a mussel-inspired chitosan/silk fibroin cryogel functionalized with near-infrared light-responsive polydopamine nanoparticles (PDA-NPs), as a multifunctional platform to regulate the wound microenvironment and enhance efficient wound healing. The cryogel has an extracellular matrix-like macroporous structure, mimicking the natural tissue environment, which allows cell attachment and tissue ingrowth. The cryogel shows high anti-oxidative activity to eliminate overproduced reactive oxygen species during inflammatory responses. Furthermore, the cryogel exhibits photothermally assisted antibacterial activity to prevent bacterial invasion. Thus, by combining the photobiostimulation of infrared light, the cryogel realizes bio-chemo-photothermal synergistic therapy for accelerating the complete skin-thickness wound healing by simultaneously suppressing adverse events due to its antibacterial activity and anti-oxidative ability, and promoting cell activities and tissue regeneration. Our work therefore presents the great promise shown by this multifunctional biopolymer cryogel as a flexible wound dressing with combinatory therapy for accelerating wound healing.

Plant-inspired adhesive and tough hydrogel based on Ag-Lignin nanoparticles-triggered dynamic redox catechol chemistry.

Adhesive hydrogels have gained popularity in biomedical applications, however, traditional adhesive hydrogels often exhibit short-term adhesiveness, poor mechanical properties and lack of antibacterial ability. Here, a plant-inspired adhesive hydrogel has been developed based on Ag-Lignin nanoparticles (NPs)triggered dynamic redox catechol chemistry. Ag-Lignin NPs construct the dynamic catechol redox system, which creates long-lasting reductive-oxidative environment inner hydrogel networks. This redox system, generating catechol groups continuously, endows the hydrogel with long-term and repeatable adhesiveness. Furthermore, Ag-Lignin NPs generate free radicals and trigger self-gelation of the hydrogel under ambient environment. This hydrogel presents high toughness for the existence of covalent and non-covalent interaction in the hydrogel networks. The hydrogel also possesses good cell affinity and high antibacterial activity due to the catechol groups and bactericidal ability of Ag-Lignin NPs. This study proposes a strategy to design tough and adhesive hydrogels based on dynamic plant catechol chemistry.

Mussel-Inspired Electroactive and Antioxidative Scaffolds with Incorporation of Polydopamine-Reduced Graphene Oxide for Enhancing Skin Wound Healing.

Wound repair and tissue regeneration are complex processes that involve many physiological signals. Thus, employing novel wound dressings with potent biological activity and physiological signal response ability to accelerate wound healing is a possible solution. Herein, inspired by mussel chemistry, we developed a polydopamine (PDA)-reduced graphene oxide (pGO)-incorporated chitosan (CS) and silk fibroin (SF) (pGO-CS/SF) scaffold with good mechanical, electroactive, and antioxidative properties as an efficient wound dressing. First, pGO with good dispersibility and cell affinity was obtained upon reduction by PDA under alkali conditions. Second, pGO was dispersed into a CS/SF mixture, and then CS and SF chains were dual-cross-linked by poly(ethylene glycol) diglycidyl ether and glutaraldehyde to obtain a pGO-incorporated gel. Finally, the gel underwent a freeze-dry process to obtain the pGO-CS/SF scaffold. Owing to PDA reduction and functionalization, pGO in the scaffold plays important roles for the performances of the scaffolds. First, the pGO acts as nanoreinforcement to enhance the mechanical properties of the scaffold by combining the dual-cross-linked CS/SF network. Second, the uniformly distributed pGO in the scaffolds comprises a well-connected electric pathway, which can provide a channel for the transmission of electrical signals in the scaffold. Moreover, pGO in the scaffolds serves as an antioxidant agent to scavenge reactive oxygen species (ROS) and therefore terminates excessive ROS oxidation. In vitro studies show that electroactive pGO-CS/SF scaffolds can respond to electrical signals and promote cytological behavior. In addition, the pGO-CS/SF scaffolds can reduce cellular oxidation by removing excessive ROS. The in vivo full-thickness skin defect model demonstrates that the electroactive and antioxidative pGO-CS/SF scaffold can efficiently enhance wound healing. In summary, the pGO-CS/SF scaffold is a promising wound dressing because of its ability to promote physiological electrical signal transmission for cell growth and reduce ROS oxidation, resulting in an improved wound regeneration effect.

An Injectable, Bifunctional Hydrogel with Photothermal Effects for Tumor Therapy and Bone Regeneration.

Significant attention has been focused on bone tumor therapy recently. At present, the treatment in clinic typically requires surgical intervention. However, a few tumor cells remain around bone defects after surgery and subsequently proliferate within several days. Thus, fabrication of biomaterials with dual functions of tumor therapy and bone regeneration is significant. Herein, the injectable hydrogel containing cisplatin (DDP) and polydopamine-decorated nano-hydroxyapatite is prepared via Schiff base reaction between the aldehyde groups on oxidized sodium alginate and amino groups on chitosan. The hydrogel exhibits sustained release properties for DDP due to the immobilization of DDP via abundant functional groups on polydopamine (PDA). Additionally, given the intense absorption of PDA in the near-infrared region, the hydrogel exhibits excellent photothermal effects when exposed to the NIR laser (808 nm). Based on the properties, the hydrogel effectively ablates tumor cells (4T1 cells) in vitro and suppresses tumor growth in vivo. Furthermore, the hydrogel promotes the adhesion and proliferation of bone mesenchymal stem cells in vitro due to the abundant functional groups on PDA and further induces bone regeneration in vivo. Therefore, the study extends research on novel biomaterials with dual functions of tumor therapy and bone regeneration.

Spectroscopic analysis of titanium surface functional groups under various surface modification and their behaviors in vitro and in vivo.

In the present study, surface functional groups of titanium surfaces gone through different treatments, including acid etched treatment (AE), nitric acid treatment (NT), heat treatment (HT), and alkali treatment (AT), and their behaviors in vitro and in vivo was thoroughly studied by spectroscopic analysis. In vitro and in vivo results revealed that the rank of bioactivity of various surfaces was AE < NT < HT < AT. XPS analysis indicated that AT greatly increased the OH group concentration on the titanium surface whereas HT reduced the OH group concentration. Thus, OH group difference could not be a good explanation of bioactivity difference. On the other hand, ToF-SIMS analysis demonstrated the TiOH+/Ti+ ratios of various surfaces correlated well with the bioactivity and the surface energies, which implied that Ti-OH could play an important role in the bioactivity. This detail investigation of the relationship between surface functional groups and surface bioactivity could help us to broaden the knowledge about the mechanism of bioactivity and to design next generation bioactive materials.

Controllable release of interleukin-4 in double-layer sol-gel coatings on TiO2 nanotubes for modulating macrophage polarization.

Classically activated M1 macrophages and alternatively activated M2 macrophages play key roles in regulating immune responses. M1 macrophages initiate angiogenesis in the early stages of wound healing or after implantation. However, their prolonged activation can lead to chronic inflammation. We speculated that biomedical implants with specific properties can induce a shift from M1 to M2 macrophages at a specific time point to promote tissue repair and wound healing. To investigate this possibility, drug-loaded double-layer sol-gel coatings were fabricated on TiO2 nanotubes (TNTs), which were used to modulate the switch from the M1 to the M2 phenotype by controlled release of interleukin (IL)-4. The lower sol-gel layer with IL-4 consisted of a carboxymethyl chitosan (CMCS) hydrogel, 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, and N-hydroxysuccinimide as a crosslinker (IL4/TNT). The upper layer fabricated on the IL4/TNT sample was another type of CMCS hydrogel that used genipin (GP) as a crosslinker (GP/IL4/TNT). We found that IL-4 was released from GP/IL4/TNTs in a controlled manner, with the greatest release occurring after 72 h. GP/IL4/TNT stimulated the polarization of macrophages from the M1 to M2 phenotype after the macrophage polarization from the M0 to M1 phenotype. This provides a template for the fabrication of biomaterials that can direct macrophage polarization and stimulate tissue regeneration following the initial inflammatory response to implants.