Proanthocyanidins-based tandem dynamic covalent cross-linking hydrogel for diabetic wound healing.

Wound healing in diabetic patients presents significant challenges in clinical wound care due to high oxidative stress, excessive inflammation, and a microenvironment prone to infection. In this study, we successfully developed a multifunctional tandem dynamic covalently cross-linked hydrogel dressing aimed at diabetic wound healing. This hydrogel was constructed using cyanoacetic acid functionalized dextran (Dex-CA), 2-formylbenzoylboric acid (2-FPBA) and natural oligomeric proanthocyanidins (OPC), catalyzed by histidine. The resulting Dex-CA/OPC/2-FPBA (DPOPC) hydrogel can be dissolved triggered by cysteine, thereby achieving "controllable and non-irritating" dressing change. Furthermore, the incorporation of OPC as a hydrogel building block endowed the hydrogel with antioxidant and anti-inflammatory properties. The cross-linked network of the DPOPC hydrogel circumvents the burst release of OPC, enhancing its biosafety. In vivo studies demonstrated that the DPOPC hydrogel significantly accelerated the wound healing process in diabetic mice compared to a commercial hydrogel, achieving an impressive wound closure rate of 98 % by day 14. The DPOPC hydrogel effectively balanced the disrupted inflammatory state during the healing process. This dynamic hydrogel based on natural polyphenols is expected to be an ideal candidate for dressings intended for chronic wounds.

Response and self-regulation of PD/A granular sludge to oxytetracycline stress.

This paper investigated the operational characteristics and self-regulation mechanism of the partial denitrification/anammox (PD/A) granular system under the stress of oxytetracycline (OTC), an emerging pollutant that accumulates in municipal wastewater treatment plants through various pathways, posing significant challenges for its future promotion in engineering applications. The results indicated that OTC concentrations below 100 mg/L intensified its short-term inhibition on the PD/A granular sludge system, decreasing functional bacterial activity, while between 150 and 300 mg/L, PD's NO3--N to NO2--N conversion ability diminished, and Anammox activity was significantly suppressed. Under long-term high OTC stress (20-30 mg/L), nitrogen removal suffered, and batch tests revealed significant inhibition of PD's NO3--N to NO2--N conversion, dropping from 73.77 % to 50.17 %. Anammox bacteria activity sharply declined from 1.81 to 0.39 mg N/gVSS/h under OTC stress. Extracellular polymeric substances (EPS) content rose from 185.39 to 210.86 mg/gVSS, indicating PD/A sludge's self-protection mechanism. However, EPS content fell due to cell lysis at high OTC (30 mg/L). The decreasing relative abundance of Candidatus_Brocadia (2.32 % to 0.93 %) and Thaure (12.63 % to 7.82 %) was a key factor in the gradual deterioration of denitrification performance. This study was expected to provide guidance for the PD/A process to cope with the interference of antibiotics and other emerging pollutants (short-term shock and long-term stress).

Novel and innovative approaches to partial denitrification coupled with anammox: A critical review.

The partial denitrification (PD) coupled with anaerobic ammonium oxidation (Anammox) (PD/A) process is a unique biological denitrification method for sewage that concurrently removes nitrate (NO3--N) and ammonium (NH4+-N) in sewage. Comparing PD/A to conventional nitrification and denitrification technologies, noticeable improvements are shown in energy consumption, carbon source demand, sludge generation and emissions of greenhouse gasses. The PD is vital to obtaining nitrites (NO2--N) in the Anammox process. This paper provided valuable insight by introduced the basic principles and characteristics of the process and then summarized the strengthening strategies. The functional microorganisms and microbial competition have been discussed in details, the S-dependent denitrification-anammox has been analyzed in this review paper. Important factors affecting the PD/A process were examined from different aspects, and finally, the paper pointed out the shortcomings of the coupling process in experimental research and engineering applications. Thus, this research provided insightful information for the PD/A process's optimization technique in later treating many types of real and nitrate-based wastewater. The review paper also provided the prospective economic and environmental position for the actual design implementation of the PD/A process in the years to come.

Biomimicking covalent organic frameworks nanocomposite coating for integrated enhanced anticorrosion and antifouling properties of a biodegradable magnesium stent.

The utilization of biodegradable magnesium (Mg) alloys in the fabrication of temporary non-vascular stents is an innovative trend in biomedical engineering. However, the heterogeneous degradation profiles of these biomaterials, together with potential bacterial colonization that could precipitate infectious or stenotic complications, are critical obstacles precluding their widespread clinical application. In pursuit of overcoming these limitations, this study applies the principles of biomimicry, particularly the hydrophobic and anti-fouling characteristics of lotus leaves, to pioneer the creation of nanocomposite coatings. These coatings integrate poly-trimethylene carbonate (PTMC) with covalent organic frameworks (COFs), to modify the stent's surface property. The strategic design of the coating's topography, porosity, and self-polishing capabilities collectively aims to decelerate degradation processes and minimize biological adhesion. The protective qualities of the coatings were substantiated through rigorous testing in both in vitro dynamic bile tests and in vivo New Zealand rabbit choledochal models. Empirical findings from these trials confirmed that the implementation of COF-based nanocomposite coatings robustly fortifies Mg implantations, conferring heightened resistance to both biocorrosion and biofouling as well as improved biocompatibility within bodily environments. The outcomes of this research elucidate a comprehensive framework for the multifaceted strategies against stent corrosion and fouling, thereby charting a visionary pathway toward the systematic conception of a new class of reliable COF-derived surface modifications poised to amplify the efficacy of Mg-based stents. STATEMENT OF SIGNIFICANCE: Biodegradable magnesium (Mg) alloys are widely utilized in temporary stents, though their rapid degradation and susceptibility to bacterial infection pose significant challenges. Our research has developed a nanocomposite coating inspired by the lotus, integrating poly-trimethylene carbonate with covalent organic frameworks (COF). The coating achieved self-polishing property and optimal surface energy on the Mg substrate, which decelerates stent degradation and reduces biofilm formation. Comprehensive evaluations utilizing dynamic bile simulations and implantation in New Zealand rabbit choledochal models reveal that the coating improves the durability and longevity of the stent. The implications of these findings suggest the potential COF-based Mg alloy stent surface treatments and a leap forward in advancing stent performance and endurance in clinical applications.

Robust rehabilitation of anammox system by granular activated carbon under long-term starvation stress: Microbiota restoration and metabolic reinforcement.

This article investigates the buffering capacity and recovery-enhancing ability of granular activated carbon (GAC) in a starved (influent total nitrogen: 20 mg/L) anaerobic ammonium oxidation (anammox) reactor. The findings revealed that anammox aggregated and sustained basal metabolism with shorter performance recovery lag (6 days) and better nitrogen removal efficiency (84.9 %) due to weak electron-repulsion and abundance redox-active groups on GAC's surface. GAC-supported enhanced extracellular polymeric substance secretion aided anammox in resisting starvation. GAC also facilitated anammox bacterial proliferation and expedited the restoration of anammox microbial community from a starved state to its initial-level. Metabolic function analyses unveiled that GAC improved the expression of genes involved in amino acid metabolism and sugar-nucleotide biosynthesis while promoted microbial cross-feeding, ultimately indicating the superior potential of GAC in stimulating more diverse metabolic networks in nutrient-depleted anammox consortia. This research sheds light on the microbial and metabolic mechanisms underlying GAC-mediated anammox system in low-substrate habitats.

Guanosine-driven hyaluronic acid-based supramolecular hydrogels with peroxidase-like activity for chronic diabetic wound treatment.

Guanosine is often used to construct supramolecular hydrogels due to its self-assembly properties, however, the high temperature and strong alkaline construction methods greatly limit its application in biomedical fields. In this work, a guanosine-driven hyaluronic acid-based supramolecular hydrogel was developed under mild condition by employing phenylboronic acid-functionalized hyaluronic acid (HA-PBA) backbone and guanosine molecules. Guanosines self-assembled into G-quartet planes under potassium ion conditions, and formed boronic ester bonds with HA-PBA, which induced rapid formation of dynamically cross-linked hydrogels. Hemin was then binding to the G-quartet plane via π-π interactions in the hydrogels, which exhibited peroxidase activity and were highly effective in killing bacteria by generating hydroxyl radicals in the presence of H2O2. Furthermore, glucose oxidase (GOx) was incorporated into the hydrogels and the HP/G@hemin@GOx hydrogels showed good antibacterial properties, modulation of wound glucose and ROS level, and good therapeutic efficacy for diabetic chronic wounds. Overall, the self-assembly of guanosine has been shown for the first time to be a feasible method for constructing natural polymer-based supramolecular hydrogels. This guanosine-driven HA-based supramolecular hydrogel can act as a potential wound dressing for chronic diabetic wound treatment. STATEMENT OF SIGNIFICANCE: Chronic wound repair remains an unsolved clinical challenge. Herein, we propose to utilize phenylboronic acid-modified hyaluronic acid and guanosine to construct supramolecular gels with peroxidase activity for chronic wound treatment. The self-assembly behavior of guanosine drives the natural macromolecular backbone to form the hydrogel, and the proposed method simplifies the gelation conditions and improves its biosafety. The G-quartets formed by the self-assembly of guanosine can act as the loading site for hemin. G-quartet/hemin complex imported peroxidase activity to the hydrogels, endowing them with the ability to kill bacteria and regulate ROS levels of cells in the wound site. This guanosine-driven supramolecular hydrogel significantly increased the rate of wound healing in diabetic mice, promising a new strategy for chronic wound treatment.

ROS-Responsive Self-Degradable DNA Nanogels for Targeted Anticancer Drug Delivery.

Here, a reactive oxygen species (ROS)-responsive targeted anticancer drug delivery system was developed by embedding a nitrophenyl tetramethyl-dioxaborolanyl benzyl carbamate (NBC)-modified deoxyribonuclease I (DNase I) in a DNase-degradable aptamer-based DNA nanogel. The DNA nanogel was formed by hybridization of three types of building blocks, namely, Y-shaped monomer 1 with three sticky ends, Y-shaped monomer 2 with two sticky ends and an aptamer end, and a DNA linker with two sticky ends. Single doxorubicin (DOX) or ribonuclease A (RNase A) as well as the combination of DOX and RNase A were effectively loaded into the nanogels, wherein DOX was embedded into DNA skeleton, while RNase A was encapsulated into nanogel matrix. The blocked enzymatic activity of DNase I due to NBC modification could be restored upon intracellular ROS-triggered NBC deprotection, resulting in self-degradation of the nanogels to release both DOX and RNase A. Consequently, the DOX and RNase A coloaded nanogels significantly inhibited the proliferation of MCF-7 cells through a synergistic effect. To sum up, this DNA-based drug delivery system with ROS-responsive self-degradation properties should be promising for application in targeted and synergistic cancer therapy.

Research on the Current Application Status of Magnesium Metal Stents in Human Luminal Cavities.

The human body comprises various tubular structures that have essential functions in different bodily systems. These structures are responsible for transporting food, liquids, waste, and other substances throughout the body. However, factors such as inflammation, tumors, stones, infections, or the accumulation of substances can lead to the narrowing or blockage of these tubular structures, which can impair the normal function of the corresponding organs or tissues. To address luminal obstructions, stenting is a commonly used treatment. However, to minimize complications associated with the long-term implantation of permanent stents, there is an increasing demand for biodegradable stents (BDS). Magnesium (Mg) metal is an exceptional choice for creating BDS due to its degradability, good mechanical properties, and biocompatibility. Currently, the Magmaris® coronary stents and UNITY-BTM biliary stent have obtained Conformité Européene (CE) certification. Moreover, there are several other types of stents undergoing research and development as well as clinical trials. In this review, we discuss the required degradation cycle and the specific properties (anti-inflammatory effect, antibacterial effect, etc.) of BDS in different lumen areas based on the biocompatibility and degradability of currently available magnesium-based scaffolds. We also offer potential insights into the future development of BDS.

Synthetic lipo-polylysine with anti-cancer activity.

Development of novel therapeutic agents that possess different anticancer mechanisms from the traditional antitumor drugs is highly attractive as no medication can cure all types of cancers. Herein, we report a rational design of antitumor lipo-polylysine polymers as synthetic mimics of biosynthetic lipopeptide surfactants featuring antimicrobial or cytotoxic activities for cancer therapy. The optimal polymer shows a wide range of anticancer activities against multiple cancer cells, including highly metastatic and drug-resistant ones, but low toxicity to normal cells. Mechanism studies show that the optimal polymer can interact with the membrane of cancer cells and induce cell necrosis by triggering cell membrane perforation, which is different from the therapeutic mechanisms of traditional anticancer drugs. In vivo studies imply that the optimal polymer efficiently inhibits tumor growth without causing obvious side effects on a C26 graft tumor model. Overall, the lipopeptide-mimicking lipo-polylysine with the advantages of easy synthesis and low cost provides a new anticancer strategy with high efficacy and biocompatibility.

Metagenomics insights into high-rate nitrogen removal from municipal wastewater by integrated nitrification, partial denitrification and Anammox at an extremely short hydraulic retention time.

To achieve high-rate nitrogen removal in municipal wastewater treatment through anaerobic ammonia oxidation (Anammox), the nitrification, partial denitrification, and Anammox processes were integrated by a step-feed strategy. An exceptional nitrogen removal load of 0.224 kg N/(m3·d) was achieved by gradient-reducing the hydraulic retention time (HRT) to 5 h. Metagenomic analysis demonstrated that Nitrosospira could express all genes encoding ammonia oxidation under low nitrogen and dissolved oxygen conditions (less than 0.5 mg/L), enabling complete nitrification. With the short of HRT, the relative abundance of Thauera increased from 2.8 % to 6.4 %. Frequent substrate exchanges at such extremely short HRT facilitated enhanced synergistic interactions among Nitrosospira, Thauera, and Candidatus Brocadia. These findings provide a comprehensive understanding of the utilization of Anammox combined processes for high-speed nitrogen removal in municipal wastewater treatment and the microbial interactions involved.

Efficient nitrogen removal from municipal wastewater by an autotrophic-heterotrophic coupled anammox system: The up-regulation of key functional genes.

The metabolic pathways based on key functional genes were innovatively revealed in the autotrophic-heterotrophic coupled anammox system for real municipal wastewater treatment. The nitrogen removal performance of the system was stabilized at 88.40 ± 3.39 % during the treatment of real municipal wastewater. The relative abundances of the nitrification functional genes ammonia oxidase (amoA/B/C), hydroxylamine oxidoreductase (hao), and nitrite oxidoreductases (nxrA/B) were increased by 1.2-2.4 times, and these three nitrification functional genes were mostly contributed by Nitrospira that dominated the efficient nitrification of the system. The relative abundance of anammox bacteria Candidatus Brocadia augmented from 0.35 % to 0.75 %, accompanied with the increased expression of hydrazine synthase (hzs) and hydrazine dehydrogenase (hdh), resulting in the major role of anammox (81.24 %) for nitrogen removal. The expression enhancement of the functional genes nitrite reductase (narG/H, napA/B) that promoted partial denitrification (PD) of the system weakened the adverse effects of the sharp decline in the population of PD microbe Thauera (from 5.7 % to 2.2 %). The metabolic module analysis indicated that the carbon metabolism pathways of the system mainly included CO2 fixation and organic carbon metabolism, and the stable enrichment of autotrophic bacteria ensured stable CO2 fixation. Furthermore, the enhanced expression of the glucokinases (glk, GCK, HK, ppgk) and the abundant pyruvate kinase (PK) achieved stable hydrolysis ability of organic carbon metabolism function of the system. This study offers research basics to practical application of the mainstream anammox process.

Microrod crystals formed via Rhein-mediated mineralization.

Herein, a Rhein-mineralized microrod crystal (H-RMM) with an ultra-high drug loading capacity was reported for anti-inflammation. Due to a dense crystal structure, the H-RMM achieved improved biocompatibility and sustained controlled release of Rhein. Also, the Rhein nanofibers released from H-RMM were favorable to be internalized by cells, leading to enhanced anti-inflammation effects.

A critical review of improving mainstream anammox systems: Based on macroscopic process regulation and microscopic enhancement mechanisms.

Full-scale anaerobic ammonium oxidation (anammox) engineering applications are vastly limited by the sensitivity of anammox bacteria to the complex mainstream ambience factors. Therefore, it is of great necessity to comprehensively summarize and overcome performance-related challenges in mainstream anammox process at the macro/micro level, including the macroscopic process variable regulation and microscopic biological metabolic enhancement. This article systematically reviewed the recent important advances in the enrichment and retention of anammox bacteria and main factors affecting metabolic regulation under mainstream conditions, and proposed key strategies for the related performance optimization. The characteristics and behavior mechanism of anammox consortia in response to mainstream environment were then discussed in details, and we revealed that the synergistic nitrogen metabolism of multi-functional bacterial genera based on anammox microbiome was conducive to mainstream anammox nitrogen removal processes. Finally, the critical outcomes of anammox extracellular electron transfer (EET) at the micro level were well presented, carbon-based conductive materials or exogenous electron shuttles can stimulate and mediate anammox EET in mainstream environments to optimize system performance from a micro perspective. Overall, this review advances the extensive implementation of mainstream anammox practice in future as well as shedding new light on the related EET and microbial mechanisms.

Highly efficient and robust treatment of low C/N actual domestic sewage via integrated fermentation, partial-nitrification, partial-denitrification and anammox (IFPNDA).

For achieving efficient and robust treatment of domestic sewage with C/N around 2.8, this study innovatively developed an integrated fermentation, partial-nitrification, partial-denitrification and anammox (IFPNDA) process based on the Anaerobic Baffled Reactor and Continuous-flow Stirred Tank Reactor (ABR-CSTR) bioreactor. Desirable N-removal efficiency of 87.5 ± 2.1% was obtained without external organics, correspondingly effluent total nitrogen (TN) concentration reached 6.1 ± 0.7 mg/L. The N-removal stability was greatly facilitated by the effective linkage between partial nitrification (PN) process and partial denitrification (PD) process in emergency. Highly enriched hydrolytic bacteria (6.9%) and acidogenic bacteria (5.7%) in A1, especially Comamonas (2.8%) and Longilinea (3.5%), induced the significant increase of volatile fatty acids (VFAs) in domestic sewage. Thauera (6.1%) in A2 and Nitrosomonas (5.4%) in A3 acted as the dominant flora of nitrite supplies for anammox in IFPNDA process. Candidatus_Brocadia (2.4%) dominated the advanced nitrogen removal. The IFPNDA process exhibited much potential for achieving energy neutrality during wastewater treatment.

Engineered hydrogels for peripheral nerve repair.

Peripheral nerve injury (PNI) is a complex disease that often appears in young adults. It is characterized by a high incidence, limited treatment options, and poor clinical outcomes. This disease not only causes dysfunction and psychological disorders in patients but also brings a heavy burden to the society. Currently, autologous nerve grafting is the gold standard in clinical treatment, but complications, such as the limited source of donor tissue and scar tissue formation, often further limit the therapeutic effect. Recently, a growing number of studies have used tissue-engineered materials to create a natural microenvironment similar to the nervous system and thus promote the regeneration of neural tissue and the recovery of impaired neural function with promising results. Hydrogels are often used as materials for the culture and differentiation of neurogenic cells due to their unique physical and chemical properties. Hydrogels can provide three-dimensional hydration networks that can be integrated into a variety of sizes and shapes to suit the morphology of neural tissues. In this review, we discuss the recent advances of engineered hydrogels for peripheral nerve repair and analyze the role of several different therapeutic strategies of hydrogels in PNI through the application characteristics of hydrogels in nerve tissue engineering (NTE). Furthermore, the prospects and challenges of the application of hydrogels in the treatment of PNI are also discussed.

AIEgen-Conjugated Phase-Separating Peptides Illuminate Intracellular RNA through Coacervation-Induced Emission.

Intrinsically disordered peptides drive dynamic liquid-liquid phase separation (LLPS) in membraneless organelles and encode cellular functions in response to environmental stimuli. Engineering design on phase-separating peptides (PSPs) holds great promise for bioimaging, vaccine delivery, and disease theranostics. However, recombinant PSPs are devoid of robust luminogen or suitable cell permeability required for intracellular applications. Here, we synthesize a peptide-based RNA sensor by covalently connecting tetraphenylethylene (TPE), an aggregation-induced emission luminogen (AIEgens), to tandem peptide repeats of (RRASL)n (n = 1, 2, 3). Interestingly, the conjugation of TPE luminogen promotes liquid-liquid phase separation of the peptide repeats, and the minimum coacervation concentration (MCC) of TPE-(RRASL)n is decreased by an order of magnitude, compared to that of the untagged, TPE-free counterparts. Moreover, the luminescence of TPE-(RRASL)n is enhanced by up to 700-fold with increasing RNA concentration, which is attributed to the constricted rotation of the TPE moiety as a result of peptide/RNA coacervates within the droplet phase. Besides, at concentrations above MCC, TPE-(RRASL)n can efficiently penetrate through human gallbladder carcinoma cells (SGC-996), translocate into the cell nucleus, and colocalize with intracellular RNA. These observations suggest that AIEgen-conjugated PSPs can be used as droplet-based biosensors for intracellular RNA imaging through a regime of coacervation-induced emission.

Synthetic neutrophil extracellular traps dissect bactericidal contribution of NETs under regulation of α-1-antitrypsin.

Deciphering the complex interplay of neutrophil extracellular traps (NETs) with the surrounding environment is a challenge with notable clinical implications. To bridge the gap in knowledge, we report our findings on the antibacterial activity against Pseudomonas aeruginosa of synthetic NET-mimetic materials composed of nanofibrillated DNA-protein complexes. Our synthetic system makes component-by-component bottom-up analysis of NET protein effects possible. When the antimicrobial enzyme neutrophil elastase (NE) is incorporated into the bactericidal DNA-histone complexes, the resulting synthetic NET-like structure exhibits an unexpected reduction in antimicrobial activity. This critical immune function is rescued upon treatment with alpha-1-antitrypsin (AAT), a physiological tissue-protective protease inhibitor. This suggests a direct causal link between AAT inhibition of NE and preservation of histone-mediated antimicrobial activity. These results help better understand the complex and, at times, contradictory observations of in vivo antimicrobial effects of NETs and AAT by excluding neutrophil, cytokine, and chemoattractant contributions.

In vitro and in vivo evaluation of effects of high-purity magnesium on tight junction of intestinal epithelial cell.

After anastomosis of sutures or pins, the restoration of intestinal barrier function can avoid several complications, such as tissue damage and inflammation. Our previous studies demonstrated the feasibility of biodegradable magnesium (Mg) pins as novel anastomosing implants to spontaneously absorb in the body, avoiding secondary removal surgery and long-term inflammation. However, the effect of Mg pins on the intestinal tight junction barrier is rarely investigated. In this study, we conducted high-purity Mg pins inserted in the intestine of rats and prepared Mg extracts cultured intestinal epithelial cell line to investigate the biological effect on the intestinal barrier associated with tight junction protein expression. We discovered that the concentration of released Mg ions over 1.7 mM was the critical threshold, above which mRNA expression of intestinal tight junction and cell apoptosis were affected considerably. Results of the immunohistochemical analysis revealed that Mg functions to stimulate ZO-1, caspase-3, occluding, and claudin-3 expressions. We offer new insight into the effectiveness of biodegradable Mg materials as the next generation of intestinal anastomosis pins, which effectively filters toxins as well as bacteria, and reduces inflammation.

Revealing the specific interactions between G-quadruplexes and ligands by surface-enhanced Raman spectroscopy.

G-quadruplexes (G4s) regulate a variety of physiological functions related to diseases and life elongation. Therefore, G4 binding ligands, such as potential drugs in gene therapy or molecular probes for biosensing and bioimaging, are receiving extensive attention. However, identifying the binding modes and interaction details between G4s and their ligands is very challenging. Recently, we demonstrated that surface-enhanced Raman scattering (SERS) could quickly provide structural details of G4s. Herein, three G4 binding ligands that interact with the separated G4 in different ways are selected as models to evaluate the feasibility of SERS analysis in studying G4-ligand interactions. As a result, adequate SERS information indicating the specific interactions between the G4s and the ligand is obtained via using Ag IANPs as substrates. The results demonstrate that SERS is a powerful tool for revealing comprehensive and specific ligand-DNA interactions with advantages such as speed, simplicity, trace sample amount requirement, and compatibility with aqueous samples.

High-purity magnesium screws modulate macrophage polarization during the tendon-bone healing process in the anterior cruciate ligament reconstruction rabbit model.

Magnesium (Mg) screws perform clinical potential in anterior cruciate ligament (ACL) reconstruction, and promote fibrocartilaginous entheses regeneration at the femoral entrance. We aim to prove that high-purity Magnesium (HP Mg) screws modulate macrophage polarization in fibrocartilage interface regeneration both in vitro and in vivo. HP Mg extracts performed good cytocompatibility and significantly promoted M2 macrophage polarization in the flow cytometry and ELISA assays. M2 macrophages stimulated fibrochondrocyte differentiation of co-cultured hBMSCs, and HP Mg extracts had synergistic effect on the process. Then we applied HP Mg screws, with Ti screws as control, in the ACL reconstruction rabbit model. In the histological and immunofluorescence analysis, HP Mg screws inhibited M1 polarization at 2 weeks and highly promoted M2 polarization at 2 and 4 weeks at the tendon-bone interface. Furthermore, regeneration of fibrocartilaginous entheses, rather than the fibrovascular scar interface, was detected in the HP Mg group at 12 weeks. For further mechanism study via RNA-seq detection and WB assays, we found that AKT1 was highly activated in M2 polarization, and HP Mg could stimulate AKT1 expression, rather than AKT2, in the early phase of tendon-bone healing. Our study elucidated macrophage polarization during tendon-bone healing process and emphasized HP Mg on M2 polarization and fibrocartilage interface regeneration via the selective activation of AKT1 and PI3K/AKT pathway.