Journal: Bioactive Materials

Article Title: Dual-engineering metalloimmunotherapy mediates Staphylococcus aureus virulence silencing and biofilm immune microenvironment reprogramming against implant-associated infections

doi: 10.1016/j.bioactmat.2026.03.008

Figure Lengend Snippet: Critical roles of bacterial virulence and host cGAS–STING signaling in IAIs. a , Schematic workflow of clinical S. aureus isolation from orthopedic patients with IAIs and subsequent laboratory virulence assessment. b , Correlation analysis of patient serum PCT levels and the hemolysis ability of infected S. aureus . c , Kaplan-Meier analysis showing the correlation of S. aureus virulence and patient infection-free survival. d , Representative X-ray and PET/CT images of patients infected with high- or low-virulence S. aureus strains. e , Experimental workflow of the murine knee periprosthetic joint infection (PJI) model. f , g , Colony-forming unit (CFU) counts of viable bacteria in peri-implant infected tissues ( f ) and extracted implants ( g ) from PJI model mice on day 14. h , Representative micro-CT images of tibiae from PJI model mice on day 28. i , Representative 3D reconstructed images of gait footprints from PJI model mice. j , o , Experimental workflow of the murine single ( j ) and mixed ( o ) infection IAIs models. k , l , p , q , CFU counts of viable bacteria in peri-implant infected tissues ( k , p ) and extracted implants ( l , q ) from IAIs model mice. m , r , Representative H&E and Giemsa staining images of peri-implant infected tissues from mice on day 14. m , r , Representative Confocal laser scanning microscopy (CLSM) 3D reconstruction images of biofilms on extracted implants from mice on day 14. t , Representative bioluminescence images of mice on day 14. u-z , Representative culture images and corresponding CFU counts of viable bacteria in peri-implant infected tissues and extracted implants from mixed infection IAIs model mice (WT-lux + Δ saeRS group, u-w ; WT + Δ saeRS -lux group, x-z ). The yellow arrow indicates the Δ saeRS colonies. Data are presented as mean ± s.d. from n biologically independent samples ( f , g , k , l , p , q , u , v , x and y , n = 5). Statistical significance was analyzed by two-way ANOVA with Tukey's test for f , g , k , l , p and q and two-tailed paired t tests for u , v , x and y .

Article Snippet: 100 μL GFP-reporting S . aureus suspension was spread on CM10 plates using the SPM method and incubated at 37 °C for 24 h. The fluorescence images of GFP-reporting S . aureus on the CM10 plates were captured using a bioluminescence system (Tanon ABL X6, Shanghai, China).

Techniques: Isolation, Infection, Positron Emission Tomography-Computed Tomography, Bacteria, Micro-CT, Staining, Confocal Laser Scanning Microscopy, Two Tailed Test

Journal: Bioactive Materials

Article Title: Dual-engineering metalloimmunotherapy mediates Staphylococcus aureus virulence silencing and biofilm immune microenvironment reprogramming against implant-associated infections

doi: 10.1016/j.bioactmat.2026.03.008

Figure Lengend Snippet: HMPF nanoparticles activate comprehensive host immunity to combat primary IAIs. a, Experimental workflow for evaluating the therapeutic efficacy against IAIs in vivo. b , Individual and average infection area growth curves of IAIs model mice in different treatment groups. c , d , CFU counts of viable bacteria in the peri-implant infected tissues ( c ) and extracted implants ( d ) from mice after different treatments on day 14. e , Representative general images, bioluminescence images, H&E and Giemsa staining images of peri-implant infected tissues from mice after different treatments on day 14. f , Representative general images, bioluminescence images, CLSM 3D reconstruction images and SEM images of biofilms on extracted implants from mice after different treatments on day 14. g , m , Representative immunofluorescent images of CD80 and CD86 signals ( g ), CD4 and CD8 signals ( m ) in infection-associated draining lymph nodes (dLNs). h-l , n-r , FACS quantification of M1-phenotype macrophages (CD80 + CD206 - cells by gating on CD11b + F4/80 + cells) ( h ), mature DCs (CD80 + CD86 + cells by gating on CD11c + MHC II + cells) ( i ), MFI of MHC II in CD11c + cells ( j ), CD4 + T cells (CD4 + cells by gating on CD3 + cells) ( k ), CD8 + T cells (CD8 + cells by gating on CD3 + cells) ( l ), NK cells (NK1.1 + cells by gating on CD45 + cells) ( n ), Treg cells (Foxp3 + cells by gating on CD3 + CD4 + cells) ( o ), M-MDSC cells (Ly6C high Ly6G − cells by gating on CD11b + cells) ( p ), PMN-MDSC cells (Ly6C low Ly6G + cells by gating on CD11b + cells) ( q ), plasma cells (CD19 − CD138 + cells by gating on CD45 + cells) ( r ) in dLNs from mice after different treatments on day 14. s , t , Quantification of IgM and IgG levels in the serum of mice after different treatments on day 14. u , Representative Western blot images and quantification of cGAS-STING pathway-related proteins of peri-infection tissues from mice after different treatments on day 5. v , Schematic diagram of IAIs models constructed using wild-type and Sting −/− mice for in vivo therapeutic effect evaluation. w , x , CFU counts of viable bacteria in the peri-implant infected tissues ( w ) and extracted implants ( x ) from wild-type and Sting −/− mice after different treatments on day 14. y , z , FACS quantification of M1-phenotype macrophages ( y ) and mature DCs ( z ) in dLNs from wild-type and Sting −/− mice after different treatments on day 14. Data are presented as mean ± s.d. from n biologically independent samples ( b , n = 6; c and d , n = 8; h - l , n - t and w - z n = 5; u , n = 3). Statistical significance was analyzed by one-way ANOVA with Tukey's multiple comparisons test for c , d , h - l and n - u , and two-way ANOVA with Tukey's test for w - z .

Article Snippet: 100 μL GFP-reporting S . aureus suspension was spread on CM10 plates using the SPM method and incubated at 37 °C for 24 h. The fluorescence images of GFP-reporting S . aureus on the CM10 plates were captured using a bioluminescence system (Tanon ABL X6, Shanghai, China).

Techniques: Drug discovery, In Vivo, Infection, Bacteria, Staining, Clinical Proteomics, Western Blot, Construct

Journal: Bioactive Materials

Article Title: Dual-engineering metalloimmunotherapy mediates Staphylococcus aureus virulence silencing and biofilm immune microenvironment reprogramming against implant-associated infections

doi: 10.1016/j.bioactmat.2026.03.008

Figure Lengend Snippet: HMPF nanoparticles exhibit excellent therapeutic efficacy in treating mice with S. aureus -indu ced PJI. a , Experimental workflow for evaluating the therapeutic efficacy against knee prosthetic joint infection (PJI) of mice in vivo. b , c , Individual ( b ) and average ( c ) infection area growth curves of PJI model mice in different treatment groups. d , Bioluminescence intensity curves of PJI model mice in different groups on days −2, 2, 6, 10, and 14. e , f , CFU counts of viable bacteria in the peri-implant infected tissues ( e ) and extracted implants ( f ) from PJI model mice after different treatments on day 14. g - k , Representative micro-CT and X-ray images ( k ) and quantification of bone volume/total volume (BV/TV) ( g ), cancellous bone mineral density (BMD) ( h ), trabecular thickness (TB. TH) ( i ) and joint angle ( j ). l , Schematic illustration of the gait analysis experiment. m , Representative footprint images of PJI mice in different groups on day 14. n - q , Quantification of gait analysis data, including support time ( n ), stride length ( o ), average intensity (average pressure on the runway) ( p ), and average speed ( q ). r , Representative 3D reconstructed images of left hind footprints from PJI mice in different treatment groups. The color bar represents the pressure intensity of each part of the mouse footprint on the ground. Data are presented as mean ± s.d. from n biologically independent samples ( b and c , n = 6; d and g - j , n = 3; e , f and n - q , n = 8). Statistical significance was analyzed by one-way ANOVA with Tukey's multiple comparisons test for e - j and n - q .

Article Snippet: 100 μL GFP-reporting S . aureus suspension was spread on CM10 plates using the SPM method and incubated at 37 °C for 24 h. The fluorescence images of GFP-reporting S . aureus on the CM10 plates were captured using a bioluminescence system (Tanon ABL X6, Shanghai, China).

Techniques: Drug discovery, Infection, In Vivo, Bacteria, Micro-CT

Journal: Bioactive Materials

Article Title: Dual-engineering metalloimmunotherapy mediates Staphylococcus aureus virulence silencing and biofilm immune microenvironment reprogramming against implant-associated infections

doi: 10.1016/j.bioactmat.2026.03.008

Figure Lengend Snippet: HMPF nanoparticles activate long-term immune memory to prevent IAIs rec urrence. a , Schematic illustration of an implant-associated secondary infection model constructed for in vivo evaluation of therapeutic efficacy against recurrent infections. b , Individual and average infection area growth curves of mice with recurrent infections in different groups. c - f , Gradient dilution bioluminescence images and CFU counts of viable bacteria in the peri-implant infected tissues ( c , d ) and extracted implants ( e , f ) from mice with recurrent infections on day 14. g , Representative general images, bioluminescence images, H&E and Giemsa staining images of peri-implant infected tissues from mice with recurrent infections on day 14. h , Representative general images, bioluminescence images, CLSM 3D reconstruction images and SEM images of biofilms on extracted implants from mice with recurrent infections on day 14. i , j , Representative FACS profiles ( i ) and quantification ( j ) of memory B cells (IgG + IgD − cells by gating on B220 + cells) in dLNs from mice with recurrent infections on day 14. k - n , Representative FACS profiles ( k , m ) and quantification ( l , n ) of memory T cells (CD44 high CD62L − cells by gating on CD3 + CD4 + and CD3 + CD8 + cells) in dLNs from mice with recurrent infections on day 14. o - q , FACS quantification of memory B cells ( o ), CD4 + memory T cells ( p ) and CD8 + memory T cells ( q ) in dLNs from wild-type and STING KO mice with recurrent infections on day 14. Data are presented as mean ± s.d. from n biologically independent samples ( b , n = 6; d and f , n = 8; j , l , n and o - q n = 5). Statistical significance was analyzed by one-way ANOVA with Tukey's multiple comparisons test for d , f , j , l and n , and two-way ANOVA with Tukey's test for o - q .

Article Snippet: 100 μL GFP-reporting S . aureus suspension was spread on CM10 plates using the SPM method and incubated at 37 °C for 24 h. The fluorescence images of GFP-reporting S . aureus on the CM10 plates were captured using a bioluminescence system (Tanon ABL X6, Shanghai, China).

Techniques: Infection, Construct, In Vivo, Drug discovery, Bacteria, Staining