Journal: Frontiers in Microbiology

Article Title: Cocktail of isobavachalcone and curcumin enhance eradication of Staphylococcus aureus biofilm from orthopedic implants by gentamicin and alleviate inflammatory osteolysis

doi: 10.3389/fmicb.2022.958132

Figure Lengend Snippet: Orthopedic implant-related infection animal model of cocktail of isobavachalcone and curcumin with gentamycin against MRSA. (A) In vivo study design. (B) Surgical procedures of distal femoral implant-related infection mouse model. 1–5 refer to: (1) Distal femoral implant using a trans-knee approach, after incise internally alongside patella and externally dislocating patella to expose the distal femoral articular surface, and the entry point, intercondylar notch, was shown by the yellow arrow. (2) 25-gauge needle was used to open a corridor into the femoral medullary canal. (3) The customized 0.6 mm diameter needle was implanted after the inoculation of 5 μl MRSA USA300 inoculum suspension. (4) The caudal end of the implant was left about 1 mm outside the femur after implantation. (5) The representative X-ray image of implant well positioned in the medullary canal and distal femur. (C) Body weight of animals throughout in vivo experiment. (D) Representative appearance of distal femurs of mice 28 days after intraperitoneal administration of different groups of antimicrobials combinations, gentamicin served as control. I.P., intraperitoneal injection; GEN, Gentamycin; ISB, Isobavachalcone; CRM, Curcumin.

Article Snippet: Gentamicin sulfate (HY-A0276), Daptomycin (HY-B0108), Rifampicin (HY-B0272), and Levofloxacin (HY-B0330) were purchased from MedChemExpress, China.

Techniques: Infection, Animal Model, In Vivo, Suspension, Control, Injection

Journal: Frontiers in Microbiology

Article Title: Cocktail of isobavachalcone and curcumin enhance eradication of Staphylococcus aureus biofilm from orthopedic implants by gentamicin and alleviate inflammatory osteolysis

doi: 10.3389/fmicb.2022.958132

Figure Lengend Snippet: Interaction of Isobavachalcone and Curcumin with Antibiotics on MSSA and MRSA strains. (A,B) Heatmaps of fractional inhibitory concentration index (FICI) and fractional bactericidal concentration index (FBCI) of combination of gentamycin with isobavachalcone or curcumin, darker orange color indicates interaction is superimposed or synergistic. (C,D) FICI and FBCI of Isobavachalcone and Curcumin with antibiotics against Staphylococcus aureus JAR strain, respectively. FICI ≤ 0.5 represents synergy, FICI > 0.5–4 represents no synergistic interaction, and FICI > 4.0 represents antagonism. FICI: fractional inhibition concentration index; FBCI: fractional bactericidal concentration index; DAP: Daptomycin with 50 μg/ml Ca2+; RIF: Rifampicin; GEN: Gentamycin; LEV: Levofloxacin; ISB: Isobavachalcone; CRM: Curcumin; MSSA: Methicillin-susceptible Staphylococcus aureus ; MRSA: Methicillin-resistant Staphylococcus aureus .

Article Snippet: Gentamicin sulfate (HY-A0276), Daptomycin (HY-B0108), Rifampicin (HY-B0272), and Levofloxacin (HY-B0330) were purchased from MedChemExpress, China.

Techniques: Concentration Assay, Inhibition

Journal: Frontiers in Microbiology

Article Title: Cocktail of isobavachalcone and curcumin enhance eradication of Staphylococcus aureus biofilm from orthopedic implants by gentamicin and alleviate inflammatory osteolysis

doi: 10.3389/fmicb.2022.958132

Figure Lengend Snippet: Interaction of isobavachalcone and curcumin with Gentamycin against Staphylococcus aureus JAR biofilm. (A) Residual S. aureus JAR on Biofilm challenged by combination of Gentamycin with Isobavachalcone or Curcumin. (B) Coefficient of Drug Interaction (CDI) of the combination of Gentamycin with isobavachalcone or curcumin. (C) Synergistic Combination of isobavachalcone with Gentamycin, and Curcumin with Gentamycin. CDI < 1, =1 or >1 indicate synergy, superposition or antagonism, respectively, CDI < 0.7 indicates significant synergism. Data were presented as mean ± SD. * represents p < 0.05, ** represents p < 0.01, *** represents p < 0.001, **** represents p < 0.001, when compared to ISB/CRM with the same Gentamycin concentration, ns represents no significance. GEN, Gentamycin; ISB, Isobavachalcone; CRM, Curcumin; CDI, Coefficient of Drug Interaction.

Article Snippet: Gentamicin sulfate (HY-A0276), Daptomycin (HY-B0108), Rifampicin (HY-B0272), and Levofloxacin (HY-B0330) were purchased from MedChemExpress, China.

Techniques: Concentration Assay

Journal: Frontiers in Microbiology

Article Title: Cocktail of isobavachalcone and curcumin enhance eradication of Staphylococcus aureus biofilm from orthopedic implants by gentamicin and alleviate inflammatory osteolysis

doi: 10.3389/fmicb.2022.958132

Figure Lengend Snippet: EUCAST checkerboard assay showed isobavachalcone and curcumin interact synergistically with presence of gentamycin against planktonic Staphylococcus aureus . (A,B) Heatmaps of fractional inhibitory concentration index (FICI) and fractional bactericidal concentration index (FBCI) of combination of isobavachalcone and curcumin, darker orange color indicates interaction is superimposed or synergistic. (C) FICI of isobavachalcone with curcumin against S. aureus JAR and USA300 strain. (D) FBCI of isobavachalcone with curcumin against S. aureus JAR and USA300 strain. FICI ≤ 0.5 represents synergy, FICI > 0.5–4 represents no synergistic interaction, and FICI > 4.0 represents antagonism. CDI < 1, =1 or >1 indicate synergy, superposition or antagonism, respectively, CDI < 0.7 indicates significant synergism. Data were presented as mean ± SD. FICI, fractional inhibition concentration index; FBCI, fractional bactericidal concentration index; GEN, Gentamycin; ISB, Isobavachalcone; CRM, Curcumin; CDI, Coefficient of Drug Interaction; MSSA, Methicillin-susceptible Staphylococcus aureus ; MRSA, Methicillin-resistant Staphylococcus aureus .

Article Snippet: Gentamicin sulfate (HY-A0276), Daptomycin (HY-B0108), Rifampicin (HY-B0272), and Levofloxacin (HY-B0330) were purchased from MedChemExpress, China.

Techniques: Concentration Assay, Inhibition

Journal: Frontiers in Microbiology

Article Title: Cocktail of isobavachalcone and curcumin enhance eradication of Staphylococcus aureus biofilm from orthopedic implants by gentamicin and alleviate inflammatory osteolysis

doi: 10.3389/fmicb.2022.958132

Figure Lengend Snippet: In vitro Eradication of Staphylococcus aureus biofilms by the GEN-ISB-CRM combination. (A,C) Quantification of residual viable cells on S. aureus JAR biofilm and USA300 biofilm, respectively. (B,D) Coefficient of Drug Interaction (CDI) shows synergy between Isobavachalcone and Curcumin with 128 μg/ml Gentamycin against S. aureus JAR biofilm and USA300 biofilm, respectively. (E) Representative characteristics of MRSA USA300 biofilm after 24 h challenge. SEM: biofilms cultured at 37°C for 24 h were treated with 128 μg/ml GEN (control), 128 μg/ml GEN + 125 μg/ml CRM, 128 μg/ml GEN + 6.25 μg/ml ISB, 128 μg/ml GEN + 125 μg/ml CRM + 6.25 μg/ml ISB for 24 h, respectively. All SEM images were captured at EHT = 10 kV, Mag = 5KX. CLSM: representative confocal laser scanning microscopy (100× oil immersion) images of LIVE/DEAD ® BacLight staining showing the synergistic effect of different drug combinations on USA300 biofilm challenged for 24 h, green: live cells; red: dead cells. Data were presented as mean ± SD. * represents p < 0.05, ** represents p < 0.01, *** represents p < 0.01 when compared to 0 μg/ml CRM with the same ISB concentration group, ns represents no significance. GEN, Gentamycin; ISB, Isobavachalcone; CRM, Curcumin; CDI, Coefficient of Drug Interaction; SEM, Scanning Electron Microscopy; CLSM, Confocal Laser Scanning Microscopy.

Article Snippet: Gentamicin sulfate (HY-A0276), Daptomycin (HY-B0108), Rifampicin (HY-B0272), and Levofloxacin (HY-B0330) were purchased from MedChemExpress, China.

Techniques: In Vitro, Cell Culture, Control, Confocal Laser Scanning Microscopy, Staining, Concentration Assay, Electron Microscopy

Journal: Frontiers in Microbiology

Article Title: Cocktail of isobavachalcone and curcumin enhance eradication of Staphylococcus aureus biofilm from orthopedic implants by gentamicin and alleviate inflammatory osteolysis

doi: 10.3389/fmicb.2022.958132

Figure Lengend Snippet: Micro Computed tomography (μCT) imaging suggests protection of cocktail therapy against osteolysis in orthopedic implant-related infections due to MRSA. (A) Representative X-ray images of distal femur of mouse with intramedullary implant. (B) Representative images of trans axial 3D reconstructed micro-architecture of distal femur around implant. (C) Bone morphometry in distal femur shows protection of combination of Isobavachalcone and Curcumin against osteolysis during orthopedic implant-related infection, related parameters including percent bone volume (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N), and trabecular separation (Tb.Sp), bone mineral density (BMD) and bone surface density (BS/TV) were measured, data were presented as mean ± SD, * represents p < 0.05 and ** represents p < 0.01 when compared to Gentamicin group; # represents p < 0.05 and ## represents p < 0.01 when GEN + ISB + CRM group compared to GEN + ISB/CRM group; GEN, 20 mg/kg/day Gentamycin; ISB, 20 mg/kg/day Isobavachalcone; CRM, 20 mg/kg/day Curcumin.

Article Snippet: Gentamicin sulfate (HY-A0276), Daptomycin (HY-B0108), Rifampicin (HY-B0272), and Levofloxacin (HY-B0330) were purchased from MedChemExpress, China.

Techniques: Micro-CT, Imaging, Infection

Journal: Frontiers in Microbiology

Article Title: Cocktail of isobavachalcone and curcumin enhance eradication of Staphylococcus aureus biofilm from orthopedic implants by gentamicin and alleviate inflammatory osteolysis

doi: 10.3389/fmicb.2022.958132

Figure Lengend Snippet: Cocktail therapy of Gentamicin with Isobavachalcone and Curcumin alleviate local tissue inflammation, enhance reduction of MDSC M1 Polarization and Eradication of MRSA biofilm in vivo . (A) HE and immunofluorescent staining of distal femur suggest reduction of tissue inflammation and M1 polarization of MDSC, green represents CD11b, a key marker to identify MDSC, while red represents CXCL10, a key marker of activated M1, he black dashed box indicates the approximate location of the implant, and the yellow dashed box range is the area where M1-polarized MDSC in the adjacent implant tissue is observed. (B) Quantification of remaining MRSA in bone with implant. (C) Flow cytometry analysis showed reduction of MDSC in peripheral blood by Gentamicin with ISB or CRM and cocktail therapy. (D) Gating strategy for flow cytometry analysis and representative analysis of frequency of MDSC in peripheral blood. All data were presented as mean ± SD, * represents p < 0.05 and ** represents p < 0.01 when compared to Gentamicin group; # represents p < 0.05 and ## represents p < 0.01 when GEN + ISB + CRM group compared to GEN + ISB/CRM group; GEN, 20 mg/kg/day Gentamycin; ISB, 20 mg/kg/day Isobavachalcone; CRM, 20 mg/kg/day Curcumin.

Article Snippet: Gentamicin sulfate (HY-A0276), Daptomycin (HY-B0108), Rifampicin (HY-B0272), and Levofloxacin (HY-B0330) were purchased from MedChemExpress, China.

Techniques: In Vivo, Staining, Marker, Flow Cytometry

Journal: Physiological Reports

Article Title: Acetate derived from the intestinal tract has a critical role in maintaining skeletal muscle mass and strength in mice

doi: 10.14814/phy2.16047

Figure Lengend Snippet: Antibiotic treatment for 2 weeks reduced grip strength in mice. (a) Experimental protocol. C57BL/6 mice were allocated to two groups and administered antibiotics (100 μg/mL neomycin, 50 μg/mL streptomycin, 100 U/mL penicillin, 50 μg/mL vancomycin, 100 μg/mL metronidazole, 125 μg/mL ciprofloxacin, 100 μg/mL ceftazidime, and 170 μg/mL gentamicin in their drinking water) or not for 2 weeks (Abx‐, n = 13; Abx+, n = 18). (b) The bodyweight of Abx‐ and Abx + mice (Abx‐, n = 13; Abx+, n = 18). (c) WAT mass of Abx‐ and Abx + mice (Abx‐, n = 13; Abx+, n = 18). (d) Skeletal muscle masses of Abx‐ and Abx + mice (Abx‐, n = 13; Abx+, n = 18). (e and f) Cecum of Abx‐ and Abx + mice (Abx‐, n = 13; Abx+, n = 18). (g) Grip strength of Abx‐ and Abx + mice (Abx‐, n = 4; Abx+, n = 11). (h) Intestinal SCFA concentrations of Abx‐ and Abx + mice (Abx‐, n = 9; Abx+, n = 13). (I) Plasma SCFA concentration of Abx‐ and Abx + mice (Abx‐, n = 9; Abx+, n = 13). Data expressed as mean; ns, not statistically significant. The lines indicate the means; analyzed using Student's t ‐tests. Abx‐, antibiotics untreated; Abx+, antibiotics treated; GAS, gastrocnemius muscle; SCFA, short‐chain fatty acid; SOL, soleus muscleTA, tibialis anterior muscle; WAT, white adipose tissue.

Article Snippet: The chemicals and reagents used were sodium acetate from Sigma (S5636), streptomycin and penicillin (Meiji Seika Pharma Co., Ltd., Tokyo, Japan), and vancomycin hydrochloride (V0252), metronidazole (M1977), ciprofloxacin (C3262), ceftazidime hydrate (C1635), gentamycin sulfate (G1658), and neomycin sulfate (N1755) from LKT laboratories, Inc. (St Paul, MN, USA) as described previously (Okamoto et al., ).

Techniques: Clinical Proteomics, Concentration Assay