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This research investigates the antibacterial, anti-inflammatory, and wound healing effects of postbiotics combined with Caesalpinia sappan extract against bovine mastitis.

April 27, 2026Open Access

Postbiotics combined with Caesalpinia sappan L. extract inhibit bovine mastitis bacterial pathogens, promote anti-inflammation and in vitro wound healing property

Puntos clave

This research investigates the antibacterial, anti-inflammatory, and wound healing effects of postbiotics combined with Caesalpinia sappan extract against bovine mastitis.
Investigated antibacterial and biofilm inhibition using Lactiplantibacillus plantarum and Lacticaseibacillus casei supernatants with C. sappan extract.
Assessed cytotoxicity on bovine endothelial and macrophage cells with varying concentrations.
Evaluated nitric oxide production and inflammatory gene expression in co-cultured CPAGE and RAW 264.7 cells.
LPC and LCC showed MIC values between 0.024–3.125 mg/mL and MBC values of 0.098–3.125 mg/mL, demonstrating significant antibacterial activity.
LPC and LCC inhibited nitric oxide production by 60.13–95.30% in LPS-induced conditions, with C. sappan extract achieving 80.33–92.79% inhibition.
Complete wound closure was observed within 12 hours for all treatment groups.

Resumen

Biological activities on inhibition of bovine mastitis-associated bacterial pathogens and bacterial biofilm, anti-inflammation, and wound healing effects of lactic acid bacteria (LAB); Lactiplantibacillus plantarum TISTR 2070 (LP) and Lacticaseibacillus casei TISTR 1340 (LC) supernatants were investigated in this study. Moreover, Caesalpinia sappan extract, and their combinations with L. plantarum (LPC) and L. casei (LCC) were determined for their biological properties. Antibacterial assays revealed that LPC and LCC exhibited the lowest MIC (0.024–3.125 mg/mL) and MBC (0.098–3.125 mg/mL) values, that showed significant antibacterial activity stronger than LP and LC. In biofilm assays, the LPC and LCC combinations (0.8–1.6 mg/mL) inhibited preformed biofilm of Streptococcus agalactiae O4 (61.97–73.49%), Staphylococcus haemolyticus (38.74–52.65%), and Streptococcus uberis (24.78–58.05%) whereas C. sappan extract alone strongly inhibited both preformed and mature biofilms of all pathogens. Cytotoxicity evaluation showed that LPC and LCC were non-toxic to bovine endothelial (CPAE) and macrophage (RAW 264.7) cells at concentration ≤ 12.5 µg/mL. Furthermore, novel co-culture of CPAE and RAW 264.7 cells was applied in this study, LPS (5 ng/mL) significantly induced nitric oxide (NO) production compared with monoculture. In this model, LPC and LCC markedly suppressed LPS-induced NO production by 60.13–77.38% and 74.64–95.30%, respectively, while C. sappan extract achieved NO inhibition by 80.33–92.79%. Furthermore, LPC and LCC significantly downregulated IL-6 and TNF-α inflammatory genes expression, with LCC showing the strongest effect. Wound healing assay demonstrated that LP, LC, C. sappan extract and the combinations promoted complete closure of wound within 12 h. Therefore, these findings indicate that the combinations of LAB postbiotics with C. sappan extract exhibit strong antibacterial, anti-biofilm, anti-inflammatory, and wound healing activities, supporting their potential as alternative therapeutics against bovine mastitis.

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