Table 1.
Critical comparison of representative human and animal evidence linking NETs to NEC.
| Study/model | Stimulus/context | NET intervention | Timing | NET readout | Main outcome | Interpretation and limitation |
|---|---|---|---|---|---|---|
| Vincent/Klinke mouse NEC; Sci Rep 2018 (54) | LPS + formula + hypoxia | PAD inhibition | Preventive/therapeutic in model protocol | Tissue H3cit/MPO; serum cfDNA/nucleosomes | Reduced NEC manifestation (4/10 vs 5/7 with severe NEC, p=0.003) and day-10 mortality (p=0.0129; day-6 survival 7/14 NEC vs 10/14 with PAD inhibition); serum cfDNA correlated with NEC score (r=0.53, p=0.001) and Chiu score (r=0.56, p<0.001). Total 76 mice; human NEC tissue n=9 | Supports pathogenic NETs in inflammatory-ischemic model; clinical samples mostly late; cfDNA nonspecific |
| Klinke/Vincent DNase mouse NEC; J Surg Res 2019 (67) | LPS + formula + hypoxia | Systemic DNase1 | After injury induction | Extracellular DNA degradation; cfDNA/H3cit/NE/MPO/TLR4 | N=44 neonatal mice; DNase1 i.p. from day 5 for 72 h reduced NEC rates (44% vs 86%, P = 0.029) and improved survival (65% vs 35%, P = 0.01), with lower cfDNA, Chiu score, MDA, GPx, NE, MPO, H3cit, and TLR4 | Targets deposited DNA, not formation; does not remove histones/proteases; timing critical |
| Chaaban dithizone/Klebsiella NEC-like model; J Cell Mol Med 2021 (68) | Chemical injury + live Gram-negative bacteria | NET inhibition | Around bacterial challenge | NET-associated signals | Cl-amidine (40 mg/kg, pan-PAD inhibitor) given 30 min before and 3 h after Klebsiella gavage nearly doubled mortality (94.7% vs 47.6%, p<0.0001) and increased bacteremia at 1 h (p=0.0074); intestinal injury scores did not differ between NEC and NEC+Cl-amidine groups, supporting a defect in bacterial clearance rather than direct tissue protection | Shows protective NETs when translocation risk is high; not equivalent to classical hypoxia/LPS NEC |
| Saha Citrobacter rodentium; Mucosal Immunol 2019 (69) | Attaching/effacing infection | PAD4 deficiency or DNase | During infection | PAD4-dependent NETs | Pad4−/− adult mice (8 weeks; histology n=6–8): CR colonization peaked at days 11–14 (vs clearance by day 14 in WT), with aggravated colonic hyperplasia, goblet cell depletion, and crypt apoptosis; DNase I (n=6) and Cl-amidine recapitulated the phenotype. No mortality endpoint; adult, non-NEC model | Mucosal defense context differs from NEC necrosis; supports organ- and phase-specific effects |
| CD177+ neutrophil-platelet aggregates; Nat Commun 2026 (25) | Human tissue/model mechanistic | Not a treatment trial | Observational/mechanistic | CD177+ aggregates, NETs, thromboinflammation | Neonatal mice: histology/flow n=12 per group, survival n=24 per group; CD177−/− or myeloid-specific CD177KO reduced intestinal NPAs and NETs and improved survival; DNase I or PAD4i (n=12/group) reduced histopathology scores; prophylactic LMWH disrupted NPAs, lowered NET burden, and improved survival. Human NEC: CD177+ NPA levels correlated with severity and thrombotic biomarkers (n=14, Spearman) | Strong mechanistic clue; requires prospective stratification and causal timing |
| Platelet exosome/GSDMD; Cell Commun Signal 2026 (26) | Platelet-derived exosomes | GSDMD inhibition | Mechanistic blockade | NET formation induced by exosomes | 5-day-old C57BL/6 pups (formula gavage + enteral LPS 5 mg/kg + hypoxia/cold stress); disulfiram reduced platelet caspase-1 activation, NET levels, MPO/ROS, IL-6 and IL-1β, ameliorated villus necrosis, and significantly improved survival; GSDMD−/− pups phenocopied. Mechanism: mitochondrial ROS and ox-mtDNA link platelet pyroptosis to NET formation | Identifies upstream node; clinical safety in neonates unknown |
| IP6K1-polyphosphate axis; Inflamm Res 2026 (70) | Platelet short-chain polyphosphates/NPA | IP6K1 inhibition | Experimental NEC | Neutrophil-platelet aggregates | Human NEC: elevated circulating NPAs and anticardiolipin IgG, reduced platelet counts with pyroptosis as dominant platelet death mode. Neonatal mouse NEC: Ip6k1−/− and TNP (IP6K1 inhibitor) attenuated NPA formation, neutrophil infiltration, and NET release, improving survival and reducing severity scores; exogenous polyP rescued NPA formation; P-selectin neutralization reduced both NPAs and NETs | Supports selective immunothrombosis targeting; not direct NET quantification of all pathways |
| IL-40 deficiency; Cell Commun Signal 2026 (71) | Experimental NEC | IL-40 deficiency | Genetic/developmental | NETosis | Human NEC (n=8; surgical controls n=5): plasma IL-40 correlated with SOFA score (r=0.9259, P<0.0001) and MPO-DNA with SOFA (r=0.678, P = 0.0007). IL-40−/− neonatal mice: higher survival, lower histopathology scores, reduced dsDNA, MPO-DNA, and H3Cit, and lower bacterial burden and plasma LPS | Cytokine-NET link; causality and reversibility need confirmation |
TLR4, Toll-like receptor 4.