Neutrophil NETosis, viability, and gene or protein expression in response to surface chemistry using isodecyl acrylate copolymer coatings
Bibliographic record
Abstract
Undesirable host responses to implants commonly lead to impaired device function. Biomaterial-based strategies focused on neutrophil behavior to improve host responses to implants have not been significantly researched. As the first immune cell to respond to inflammation, neutrophil activation resulting in the release of antimicrobials and neutrophil extracellular traps primes this microenvironment for macrophages and other infiltrating cells. Methacrylic acid (MAA) and methyl methacrylate (MM) based polymers modulate healing responses through altered macrophage behavior and vascularization. This project investigates the neutrophil response to MAA and MM copolymerized with isodecyl acrylate (IDA) (MAAcoIDA and MMcoIDA) and hexamethylenediamine-functionalized MMcoIDA coatings (HMD-MMcoIDA). In this research, MAAcoIDA and MMcoIDA were coated onto glass coverslips and MMcoIDA. MAAcoIDA coatings had 4975 ± 580 nmol/cm2 of carboxyl groups at the surfaces while MMcoIDA was further functionalized to add 58 ± 18 nmol/cm2 of amine groups to the surfaces via aminolysis with HMD. The HL60 cell line and murine bone marrow derived neutrophils (BMDN) were then incubated with the coatings to study neutrophil-material interactions. HL60 gene expression and viability were assessed to determine inflammatory and anti-inflammatory characteristics induced by the different surface chemistries. NETosis, indicated by citrullinated histone H3 (citH3), elicited by the coatings was visualized for both HL60 cells and murine BMDN, and protein synthesis was additionally quantified for BMDN. HL60 cells demonstrated increased viability when incubated with MAAcoIDA relative to MMcoIDA. HL60 viability on HMD-MMcoIDA decreased after 48 hours, coinciding with increased citH3. HL60 cells exhibited increased expression of ICAM-1 and TNF-α when incubated on aminated coatings compared to those with MMcoIDA or MAAcoIDA after 48 hours. Murine BMDN displayed lower ratios of viable cells on the coatings than uncoated glass, and cells with MAAcoIDA coatings underwent less histone citrullination while HMD-MMcoIDA elicited increased histone citrullination, relative to MMcoIDA. BMDN secreted varied concentrations of MIP-1α and MIP-2 in response to coatings with no added stimulus after 2 hours and increased over 16 hours, although no significant differences were noted between surface chemistries. Future work looks to characterize these coatings and examine protein-material interactions, as well as follow up studies with these materials in vivo.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.001 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".