Dimethylformamide

Tin(ii) bromide, tin(ii) iodide, antimony trichloride, N,N′-dimethylethylenediamine (99%), 1-butyl-1-methylpyrrolidinium chloride, γ-butyrolactone (GBL, ≥99%), hydrobromic acid (48 wt% in H₂O) and hydriodic acid (55%) were purchased from Sigma-Aldrich. Dichloromethane (DCM, 99.9%), dimethylformamide (DMF, 99.8%), toluene (anhydrous, 99.8%) and ethyl ether (stabilized with ∼1 ppm BHT) were purchased from VWR. Acetone (HPLC grade) was purchased from EMD Millipore. All reagents and solvents were used without further purification unless otherwise stated.

The amino acids and the PEG resin were provided by Novabiochem^®.
Hydroxybenzotriazole (HOBt), piperidine, formic acid,
O-Benzotriazol-1-yl)-N,N,N,N-tetramethyluronium hexafluorophosphate (HBTU),
N-methyl- morpholine (NMM), 1,2-ethanedithiol (EDT), phenol, thioanisol,
N,N′-Diisopropylcarbodiimide (DIC), Triisopropylsilane (TIS), Trifluoroacetic
acid (TFA), ether, human trypsin and trypsin substrate Nα-Benzoyl-DL-arginine
4-nitroanilide hydrochloride (BAPNA) were provided by Sigma Aldrich^®.
VWR Chemicals^® provided the Dimethylformamide (DMF) and acetonitrile
(ACN). Fmoc-Rink MBHA resin and reagent Oxyma Pure were provided by Iris
Biotech. Human factor Xa, thrombin, and their chromogenic substrates, Bz-Ile-Glu
(γ-OCH₃)-Gly-Arg-pNA [CS-11(22)] and H-D-Phe-Pip-Arg-pNa, 2HCl
[CS-01(38)], respectively, were provided by Hyphen. Batroxobin was provided by
Pentapharm. Bothrops jararaca venom (BjV) was supplied by the
Herpetology Laboratory from Butantan Institute (IB), São Paulo, Brazil. The
fluorogenic substrate Abz-Ser (Abz-RPPGFSPFRQ-EDDnp) was provided by Dr.
Fernanda C.V. Portaro.

Out of the HCA standards MeIQx, 4,8-DiMeIQx and PhIP were purchased from Toronto Research Chemicals (North York, ON, Canada), HAR and NOR from Sigma-Aldrich (Saint Loius, MO, USA), as well as caffeine used as internal standard. Other chemicals such as acetonitrile, dimethylformamide, methanol, formic acid, acetic acid and sodium hydroxide were purchased from VWR International (Radnor, PA, USA). Type I ultra-pure water was produced by SUEZ Environment^® Water Purification System (Paris, France).

A PCL solution was prepared from medical-grade PCL granules (CAPA^® 6800, average 80,000 MW, Perstorp, Sweden) dissolved at a 12% w/v concentration in a 65:35 solvent mixture of Chloroform (99.2% purity, stabilized with 0.6% ethanol, VWR, Radnor, PA, USA) and Dimethylformamide (100% purity, VWR, Radnor, PA, USA) under mild stirring overnight at room temperature. The supplier reported a theoretical density for the PCL of ρ0 = 1.145 g/cm³.
A PEO solution was prepared from PEO granules (35,000, FLUKA Chemicals, Buchs, Switzerland) dissolved at a 30% w/v concentration in distilled water under mild stirring at 50 °C for two days. The supplier reported a theoretical density for the PEO of ρ0 = 0.45 g/cm³.
1-PWD formulation, obtained by API spray dispensing directly from its container, was then mixed with the PEO solution (30 wt%) under magnetic stirring until the incorporation of a 4% product.
Biomedical scaffolds were fabricated with electrospinning using a pilot scale electrospinning station (Fluidnatek LE100, Bioinicia, Valencia, Spain) using different variations of a process recipe, provided in-kind by the company Nanofaber srl (Rome, Italy), varying the applied voltage and flow rate (FR). The relative humidity (RH) and ambient temperature during the deposition process were recorded.

Polycaprolactone (PCL, MW = 80,000) was purchased from Perstorp (Lund, Sweden), and PEO or poly(ethylene glycol) with an Mw of 35,000 was provided by Fluka (Buchs, Switzerland). Chloroform and Dimethylformamide were obtained by VWR (Radnor, PA, USA).
The patented plant-derived formulation 1 Primary Wound Dressing© (1-PWD), composed of neem oil (Azadirachta indica A. Juss.) and the oily extract of Hypericum perforatum (L.) flowers, was supplied by the ENEA (Italian National Agency for New Technologies, Energy, and Sustainable Development).

A PCL solution was prepared from medical grade PCL granules (CAPA^® 6500, average 50,000 MW, Perstorp, Sweden) dissolved at 12% w/v concentration in a 65:35 solvent mixture of Chloroform (99.2% purity, stabilized with 0.6% ethanol, VWR, Radnor, PA, USA) and Dimethylformamide (100% purity, VWR, Radnor, PA, USA) under mild stirring overnight at room temperature. The supplier reported a theoretical density for PCL of ρ₀ = 1.145 g/cm³. Non-woven scaffolds were fabricated with electrospinning using a pilot scale electrospinning station (Fluidnatek LE100, Bioinicia, Spain) using different variations of a process recipe, kindly provided by the Company Nanofaber srl (Rome, Italy) as indicated in Table 4, and varying the applied voltage and flow rate (FR).