Succinylation of Polyallylamine: Influence on Biological Efficacy and the Formation of Electrospun Fibers

Abstract

Succinylation of proteins is a commonly encountered reaction in biology and introduces negatively charged carboxylates on previously basic primary amine groups of amino acid residues. In analogy, this work investigates the succinylation of primary amines of the synthetic polyelectrolyte polyallylamine (PAA). It investigates the influence of the degree of succinylation on the cytotoxicity and antibacterial activity of the resulting polymers. Succinylation was performed in water with varying amounts of succinic anhydride and at different pH values. The PAA derivatives were analyzed in detail with respect to molecular structure using nuclear magnetic resonance and infrared absorbance spectroscopy. Polyelectrolyte and potentiometric charge titrations were used to elucidate charge ratios between primary amines and carboxylates in the polymers. The obtained materials were then evaluated with respect to their minimum inhibitory concentration against Staphylococcus aureus and Pseudomonas aeruginosa. The biocompatibility was assessed using mouse L929 fibroblasts. The degree of succinylation decreased cytotoxicity but more significantly reduced antibacterial efficacy, demonstrating the sensitivity of the fibroblast cells against this type of ampholytic polyelectrolytes. The obtained polymers were finally electrospun into microfiber webs in combination with neutral water-soluble polyvinyl alcohol. The resulting non-woven could have the potential to be used as wound dressing materials or coatings.

Cytotoxicity and Antibacterial Efficacy of Betaine- and Choline-Substituted Polymers

“…The biological reactivity and potential cytotoxic activity of HEC, PVA, and their betaine and choline derivatives were evaluated on mouse fibroblasts (NCTC clone 929: CCL 1; L929; American Type Culture Collection, ATCC, Manassas, VA, USA) in vitro in accordance with the International Standard ISO 10993-5:2009, Biological evaluation of medical devicesPart 5: Tests for in vitro cytotoxicity, as previously described . Biological reactivity was evaluated by light microscopy, following exposure to the samples for 24 ± 1 and 48 ± 1 h, and rated on a scale of 0–4 as described in ISO 10993-5:2009.…”

“…Three independent experiments were performed in five replicates. The program GraphPad Prism 9 (GraphPad Software, San Diego, CA, USA) was used for statistical analysis and calculation of half-maximal inhibitory concentration (IC 50 ) values, as previously described . Light microscopy images of the cells after exposure to graded concentrations of the tested samples are shown in the Supporting Information (Figures S24–30).…”

“…The biological reactivity and potential cytotoxic activity of HEC, PVA, and their betaine and choline derivatives were evaluated on mouse fibroblasts (NCTC clone 929: CCL 1; L929; American Type Culture Collection, ATCC, Manassas, VA, USA) in vitro in accordance with the International Standard ISO 10993-5:2009, Biological evaluation of medical devices—Part 5: Tests for in vitro cytotoxicity, as previously described. 5 Biological reactivity was evaluated by light microscopy, following exposure to the samples for 24 ± 1 and 48 ± 1 h, and rated on a scale of 0–4 as described in ISO 10993-5:2009. The samples were dissolved in complete growth medium (MEM supplemented with 10% FBS, 4 mM l -glutamine, 0.11 mg/mL sodium-pyruvate, 100 IU/mL penicillin, and 100 μg/mL streptomycin) at the following concentrations: 20 mg/mL for HEC, b-HEC, q-HEC, BET HCl and Ch-Cl, and 2 mg/mL for c-HEC and b-PVA.…”

“…Cationic polymers are ubiquitous and extensively used in cosmetics (polyquaterniums) and other industries including medical research . Many cationic polymers based on amines or quaternary ammonium salts (QAS) may exhibit non-selective antimicrobial or other cytotoxic properties. − Generally, QAS and especially protonated (poly-) amines are expected to bind ionically to negatively charged cell membranes, causing their destabilization. This may lead to membrane rupture and consequently to cell penetration or lysis .…”

Acrylic thiazole copolymer microspheres with “cluster effect” for long‐lasting antimicrobial activity and plant growth promotion

“…24 The polymer antibacterial material (PAM) that is not easily leached and stable has become one of the most potential antibacterial agents. 25 However, the performance of PAM is limited by the phenomenon of agglomeration, and materials with special properties are aggregated, masking and burying the effective active sites, which greatly reduces the functional value of the material. However, the "cluster effect" perfectly reflects the value of each department.…”