The design and preparation of wound dressings that redress the protease imbalance in chronic wounds is an important goal of wound healing and medical materials science. Chronic wounds contain high levels of tissue and cytokine-destroying proteases including matrix metalloprotease and neutrophil elastase. Thus, the lowering of excessive protease levels in the wound environment by wound dressing sequestration prevents the breakdown of extracellular matrix proteins and growth factors necessary for wound healing. Phosphorylated cotton wound dressings were prepared to target sequestration of proteases from chronic wound exudate through a cationic uptake binding mechanism involving salt bridge formation of the positively charged amino acid side chains of proteases with the phosphate counterions of the wound dressing fiber. Dressings were prepared by applying sodium hexametaphosphate and diammonium phosphate in separate formulations to cotton gauze by pad/dry/cure methods. Phosphorylated cotton dressings were assessed for their ability to lower elastase and collagenase activity. The phosphorylated cotton dressings lowered elastase and collagenase activity 40-80% more effectively than the untreated cotton wound dressings under conditions that mimic chronic wound exudate. Efficacy of the phosphorylated cotton was found to be related to the level of phosphorylation and a lower pH due to protonated phosphate at the surface of the dressing. The capacity of the modified gauze to sequester continued elastase secretions similar to that found in a chronic wound over a 24-h period was retained within a 80% retention of elastase sequestration and was dose-dependent.

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Celluloses from a variety of common sources were analyzed for availabilities of O(2)H, O(3)H, and O(6)H in order to estimate the extent of hydrogen bonding on accessible fibrillar surfaces. Celluloses from flax, ramie, sisal, and wood (both cellulose I and II from wood) together with liquid NH3‐swollen cotton and NaOH‐swollen cotton (cellulose II) had relative availabilities similar to those of native cotton. Celluloses from Valonia centricosa and in rayon samples stood apart from each other and from the “cotton family.” The difference between Valonia and cotton celluloses appears to result, in addition to the accepted smaller, less perfect crystallites in cotton, from an O(2)H hydrogen bond which is likely the intramolecular bond between O(2)H and O(6′)H that is present in Valonia and absent in cotton. Rayon samples also showed evidence of similar bonds involving O(2)H on accessible surfaces. Since the regenerated rayons had relative availabilities different from those of mercerized cotton and wood cellulose samples, it is proposed that chain packing arrangements are not the same in these two types of cellulose II.

Cotton fabric is treated with a total cellulase preparation in either a Launderometer or a reciprocal water bath/shaker for times ranging from 10 to 180 minutes. Control fabrics, buffer-only treated fabrics, and cellulase-treated fabrics are analyzed by gel permeation chromatography to determine peak, weight-average, and number-average molecular weights. The GPC data are compared to weight loss and breaking load mea surements on control and treated fabrics. Despite considerable weight loss and breaking load reduction with the total cellulase treatment in the Launderometer, GPC analysis does not show a reduction in the molecular weight of the cellulose.

A phosphorylated form of cotton gauze for treatment of chronic wounds was designed to improve the wound dressing's capacity to remove harmful proteases from the wound and facilitate healing. Development of the fabric finishing chemistry of the wound dressing with a process suitable for textile mill production required adapting the stationary finishing chemistry of the cotton phosphorylation from a batch-type pad-dry-cure finishing treatment to a continuous pilot scale finishing process. Issues in optimizing the cotton finishing process took into consideration dressing sterilization, the effect of city water versus de-ionized water, retention of the fabric whiteness index and protease sequestration capacity of the dressing, which is the index of the dressing's efficacy. Three types of sterilization approaches were assessed, including gamma ray, ethylene oxide and steam sterilization to determine the effect of sterilization on the phosphorylated cotton dressing and the subsequent efficacy of the sterilized dressing to remove proteases from the wound. Two phosphorylation reagents were compared for their ability to phosphorylate cotton in a urea-based formulation and yield an active, effective dressing, with a high whiteness index. Phosphorylation with a diammonium phosphate (DAP) : urea formulation generally gave a more effective dressing as an active protease sequestrant, and phosphorylation with sodium hexametaphosphate (SMP) : urea gave a higher whiteness index. Finishing formulations combining the Downloaded from two phosphorylating reagents, DAP and SMP : urea, were assessed to improve both whiteness and efficacy. However, sterilization of DAP treated cotton with ethylene oxide eradicated activity through apparent masking of the cellulose phosphate hydroxyls. Side reactions that may occur during ethylene oxide treatment were discussed as the possible origin of the phosphate hydroxyl masking. On the other hand, sterilization with gamma irradiation produced significant yellowing of the dressing. A SMP : urea (16 : 30) formulation was employed in the continuous process finishing treatment, and found to be most optimal for whiteness, efficacy and ease of sterilization, when adapted to industrial scale production of the cotton chronic wound dressing.

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Recent developments in cellulose wound dressings targeted to different stages of wound healing have been based on structural and charge modifications that function to modulate events in the complex inflammatory and hemostatic phases of wound healing. Hemostasis and inflammation comprise two overlapping but distinct phases of wound healing wherein different dressing material properties are required to bring pathological events under control when they present as a result of trauma or chronic wounds. Thus, we have designed cellulose wound dressings with properties that function through modified fiber surface properties to lower protease levels in the chronic wound and promote clotting in hemorrhaging wounds. With this in mind three finishing chemistries utilizing traditional paddry-cure approaches were explored for their potential to confer charged properties to cotton dressings. Cellulose dressings designed to remove cationic serine proteases from highly exudative chronic wounds were created to present negatively charged fibers as an ion exchange mechanism of proteaselowering. Phosphorylated cotton and polycarboxylic acid crosslinked cotton were prepared to examine their ability to remove human neutrophil elastase (HNE) from surrogate wound fluid. A cellulose phosphorylation reaction utilizing sodium hexametaphosphate: urea was explored to optimize cellulose phosphorylation as a function of HNE sequestration efficacy. Acid catalyzed cross linking of cellulose with butane-tetracarboxylic acid also resulted in a negatively charged dressing that removed HNE from solution more effectively than phosphorylated cellulose. Collagenase sequestration was also assessed with phosphorylated cellulose and polycarboxylic acid cross linked cellulose derivatives. Butanetetracarboxylic acid and phosphorylated cellulose functioned to remove collagenase from solution most effectively. Cellulose dressings designed to accelerate thrombosis and aggregation of blood platelets were prepared with a view to examining derivatized cotton fibers bearing a net positive charge to promote hemostasis. Cellulose and chitosan dressings bearing an aminoglucan functionality were created by grafting chitosan on cotton and preparing aminized cotton. The preparation of chitosan-grafted cotton dressings was completed with a citric acid grafting onto cellulose. Aminized cotton was functionalized as an ethylamino-ether cellulose derivative. The chitosangrafted and aminized cotton demonstrated a dose response gelling of citrated sheep blood.