In vitro cellular viability studies on a concentrated surfactant-based wound dressing

Rui Chen, Anne-Marie Salisbury, Steven L Percival, Rui Chen, Anne-Marie Salisbury, Steven L Percival

Abstract

In this study, three cellular cytotoxic assays (direct contact assay, extraction assay, and cell insert assay) were applied to evaluate the effects of a concentrated surfactant gel preserved with antimicrobials and a concentrated surfactant gel with 1% silver sulfadiazine on both the mouse fibroblast cell line L929 and human dermal fibroblasts (HDFa). Also, the in vitro wound model was wounded by a 100 μL pipette tip and used to assess cell migration and wound closure after treatment with both gels. A needle-scratched membrane disruption model was used to preliminarily evaluate membrane stabilisation and the membrane-resealing effects of concentrated surfactant gels. It was demonstrated that the concentrated surfactant gel preserved with antimicrobials was not toxic to both L929 and HDFa. However, the concentrated surfactant gel with 1% silver sulfadiazine demonstrated a degree of cytotoxicity to both cell types. After treatment with a concentrated surfactant gel preserved with antimicrobials, cell movement to close the scratch gap was enhanced at 24 and 48 hours. The results also showed that cells treated with the concentrated surfactant gel preserved with antimicrobials decreased cell necrosis and improved cell resistance of the f-actin rearrangement after a needle scratch. The results demonstrated that a concentrated surfactant gel preserved with antimicrobials is non-cytotoxic and has ability to accelerate wound closure by enhancing cell mobility. Furthermore, the concentrated surfactant gel appeared to stabilise the plasma membrane and demonstrated a resealing ability and helped to retain the plasma membrane integrity and enhanced wound healing.

Keywords: cell membrane stabilisation and resealing; cytotoxicity; surfactant; wound closure; wound dressing.

© 2019 Medicalhelplines.com Inc and John Wiley & Sons Ltd.

Figures

Figure 1
Figure 1
Cell images after concentrated surfactant gel preserved with antimicrobials (CSG) and concentrated surfactant gel with 1% silver sulphadiazine (CSG‐SSD) treatment at day 1. Scale bar: 50 μm
Figure 2
Figure 2
Cell viability of L929 (left) and human dermal fibroblasts (HDFa) (right) in different cytotoxic tests after concentrated surfactant gel preserved with antimicrobials (CSG) and concentrated surfactant gel with 1% silver sulphadiazine (CSG‐SSD) treatment at day 1
Figure 3
Figure 3
Cell images after concentrated surfactant gel preserved with antimicrobials (CSG) and concentrated surfactant gel with 1% silver sulphadiazine (CSG‐SSD) treatment at day 7. Scale bar: 50 μm
Figure 4
Figure 4
Cell viability of L929 (left) and human dermal fibroblasts (HDFa) (right) in different cytotoxic tests after concentrated surfactant gel preserved with antimicrobials and concentrated surfactant gel with 1% silver sulphadiazine treatment at day 7. **P < 0.01 vs. Control by one‐way analysis of variance
Figure 5
Figure 5
Representative L929 cell images after tip scratch with/without pre‐treatment with concentrated surfactant gel preserved with antimicrobials at 0, 24 and 48 hours. Scale bar: 50 μm
Figure 6
Figure 6
Representative human dermal fibroblast (HDF) images after tip scratch with/without pre‐treatment with concentrated surfactant gel preserved with antimicrobials at 0, 24, and 48 hours. Scale bar: 50 μm
Figure 7
Figure 7
Live and needle‐scratch killed L929 cells and human dermal fibroblasts (HDFa) stained with the LIVE/DEAD cell viability/cytotoxicity assay kit L3224. Live cells fluorescence bright green, whereas dead cells with disrupted membranes fluoresce red‐orange. Scale bar: 50 μm
Figure 8
Figure 8
LDH in supernatants versus the total LDH released by HDFa. LDH Wound dressing gels, Plurogel PSSD increase LDH release of HDFa cells. The figure showed LDH released in supernatants vs. the total LDH released by the cells on the coverslips and in supernatants. P < 0.05 vs. CSG; **P < 0.01 versus control by one way analysis of variance
Figure 9
Figure 9
Confocal imaging of needle‐scratched L929 cells and HDFa after rapid fixed, actin was stained using the ActinGreenTM 488 ReadyProbes reagent. Scale bar: 50 μm

References

    1. Percival SL, Chen R, Mayer D, Salisbury AM. Mode of action of poloxamer‐based surfactants in wound care and efficacy on biofilms. Int Wound J. 2018;15:749‐755.
    1. Lipsky BA, Hoey C. Topical antimicrobial therapy for treating chronic wounds. Clin Infect Dis. 2009;49(10):1541‐1549.
    1. Roberts CD, Leaper DJ, Assadian O. The role of topical antiseptic agents within antimicrobial stewardship strategies for prevention and treatment of surgical site and chronic open wound infection. Adv Wound Care. 2017;6(2):63‐71.
    1. Ip M, Lui SL, Poon VK, Lung I, Burd A. Antimicrobial activities of silver dressings: an in vitro comparison. J Med Microbiol. 2006;55(Pt 1:59‐63.
    1. Percival SL, Mayer D, Salisbury AM. Efficacy of a surfactant‐based wound dressing on biofilm control. Wound Repair Regen. 2017;25(5):767‐773.
    1. Percival SL, Thomas J, Linton S, Okel T, Corum L, Slone W. The antimicrobial efficacy of silver on antibiotic‐resistant bacteria isolated from burn wounds. Int Wound J. 2012;9(5):488‐493.
    1. Dover R, Otto WR, Nanchahal J, Riches DJ. Toxicity testing of wound dressing materials in vitro. Br J Plast Surg. 1995;48(4):230‐235.
    1. Hajska M, Dragunova J, Koller J. Cytotoxicity testing of burn wound dressings: first results. Cell Tissue Bank. 2017;18(2):143‐151.
    1. Kempf M, Kimble RM, Cuttle L. Cytotoxicity testing of burn wound dressings, ointments and creams: a method using polycarbonate cell culture inserts on a cell culture system. Burns. 2011;37(6):994‐1000.
    1. Paddle‐Ledinek JE, Nasa Z, Cleland HJ. Effect of different wound dressings on cell viability and proliferation. Plast Reconstr Surg. 2006;117(7 Suppl):110S‐118S. discussion 9S‐20S.
    1. Demirci S, Dogan A, Karakus E, et al. Boron and Poloxamer (F68 and F127) containing hydrogel formulation for burn wound healing. Biol Trace Elem Res. 2015;168(1):169‐180.
    1. Harries FJ, Begg PA. Non‐rinse skin cleansers: the way forward in preventing incontinence related moisture lesions? J Wound Care. 2016;25(5):268‐276.
    1. Metcalf D, Parsons D, Bowler IP. Development of a next‐generation antimicrobial wound dressing. Acta Med Croatica. 2016;70(1):49‐56.
    1. Palumbo FP, Harding KG, Abbritti F, et al. New surfactant‐based dressing product to improve wound closure rates of nonhealing wounds: a European multicenter study including 1036 patients. Wounds. 2016;28(7):233‐240.
    1. Mayer D, Armstrong D, Schultz G, et al. Cell salvage in acute and chronic wounds: a potential treatment strategy. Experimental data and early clinical results. J Wound Care. 2018;27(9):594‐605.
    1. Yang QP, Larose C, Della Porta AC, Schultz GS, Gibson DJ. A surfactant‐based wound dressing can reduce bacterial biofilms in a porcine skin explant model. Int Wound J. 2017;14(2):408‐413.
    1. Fowler EB, Cuenin MF, Hokett SD, et al. Evaluation of pluronic polyols as carriers for grafting materials: study in rat calvaria defects. J Periodontol. 2002;73(2):191‐197.
    1. Kant V, Gopal A, Kumar D, et al. Topical pluronic F‐127 gel application enhances cutaneous wound healing in rats. Acta Histochem. 2014;116(1):5‐13.
    1. Dunnill CW, Parkin IP. Antimicrobial coatings for 'self‐sterilisation'. Sterilisation of Biomaterials and Medical Devices. Cambridge, UK: Woodhead Publishing Limited; 2012:240‐260.
    1. Gabbiani G, Gabbiani F, Heimark RL, Schwartz SM. Organization of actin cytoskeleton during early endothelial regeneration in vitro. J Cell Sci. 1984;66(Mar):39‐50.
    1. Monsuur HN, Boink MA, Weijers EM, et al. Methods to study differences in cell mobility during skin wound healing in vitro. J Biomech. 2016;49(8):1381‐1387.
    1. Lin A, Hokugo A, Nishimura I. Wound closure and wound management a new therapeutic molecular target. Cell Adh Migr. 2010;4(3):396‐399.
    1. Padanilam JT, Bischof JC, Lee RC, et al. Effectiveness of poloxamer‐188 in arresting calcein leakage from thermally damaged isolated skeletal‐muscle cells. Ann N Y Acad Sci. 1994;720:111‐123.
    1. Merchant FA, Holmes WH, Capelli‐Schellpfeffer M, Lee RC, Toner M. Poloxamer 188 enhances functional recovery of lethally heat‐shocked fibroblasts. J Surg Res. 1998;74(2):131‐140.
    1. Swanson JA, McNeil PL. Nuclear reassembly excludes large macromolecules. Science. 1987;238(4826):548‐550.
    1. Miyake K, McNeil PL, Suzuki K, Tsunoda R, Sugai N. An actin barrier to resealing. J Cell Sci. 2001;114(Pt 19:3487‐3494.

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