Chitosan/PVA/Doxycycline Film and Nanofber Accelerate Diabetic Wound Healing in a Rat Model

Document Type : Research article

Authors

1 Department of Physiology and Pharmacology, School of Medicine, Alborz University of Medical Sciences, Karaj, Iran. Cardiovascular Research Center, Alborz University of Medical Sciences, Karaj, Iran.

2 Polymer and Color Engineering Department Amirkabir University of Technology (Tehran Polytechnic) Tehran, Iran.

3 Department of Biology and Anatomical Sciences, School of Medicine, ShahidBeheshtiUniversity of Medical Sciences, Tehran,Iran.

4 Department of Immunology, School of Medicine, Shahid BeheshtiUniversity of Medical Sciences, Tehran, Iran.

5 Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

6 Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran,Iran.

7 Department of Immunology, Faculty of Medicine, Ahvaz Jundishapur University of Medical Sciences, Ahvaz, Iran.

Abstract

In this study, we evaluated the effects of nanofiber and film polymers with doxycycline for treating a wound in a diabetic rat model. 108 male rats were divided into six groups, the control group, the diabetic control, and the groups were diabetic rats receiving different wound dressing. At the 3rd, 7th, and 14th days, macroscopic/histologic imaging and tissue sampling were performed. Tissues were analyzed for IL-1β, TNF-α, IL-10, TIMP-1, and MMP-2 by using ELISA. Dressings of chitosan, polyvinyl alcohol and doxycycline increased the rate of wound closure, the volume of collagen, dermal, and epidermis; in addition, it increased the number of fibroblasts and basal cell epidermis cells, vascular length, and decreased the number of neutrophil cells. Inflammatory cytokines and MMP-2 were decreased, and anti-inflammatory IL-10 and TIMP-1 were increased. It was ultimately attained that the combination of chitosan/ polyvinyl alcohol /doxycycline could be a useful dressing for the healing of diabetic wounds.

Graphical Abstract

Chitosan/PVA/Doxycycline Film and Nanofber Accelerate Diabetic Wound Healing in a Rat Model

Keywords


(1) Kolarsick PA, Kolarsick MA and Goodwin C. Anatomy and physiology of the skin. J. Dermatol. Nurses Assoc. (2011) 3: 203-13.
(2) Moura LI, Dias AM, Carvalho E and de Sousa HC. Recent advances on the development of wound dressings for diabetic foot ulcer treatment—a review. Acta Biomaterialia. (2013) 9: 7093-114.
(3) Berlanga Acosta J, Fernández Montequín J, Valdés Pérez C, Savigne Gutiérrez W, Mendoza Marí Y, García-Ojalvo A, Falcón Cama V, García del Barco Herrera D, Fernández Mayola M, Pérez Saad H, Pimentel Vázquez E, Urquiza Rodríguez A, Kulikovsky M and Guillén-Nieto G. Diabetic Foot Ulcers and Epidermal Growth Factor: Revisiting the Local Delivery Route for a Successful Outcome. BioMed. Res. Int. (2017) 2017: 1-10.
(4) McCarty SM and Percival SL. Proteases and Delayed Wound Healing. Adv. Wound Care (New Rochelle). (2013) 2: 438-47.
(5) Ayuk SM, Abrahamse H and Houreld NN. The Role of Matrix Metalloproteinases in Diabetic Wound Healing in relation to Photobiomodulation. J. Diabetes Res. (2016) 2016: 1-9.
(6) Muller M, Trocme C, Lardy B, Morel F, Halimi S and Benhamou PY. Matrix metalloproteinases and diabetic foot ulcers: the ratio of MMP‐1 to TIMP‐1 is a predictor of wound healing. Diabetic Med. (2008) 25: 419-26.
(7) Gill SE and Parks WC. Metalloproteinases and their inhibitors: regulators of wound healing. Int. J. Biochem. Cell Biol. (2008) 40: 1334-47.
(8) Martins VL, Caley M and O'Toole EA. Matrix metalloproteinases and epidermal wound repair. Cell Tissue Res. (2013) 351: 255-68.
(9) Peranteau WH, Zhang L, Muvarak N, Badillo AT, Radu A, Zoltick PW and Liechty KW. IL-10 overexpression decreases inflammatory mediators and promotes regenerative healing in an adult model of scar formation. J. Invest. Dermatol. (2008) 128: 1852-60.
(10) Stearns ME, Rhim J and Wang M. Interleukin 10 (IL-10) inhibition of primary human prostate cell-induced angiogenesis: IL-10 stimulation of tissue inhibitor of metalloproteinase-1 and inhibition of matrix metalloproteinase (MMP)-2/MMP-9 secretion. Clin. Cancer Res. (1999) 5: 189-96.
(11) Yao K, Li J, Yao F and Yin Y. Chitosan-based hydrogels: functions and applications, CRC Press2011.
(12) Ali A and Ahmed S. A review on chitosan and its nanocomposites in drug delivery. Int. J. Biol. Macromol. (2018) 109: 273-86.
(13) Dai T, Tanaka M, Huang YY and Hamblin MR. Chitosan preparations for wounds and burns: antimicrobial and wound-healing effects. Expert. Rev. Anti. Infect. Ther. (2011) 9: 857-79.
(14) Sarmento B and das Neves J. Chitosan-based systems for biopharmaceuticals: delivery, targeting and polymer therapeutics, John Wiley & Sons (2012).
(15) Minagawa T, Okamura Y, Shigemasa Y, Minami S and Okamoto Y. Effects of molecular weight and deacetylation degree of chitin/chitosan on wound healing. Carbohydr. Polym. (2007) 67: 640-44.
(16) Alsarra I A. Chitosan topical gel formulation in the management of burn wounds. Int. J. Biol. Macromol. (2009) 45: 16-21.
(17) Moeini A, Pedram P, Makvandi P, Malinconico M and Gomez d'Ayala G. Wound healing and antimicrobial effect of active secondary metabolites in chitosan-based wound dressings: A review. Carbohydr. Polym. (2020) 233: 115839.
(18) Baker MI, Walsh SP, Schwartz Z and Boyan BD. A review of polyvinyl alcohol and its uses in cartilage and orthopedic applications. J. Biomed. Mater. Res. B Appl. Biomater. (2012) 100: 1451-7.
(19) Shanmuganathan S, Shanumugasundaram N, Adhirajan N, Lakshmi TR and Babu M. Preparation and characterization of chitosan microspheres for doxycycline delivery. Carbohydr. Polym. (2008) 73: 201-11.
(20) Tort S, Acarturk F and Besikci A. Evaluation of three-layered doxycycline-collagen loaded nanofiber wound dressing. Int. J. Pharm. (2017) 529: 642-53.
(21) Sapadin A N and Fleischmajer R. Tetracyclines: nonantibiotic properties and their clinical implications. J. Am. Acad. Dermatol. (2006) 54: 258-65.
(22) Federici T J. The non-antibiotic properties of tetracyclines: clinical potential in ophthalmic disease. Pharmacol. Res. (2011) 64: 614-23.
(23) Gu Y, Lee HM, Sorsa T, Salminen A, Ryan ME, Slepian MJ and Golub LM. Non-antibacterial tetracyclines modulate mediators of periodontitis and atherosclerotic cardiovascular disease: a mechanistic link between local and systemic inflammation. Pharmacol. Res. (2011) 64: 573-79.
(24) Tilakaratne A and Soory M. Anti-inflammatory Actions of Adjunctive Tetracyclines and Other Agents in Periodontitis and Associated Comorbidities. Open Dent. J. (2014) 8: 109-24.
(25) Zehtabi F, Ispas-Szabo P, Djerir D, Sivakumaran L, Annabi B, Soulez G, Mateescu M A and Lerouge S. Chitosan-doxycycline hydrogel: An MMP inhibitor/sclerosing embolizing agent as a new approach to endoleak prevention and treatment after endovascular aneurysm repair. Acta Biomater. (2017) 64: 94-105.
(26) Solomon A, Rosenblatt M, Li DQ, Liu Z, Monroy D, Ji Z, Lokeshwar BL and Pflugfelder SC. Doxycycline inhibition of interleukin-1 in the corneal epithelium. Invest. Ophthalmol. Vis. Sci. (2000) 41: 2544-57.
(27) Bernardino AL, Kaushal D and Philipp MT. The antibiotics doxycycline and minocycline inhibit the inflammatory responses to the Lyme disease spirochete Borrelia burgdorferi. J. Infect. Dis. (2009) 199: 1379-88.
(28) Jantzie LL and Todd KG. Doxycycline inhibits proinflammatory cytokines but not acute cerebral cytogenesis after hypoxia–ischemia in neonatal rats. J. Psychiatry Neurosci. (2010) 35: 20-32.
(29) Chew SY, Wen Y, Dzenis Y and Leong KW. The role of electrospinning in the emerging field of nanomedicine. Curr. Pharm. Des. (2006) 12: 4751-70.
(30) Bhardwaj N and Kundu SC. Electrospinning: a fascinating fiber fabrication technique. Biotechnol. Adv. (2010) 28: 325-47.
(31) Hedayatyanfard K, Bagheri Khoulenjani S, Hashemi A and Ziai S A. Semi-IPN Films and Electrospun Nanofibers Based on Chitosan/PVA as an Antibacterial Wound Dressing. Iran. J. Pharm. Res. (2019) 18: 1156-67.
(32) Gundersen H, Bendtsen TF, Korbo L, Marcussen N, Møller A, Nielsen K, Nyengaard J, Pakkenberg B, Sørensen F B and Vesterby A. Some new, simple and efficient stereological methods and their use in pathological research and diagnosis. Apmis (1988) 96: 379-94.
(33) Mühlfeld C, Nyengaard JR and Mayhew TM. A review of state-of-the-art stereology for better quantitative 3D morphology in cardiac research. Cardiovas Pathol. (2010) 19: 65-82.
(34) Velnar T, Bailey T and Smrkolj V. The wound healing process: an overview of the cellular and molecular mechanisms. J. Int. Med. Res. (2009) 37: 1528-42.
(35) Guo S and Dipietro L A. Factors affecting wound healing. J. Dent. Res. (2010) 89: 219-29.
(36) Adhirajan N, Shanmugasundaram N, Shanmuganathan S and Babu M. Collagen-based wound dressing for doxycycline delivery: in-vivo evaluation in an infected excisional wound model in rats. J. Pharm. Pharmacol. (2009) 61: 1617-23.
(37) Moore AL, desJardins-Park HE, Duoto BA, Mascharak S, Murphy MP, Irizarry DM, Foster DS, Jones RE, Barnes LA, Marshall CD, Ransom RC, Wernig G and Longaker M T. Doxycycline Reduces Scar Thickness and Improves Collagen Architecture. Ann. Surg. (2020) 272: 183-93.
(38) Tian M, Qing C, Niu Y, Dong J, Cao X, Song F, Ji X and Lu S. Aminoguanidine cream ameliorates skin tissue microenvironment in diabetic rats. Arch. Med. Sci. (2016) 12: 179-87.
(39) Tian M, Qing C, Niu Y, Dong J, Cao X, Song F, Ji X and Lu S. Effect of aminoguanidine intervention on neutrophils in diabetes inflammatory cells wound healing. Exp. Clin. Endocrinol. Diabetes (2013) 121: 635-42.
(40) Tian M, Qing C, Niu Y, Dong J, Cao X, Song F, Ji X and Lu S. The relationship between inflammation and impaired wound healing in a diabetic rat burn model. J. Burn Care Res. (2016) 37: e115-e24.
(41) Lerman OZ, Galiano RD, Armour M, Levine J P and Gurtner G C. Cellular dysfunction in the diabetic fibroblast: impairment in migration, vascular endothelial growth factor production, and response to hypoxia. Am. J. Pathol. (2003) 162: 303-12.
(42) Okonkwo U A and DiPietro L A. Diabetes and Wound Angiogenesis, Int. J. Mol. Sci. (2017) 18: 1419.
(43) Mirza RE, Fang MM, Ennis WJ and Koh TJ. Blocking interleukin-1β induces a healing-associated wound macrophage phenotype and improves healing in type 2 diabetes. Diabetes (2013) 62: 2579-87.
(44) Han L, Su W, Huang J, Zhou J, Qiu S and Liang D. Doxycycline inhibits inflammation-induced lymphangiogenesis in mouse cornea by multiple mechanisms. PLoS One (2014) 9: e108931.
(45) Valentín S, Morales A, Sánchez J L and Rivera A. Safety and efficacy of doxycycline in the treatment of rosacea. Clin. Cosmet. Investig. Dermatol. (2009) 2: 129-40.
(46) Castro JE, Vado Solis I, Perez Osorio C and Fredeking TM. Modulation of cytokine and cytokine receptor/antagonist by treatment with doxycycline and tetracycline in patients with dengue fever. Clin. Dev. Immunol. (2011) 2011: 370872.
(47) Xu F, Zhang C and Graves D T. Abnormal cell responses and role of TNF-in impaired diabetic wound healing. Biomed. Res. Int. (2013) 2013.
(48) Goren I, Müller E, Schiefelbein D, Christen U, Pfeilschifter J, Mühl H and Frank S. Systemic anti-TNFα treatment restores diabetes-impaired skin repair in ob/ob mice by inactivation of macrophages. J. Invest. Dermatol. (2007) 127: 2259-67.
(49) Di Caprio R, Lembo S, Di Costanzo L, Balato A and Monfrecola G. Anti-inflammatory properties of low and high doxycycline doses: an in-vitro study. Mediators. Inflamm. (2015) 2015: 329418.
(50) Sun J, Shigemi H, Tanaka Y, Yamauchi T, Ueda T and Iwasaki H. Tetracyclines downregulate the production of LPS-induced cytokines and chemokines in THP-1 cells via ERK, p38, and nuclear factor-κB signaling pathways. Biochem. Biophys. Reports. (2015) 4: 397-404.
(51) Mostafa Mtairag E, Chollet Martin S, Oudghiri M, Laquay N, Jacob MP, Michel JB and Feldman LJ. Effects of interleukin-10 on monocyte/endothelial cell adhesion and MMP-9/TIMP-1 secreti. Cardiovasc. Res. (2001) 49: 882-90.
(52) Silvestre JS, Mallat Z, Tamarat R, Duriez M, Tedgui A and Levy BI. Regulation of matrix metalloproteinase activity in ischemic tissue by interleukin-10: role in ischemia-induced angiogenesis. Circ. Res. (2001) 89: 259-64.
(53) Bostanci N, Akgul B, Tsakanika V, Allaker RP, Hughes FJ and McKay IJ. Effects of low-dose doxycycline on cytokine secretion in human monocytes stimulated with Aggregatibacter actinomycetemcomitans. Cytokine (2011) 56: 656-61.
(54) Li Z, Guo S, Yao F, Zhang Y and Li T. Increased ratio of serum matrix metalloproteinase-9 against TIMP-1 predicts poor wound healing in diabetic foot ulcers, J. Diabetes Complications. (2013) 27: 380-2.
(55) Liu Y, Min D, Bolton T, Nubé V, Twigg S M, Yue D K and McLennan S V. Increased matrix metalloproteinase-9 predicts poor wound healing in diabetic foot ulcers. Diabetes Care (2009) 32: 117-19.
(56) Zhao S, Choksuchat C, Zhao Y, Ballagh SA, Kovalevsky GA and Archer DF. Effects of doxycycline on serum and endometrial levels of MMP-2, MMP-9 and TIMP-1 in women using a levonorgestrel-releasing subcutaneous implant. Contraception (2009) 79: 469-78.
(57) Gabriele S, Buchanan B, Kundu A, Dwyer HC, Gabriele J P, Mayer P and Baranowski DC. Stability, Activity, and Application of Topical Doxycycline Formulations in a Diabetic Wound Case Study. Wounds (2019) 31: 49-54.
(58) Choi DH, Moon IS, Choi BK, Paik JW, Kim YS, Choi SH and Kim CK. Effects of sub-antimicrobial dose doxycycline therapy on crevicular fluid MMP-8, and gingival tissue MMP-9, TIMP-1 and IL-6 levels in chronic periodontitis. J. Periodontal. Res. (2004) 39: 20-6.
(59) Qing C. The molecular biology in wound healing & non-healing wound. Chin. J. Traumatol. (2017) 20: 189-93.
(60) Serra R, Gallelli L, Buffone G, Molinari V, Stillitano D M, Palmieri C and de Franciscis S. Doxycycline speeds up healing of chronic venous ulcers. Int. Wound J. (2015) 12: 179-84.