Chitosan hydrogel encapsulated with LL-37 peptide promotes deep tissue injury healing in a mouse model

Xu Yang; Jing-Lin Guo; Jing Han; Rui-Juan Si; Pan-Pan Liu; Zi-Rui Zhang; Ai-Min Wang; Ju Zhang

doi:10.1186/s40779-020-00249-5

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Chitosan hydrogel encapsulated with LL-37 peptide promotes deep tissue injury healing in a mouse model

RESEARCH | Updated：2023-06-05

- Chitosan hydrogel encapsulated with LL-37 peptide promotes deep tissue injury healing in a mouse model
- Military Medical Research Vol. 7, Issue 4, Pages: 409-417(2020)
- Affiliations：
  
  School of Nursing, Qingdao University, Qingdao 266021, China
- Author bio：
  
  * zhangju111@qdu.edu.cn
- Funds：
- DOI：10.1186/s40779-020-00249-5
  CLC：
- Published：2020-12
- Accepted：
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Yang et al.: Chitosan hydrogel encapsulated with LL-37 peptide promotes deep tissue injury healing in a mouse model. Mil Med Res, 2020, 7: 20.
DOI：

Yang et al.: Chitosan hydrogel encapsulated with LL-37 peptide promotes deep tissue injury healing in a mouse model. Mil Med Res, 2020, 7: 20. DOI： 10.1186/s40779-020-00249-5.

摘要

Abstract

Background:

LL-37 peptide is a member of the human cathelicidin family

and has been shown to promote the healing of pressure ulcers. However

the low stability of this peptide within the wound environment limits its clinical use. Chitosan (CS) hydrogel is commonly used as a base material for wound dressing material.

Methods:

CS hydrogel (2.5% w/v) was encapsulated with LL-37. Cytotoxicity of the product was examined in cultured NIH3T3 fibroblasts. Effects on immune response was examined by measuring tumor necrosis factor-α (TNF-α) release from RAW 264.7 macrophages upon exposure to lipopolysaccharides. Antibacterial activity was assessed using Staphylococcus aureus. Potential effect on pressure ulcers was examined using a mouse model. Briefly

adult male C57BL/6 mice were subjected to skin pressure using magnets under a 12/12 h schedule for 21 days. Mice were randomized to receive naked LL-37 (20 μg)

chitosan gel containing 20 μg LL-37 (LL-37/CS hydrogel) or hydrogel alone under the ulcer bed (

=6). A group of mice receiving no intervention was also included as a control.

Results:

LL-37/CS hydrogel did not affect NIH3T3 cell viability. At a concentration of 1–5 μg/ml

LL-37/CS inhibited TNF-α release from macrophage. At 5 μg/ml

LL-37/CS inhibited the growth of

Staphylococcus aureus

. The area of the pressure ulcers was significantly lower in mice receiving LL-37/CS hydrogel in comparison to all other 3 groups on days 11 (84.24%±0.25%)

13 (56.22%±3.91%) and 15 (48.12%±0.28%). Histological examination on days 15 and 21 showed increased epithelial thickness and density of newly-formed capillary with naked LL-37 and more so with LL-37/CS. The expression of key macromolecules in the process of angiogenesis (i.e.

hypoxia inducible factor-1α(HIF-1α) and vascular endothelial growth factor-A (VEGF-A)) in wound tissue was increased at both the mRNA and protein levels.

Conclusion:

Chitosan hydrogel encapsulated with LL-37 is biocompatible and could promote the healing of pressure ulcers.

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references

Artico M , Dante A , D'Angelo D , Lamarca L , Mastroianni C , Petitti T , et al . Prevalence, incidence and associated factors of pressure ulcers in home palliative care patients: a retrospective chart review . Palliat Med . 2018 ; 32 ( 1 ): 299-307 .

Demarré L , van Lancker A , van Hecke A , Verhaeghe S , Grypdonck M , Lemey J , et al . The cost of prevention and treatment of pressure ulcers: a systematic review . Int J Nurs Stud . 2015 ; 52 ( 11 ): 1754-74 .

Coyer F , Miles S , Gosley S , Fulbrook P , Sketcher-Baker K , Cook JL , et al . Pressure injury prevalence in intensive care versus non-intensive care patients: a state-wide comparison . Aust Crit Care . 2017 ; 30 ( 5 ): 244-50 .

Zhao R , Liang H , Clarke E , Jackson C , Xue M . Inflammation in chronic wounds . Int J Mol Sci . 2016 ; 17 ( 12 ): 2085 .

Peart J . The aetiology of deep tissue injury: a literature review . Br J Nurs . 2016 ; 25 ( 15 ): 840-3 .

Atkinson RA , Cullum NA . Interventions for pressure ulcers: a summary of evidence for prevention and treatment . Spinal Cord . 2018 ; 56 ( 3 ): 186-98 .

Espejo E , Andrés M , Borrallo RM , Padilla E , Garcia-Restoy E , Bella F . Bacteremia associated with pressure ulcers: a prospective cohort study . Eur J Clin Microbiol Infect Dis . 2018 ; 37 ( 5 ): 969-75 .

Lowry D , Saeed M , Narendran P , Tiwari A . The difference between the healing and the nonhealing diabetic foot ulcer: a review of the role of the microcirculation . J Diabetes Sci Technol . 2017 ; 11 ( 5 ): 914-23 .

Xhindoli D , Pacor S , Benincasa M , Scocchi M , Gennaro R , Tossi A . The human cathelicidin LL-37--a pore-forming antibacterial peptide and hostcell modulator . Biochim Biophys Acta . 2016 ; 1858 ( 3 ): 546-66 .

Fumakia M , Ho EA . Nanoparticles encapsulated with LL37 and serpin A1 promotes wound healing and synergistically enhances antibacterial activity . Mol Pharm . 2016 ; 13 ( 7 ): 2318-31 .

Agier J , Brzezińska-Błaszczyk E , Żelechowska P , Wiktorska M , Pietrzak J , Różalska S . Cathelicidin LL-37 affects surface and intracellular toll-like receptor expression in tissue mast cells . J Immunol Res . 2018 ; 2018 : 7357162 .

Adase CA , Borkowski AW , Zhang LJ , Williams MR , Sato E , Sanford JA , et al . Non-coding double-stranded RNA and antimicrobial peptide LL-37 induce growth factor expression from keratinocytes and endothelial cells . J Biol Chem . 2016 ; 291 ( 22 ): 11635-46 .

Hou S , Sun X , Dong X , Lin H , Tang L , Xue M , et al . Chlamydial plasmid encoded virulence factor Pgp3 interacts with human cathelicidin peptide LL-37 to modulate immune response . Microbes Infect . 2019 ; 21 ( 1 ): 50-5 .

Steinstraesser L , Lam MC , Jacobsen F , Porporato PE , Chereddy KK , Becerikli M , et al . Skin electroporation of a plasmid encoding hCAP-18/LL-37 host defense peptide promotes wound healing . Mol Ther . 2014 ; 22 ( 4 ): 734-42 .

Grönberg A , Mahlapuu M , Ståhle M , Whately-Smith C , Rollman O . Treatment with LL-37 is safe and effective in enhancing healing of hard-to-heal venous leg ulcers: a randomized, placebo-controlled clinical trial . Wound Repair Regen . 2014 ; 22 ( 5 ): 613-21 .

Garcia-Orue I , Gainza G , Girbau C , Alonso R , Aguirre JJ , Pedraz JL , et al . LL37 loaded nanostructured lipid carriers (NLC): a new strategy for the topical treatment of chronic wounds . Eur J Pharm Biopharm . 2016 ; 108 : 310-6 .

Boateng J , Catanzano O . Advanced therapeutic dressings for effective wound healing--a review . J Pharm Sci . 2015 ; 104 ( 11 ): 3653-80 .

Desfrançois C , Auzély R , Texier I . Lipid nanoparticles and their hydrogel composites for drug delivery: a review . Pharmaceuticals(Basel) . 2018 ; 11 ( 4 ): E118 .

Das S , Baker AB . Biomaterials and nanotherapeutics for enhancing skin wound healing . Front Bioeng Biotechnol . 2016 ; 4 : 82 .

Naseri-Nosar M , Ziora ZM . Wound dressings from naturally-occurring polymers: a review on homopolysaccharide-based composites . Carbohydr Polym . 2018 ; 189 : 379-98 .

Gao J , Zheng W , Zhang J , Guan D , Yang Z , Kong D , et al . Enzyme controllable delivery of nitric oxide from a molecular hydrogel . Chem Commun (Camb) . 2013 ; 49 ( 80 ): 9173-5 .

Cong Z , Shi Y , Wang Y , Wang Y , Niu J , Chen N , et al . A novel controlled drug delivery system based on alginate hydrogel/chitosan micelle composites . Int J Biol Macromol . 2018 ; 107 ( Pt A ): 855-64 .

Frade ML , de Annunzio SR , Calixto GMF , Victorelli FD , Chorilli M , Fontana CR . Assessment of chitosan-based hydrogel and photodynamic inactivation against Propionibacterium acnes . Molecules . 2018 ; 23 ( 2 ): E473 .

Xu J , Tam M , Samaei S , Lerouge S , Barralet J , Stevenson MM , et al . Mucoadhesive chitosan hydrogels as rectal drug delivery vessels to treat ulcerative colitis . Acta Biomater . 2017 ; 48 : 247-57 .

Karri VV , Kuppusamy G , Talluri SV , Mannemala SS , Kollipara R , Wadhwani AD , et al . Curcumin loaded chitosan nanoparticles impregnated into collagen-alginate scaffolds for diabetic wound healing . Int J Biol Macromol . 2016 ; 93 ( Pt B ): 1519-29 .

Vignesh S , Sivashanmugam A , Annapoorna M , Janarthanan R , Subramania I , Nair Shantikumar V , et al . Injectable deferoxamine nanoparticles loaded chitosan-hyaluronic acid coacervate hydrogel for therapeutic angiogenesis . Colloids Surf B Biointerfaces . 2018 ; 161 : 129-38 .

Chen X , Cao X , Jiang H , Che X , Xu X , Ma B , et al . SIKVAV-modified chitosan hydrogel as a skin substitutes for wound closure in mice . Molecules . 2018 ; 23 ( 10 ): E2611 .

Furuike T , Komoto D , Hashimoto H , Tamura H . Preparation of chitosan hydrogel and its solubility in organic acids . Int J Biol Macromol . 2017 ; 104 ( Pt B ): 1620-5 .

Sun Y , Liu Y , Liu W , Lu C , Wang L . Chitosan microparticles ionically crosslinked with poly(γ-glutamic acid) as antimicrobial peptides and nitric oxide delivery systems . Biochem Eng J . 2015 ; 95 : 78-85 .

Gopal A , Kant V , Gopalakrishnan A , Tandan SK , Kumar D . Chitosan-based copper nanocomposite accelerates healing in excision wound model in rats . Eur J Pharmacol . 2014 ; 731 : 8-19 .

Cui FF , Pan YY , Xie HH , Wang XH , Shi HX , Xiao J , et al . Pressure combined with ischemia/reperfusion injury induces deep tissue injury via endoplasmic reticulum stress in a rat pressure ulcer model . Int J Mol Sci . 2016 ; 17 ( 3 ): 284 .

Makrantonaki E , Wlaschek M , Scharffetter-Kochanek K . Pathogenesis of wound healing disorders in the elderly . J Dtsch Dermatol Ges . 2017 ; 15 ( 3 ): 255-75 .

Arpino JM , Nong Z , Li F , Yin H , Ghonaim N , Milkovich S , et al . Four-dimensional microvascular analysis reveals that regenerative angiogenesis in ischemic muscle produces a flawed microcirculation . Circ Res . 2017 ; 120 ( 9 ): 1453-65 .

Al Salihi MO , Kobayashi M , Tamari K , Miyamura T , Takeuchi K . Tumor necrosis factor-alpha antagonist suppresses local inflammatory reaction and facilitates olfactory nerve recovery following injury . Auris Nasus Larynx . 2017 ; 44 ( 1 ): 70-8 .

Hirosaki H , Maeda Y , Shimojima M , Maeda K , Iwata H , Takeyoshi M . Effects of soluble tumor necrosis factor (TNF) on antibody-dependent cellular cytotoxicity of therapeutic anti-TNF-alpha antibody . Immunol Investig . 2019 ; 48 ( 5 ): 441-50 .

Jordan SC , Choi J , Kim I , Wu G , Toyoda M , Shin B , et al . Interleukin-6, a cytokine critical to mediation of inflammation, autoimmunity and allograft rejection: therapeutic implications of IL-6 receptor blockade . Transplantation . 2017 ; 101 ( 1 ): 32-44 .

Yang G , Gu M , Chen W , Liu W , Xiao Y , Wang H , et al . SPHK-2 promotes the particle-induced inflammation of RAW264.7 by maintaining consistent expression of TNF-α and IL-6 . Inflammation . 2018 ; 41 ( 4 ): 1498-507 .

Yu X , Quan J , Long W , Chen H , Wang R , Guo J , et al . LL-37 inhibits LPS induced inflammation and stimulates the osteogenic differentiation of BMSCs via P2X7 receptor and MAPK signaling pathway . Exp Cell Res . 2018 ; 372 ( 2 ): 178-87 .

Sun L , Wang W , Xiao W , Yang H . The roles of cathelicidin LL-37 in inflammatory bowel disease . Inflamm Bowel Dis . 2016 ; 22 ( 8 ): 1986-91 .

Zhao X , Liu HQ , Li J , Liu XL . Endothelial progenitor cells promote tumor growth and progression by enhancing new vessel formation . Oncol Lett . 2016 ; 12 ( 2 ): 793-9 .

Jia J , Ye T , Cui P , Hua Q , Zeng H , Zhao D . AP-1 transcription factor mediates VEGF-induced endothelial cell migration and proliferation . Microvasc Res . 2016 ; 105 : 103-8 .

Park K , Lee HE , Lee SH , Lee D , Lee T , Lee YM . Molecular and functional evaluation of a novel HIF inhibitor, benzopyranyl 1,2,3-triazole compound . Oncotarget . 2017 ; 8 ( 5 ): 7801-13 .

Jarad M , Kuczynski EA , Morrison J , Viloria-Petit AM , Coomber BL . Release of endothelial cell associated VEGFR2 during TGF-β modulated angiogenesis in vitro . BMC Cell Biol . 2017 ; 18 ( 1 ): 10 .

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