Evaluation of debridement effects of bromelain-loaded sodium alginate nanoparticles incorporated into chitosan hydrogel in animal models

Document Type : Original Article

Authors

1 Scool of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

2 Pharmaceutical Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

3 Department of Pharmaceutics, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

4 Department of Neurology and Neurological Science, Stanford University, Stanford, CA, USA

5 Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

6 Nanotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran

7 Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s): Bromelain, a mixture of proteolytic enzymes from pineapple (Ananas comosus) is known as a potential debriding agent in burn treatment. In this research, the debridement efficiency of chitosan hydrogel loaded by sodium alginate-chitosan nanoparticles (NPs) containing bromelain (Br 10%-AG-CS NPs) was evaluated in animal models. 
Materials and Methods: The NPs were prepared using the ionic gelation technique and their properties were identified. Then, the debridement effect of bromelain NPs incorporated into chitosan hydrogel was evaluated 4 hr after wound treatment in animal models. 
Results: The particle size of positively charged Br-AG-Cs NPs was about 390±25 nm. The encapsulation efficiency of bromelain into AG-CS NPs was about 92%. The in vitro release profile showed that the maximum release of bromelain from NPs occurred during the first 4 hr (70%). The hydrogel structure did not significantly affect the profile release of bromelain in the formulation. After 6 months of storage at 4 and 25 °C, the synthesized NPs indicated no significant changes in bromelain activity. It was found that Br 10%-Ag-Cs NPs-CS hydrogel had the most beneficial effects on reducing necrotic tissues and resulted in re-epithelialization compared with other treated groups (negative and positive control, CS hydrogel, and Br 10%-CS hydrogel). 
Conclusion: Therefore, using this novel formulation can be considered a potential debridement agent.

Keywords


1. Evdokiou A, O Kanisicak, S Gierek, A Barry, MJ Ivey, X Zhang, et al. Characterization of Burn Eschar Pericytes. J Clin Med 2020; 9: 606-628.
2.    Cinat ME and Smith MM. Acute burn management. Achauer and Sood’s Burn Surgery Reconstruction and rehabilitation, Philadelphia: Saunders 2006; 1: 50-76.
3.    Hirche C, Citterio A, Hoeksema H, Koller J, Lehner M, Martinez JR, et al. Eschar removal by bromelain based enzymatic debridement (Nexobrid®) in burns: An European consensus. Burns 2017; 43: 1640-1653.
4.    Rosenberg L, Krieger Y, Bogdanov-Berezovski A, Silberstein E, Shoham Y, and Singer AJ. A novel rapid and selective enzymatic debridement agent for burn wound management: a multi-center RCT. Burns 2014; 40: 466-474.
5.    Singer AJ, Taira BR, Anderson R, McClain SA, and Rosenberg L. Reepithelialization of mid-dermal porcine burns after rapid enzymatic debridement with Debrase®. J Burn Care Res 2011; 32:  647-653.
6.    Singer AJ, McClain SA, Taira BR, Rooney J, Steinhauff N, Rosenberg L. Rapid and selective enzymatic debridement of porcine comb burns with bromelain-derived Debrase®: acute-phase preservation of noninjured tissue and zone of stasis. J Burn Care Res 2010; 31: 304-309.
7.    Silvestre MPC, Carreira RL, Silva MR, Corgosinho FC, Monteiro MRP,  Morais HA. Effect of pH and temperature on the activity of enzymatic extracts from pineapple peel. Food Bioproc Tech 2012; 5: 1824-1831.
8.    Ataide JA, Gérios EF, Mazzola PG, Sout EB., Bromelain-loaded nanoparticles: A comprehensive review of the state of the art. Adv Colloid Interface Sci 2018; 254: 48-55.
9.    Schulz A, Fuchs PC, Oplaender C, Valdez LB, Schiefer JL. Effect of Bromelain-Based Enzymatic Debridement on Skin Cells. J Burn Care Res 2018; 39: 527-535.
10.    Bayat S, Amiri N, Pishavar E, Kalalinia F, Movaffagh J, Hahsemi M. Bromelain-loaded chitosan nanofibers prepared by electrospinning method for burn wound healing in animal models. Life Sci 2019; 229: 57-66.
11.    Wang W, Lu K-j, Yu C-h, Huang Q-l,  DuY -Z. Nano-drug delivery systems in wound treatment and skin regeneration. J Nanobiotechnology 2019 ;17:82. 
12.    Du J and Wong KK, Nanomaterials for Wound Healing: Scope and Advances, in Theranostic Bionanomaterials. 2019, Elsevier. p. 211-230.
13.    El-Feky GS, Banna ST El-, El-Bahy GS, Abdelrazek E, Kamal M. Alginate coated chitosan nanogel for the controlled topical delivery of Silver sulfadiazine. Carbohydr Polym 2017; 177: 194-202.
14.    Ataide JA, Cefali LC, Rebelo MA, Spir LG, Tambourgi EB, Jozala AF, et al. Bromelain Loading and Release from a Hydrogel Formulated Using Alginate and Arabic Gum. Planta Med 2017; 83:  870-876.
15.    Choukaife H, Doolaanea AA, Alfatama M. Alginate nanoformulation: Influence of process and selected variables. Pharmaceuticals (Basel) 2020; 13:335.
16.    Kim HS, Sun X, LeeJ -H, Kim H-W, Fu X,  Leong KW. Advanced drug delivery systems and artificial skin grafts for skin wound healing. Adv Drug Deliv Rev 2019; 146: 209-239.
17.    Ji M, Sun X, Guo X, Zhu W, Wu J, Chen L, et al. Green synthesis, characterization and in vitro release of cinnamaldehyde/sodium alginate/chitosan nanoparticles. Food Hydrocoll 2019; 90:  515-522.
18.    Liu H, Wang C, Li C, Qin Y, Wang Z, Yang F, et al. A functional chitosan-based hydrogel as a wound dressing and drug delivery system in the treatment of wound healing. RSC Adv 2018; 8: 7533-7549.
19.    Grebinişan D, Holban M, Şunel V, Popa M, Desbrieres J, Lionte C. Novel acyl derivatives of N-(p-aminobenzoyl)-L-glutamine encapsulated in polymeric nanocapsules with potential antitumoral activity. Cellul Chem Technol 2011; 45: 571-577.
20.    Li P, Dai Y-N, Zhang J-P, Wang A-Q, and Wei Q. Chitosan-alginate nanoparticles as a novel drug delivery system for nifedipine. Int J Biomed Sci 2008; 4:221-228.
21.    Dubey R, Reddy S, Murthy N. Optimization of activity of bromelain. Asian J Chem 2012. 24: 1429-1431.
22.    Mudshinge SR, Deore AB, Patil S,  Bhalgat CM. Nanoparticles: emerging carriers for drug delivery. Saudi Pharm J 2011;19: 129-141.
23.    Demetzos C and Pippa N. Advanced drug delivery nanosystems (aDDnSs): a mini-review. Drug deliv 2014; 21: 250-257.
24.    Siafaka PI, Ustundag Okur N, Karavas E, Bikiaris DN. Surface Modified Multifunctional and Stimuli Responsive Nanoparticles for Drug Targeting: Current Status and Uses. Int J Mol Sci 2016;17:1440.
25.    Aderibigbe BA and Buyana B. Alginate in wound dressings.  Pharmaceutics 2018; 10: 42-60.
26.    Szekalska M, Puciłowska A, Szymańska E, Ciosek P, Winnicka K, Alginate: current use and future perspectives in pharmaceutical and biomedical applications. Int J Polym Sci  2016; 8: 1-17.
27.    Nagpal K, Singh SK, Mishra DN. Chitosan nanoparticles: a promising system in novel drug delivery. Chem Pharm Bull 2010; 58: 1423-1430.
28.    Charernsriwilaiwat N, Rojanarata T, Ngawhirunpat T, Opanasopit P. Electrospun chitosan/polyvinyl alcohol nanofibre mats for wound healing. Int Wound J 2014; 11: 215-222.
29.    Hamidi M, Azadi A, Rafiei P. Hydrogel nanoparticles in drug delivery. Adv Drug Deliv Rev 2008; 60: 1638-1649.
30.    Ratner BD, Hoffman AS, Schoen FJ, Lemons JE, Biomaterials science: an introduction to materials in medicine. 2004: Elsevier.
31.    Rachmawati H, Sulastri E, Immaculata Iwo M, Safitri D, Rahma A. Bromelain Encapsulated in Self Assembly Nanoemulsion Exhibits Better Debridement Effect in Animal Model of Burned Skin. J Nano Res 2016; 40: 158-166.
32.    Shoham Y, Krieger Y, Tamir E, Silberstein E, Bogdanov‐Berezovsky A, Haik J, et al. Bromelain‐based enzymatic debridement of chronic wounds: A preliminary report. Int Wound J.2018. 15: 769-775.
33.    Honardar S, Kordestani S, Daliri M, NayebHabib F. The effect of chitosan-based gel on second degree burn wounds. J Wound Care 2016; 25: 488-494.
34.    Sorasitthiyanukarn FN, Muangnoi C, Bhuket PRN, Rojsitthisak P, Rojsitthisak P. Chitosan/alginate nanoparticles as a promising approach for oral delivery of curcumin diglutaric acid for cancer treatment. Mater Sci Eng C 2018; 93: 178-190.
35.    Motwani SK, Chopra S, Talegaonkar S, Kohli K, Ahmad FJ, Khar RK. Chitosan–sodium alginate nanoparticles as submicroscopic reservoirs for ocular delivery: Formulation, optimisation and in vitro characterisation. Eur J Pharm Biopharm. 2008; 68: 513-525.
36.    Panyam J and Labhasetwar V, Biodegradable nanoparticles for drug and gene delivery to cells and tissue. Adv Drug Deliv Rev 2003; 55: 329-347.
37.    Liu X, Shen B, Shen C, Zhong R, Wang X, Yuan H. Nanoparticle-loaded gels for topical delivery of nitrofurazone: effect of particle size on skin permeation and retention. J Drug Deliv Sci Technol 2018; 45: 367-372.
38.    Mahmoud AA, El-Feky GS, Kamel R, Awad GE. Chitosan/sulfobutylether-β-cyclodextrin nanoparticles as a potential approach for ocular drug delivery. Int J Pharm 2011; 413: 229-236.
39.    Hoffmann F, Cinatl J, Kabičková H, Kreuter J, Stieneker F. Preparation, characterization and cytotoxicity of methylmethacrylate copolymer nanoparticles with a permanent positive surface charge. Int J Pharm 1997; 157: 189-198.
40.    Kim JH, Kim YS, Kim S, Park JH, Kim K, Choi K, et al. Hydrophobically modified glycol chitosan nanoparticles as carriers for paclitaxel. J Control Release 2006; 111: 228-234.
41.    Pal K, Paulson AT, Rousseau D, Biopolymers in controlled-release delivery systems, In book: Modern Biopolymer Science 2009, Elsevier. p. 519-557.
42.    Bhatnagar P, Pant AB, Shukla Y, Chaudhari B, Kumar P, Gupta KC. Bromelain nanoparticles protect against 7,12-dimethylbenz[a]anthracene induced skin carcinogenesis in mouse model. Eur J Pharm Biopharm 2015; 91: 35-46.
43.    Percival SL, McCarty S, Hunt JA, Woods EJ. The effects of pH on wound healing, biofilms, and antimicrobial efficacy. Wound Repair Regen 2014; 22: 174-186.
44.    Amid A, Ismail NA, Yusof F, Salleh HM. Expression, purification, and characterization of a recombinant stem bromelain from Ananas comosus. Process Biochem 2011; 46: 2232-2239.
45.    Rosenberg L, Krieger Y, Silberstein E, Arnon O, Sinelnikov IA, Bogdanov-Berezovsky A, et al. Selectivity of a bromelain based enzymatic debridement agent: a porcine study. Burns 2012; 38: 1035-1040.
46.    NexoBrid | European Medicines Agency. Assessment report. Procedure No. EMEA/H/C/002246. 2012.
47.    Hirche C, Almeland SK, Dheansa B, Fuchs P, Governa M, Hoeksema H, et al. Eschar removal by bromelain based enzymatic debridement (Nexobrid®) in burns: European consensus guidelines update. Burns 2020; 46: 782-796.
48.    Gunde M and AmnerNar N. An Investigation on Burn Wound Healing in Rats with Chitosan gel Formulation containing Plant Enzymes. Burns 2006; 32: 319-327.