Evaluation of Effect of Biologically Synthesized Ethanolic Extract of Propolis-Loaded Poly(-Lactic-co-Glycolic Acid) Nanoparticles on Wound Healing in Diabetic Rats

Abstract

Wound healing is interaction of a complex cascade of cellular/biochemical actions leading to restoration of structural and functional integrity with regain of injured tissues strength. This study was aimed at evaluation of application of ethanolic extract of propolis-loaded poly(-lactic-co-glycolic acid) nanoparticles (EEP-PLGA NPs) on wound healing in diabetic rats. Sixty rats were randomized into four groups of 15 rats each: In control group (Control) diabetic wound was treated with normal saline. In Carrier 1 group diabetic wound was treated with PLGA nanoparticles based solution. In Carrier 2 group the diabetic wound was treated with EEP. In Treatment group animals received EEP-PLGA NPs on the wound. Wound size was measured on 7, 14 and 21 days after surgery. The expression of p53, bcl-2, Caspase III, were evaluated using reverse-transcription PCR and Immunohistochemical staining. The Treatment group had significantly reduced the wound size compared to other groups (P = 0.001). histological and morphometric studies, and mean rank of the qualitative studies demonstrated that there was significant difference between Treatment group and other groups (P < .05). Observations demonstrated that ethanolic extract of propolis-loaded PLGA nanoparticles significantly shortened the inflammatory phase and accelerated the cellular proliferation. Accordingly, the animals in Treatment group revealed significantly (P < .05) higher fibroblast distribution/one mm² of wound area and rapid re epithelialization. The mRNA levels of bcl-2, p53 and caspase III were remarkably (P < .05) higher in Treatment group compared to control and animals. The immunohistochemical analyzes confirmed the RT-PCR findings. EEP-PLGA NPs offered potential advantages in wound healing acceleration and improvement through angiogenesis stimulation, fibroblast proliferation and granulation tissue formation in early days of healing phases, acceleration in diabetic wound repair associated with earlier wound contraction and stability of damaged area by rearrangement of granulation tissue and collagen fibers.

Keywords

propolis PLGA nanoparticles diabetic wound rat

Get full access to this article

View all access options for this article.

References

American Diabetes Association. Diagnosis and classification of diabetes mellitus. Diabetes Care. 2014;37 Suppl 1(Supplement_1):S81‐S90.

Komesu

Tanga

Buttros

, et al. Effects of acute diabetes on rat cutaneous wound healing. Pathophysiology. 2004;11:(2)63‐67.

Gosain

DiPietro

. Aging and wound healing. World J Surg. 2004;28(3):321‐326.

Mathieu

Linke

J-C

Wattel

. Non-Healing wounds. In: Handbook on Hyperbaric Medicine. Springer-Verlag; 2006:401‐428.

Guo

Dipietro

. Factors affecting wound healing. J Dent Res. 2010;89(3):219‐229.

Dumville

Lipsky

Hoey

, et al. Topical antimicrobial agents for treating foot ulcers in people with diabetes. Cochrane Database Syst Rev. 2017;6(6):CD011038.

El-Guendouz

Al-Waili

Aazza

, et al. Antioxidant and diuretic activity of co-administration of capparis spinosa honey and propolis in comparison to furosemide. Asian Pac J Trop Med. 2017;10(10):974‐980.

Park

Lee

Seo

, et al. Evaluation of anti-allergic properties of caffeic acid phenethyl ester in a murine model of systemic anaphylaxis. Toxicol Appl Pharmacol. 2008;226(1):22‐29.

Sforcin

. Biological properties and therapeutic applications of propolis. Phytother Res. 2016;30(6):894‐905.

10.

Barbosa

Nunes

, et al. Hydroalcoholic extract of red propolis promotes functional recovery and axon repair after sciatic nerve injury in rats. Pharm Biol. 2016;54(6):993‐1004.

11.

Chen

Kuo

Chen

. Role of CAPE on cardiomyocyte protection via connexin 43 regulation under hypoxia. Int J Med Sci. 2016;13(10):754‐758.

12.

Kabała-Dzik

Rzepecka-Stojko

Kubina

, et al. Migration rate inhibition of breast cancer cells treated by caffeic acid and caffeic acid phenethyl ester: an In vitro comparison study. Nutrients. 2017;9(10):1144.

13.

Nie

Chang

, et al. Caffeic acid phenethyl ester (propolis extract) ameliorates insulin resistance by inhibiting JNK and NF-κB inflammatory pathways in diabetic mice and HepG2 cell models. J Agric Food Chem. 2017;65(41):9041‐9053.

14.

Kubiliene

Laugaliene

Pavilonis

, et al. Alternative preparation of propolis extracts: comparison of their composition and biological activities. BMC Complement Altern Med. 2015;15:156.

15.

Bamrungsap

Zhao

Chen

, et al. Nanotechnology in therapeutics: a focus on nanoparticles as a drug delivery system. Nanomedicine (Lond). 2012;7(8):1253‐1271.

16.

Jiang

Greene

Insua

, et al. Clearance of intracellular klebsiella pneumoniae infection using gentamicin-loaded nanoparticles. J Control Release. 2018;279:316‐325.

17.

Kesavan

Kotla

Ayyanaar

, et al. A theranostic nanocomposite system based on iron oxide-drug nanocages for targeted magnetic field responsive chemotherapy. Nanomedicine. 2018;14(5):1643‐1654.

18.

Jose

Netto

. Role of solid lipid nanoparticles as photoprotective agents in cosmetics. J Cosmet Dermatol. 2019;18(1):315‐321.

19.

Bonifácio

Silva

Ramos

, et al. Nanotechnology-based drug delivery systems and herbal medicines: a review. Int J Nanomedicine. 2014;9:1‐15.

20.

Kapoor

Bhatia

Kaur

, et al. PLGA: a unique polymer for drug delivery. Ther Deliv. 2015;6(1):41‐58.

21.

El-Say

El-Sawy

. Polymeric nanoparticles: promising platform for drug delivery. Int J Pharm. 2017;528(1-2):675‐691.

22.

Hickey

Santos

Williford

, et al. Control of polymeric nanoparticle size to improve therapeutic delivery. J Control Release. 2015;219:536‐547.

23.

Kumari

Yadav

. Biodegradable polymeric nanoparticles based drug delivery systems. Colloids Surf B Biointerfaces. 2010;75:1‐18.

24.

Danhier

Ansorena

Silva

, et al. PLGA-based nanoparticles: an overview of biomedical applications. J Control Release. 2012;161(2):505‐522.

25.

Derman

. Caffeic acid phenethyl ester loaded PLGA nanoparticles: effect of various process parameters on reaction yield, encapsulation efficiency, and particle size. J Nanomaterials. 2015;2015(1):341848.

26.

Menéndez-Manjón

Wagener

Barcikowski

. Transfer-matrix method for efficient ablation by pulsed laser ablation and nanoparticle generation in liquids. J Phys Chem C. 2011;115(12):5108‐5114.

27.

Taghavifar

Afroughi

Saadati Keyvan

. Curcumin nanoparticles improved diabetic wounds infected with methicillin-resistant staphylococcus aureus sensitized with HAMLET [published online ahead of print, 2020 Jun 28]. Int J Low Extrem Wounds. 2020;1534734620933079.

28.

Martin

Zenilman

Lazarus

. Molecular microbiology: new dimensions for cutaneous biology and wound healing. J Invest Dermatol. 2010;130(1):38‐48.

29.

McLaughlin

Podrebarac

Ruel

, et al.

Nano-engineered biomaterials for tissue regeneration: what has been achieved so far?

Front Mater. 2016;3(27).

30.

Wang

. Nanoparticles and their applications in cell and molecular biology. Integr Biol. 2016;6(1):9‐26.

31.

Aziz

. Abdul rasool hassan B. The effects of honey compared to silver sulfadiazine for the treatment of burns: a systematic review of randomized controlled trials. Burns. 2017;43(1):50‐57.

32.

Tan

Zhang

Lei

, et al. The roles of autophagy and apoptosis in burn wound progression in rats. Burns. 2013;39:1551‐1556.

33.

Tang

Xiang

, et al. Largely improved tensile properties of chitosan film via unique synergistic reinforcing effect of carbon nanotube and clay. J Phys Chem B. 2008;112(13):3876‐3881.

34.

Gopal

Kant

Gopalakrishnan

Tandan

Kumar

. Chitosan-based copper nanocomposite accelerates healing in excision wound model in rats. Eur J Pharmacol. 2014;731:8‐19.

35.

desJardins-Park

Foster

Longaker

. Fibroblasts and wound healing: an update. Regen Med. 2018;13(5):491‐495.

36.

Diegelmann

Evans

. Wound healing: an overview of acute, fibrotic and delayed healing. Front Biosci. 2004;9:283‐289.

37.

Baum

Arpey

. Normal cutaneous wound healing: clinical correlation with cellular and molecular events. Dermatol Surg. 2005;31(6):674‐686.

38.

Chen

Hao

, et al.

Insight into reepithelialization: how Do mesenchymal stem cells perform?

Stem Cells Int. 2016;2016:6120173.

39.

Kogan

Halsey

Agag

. Biologics in acute burn injury. Ann Plast Surg. 2019;83(1):26‐33.

40.

Salerian

. Burn wound infections and pseudomonas aeruginosa. Burns. 2020;46(1):257‐258.

41.

Chen

Wang

, et al. Functionalization of silver/titanium dioxide composites in chitosan-based coatings and their Egg preservation performances. J Vis Exp. 2021;173, 10.3791/61850.

42.

Umar

Sriwidodo

Maksum

, et al. Film-Forming spray of water-soluble chitosan containing liposome-coated human epidermal growth factor for wound healing. Molecules. 2021;26(17):5326.

43.

Riwaldt

Corydon

Pantalone

, et al. Role of apoptosis in wound healing and apoptosis alterations in microgravity. Front Bioeng Biotechnol. 2021;9:679650.

44.

Bhan

Mitra

Arya

, et al. A study on evaluation of apoptosis and expression of bcl-2-related marker in wound healing of streptozotocin-induced diabetic rats. ISRN Dermatol. 2013;2013:739054.

45.

Behroozi-Lak

Ebrahimpour

Zarei

, et al. Systemic administration of curcumin nanoparticles protects ischemia-reperfusion injury in ovaries: an animal model study. Rev Assoc Med Bras (1992). 2018;64(1):22‐31.