Formulation of Zinc-Based Nanomaterials using the Eucommia ulmoides Bark Extract and their Wound Healing Potential

Summary

Here, we present a protocol to synthesize the zinc oxide (ZnO) nanoparticles using the polyisoprene-rich aqueous extract obtained from Eucommia ulmoides tree bark. The wound-healing potential exhibited by the synthesized ZnO nanoparticles on human umbilical vein endothelial cells (HUVECs) was evaluated using scratch assay, a simple, cost-effective, and efficient method.

Abstract

The aqueous extract from the bark of Eucommia ulmoides serves as a rich source of bioactive compounds with numerous health benefits. The protocol here aims to explore the preparation of zinc oxide (ZnO) nanoparticles using the Eucommia ulmoides bark-mediated polyisoprene-rich aqueous extract. Meanwhile, the proposed protocol is associated with the preparation of wound healing material by easing the process. In addition, the wound-healing potential of the synthesized nanoparticles (Eu-ZnO-NPs) was evaluated using a simple scratch assay on a human umbilical vein endothelial cell (HUVEC) monolayer. After 24 h of treatment with Eu-ZnO-NPs, the cell proliferation and migration of HUVEC cells were assessed. At the end of the study, cell proliferation and migration were observed in scratched monolayer treated with different concentrations of Eu-ZnO-NPs, whereas poor cell migration and proliferation rates were observed in control cells. Of the chosen concentrations, 20 µg/mL Eu-ZnO nanomaterials showed better cell migration and enhanced wound healing potential.

Introduction

Medicinal plants and plant-derived compounds have been shown to exhibit numerous health benefits¹. The World Health Organization (WHO) reported that 80% of the global population depends upon traditional medicinal plants for primary health care. China is well-recognized and popular for its Traditional Chinese Medicine (TCM) practices. Chinese medicinal herbs have been reported to treat various diseases and utilized for their biological potential. Medicinal plants serve as reservoirs for bioactive compounds and multiple therapeutic roles. Medicinal plants have also been utilized to treat the wounds. There are several types of approaches applied t....

Protocol

NOTE: Before preparing the extract, the obtained bark material was washed twice using deionized water and dried in a shaded place. The shade-dried barks were stored in an airtight container.

1. Preparation of Eucommia ulmoides bark extract

1. Chop the bark collected from the Eucommia ulmoides tree into small pieces using scissors.
2. Wash the chopped bark materials twice with double distilled water.
3. .......

Discussion

The E. ulmoides bark, seed, and leaves are believed to exhibit numerous health benefits. Our results have shown that the synthesis of EU-ZnO nanoparticles was achieved using a simple and cost-effective approach. The aqueous extract was utilized to synthesize the nanoparticles. Heating the bark material at high temperatures may cause degradation of some phytoconstituents and lower the extraction efficiency. The phytoconstituents present in the aqueous herbal extract can reduce the metal ions to form nanoparticles.......

Materials

Name	Company	Catalog Number	Comments
12 well plate	NEST	703011	Used for cell culture and assay
Centrifuge	SCILOGEX	SC1406	Used to separate the nanoparticles from the colloidal mixture
Centrifuge Tube – 15 mL	BIOFIL	CFT-312150	To centrifuge the synthesized solution
Centrifuge tube – 5 mL	Biosharp	BS-50-CM-S	To store the nanoparticles
CO2 incubator	Thermo scientific	3010	To culture the HUVEC cells
Denoised water	Millipore	Not applicable	For preparation of the extract
DMEM medium	Cytiva	SH30243.01	Used for cell culture work
Funnel	Thermo scientific	42600060	To hold the filter paper during the filtration
Glass beakers	Borosilicate	1102-50	Used to prepare the aqueous extract
Hot air oven	Genetimes	Not Applicable	Used to dry the nanoparticles and collect in the powder form
Magnetic stirrer	KYLIN-BELL	GL-5250-A	Used for nanoparticles synthesis
Microscope	Nikon Eclipse	Ts2	Used to take microphotographs
Petri dish	NEST	753001	Used to collect the nanoparticles
Pipette 1 mL	Lab Science	YEA17AD0055580	To take/add the specific volume of solution/extract
Pipette tips 1 mL	SAINING	3014200-T	To take/add the specific volume of solution/extract
PTFE Magnetic Mixer Stir Bars	LAN RAN	Not applicable	Used for nanomaterial synthesis process
Sodium hydroxide	Sigma Alrich	71690	Used to adjust pH during the synthesis
Stainless Scissor	Deli	6034	Used for chopping the bark materials
T25 tissue culture flask	NEST	707001	Used to maintain the cells
Weighing Balance	Radwag	AS220R2	Used to weigh the chemicals
Whatman filter paper No.1	Newstar	GB/T1914-2017	Used to filter the extract for synthesis
Zinc nitrate	Sigma Alrich	13778-30-8	Used as precursor for the nanoparticle’s synthesis