Synthesis of Black Seed Oil-loaded-, Alginate-based, pH-sensitive Beads Using Electrospraying Technique

Summary

A technique employing high electric voltage and a targeted, active ingredient-loaded emulsion to fabricate pH-responsive, uniform microbeads is presented.

Abstract

Black seed oil (BSO), derived from the seeds of the Nigella sativa plant, has garnered attention for its potential anti-cancer properties, particularly in the context of colon cancer. Its active compound, thymoquinone, may help inhibit cancer cell growth and induce apoptosis in colon cancer cells. Additionally, black seed oil's anti-inflammatory and antioxidant effects could contribute to a healthier gut environment, potentially reducing cancer risk. Therefore, this study synthesized pH-sensitive alginate beads to deliver BSO into the colon in a controlled-release manner without releasing the drug at pH 1.2 (stomach), thus providing a well-defined release pattern at pH 6.8. The use of electrospray technology improves process performance by making it easier to formulate small, homogeneous beads with a higher rate of swelling and diffusion in the gastrointestinal medium.

The formulated beads were characterized by an ex-vivo mucoadhesive strength test, bead size, sphericity factor (SF), encapsulation efficiency (EE), scanning electron microscope (SEM), in vitro swelling behavior (SB), and in vitro drug release in acidic and buffer media. All these manufactured beads demonstrated modest sizes of 0.58 ± 0.01 mm and spherical shape of 0.03 ± 0.00 mm in this testing. The formulation showed promising floating and releasing properties in vitro. With a very low cumulative percentage of beads, the oil EE of 90.13% ± 0.93% was high, and the release study demonstrated more than 90% in pH 6.8 with good floating nature in the stomach. Additionally, the beads were evenly spaced throughout the intestine. The electrospraying approach used in this protocol can be reproducible, yielding consistent outcomes. Therefore, this protocol can be used for large-scale production for commercialization purposes.

Introduction

Black seed, and more especially BSO, has been used for ages to cure a wide range of illnesses due to its well-established medicinal properties. Thymoquinone is perhaps one of the most important phytochemicals found in BSO¹. In recent years, researchers have studied thymoquinone's potential therapeutic benefits in vivo and in vitro, producing empirical evidence to support the use of BSO. Antihypertensive, antibacterial, antihistaminic, antifungal, analgesic, antidiabetic, lipid-lowering, and anti-inflammatory properties have all been demonstrated by these studies for BSO, which may be used to treat symptoms such as eczema, ....

Protocol

1. Preparation of alginate-BSO emulsion

1. Disperse 10% w/v BSO in 1% w/v sodium alginate solution containing 1, 3, and 5% w/v lecithin in a 50 mL beaker.
2. Obtain a nano-emulsion by using an ultrasonic homogenizer. Set the power level to 20%. Run the homogenizer for 55 s by clicking the start button to complete the process.

2. Characterization of alginate-BSO emulsion

1. Analy.......

Discussion

Using the EHDA process, BSO-loaded alginate microbeads were created as a pH-sensitive carrier. The beads' network exhibited pH-dependent swelling and drug-release behavior due to the abundant presence of carboxylic acid groups. The strong intermolecular hydrogen bonding between the polymer chains was revealed to be the reason behind the decreased swelling character of BSO-loaded beads at pH 1.2. BSO-loaded beads may benefit from this reduced SB for pH-sensitive medication administration. pH had an impact on the BSO r.......

Materials

Name	Company	Catalog Number	Comments
10 mL Centrifuge Tubes	Globe Scientific	22-171-624
22 G needle	Sigma-Aldrich (St.Louis, Missouri, USA).	CAD4172
3 mL quartz-cuvette	Sigma-Aldrich (St.Louis, Missouri, USA).	Z276669
50 mL beaker
Aluminum stubs
An electronic analytical balance
ATR-FTIR	Bruker Malaysia Sdn Bhd, Kawasan Perindustrian Temasya, 40150 Shah Alam, Selangor, Malaysia.
Black seed oil	IKOP Pharmaceutical Ltd. (IKOP, Faculty of Pharmacy, IIUM, 25200 Kuantan, Pahang, Malaysia	B182111	Active ingredient
Calcium chloride dehydrate, CaCl2 · 2H2O	Sigma-Aldrich (St.Louis, Missouri, USA).	21074	Gelling agent
Carbon adhesive tapes
Centrifuge
Differential scanning calorimetry
Digital   camera
Grounded beaker
High guluronic acid content Sodium alginate (mw. 97,000) with medium viscosity (40 – 100 mPa s)	Sigma-Aldrich (St.Louis, Missouri, USA).	W201502	Polymer
High voltage power supply
Isopropyl alcohol	Sigma-Aldrich (St.Louis, Missouri, USA).	W292912	ATR-FTIR cleaning purpose
Lecithin	Sigma-Aldrich (St.Louis, Missouri, USA).	P7568	Surfactant
Microscope
Paper towel
Scanning electron microscopy
Simulated gastric fluid	Sigma-Aldrich (St.Louis, Missouri, USA).	1651	Release media and swelling media
Simulated intestinal fluid	Sigma-Aldrich (St.Louis, Missouri, USA).	84082-64-4	Release media and swelling media
Spectroscopy software
Stainless-steel filter
Syringe pump	SEN JIN SDN BHD Malaysia, Taman Desaria, 46150 Petaling Jaya, Selangor Darul Ehsan Malaysia
Ultrapure distilled water	Supplied by institutional lab
Ultrasonic homogenizer	SEN JIN SDN BHD Malaysia, Taman Desaria, 46150 Petaling Jaya, Selangor Darul Ehsan Malaysia
UV-vis spectrophotometer.
Vacuum evaporator	SEN JIN SDN BHD Malaysia, Taman Desaria, 46150 Petaling Jaya, Selangor Darul Ehsan Malaysia
Voltage accelerator	SEN JIN SDN BHD Malaysia, Taman Desaria, 46150 Petaling Jaya, Selangor Darul Ehsan Malaysia
Zetasizer Nano-ZS	(Malvern Zetasizer Nano series Nano-S and Nano-Z, Malvern Instruments Ltd., Worcestershire, UK)