Name: Auteur in de schijnwerpers: Ontwikkeling en karakterisering van milieuvriendelijke microdeeltjes op basis van lignine voor een verbeterde afgifte van bioflavonoïden
Uploaded: 2024-03-01T14:05:00
Description: Watch this Scientific Journal Video about Author Spotlight: Development and Characterization of Eco-Friendly Lignin-Based Microparticles for Enhanced Delivery of Bioflavonoids at JoVE.com

Deze studie werd ondersteund door het Bulgaarse Wetenschappelijk Fonds in het kader van contract nr. KΠ-06 H59/3 en door wetenschappelijk project nr. 07/2023 FVM, Trakia University.

1. Synthese van lignine microdeeltjes
<ol><li>Bereid een waterige oplossing van 50 mg/ml alkalische lignine door 2,5 g alkalische lignine op te lossen in 50 ml ultrapuur water op een magnetische roerder.</li>
	<li>Bereid 1% Tween 80-oplossing door 1 ml Tween 80 op te lossen in 100 ml ultrapuur water.</li>
	<li>Bereid een 2 M-oplossing van HNO3 door 6,65 ml 67% HNO3 (dichtheid = 1,413 g/ml) te verdunnen met ultrapuur water tot een eindvolume van 50 ml.</li>
	<li>Voeg langzaam 15 ml...

Synthese van lignine-, micro-/submicrondeeltjes voor de afgifte van bioflavonoïden

<ol>
	<li>Yu, X., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lignin+nanoparticles+with+high+phenolic+content+as+efficient+antioxidant+and+sun-blocker+for+food+and+cosmetics.">Lignin nanoparticles with high phenolic content as efficient antioxidant and sun-blocker for food and cosmetics.</a> ACS Sustainable Chem. Eng. 11 (10), 4082-4092 (2023).</li><li>Boarino, A., Klok, H. -. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Opportunities+and+challenges+for+lignin+valorization+in+food+packaging,+antimicrobial,+and+agricultural+applications.">Opportunities and challenges for lignin valorization in food packaging, antimicrobial, and agricultural applications.</a> Biomacromolecules. 24 (3), 1065-1077 (2023).</li><li>Aadil, K., Barapatre, A., Jha, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+and+characterization+of+Acacia+lignin-gelatin+film+for+its+possible+application+in+food+packaging.">Synthesis and characterization of Acacia lignin-gelatin film for its possible application in food packaging.</a> Bioresour. Bioprocess. 3 (27), 1-11 (2016).</li><li>Sharma, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Valorization+of+lignin+into+nanoparticles+and+nanogel:+characterization+and+application.">Valorization of lignin into nanoparticles and nanogel: characterization and application.</a> Bioresour. Technol. Reports. 18, 101041 (2022).</li><li>Zadeh, E. M., O'Keefe, S. F., Kim, Y. -. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Utilization+of+lignin+in+biopolymeric+packaging+films.">Utilization of lignin in biopolymeric packaging films.</a> ACS Omega. 3 (7), 7388-7398 (2018).</li><li>Beaucamp, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lignin+for+energy+applications+-+state+of+the+art,+life+cycle,+technoeconomic+analysis+and+future+trends+(Critical+Review).">Lignin for energy applications - state of the art, life cycle, technoeconomic analysis and future trends (Critical Review).</a> Green Chem. 24, 8193-8226 (2022).</li><li>Antunes, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+sugarcane+to+skin:+Lignin+as+a+multifunctional+ingredient+for+cosmetic+application.">From sugarcane to skin: Lignin as a multifunctional ingredient for cosmetic application.</a> Int J Biol Macromol. 234, 123592 (2023).</li><li>Garg, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+lignin+nanoparticles+for+cancer+drug+delivery:+An+update.">Applications of lignin nanoparticles for cancer drug delivery: An update.</a> Materials Letters. 311, 131573 (2022).</li><li>Anushikha, K. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lignin+as+a+UV+blocking,+antioxidant,+and+antimicrobial+agent+for+food+packaging+applications.">Lignin as a UV blocking, antioxidant, and antimicrobial agent for food packaging applications.</a> Biomass Conv. Bioref. , 1-14 (2023).</li><li>Freitas, F. M. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=synthesis+of+lignin+nano-+and+micro-particles:+Physicochemical+characterization,+bioactive+properties+and+cytotoxicity+assessment.">synthesis of lignin nano- and micro-particles: Physicochemical characterization, bioactive properties and cytotoxicity assessment.</a> Int J Biol Macromol. 163, 1798-1809 (2020).</li><li>Rismawati, R., Nurdin, I. A., Pradiptha, M. N., Maulidiyah, A., Mubarakati, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preparation+and+characterization+of+lignin+nanoparticles+from+rice+straw+after+biosynthesis+using+Lactobacillus+bulgaricus.">Preparation and characterization of lignin nanoparticles from rice straw after biosynthesis using Lactobacillus bulgaricus.</a> Journal of Physics: Conference Series. 9th International Seminar on New Paradigm and Innovation of Natural Sciences and its Application. 1524, 012070 (2020).</li><li>Worku, L. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+lignin+nanoparticles+from+Oxytenanthera+abyssinica+by+nanoprecipitation+method+followed+by+ultrasonication+for+the+nanocomposite+application.">Synthesis of lignin nanoparticles from Oxytenanthera abyssinica by nanoprecipitation method followed by ultrasonication for the nanocomposite application.</a> Journal of King Saud University - Science. 35 (7), 102793 (2023).</li><li>Gala Morena, A., Tzanov, T. z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibacterial+lignin-based+nanoparticles+and+their+use+in+composite+materials.">Antibacterial lignin-based nanoparticles and their use in composite materials.</a> Nanoscale Adv. 4, 4447-4469 (2022).</li><li>Ivanova, D., Nikolova, G., Karamalakova, Y., Marutsova, V., Yaneva, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Water-soluble+alkali+lignin+as+a+natural+radical+scavenger+and+anticancer+alternative.">Water-soluble alkali lignin as a natural radical scavenger and anticancer alternative.</a> Int J Mol Sci. 24 (16), 12705 (2023).</li><li>Ivanova, D., Toneva, M., Simeonov, E., Antov, G., Yaneva, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Newly+synthesized+lignin+microparticles+as+bioinspired+oral+drug-delivery+vehicles:+Flavonoid-carrier+potential+and+in+vitro+radical-scavenging+activity.">Newly synthesized lignin microparticles as bioinspired oral drug-delivery vehicles: Flavonoid-carrier potential and in vitro radical-scavenging activity.</a> Pharmaceutics. 15 (4), 1067 (2023).</li><li>Yaneva, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antimicrobial+potential+of+conjugated+lignin/morin/chitosan+combinations+as+a+function+of+system+complexity.">Antimicrobial potential of conjugated lignin/morin/chitosan combinations as a function of system complexity.</a> Antibiotics. 11, 650 (2022).</li><li>Handral, H. K., Wyrobnik, T. A., Lam, A. T. -. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Emerging+trends+in+biodegradable+microcarriers+for+therapeutic+applications.">Emerging trends in biodegradable microcarriers for therapeutic applications.</a> Polymers. 15 (6), 1487 (2023).</li><li>Figueiredo, P., Lintinen, K., Hirvonen, J. T., Kostiainen, M. A., Santos, H. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Properties+and+chemical+modifications+of+lignin:+Towards+lignin-based+nanomaterials+for+biomedical+applications.">Properties and chemical modifications of lignin: Towards lignin-based nanomaterials for biomedical applications.</a> Prog. Mater. Sci. 93, 233-269 (2018).</li><li>Tang, Q., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lignin-based+nanoparticles:+a+review+on+their+preparations+and+applications.">Lignin-based nanoparticles: a review on their preparations and applications.</a> Polymers. 12 (11), 2471 (2020).</li><li>Zhao, W., Simmons, B., Singh, S., Ragauskas, A., Cheng, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+lignin+association+to+nano-/micro-particle+preparation:+extracting+higher+value+of+lignin.">From lignin association to nano-/micro-particle preparation: extracting higher value of lignin.</a> Green Chemistry. 18 (21), 5693-5700 (2016).</li><li>Stewart, H., Golding, M., Matia-Merino, L., Archer, R., Davies, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Manufacture+of+lignin+microparticles+by+anti-solvent+precipitation:+Effect+of+preparation+temperature+and+presence+of+sodium+dodecyl+sulfate.">Manufacture of lignin microparticles by anti-solvent precipitation: Effect of preparation temperature and presence of sodium dodecyl sulfate.</a> Food Res Int. 66, 93-99 (2014).</li><li>Beisl, S., Friedl, A., Miltner, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lignin+from+micro-+to+nanosize:+Applications.">Lignin from micro- to nanosize: Applications.</a> Int. J. Mol. Sci. 18, 2367 (2017).</li><li>Mishra, P. K., Ekielski, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+method+to+synthesize+lignin+nanoparticles.">A simple method to synthesize lignin nanoparticles.</a> Colloids Interfaces. 3, 52 (2019).</li><li>Qian, Y., Deng, Y., Qiu, X., Li, H., Yang, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Formation+of+uniform+colloidal+spheres+from+lignin,+a+renewable+resource+recovered+from+pulping+spent+liquor.">Formation of uniform colloidal spheres from lignin, a renewable resource recovered from pulping spent liquor.</a> Green Chem. 16, 2156-2163 (2014).</li><li>Tardy, B. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lignin+nano-+and+microparticles+as+template+for+nanostructured+materials:+formation+of+hollow+metal-phenolic+capsules.">Lignin nano- and microparticles as template for nanostructured materials: formation of hollow metal-phenolic capsules.</a> Green Chem. 20, 1335-1344 (2018).</li><li>Silva, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Paraquat-loaded+alginate/chitosan+nanoparticles:+preparation,+characterization+and+soil+sorption+studies.">Paraquat-loaded alginate/chitosan nanoparticles: preparation, characterization and soil sorption studies.</a> J Haz Mat. 190 (1-3), 366-374 (2011).</li><li>Georgieva, N., Yaneva, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+evaluation+of+natural+and+acid-modified+layered+mineral+materials+as+rimifon-carriers+using+UV/VIS,+FTIR,+and+equilibrium+sorption+study.">Comparative evaluation of natural and acid-modified layered mineral materials as rimifon-carriers using UV/VIS, FTIR, and equilibrium sorption study.</a> Cogent Chem. 1 (1), 1-16 (2015).</li><li>Zhang, P., Chen, D., Li, L., Sun, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Charge+reversal+nano-systems+for+tumor+therapy.">Charge reversal nano-systems for tumor therapy.</a> J Nanobiotechnol. 20, 31 (2022).</li><li>Yaneva, Z. L., Georgieva, N. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Removal+of+diazo+dye+from+the+aqueous+phase+by+biosorption+onto+ball-milled+maize+cob+(BMMC)+biomass+of+Zea+mays.">Removal of diazo dye from the aqueous phase by biosorption onto ball-milled maize cob (BMMC) biomass of Zea mays.</a> Maced. J. Chem. Chem. Eng. 32 (1), 133-149 (2013).</li><li>Zatorska, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Drug-loading+capacity+of+polylactide-based+micro-+and+nanoparticles+-+Experimental+and+molecular+modeling+study.">Drug-loading capacity of polylactide-based micro- and nanoparticles - Experimental and molecular modeling study.</a> Int J Pharmaceutics. 591, 120031 (2020).</li><li>Yaneva, Z., Georgieva, N., Grumezescu, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chapter+5+-+Physicochemical+and+morphological+characterization+of+pharmaceutical+nanocarriers+and+mathematical+modeling+of+drug+encapsulation/release+mass+transfer+processes.">Chapter 5 - Physicochemical and morphological characterization of pharmaceutical nanocarriers and mathematical modeling of drug encapsulation/release mass transfer processes.</a> Nanoscale Fabrication, Optimization, Scale-Up and Biological Aspects of Pharmaceutical Nanotechnology. , 173-218 (2018).</li><li>Yaneva, Z., Georgieva, N., Staleva, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+d,l-&#945;-tocopherol+acetate/zeolite+carrier+system:+equilibrium+study.">Development of d,l-&#945;-tocopherol acetate/zeolite carrier system: equilibrium study.</a> Monatshefte fur Chemie Chemical Monthly. 147 (7), 1167-1175 (2016).</li><li>Yaneva, Z., Georgieva, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Study+on+the+physical+chemistry,+equilibrium,+and+kinetic+mechanism+of+Azure+A+biosorption+by+Zea+mays+biomass.">Study on the physical chemistry, equilibrium, and kinetic mechanism of Azure A biosorption by Zea mays biomass.</a> Journal of Dispersion Science and Technology. 35 (2), 193-204 (2014).</li></ol>

Een van de belangrijkste kritieke kwesties van moderne synthesemethodologieën voor het ontwerp van formuleringen van geneesmiddeldragers op basis van biopolymeren is de toepassing van gevaarlijke organische reagentia - vluchtige en ontvlambare oplosmiddelen, zoals tetrahydrofuraan, aceton, methanol en zelfs DMSO in hoge concentraties - die hun toepasbaarheid in de biogeneeskunde, de farmaceutische industrie en de voedingstechnologie beperkt vanwege de manifestatie van mogelijke toxische effecten20</sup...

Tegenwoordig is de neiging naar biopolymeren zoals cellulose, chitosan, collageen, dextran, gelatine en lignine als voorlopers voor het ontwerp van micro-/submicrondragers met aanpasbare grootte, fysisch-chemische eigenschappen en biofunctionaliteiten toegenomen in de biomedische, farmaceutische en voedingstechnologie-industrieën vanwege hun toepasbaarheid in weefselmanipulatie, 3D-bioprinten, in vitro Platforms voor ziektemodellering, verpakkingsindustrie, emulsiepreparaat en afgifte van voedingsstoffen onder andere 1,2,3.
Nieuwe studies benadrukken de aspecten van hydrogels op basis van lignine en micro- en nanoformuleringen4 als voordelige voertuigen die worden gebruikt voor voedselverpakkingsmaterialen5, energieopslag6, cosmetica7, thermische/lichte stabilisatoren, versterkte materialen en medicijndragermatrices8 voor de afgifte van hydrofobe moleculen, verbetering van UV-barrières9, als versterkende middelen in nanocomposieten, en als alternatief voor anorganische nanodeeltjes vanwege enkele recente veiligheidsproblemen 10,11,12. De reden achter deze tendens is de biocompatibiliteit, biologische afbreekbaarheid en niet-toxiciteit van het natuurlijke hetero-biopolymeer, evenals de bewezen bio-activiteiten van lignine-antioxidantpotentieel en radicalenopruimende, antiproliferatieve en antimicrobiële activiteiten 13,14,15,16,17.
In de wetenschappelijke literatuur worden verschillende methoden voor synthese beschreven (zelfassemblage, anti-solvent precipitatie, zure precipitatie en solvent shifting)18 en karakterisering van op lignine gebaseerde formuleringen op micro-/nanoschaal, waaronder de toepassing van dure of schadelijke oplosmiddelen zoals tetrahydrofuraan (THF), dimethylsulfoxide (DMSO), N,N-dimethylformamide (DMF) en aceton, en gecompliceerde, indirecte en vervelende processen waarbij veel apparatuur en giftige stoffen worden gebruikt12,19,20.
Om deze laatste nadelen te ondervangen, presenteren de volgende protocollen nieuwe methodologieën voor de synthese van op lignine gebaseerde micro-/submicrondeeltjes met behulp van goedkope en groene oplosmiddelen; Eenvoudige, ongecompliceerde, snelle en gevoelige processen die weinig apparatuur, niet-toxische stoffen en eenvoudige methoden vereisen voor hun karakterisering en de bepaling van de inkapselingscapaciteit voor slecht in water oplosbare bioactieve stoffen en het in vitro afgiftepotentieel van de ligninematrices. De gepresenteerde productiemethoden op laboratoriumschaal zijn voordelig voor de vervaardiging van functionele ligninedragers met afstembare maten, hoge inkapselingscapaciteit en duurzaam in-vitro-afgiftegedrag met behulp van eenvoudige karakteriseringsprocedures en milieuvriendelijke chemicaliën die toepassing kunnen vinden in verschillende gebieden van de biomedische wetenschappen en voedseltechnologie. Twee flavonoïden werden toegepast als doelmoleculen ingekapseld in de ligninedeeltjes: morin in de microdeeltjes en quercetine in de submicrondeeltjes. Het verschil in de structuren van beide flavonoïden is alleen de positie van de tweede -OH-groep in de B-aromatische ring: de -OH-groep bevindt zich op de 2'-positie in morin en op de 3'-positie in quercetine, dus beide organische verbindingen zijn positionele isomeren. Dit laatste feit veronderstelt vergelijkbaar gedrag van beide bioactieve natuurlijke verbindingen in de processen van inkapseling en/of afgifte.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>automatic-cell counter</td>
			<td>EVE, NanoEnTek</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Citric acid</td>
			<td>Sigma</td>
			<td>251275</td>
			<td>&#160;ACS reagent, &#8805;99.5%</td>
		</tr>
		<tr>
			<td>digital water bath</td>
			<td>Memmert</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Eppendorf tubes, 1.5-2 mL</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Sigma</td>
			<td>34852-M</td>
			<td>absolute, suitable for HPLC, &#8805;99.8%</td>
		</tr>
		<tr>
			<td>Folin&#8211;Ciocalteu&#8217;s phenol reagent</td>
			<td>Sigma</td>
			<td>F9252</td>
			<td></td>
		</tr>
		<tr>
			<td>&#160;freeze dryer</td>
			<td>Biobase</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>gallic acid</td>
			<td>Sigma-</td>
			<td>BCBW7577</td>
			<td>monohydrate</td>
		</tr>
		<tr>
			<td>HCl</td>
			<td>Sigma</td>
			<td>258148</td>
			<td>ACS reagent, 37%</td>
		</tr>
		<tr>
			<td>HNO3</td>
			<td>Sigma</td>
			<td>438073</td>
			<td>&#160;ACS reagent, 70%</td>
		</tr>
		<tr>
			<td>lignin, alkali</td>
			<td>Sigma</td>
			<td>370959</td>
			<td></td>
		</tr>
		<tr>
			<td>morin</td>
			<td>Sigma</td>
			<td>PHL82601</td>
			<td></td>
		</tr>
		<tr>
			<td>NaCl</td>
			<td>Sigma</td>
			<td>S9888</td>
			<td>ACS reagent, &#8805;99.0%</td>
		</tr>
		<tr>
			<td>Na2CO3</td>
			<td>Sigma</td>
			<td>223530</td>
			<td>powder, &#8805;99.5%, ACS reagent</td>
		</tr>
		<tr>
			<td>NaOH</td>
			<td>Sigma</td>
			<td>655104</td>
			<td>reagent grade, 97%, powder</td>
		</tr>
		<tr>
			<td>orbital shaker</td>
			<td>IKA</td>
			<td>KS 130 basic</td>
			<td></td>
		</tr>
		<tr>
			<td>pH-meter</td>
			<td>Consort</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>phosphate-buffered saline (PBS)</td>
			<td>Sigma</td>
			<td>RNBH7571</td>
			<td></td>
		</tr>
		<tr>
			<td>Quercetin hydrate</td>
			<td>Sigma</td>
			<td>STBG3815V</td>
			<td></td>
		</tr>
		<tr>
			<td>statistical software for Excel</td>
			<td>Microsoft Corporation</td>
			<td></td>
			<td>XLSTAT&#160; Version 2022.4.5.</td>
		</tr>
		<tr>
			<td>Tween 80</td>
			<td>Sigma</td>
			<td>P8074</td>
			<td>BioXtra, viscous liquid</td>
		</tr>
		<tr>
			<td>ultracentrifuge</td>
			<td>Hermle</td>
			<td>Z 326 K</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrapure water system</td>
			<td>Adrona</td>
			<td>INTEGRITY+</td>
			<td></td>
		</tr>
		<tr>
			<td>ultrasound homogenizer</td>
			<td>Bandelin Sonopuls</td>
			<td>HD 2070</td>
			<td></td>
		</tr>
		<tr>
			<td>UV/Vis spectrophotometer</td>
			<td>Hach-Lange</td>
			<td>DR 5000</td>
			<td></td>
		</tr>
	</tbody>
</table>

green synthesis, characterization, encapsulation, and measurement of the release potential of novel alkali lignin micro-/submicron particles

De toepasbaarheid van biopolymeer micro-/nanotechnologie in de humane, diergeneeskunde, farmaceutica en voedingstechnologie neemt snel toe vanwege het grote potentieel van op biopolymeren gebaseerde deeltjes als effectieve draagsystemen. Het gebruik van lignine als een basis heteropolymeer biomatrix voor het ontwerp van innovatieve micro-/submicronformuleringen maakt het mogelijk om een verhoogde biocompatibiliteit te bereiken en biedt verschillende actieve functionele groepen die mogelijkheden bieden voor het aanpassen van de fysisch-chemische eigenschappen en bio-activiteiten van de formuleringen voor diverse toepassingen. Het doel van deze studie was het ontwikkelen van een eenvoudige en milieuvriendelijke methodologie voor de synthese van ligninedeeltjes met micro- en submicrongrootte; om hun fysisch-chemische, spectrale en structurele kenmerken te evalueren; en om hun vermogen voor inkapseling van biologisch actieve moleculen en het potentieel voor in vitro afgifte van bioflavonoïden in gesimuleerde gastro-intestinale media te onderzoeken. De gepresenteerde methodologieën maken gebruik van goedkope en groene oplosmiddelen; Eenvoudige, ongecompliceerde, snelle en gevoelige processen die weinig apparatuur, niet-toxische stoffen en eenvoudige methoden voor hun karakterisering vereisen, de bepaling van de inkapselingscapaciteit voor de slecht in water oplosbare bioactieve stoffen morin en quercetine, en het in vitro afgiftepotentieel van de ligninematrices.

Nowadays inclination towards biopolymers such as cellulose, chitosan, collagen, dextran, gelatin, and lignin as precursors for the design of micro-/submicron carriers with customizable size, physicochemical properties, and biofunctionalities has increased in the biomedical, pharmaceutical, and food technology industries due to their applicability in tissue engineering, 3D bioprinting, in vitro disease modeling platforms, packaging industry, emulsion preparation, and nutrient delivery among others1,2,3.
Novel studies highlight the aspects of lignin-based hydrogels as well as micro- and nano- formulations4 as advantageous vehicles used for food packaging materials5, energy storage6, cosmetics7, thermal/light stabilizers, reinforced materials, and drug-carrier matrices8 for the delivery of hydrophobic molecules, improvement of UV barriers9, as reinforcing agents in nanocomposites, and as an alternative to inorganic nanoparticles due to some recent safety issues10,11,12. The reason behind this tendency is the biocompatibility, biodegradability, and non-toxicity of the natural hetero biopolymer, as well as its proven bioactivities of lignin-antioxidant potential and radical scavenging, anti-proliferative, and antimicrobial activities13,14,15,16,17.
Scientific literature reports various methods for synthesis (self-assembly, anti-solvent precipitation, acid precipitation, and solvent shifting)18 and characterization of lignin-based micro-/nano- scaled formulations, including the application of expensive or harmful solvents such as tetrahydrofuran (THF), dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF), and acetone, and complicated, indirect, and tedious processes that use a lot of equipment and toxic substances12,19,20.
To overcome the latter disadvantages, the following protocols present novel methodologies for the synthesis of lignin-based micro-/submicron particles using cheap and green solvents; easy, straightforward, quick, and sensitive processes requiring little equipment, non-toxic substances, and simple methods for their characterization and the determination of encapsulation capacity towards poorly water-soluble bioactive compounds and in vitro release potential of the lignin matrices. The presented lab-scale production methods are advantageous for the manufacture of functional lignin carriers with tunable sizes, high encapsulation capacity, and sustainable in vitro release behavior utilizing simple characterization procedures and eco-friendly chemicals that can find application in various areas of biomedical sciences and food technology. Two flavonoids were applied as target molecules encapsulated into the lignin particles: morin-into the microparticles, and quercetin-into the submicron particles. The difference in the structures of both flavonoids Is only the position of the second -OH group in the B-aromatic ring: the -OH group is on the 2&#39; position in morin and on the 3&#39; position in quercetin, thus both organic compounds are positional isomers. The latter fact presumes similar behavior of both bioactive natural compounds in the processes of encapsulation and/or release.

Auteur in de schijnwerpers: Ontwikkeling en karakterisering van milieuvriendelijke microdeeltjes op basis van lignine voor een verbeterde afgifte van bioflavonoïden

Groene synthese, karakterisering, inkapseling en meting van het afgiftepotentieel van nieuwe alkali lignine micro-/submicron deeltjes

The objective of this research was to create a straightforward and environmentally friendly process to produce small lignin particles. The study aimed to analyze their physical, chemical, and structural properties, as well as investigate their ability to encapsulate and release bioflavonoids in stimulated gastrointestinal environments. Our method synthesizes lignin based particles using cheap and green solvents.The process is easy, quick, and requires minimal equipment and non-toxic substances. It also has simple methods for characterization, encapsulation, capacity determination, and in-vitro release potential of poorly water soluble bioactive compounds. The innovative biopolymer microparticles are suitable for oral administration due to the lower risk of gastric irritation.The submicron particles, due to the small size and significant release potential, could be applied as injectable formulations.

Er werd een anti-solvent precipitatietechniek uitgevoerd om alkalische lignine micro-/submicrondeeltjes te produceren. Een waterige oplossing van verdund anorganisch zuur-salpeterzuur/organisch zuur-citroenzuur werd gedispergeerd in een alkalische lignine-waterige oplossing, verrijkt met een milieuvriendelijke oppervlakteactieve stof/ethanol, wat resulteerde in de geleidelijke precipitatie van de biopolymeer opgeloste stof en, na sonicatie, werd uiteindelijk een suspensie van compacte micro-/submicrondeeltjes geproduceer...

Watch this Scientific Journal Video about Author Spotlight: Development and Characterization of Eco-Friendly Lignin-Based Microparticles for Enhanced Delivery of Bioflavonoids at JoVE.com

We beschrijven nieuwe, eenvoudige methodologieën voor synthese en karakterisering van biocompatibele lignine micro- en submicrondeeltjes. Deze formuleringen bieden een gemakkelijke benadering voor het gebruik van het heteropolymeer, evenals een alternatief voor het rationele ontwerp van multifunctionele draagmatrices met potentiële toepasbaarheid in de biogeneeskunde, farmaceutische technologie en de voedingsindustrie.

The applicability of biopolymer micro-/nano- technology in human, veterinary medicine, pharmaceutical, and food technology is rapidly growing due to the ...

Het doel van dit onderzoek was om een eenvoudig en milieuvriendelijk proces te creëren om kleine ligninedeeltjes te produceren. De studie was gericht op het analyseren van hun fysische, chemische en structurele eigenschappen, en op het onderzoeken van hun vermogen om bioflavonoïden in te kapselen en af te geven in gestimuleerde gastro-intestinale omgevingen. Onze methode synthetiseert deeltjes op basis van lignine met behulp van goedkope en groene oplosmiddelen.Het proces is eenvoudig, snel en vereist minimale apparatuur en niet-giftige stoffen. Het heeft ook eenvoudige methoden voor karakterisering, inkapseling, capaciteitsbepaling en in-vitro-afgiftepotentieel van slecht in water oplosbare bioactieve stoffen. De innovatieve biopolymeer microdeeltjes zijn geschikt voor orale toediening vanwege het lagere risico op maagirritatie.De submicrondeeltjes kunnen, vanwege de kleine omvang en het aanzienlijke afgiftepotentieel, worden toegepast als injecteerbare formuleringen.

green-synthesis-characterization-encapsulation-measurement-release

Research

JoVE Journal

Chemistry

Green Synthesis, Characterization, Encapsulation, and Measurement of the Release Potential of Novel Alkali Lignin Micro-/Submicron Particles

The applicability of biopolymer micro-/nano- technology in human, veterinary medicine, pharmaceutical, and food technology is rapidly growing due to the great potential of biopolymer-based particles as effective carrier systems. The use of lignin as a basic heteropolymer biomatrix for the design of innovative micro-/submicron formulations allows the achievement of increased biocompatibility and offers various active functional groups presenting opportunities for customization of the physicochemical properties and bioactivities of the formulations for diverse applications. The aim of the present study was to develop a simple and ecofriendly methodology for the synthesis of lignin particles with micro- and submicron size; to evaluate their physicochemical, spectral, and structural characteristics; and to examine their capacity for encapsulation of biologically active molecules and potential for in vitro release of bioflavonoids in simulated gastrointestinal media. The presented methodologies apply cheap and green solvents; easy, straightforward, quick, and sensitive processes requiring little equipment, non-toxic substances, and simple methods for their characterization, the determination of encapsulation capacity towards the poorly water-soluble bioactive compounds morin and quercetin, and the in vitro release potential of the lignin matrices.

We describe novel, simple methodologies of synthesis and characterization of biocompatible lignin micro- and submicron particles. These formulations provide a facile approach for the utilization of the heteropolymer, as well as an alternative for the rational design of multifunctional carrier matrices with potential applicability in biomedicine, pharmaceutical technology, and the food industry.