The publications of the members of Eng4Life.
2024
Caccavo, Diego; Lamberti, Gaetano; Barba, Anna Angela
Analysis and simulation of wet-granulation processes Journal Article
In: Journal of the Taiwan Institute of Chemical Engineers, vol. 159, iss. 105455, 2024.
Abstract | Links | BibTeX | Tags: Dynamic image analysis, Modeling, Particle size distribution, Population balance equation, Wet-granulation
@article{Caccavo2024,
title = {Analysis and simulation of wet-granulation processes},
author = {Diego Caccavo and Gaetano Lamberti and Anna Angela Barba},
url = {https://doi.org/10.1016/j.jtice.2024.105455},
year = {2024},
date = {2024-03-22},
urldate = {2024-03-22},
journal = {Journal of the Taiwan Institute of Chemical Engineers},
volume = {159},
issue = {105455},
abstract = {Size enlargement by wet-granulation is one of the most important unit operation in several industrial fields: pharmaceutics, food processing, animal nutrition, agronomics, and so on. A lot of techniques have been used in order to measure the particle size distributions (PSD) and a few different approaches are available to model the PSD evolution during the process, mainly based on population balance equations (PBE). However, difficulties in both the activities (reliable measurement and robust modeling) have hindered their diffusion in industrial practice, keeping the granulation closer to an art than to a science.Size enlargement by wet-granulation is one of the most important unit operation in several industrial fields: pharmaceutics, food processing, animal nutrition, agronomics, and so on. A lot of techniques have been used in order to measure the particle size distributions (PSD) and a few different approaches are available to model the PSD evolution during the process, mainly based on population balance equations (PBE). However, difficulties in both the activities (reliable measurement and robust modeling) have hindered their diffusion in industrial practice, keeping the granulation closer to an art than to a science.},
keywords = {Dynamic image analysis, Modeling, Particle size distribution, Population balance equation, Wet-granulation},
pubstate = {published},
tppubtype = {article}
}
Size enlargement by wet-granulation is one of the most important unit operation in several industrial fields: pharmaceutics, food processing, animal nutrition, agronomics, and so on. A lot of techniques have been used in order to measure the particle size distributions (PSD) and a few different approaches are available to model the PSD evolution during the process, mainly based on population balance equations (PBE). However, difficulties in both the activities (reliable measurement and robust modeling) have hindered their diffusion in industrial practice, keeping the granulation closer to an art than to a science.Size enlargement by wet-granulation is one of the most important unit operation in several industrial fields: pharmaceutics, food processing, animal nutrition, agronomics, and so on. A lot of techniques have been used in order to measure the particle size distributions (PSD) and a few different approaches are available to model the PSD evolution during the process, mainly based on population balance equations (PBE). However, difficulties in both the activities (reliable measurement and robust modeling) have hindered their diffusion in industrial practice, keeping the granulation closer to an art than to a science.
2023
Apicella, Marco; Amato, Giuseppe; de Bartolomeis, Pietro; Barba, Anna Angela; De Feo, Vincenzo
Natural Food Resource Valorization by Microwave Technology: Purslane Stabilization by Dielectric Heating Journal Article
In: Foods , vol. 12, iss. 23, no. 4247, 2023.
Abstract | Links | BibTeX | Tags: dielectric heating, fatty acids, food recovery, future food, microwave drying technology, phytosterols, Portulaca oleracea L., valorization
@article{Apicella2023,
title = {Natural Food Resource Valorization by Microwave Technology: Purslane Stabilization by Dielectric Heating},
author = {Marco Apicella and Giuseppe Amato and {de Bartolomeis, Pietro} and Anna Angela Barba and {De Feo, Vincenzo}},
url = {https://doi.org/10.3390/foods12234247},
doi = {10.3390/foods12234247},
year = {2023},
date = {2023-11-24},
urldate = {2023-11-24},
journal = {Foods },
volume = {12},
number = {4247},
issue = {23},
abstract = {The application of microwave-assisted drying is a promising technique due to the features of process sustainability that are usable for responsible productions. It is largely applied for the stabilization of food products, especially in the agro-food sector. In this study, the weed Portulaca oleracea L. (purslane), with its richness in antioxidant components in addition to its recognized pharmacological properties, has been considered due to its potential to be a natural, well-accepted future food. Attention was focused on the role of the heat and mass transfer rates involved in the drying processes on the nutritional profile of the dried products. For this purpose, different drying protocols (convective, microwave irradiation, microwave-vacuum irradiation) were applied to different parts of purslane herb (apical, twigs, entire structures) and chemical characterizations were performed by a GC/MS analysis of the extracts of the dried products. The results show that microwave treatments can assure a better preservation of fatty acids such as SFAs, MUFAs, and PUFAs (which constitute over 90% of the total components in the apical part, 65% in twigs, and 85% in microwave-vacuum-dried entire purslane samples) and phytosterols (their highest preservation was found in microwave-dried twigs) compared with convective treatments. The chemical composition variability as well as treatment times depend on the drying rates (in microwave treatments, the times are on a minute scale and the rates are up to three orders of magnitude greater than convective ones), which in turn depend on the heating transport phenomena. This variability can lead towards products that are diversified by properties that transform a weed into a valorized food source.},
keywords = {dielectric heating, fatty acids, food recovery, future food, microwave drying technology, phytosterols, Portulaca oleracea L., valorization},
pubstate = {published},
tppubtype = {article}
}
The application of microwave-assisted drying is a promising technique due to the features of process sustainability that are usable for responsible productions. It is largely applied for the stabilization of food products, especially in the agro-food sector. In this study, the weed Portulaca oleracea L. (purslane), with its richness in antioxidant components in addition to its recognized pharmacological properties, has been considered due to its potential to be a natural, well-accepted future food. Attention was focused on the role of the heat and mass transfer rates involved in the drying processes on the nutritional profile of the dried products. For this purpose, different drying protocols (convective, microwave irradiation, microwave-vacuum irradiation) were applied to different parts of purslane herb (apical, twigs, entire structures) and chemical characterizations were performed by a GC/MS analysis of the extracts of the dried products. The results show that microwave treatments can assure a better preservation of fatty acids such as SFAs, MUFAs, and PUFAs (which constitute over 90% of the total components in the apical part, 65% in twigs, and 85% in microwave-vacuum-dried entire purslane samples) and phytosterols (their highest preservation was found in microwave-dried twigs) compared with convective treatments. The chemical composition variability as well as treatment times depend on the drying rates (in microwave treatments, the times are on a minute scale and the rates are up to three orders of magnitude greater than convective ones), which in turn depend on the heating transport phenomena. This variability can lead towards products that are diversified by properties that transform a weed into a valorized food source.
Piano, Raffaelle De; Caccavo, Diego; Barba, Anna Angela; Lamberti, Gaetano
Polyelectrolyte hydrogels in biological systems: Modeling of swelling and deswelling behavior Journal Article
In: Chemical Engineering Science, vol. 279, 2023.
Abstract | Links | BibTeX | Tags: Equilibrium, Hydrogels, Modeling, Polyelectrolytes
@article{Piano}2023,
title = {Polyelectrolyte hydrogels in biological systems: Modeling of swelling and deswelling behavior},
author = {Raffaelle {De Piano} and Diego Caccavo and Anna Angela Barba and Gaetano Lamberti},
doi = {https://doi.org/10.1016/j.ces.2023.118959},
year = {2023},
date = {2023-06-05},
urldate = {2023-06-05},
journal = {Chemical Engineering Science},
volume = {279},
abstract = {Polyelectrolyte hydrogels are a particular class of hydrogel whose behavior is connected to the variation of pH in the surrounding solution. Their behavior is influenced by the ionizable groups present on their chain. These groups could be acid or basic and polyelectrolytes could be anionic or cationic. To fully understand their behavior mathematical modeling has been widely used over many years. In this work a model based on a monophasic approach will be used to describe a general behavior of anionic hydrogels in a steady state condition at pH lower (or equal) to seven. Free swelling experiments and constrained swelling experiments have been simulated varying the parameters of the model to highlight the properties of the material. From a comparison with experimental data, it results that the proposed model can describe the general behavior of the system as described in the literature.},
keywords = {Equilibrium, Hydrogels, Modeling, Polyelectrolytes},
pubstate = {published},
tppubtype = {article}
}
Polyelectrolyte hydrogels are a particular class of hydrogel whose behavior is connected to the variation of pH in the surrounding solution. Their behavior is influenced by the ionizable groups present on their chain. These groups could be acid or basic and polyelectrolytes could be anionic or cationic. To fully understand their behavior mathematical modeling has been widely used over many years. In this work a model based on a monophasic approach will be used to describe a general behavior of anionic hydrogels in a steady state condition at pH lower (or equal) to seven. Free swelling experiments and constrained swelling experiments have been simulated varying the parameters of the model to highlight the properties of the material. From a comparison with experimental data, it results that the proposed model can describe the general behavior of the system as described in the literature.
Piano, Raffaelle De; Caccavo, Diego; Lamberti, Gaetano; Remaut, Katrien; Seynaeve, Hanne; Barba, Anna Angela
A New Productive Approach and Formulative Optimization for Curcumin Nanoliposomal Delivery Systems Journal Article
In: Pharmaceutics, vol. 15, iss. 3, no. 959, 2023, ISSN: 19994923.
Abstract | Links | BibTeX | Tags: anticancer, antioxidant activity, curcumin, nanoliposomes, nanotechnologies, nutraceuticals, simil-microfluidic technology
@article{Piano}2023b,
title = {A New Productive Approach and Formulative Optimization for Curcumin Nanoliposomal Delivery Systems},
author = {Raffaelle {De Piano} and Diego Caccavo and Gaetano Lamberti and Katrien Remaut and Hanne Seynaeve and Anna Angela Barba},
url = {https://www.scopus.com/inward/record.uri?eid=2-s2.0-85151381874&doi=10.3390%2fpharmaceutics15030959&partnerID=40&md5=fceef78253390aceb67f23b8ff583eb8},
doi = {10.3390/pharmaceutics15030959},
issn = {19994923},
year = {2023},
date = {2023-03-16},
urldate = {2023-03-16},
journal = {Pharmaceutics},
volume = {15},
number = {959},
issue = {3},
abstract = {The use of natural resources and the enhancing of technologies are outlining the strategies of modern scientific-technological research for sustainable health products manufacturing. In this context, the novel simil-microfluidic technology, a mild production methodology, is exploited to produce liposomal curcumin as potential powerful dosage system for cancer therapies and for nutraceutical purposes. Through simil-microfluidic technology, based on interdiffusion phenomena of a lipid-ethanol phase in an aqueous flow, massive productions of liposomes at nanometric scale can be obtained. In this work, studies on liposomal production with useful curcumin loads were performed. In particular, process issues (curcumin aggregations) were elucidated and formulation optimization for curcumin load was performed. The main achieved result has been the definition of operative conditions for nanoliposomal curcumin production with interesting loads and encapsulation efficiencies.},
keywords = {anticancer, antioxidant activity, curcumin, nanoliposomes, nanotechnologies, nutraceuticals, simil-microfluidic technology},
pubstate = {published},
tppubtype = {article}
}
The use of natural resources and the enhancing of technologies are outlining the strategies of modern scientific-technological research for sustainable health products manufacturing. In this context, the novel simil-microfluidic technology, a mild production methodology, is exploited to produce liposomal curcumin as potential powerful dosage system for cancer therapies and for nutraceutical purposes. Through simil-microfluidic technology, based on interdiffusion phenomena of a lipid-ethanol phase in an aqueous flow, massive productions of liposomes at nanometric scale can be obtained. In this work, studies on liposomal production with useful curcumin loads were performed. In particular, process issues (curcumin aggregations) were elucidated and formulation optimization for curcumin load was performed. The main achieved result has been the definition of operative conditions for nanoliposomal curcumin production with interesting loads and encapsulation efficiencies.
2021
Bochicchio, Sabrina; Dalmoro, Annalisa; Lamberti, Gaetano; Barba, Anna Angela
Nanoliposomi in cosmetica e cosmeceutica Journal Article
In: ICF – Rivista dell’industria chimica e farmaceutica, vol. Febbraio/Marzo 2021, no. 1, pp. 66-71, 2021.
Abstract | Links | BibTeX | Tags: cosmeceutics, Liposome bioadhesive, nanoliposomes
@article{Bochicchio2021,
title = {Nanoliposomi in cosmetica e cosmeceutica},
author = {Sabrina Bochicchio and Annalisa Dalmoro and Gaetano Lamberti and Anna Angela Barba},
url = {https://interprogettied.com/icf-rivista-dellindustria-chimica-e-farmaceutica-n1-febbraio-marzo-2021/},
year = {2021},
date = {2021-03-26},
journal = {ICF – Rivista dell’industria chimica e farmaceutica},
volume = {Febbraio/Marzo 2021},
number = {1},
pages = {66-71},
abstract = {L’uso di nanotecnologie per potenziare le performance dei prodotti cosmetici e cosmeceutici ha consentito una rapida crescita di questo settore industriale. Registrano un’ampia richiesta di mercato i nanocosmeceutici per la cura della pelle, dei capelli, delle unghie e delle labbra, oltre a quelli per contrastare rughe, fotoinvecchiamento, iperpigmentazione, forfora e danni ai capelli.},
keywords = {cosmeceutics, Liposome bioadhesive, nanoliposomes},
pubstate = {published},
tppubtype = {article}
}
L’uso di nanotecnologie per potenziare le performance dei prodotti cosmetici e cosmeceutici ha consentito una rapida crescita di questo settore industriale. Registrano un’ampia richiesta di mercato i nanocosmeceutici per la cura della pelle, dei capelli, delle unghie e delle labbra, oltre a quelli per contrastare rughe, fotoinvecchiamento, iperpigmentazione, forfora e danni ai capelli.
2020
Simone, Veronica De; Dalmoro, Annalisa; Bochicchio, Sabrina; Caccavo, Diego; Lamberti, Gaetano; Bertoncin, Paolo; Barba, Anna Angela
Nanoliposomes in polymeric granules: Novel process strategy to produce stable and versatile delivery systems Journal Article
In: Journal of Drug Delivery Science and Technology, vol. 59, pp. 7, 2020.
Abstract | Links | BibTeX | Tags: Compressibility index, Lipid-polymeric, Simil-microfluidic method, Texture analyses, Wet granulation
@article{Simone}2020,
title = {Nanoliposomes in polymeric granules: Novel process strategy to produce stable and versatile delivery systems},
author = {Veronica {De Simone} and Annalisa Dalmoro and Sabrina Bochicchio and Diego Caccavo and Gaetano Lamberti and Paolo Bertoncin and Anna Angela Barba},
url = {https://www.sciencedirect.com/science/article/abs/pii/S1773224720311679?via%3Dihub},
doi = {10.1016/j.jddst.2020.101878},
year = {2020},
date = {2020-07-09},
journal = {Journal of Drug Delivery Science and Technology},
volume = {59},
pages = {7},
abstract = {Liposomes, due to their mimetic cellular composition, are gaining great attention as release systems for lipophilic and hydrophilic molecules. However, liposomes can present a high tendency to degrade and aggregate into biological fluids and under storage conditions. To overcome these limitations, in this work, a stabilizing strategy consisting in liposomes incorporation into polymeric granules was studied. Wet granulation was adopted to produce granules of hydroxypropyl methylcellulose (HPMC) and liposomal suspensions were used as the binder phase. In particular, in this study, three different percentages of liposome load in HPMC granules were investigated (1%, 5% and 10% w/w) focusing the attention on several relevant technological characteristics of the achieved solid particulates: size, flow index, mechanical strength (granules without liposomal inclusions were used as a control). Morphological observations (by TEM) confirmed the presence of intact liposomes in dry HPMC granules; moreover, it was found that the binder phase with the lower liposome concentrations (1%, 5%) did not significantly affect size, flowability and hardness of the lipid-polymer granules. Instead, the granules containing the highest percentage of liposomes (10% w/w) have larger dimensions, harder structure and reduced flowability. Therefore, the followed process strategy, under liposomal concentration restrictions, allowed to obtain both the liposomes stabilization, a not trivial technological issue, and the production of particulates with good solid state properties, useful as a versatile dosage form (lipid carriers in polymer carriers).},
keywords = {Compressibility index, Lipid-polymeric, Simil-microfluidic method, Texture analyses, Wet granulation},
pubstate = {published},
tppubtype = {article}
}
Liposomes, due to their mimetic cellular composition, are gaining great attention as release systems for lipophilic and hydrophilic molecules. However, liposomes can present a high tendency to degrade and aggregate into biological fluids and under storage conditions. To overcome these limitations, in this work, a stabilizing strategy consisting in liposomes incorporation into polymeric granules was studied. Wet granulation was adopted to produce granules of hydroxypropyl methylcellulose (HPMC) and liposomal suspensions were used as the binder phase. In particular, in this study, three different percentages of liposome load in HPMC granules were investigated (1%, 5% and 10% w/w) focusing the attention on several relevant technological characteristics of the achieved solid particulates: size, flow index, mechanical strength (granules without liposomal inclusions were used as a control). Morphological observations (by TEM) confirmed the presence of intact liposomes in dry HPMC granules; moreover, it was found that the binder phase with the lower liposome concentrations (1%, 5%) did not significantly affect size, flowability and hardness of the lipid-polymer granules. Instead, the granules containing the highest percentage of liposomes (10% w/w) have larger dimensions, harder structure and reduced flowability. Therefore, the followed process strategy, under liposomal concentration restrictions, allowed to obtain both the liposomes stabilization, a not trivial technological issue, and the production of particulates with good solid state properties, useful as a versatile dosage form (lipid carriers in polymer carriers).
Bochicchio, Sabrina; Dalmoro, Annalisa; Lamberti, Gaetano; Barba, Anna Angela
Advances in Nanoliposomes Production for Ferrous Sulfate Delivery Journal Article
In: Pharmaceutics, vol. 12, no. 5, pp. 445, 2020.
Abstract | Links | BibTeX | Tags: drug delivery, ferrous sulfate, nanoliposome, simil-microfluidic apparatus, sonication
@article{Bochicchio2020b,
title = {Advances in Nanoliposomes Production for Ferrous Sulfate Delivery},
author = {Sabrina Bochicchio and Annalisa Dalmoro and Gaetano Lamberti and Anna Angela Barba},
editor = {MDPI},
url = {https://www.mdpi.com/1999-4923/12/5/445/pdf},
doi = {10.3390/pharmaceutics12050445},
year = {2020},
date = {2020-05-11},
journal = {Pharmaceutics},
volume = {12},
number = {5},
pages = {445},
abstract = {In this study, a continuous bench scale apparatus based on microfluidic fluid dynamic principles was used in the production of ferrous sulfate-nanoliposomes for pharmaceutical/nutraceutical applications, optimizing their formulation with respect to the products already present on the market. After an evaluation of its fluid dynamic nature, the simil-microfluidic (SMF) apparatus was first used to study the effects of the adopted process parameters on vesicles dimensional features by using ultrasonic energy to enhance liposomes homogenization. Subsequently, iron-nanoliposomes were produced at different weight ratios of ferrous sulfate to the total formulation components (0.06, 0.035, 0.02, and 0.01 w/w) achieving, by using the 0.01 w/w, vesicles of about 80 nm, with an encapsulation efficiency higher than 97%, an optimal short- and long-term stability, and an excellent bioavailability in Caco-2 cell line. Moreover, a comparison realized between the SMF method and two more conventional production techniques showed that by using the SMF setup the process time was drastically reduced, and the process yield increased, achieving a massive nanoliposomes production. Finally, duty-cycle sonication was detected to be a scalable technique for vesicles homogenization. },
keywords = {drug delivery, ferrous sulfate, nanoliposome, simil-microfluidic apparatus, sonication},
pubstate = {published},
tppubtype = {article}
}
In this study, a continuous bench scale apparatus based on microfluidic fluid dynamic principles was used in the production of ferrous sulfate-nanoliposomes for pharmaceutical/nutraceutical applications, optimizing their formulation with respect to the products already present on the market. After an evaluation of its fluid dynamic nature, the simil-microfluidic (SMF) apparatus was first used to study the effects of the adopted process parameters on vesicles dimensional features by using ultrasonic energy to enhance liposomes homogenization. Subsequently, iron-nanoliposomes were produced at different weight ratios of ferrous sulfate to the total formulation components (0.06, 0.035, 0.02, and 0.01 w/w) achieving, by using the 0.01 w/w, vesicles of about 80 nm, with an encapsulation efficiency higher than 97%, an optimal short- and long-term stability, and an excellent bioavailability in Caco-2 cell line. Moreover, a comparison realized between the SMF method and two more conventional production techniques showed that by using the SMF setup the process time was drastically reduced, and the process yield increased, achieving a massive nanoliposomes production. Finally, duty-cycle sonication was detected to be a scalable technique for vesicles homogenization.
Bochicchio, Sabrina; Dalmoro, Annalisa; Simone, Veronica De; Bertoncin, Paolo; Lamberti, Gaetano; Barba, Anna Angela
Simil-Microfluidic Nanotechnology in Manufacturing of Liposomes as Hydrophobic Antioxidants Skin Release Systems Journal Article
In: Cosmetics, vol. 7, no. 22, pp. 13, 2020.
Abstract | Links | BibTeX | Tags: antioxidants, cosmeceutics, nanoliposomes, simil-microfluidic technology, transdermal delivery
@article{Bochicchio2020,
title = {Simil-Microfluidic Nanotechnology in Manufacturing of Liposomes as Hydrophobic Antioxidants Skin Release Systems},
author = {Sabrina Bochicchio and Annalisa Dalmoro and Veronica De Simone and Paolo Bertoncin and Gaetano Lamberti and Anna Angela Barba},
editor = {MDPI},
url = {https://www.mdpi.com/2079-9284/7/2/22/pdf},
doi = {10.3390/cosmetics7020022},
year = {2020},
date = {2020-04-03},
journal = {Cosmetics},
volume = {7},
number = {22},
pages = {13},
abstract = {Novel nanotechnologies represent the most attractive and innovative tools to date exploited by cosmetic companies to improve the effectiveness of their formulations. In this context, nanoliposomes have had a great impact in topical preparations and dermocosmetics, allowing the transcutaneous penetration and absorption of several active ingredients and improving the stability of sensitive molecules. Despite the recent boom of this class of delivery systems, their industrial production is still limited by the lack of easily scalable production techniques. In this work, nanoliposomes for the topical administration of vitamin D3, K2, E, and curcumin, molecules with high antioxidant and skin curative properties but unstable and poorly absorbable, were produced through a novel simil-microfluidic technique. The developed high-yield semi continuous method is proposed as an alternative to face the problems linked with low productive conventional methods in order to produce antioxidant formulations with improved features. The novel technique has allowed to obtain a massive production of stable antioxidant vesicles of an 84–145 nm size range, negatively charged, and characterized by high loads and encapsulation efficiencies. The obtained products as well as the developed high-performance technology make the achieved formulations very interesting for potential topical applications in the cosmetics/cosmeceutical field. },
keywords = {antioxidants, cosmeceutics, nanoliposomes, simil-microfluidic technology, transdermal delivery},
pubstate = {published},
tppubtype = {article}
}
Novel nanotechnologies represent the most attractive and innovative tools to date exploited by cosmetic companies to improve the effectiveness of their formulations. In this context, nanoliposomes have had a great impact in topical preparations and dermocosmetics, allowing the transcutaneous penetration and absorption of several active ingredients and improving the stability of sensitive molecules. Despite the recent boom of this class of delivery systems, their industrial production is still limited by the lack of easily scalable production techniques. In this work, nanoliposomes for the topical administration of vitamin D3, K2, E, and curcumin, molecules with high antioxidant and skin curative properties but unstable and poorly absorbable, were produced through a novel simil-microfluidic technique. The developed high-yield semi continuous method is proposed as an alternative to face the problems linked with low productive conventional methods in order to produce antioxidant formulations with improved features. The novel technique has allowed to obtain a massive production of stable antioxidant vesicles of an 84–145 nm size range, negatively charged, and characterized by high loads and encapsulation efficiencies. The obtained products as well as the developed high-performance technology make the achieved formulations very interesting for potential topical applications in the cosmetics/cosmeceutical field.
2019
Barba, Anna Angela; Bochicchio, Sabrina; Bertoncin, Paolo; Lamberti, Gaetano; Dalmoro, Annalisa
Coating of Nanolipid Structures by a Novel Simil-Microfluidic Technique: Experimental and Theoretical Approaches Journal Article
In: Coatings, vol. 9, no. 8, pp. 491, 2019.
Abstract | Links | BibTeX | Tags: Chitosan, Coated nanoliposomes, Guar guam, Liposome bioadhesive, Liposomi bioadesivi, Muchoadesiveness, Simil-microfluidic technique
@article{Barba2019,
title = {Coating of Nanolipid Structures by a Novel Simil-Microfluidic Technique: Experimental and Theoretical Approaches },
author = {Anna Angela Barba and Sabrina Bochicchio and Paolo Bertoncin and Gaetano Lamberti and Annalisa Dalmoro},
url = {https://www.mdpi.com/2079-6412/9/8/491/htm},
doi = {10.3390/coatings9080491},
year = {2019},
date = {2019-08-02},
journal = {Coatings},
volume = {9},
number = {8},
pages = {491},
abstract = {Nanolipid vesicular structures are ideal candidates for the controlled release of various ingredients, from vitamins for nutraceutical purposes to chemoterapic drugs. To improve their stability, permeability, and some specific surface properties, such as mucoadhesiveness, these structures can require a process of surface engineering. The interaction of lipid vesicles with oppositely charged polyelectrolytes seems to be an interesting solution, especially when the negatively charged liposomes are complexed with the cationic chitosan. In this work, a novel simil-microfluidic technique was used to produce both chitosan-coated vesicles and a vegan alternative composed of cholesterol-free liposomes coated by Guar Hydroxypropyltrimonium Chloride (Guar-HC). The combination between the experimental approach, based on experimental observations in terms of Z-potential, and size evolutions, and the theoretical approach, based on concepts of saturation, was the methodology applied to define the best polycation concentration to fairly cover (vegan or not) liposomes without aggregation. The smart production of coated nanolipid structures was confirmed by characterizations of morphology, mucoadhesiveness, and stability.},
keywords = {Chitosan, Coated nanoliposomes, Guar guam, Liposome bioadhesive, Liposomi bioadesivi, Muchoadesiveness, Simil-microfluidic technique},
pubstate = {published},
tppubtype = {article}
}
Nanolipid vesicular structures are ideal candidates for the controlled release of various ingredients, from vitamins for nutraceutical purposes to chemoterapic drugs. To improve their stability, permeability, and some specific surface properties, such as mucoadhesiveness, these structures can require a process of surface engineering. The interaction of lipid vesicles with oppositely charged polyelectrolytes seems to be an interesting solution, especially when the negatively charged liposomes are complexed with the cationic chitosan. In this work, a novel simil-microfluidic technique was used to produce both chitosan-coated vesicles and a vegan alternative composed of cholesterol-free liposomes coated by Guar Hydroxypropyltrimonium Chloride (Guar-HC). The combination between the experimental approach, based on experimental observations in terms of Z-potential, and size evolutions, and the theoretical approach, based on concepts of saturation, was the methodology applied to define the best polycation concentration to fairly cover (vegan or not) liposomes without aggregation. The smart production of coated nanolipid structures was confirmed by characterizations of morphology, mucoadhesiveness, and stability.
Barba, Anna Angela; Bochicchio, Sabrina; Dalmoro, Annalisa; Lamberti, Gaetano
Lipid Delivery Systems for Nucleic-Acid-Based-Drugs: From Production to Clinical Applications Journal Article
In: Pharmaceutics, vol. 11, no. 360, 2019.
Abstract | Links | BibTeX | Tags: clinical trials, liposomes, Micro and Nano Vectors, NABDs, siRNA
@article{Barba2019b,
title = {Lipid Delivery Systems for Nucleic-Acid-Based-Drugs: From Production to Clinical Applications},
author = {Anna Angela Barba and Sabrina Bochicchio and Annalisa Dalmoro and Gaetano Lamberti},
url = {https://www.mdpi.com/1999-4923/11/8/360},
doi = {10.3390/pharmaceutics11080360},
year = {2019},
date = {2019-07-24},
journal = {Pharmaceutics},
volume = {11},
number = {360},
abstract = {In the last years the rapid development of Nucleic Acid Based Drugs (NABDs) to be used in gene therapy has had a great impact in the medical field, holding enormous promise, becoming “the latest generation medicine” with the first ever siRNA-lipid based formulation approved by the United States Food and Drug Administration (FDA) for human use, and currently on the market under the trade name Onpattro™. The growth of such powerful biologic therapeutics has gone hand in hand with the progress in delivery systems technology, which is absolutely required to improve their safety and effectiveness. Lipid carrier systems, particularly liposomes, have been proven to be the most suitable vehicles meeting NABDs requirements in the medical healthcare framework, limiting their toxicity, and ensuring their delivery and expression into the target tissues. In this review, after a description of the several kinds of liposomes structures and formulations used for in vitro or in vivo NABDs delivery, the broad range of siRNA-liposomes production techniques are discussed in the light of the latest technological progresses. Then, the current status of siRNA-lipid delivery systems in clinical trials is addressed, offering an updated overview on the clinical goals and the next challenges of this new class of therapeutics which will soon replace traditional drugs},
keywords = {clinical trials, liposomes, Micro and Nano Vectors, NABDs, siRNA},
pubstate = {published},
tppubtype = {article}
}
In the last years the rapid development of Nucleic Acid Based Drugs (NABDs) to be used in gene therapy has had a great impact in the medical field, holding enormous promise, becoming “the latest generation medicine” with the first ever siRNA-lipid based formulation approved by the United States Food and Drug Administration (FDA) for human use, and currently on the market under the trade name Onpattro™. The growth of such powerful biologic therapeutics has gone hand in hand with the progress in delivery systems technology, which is absolutely required to improve their safety and effectiveness. Lipid carrier systems, particularly liposomes, have been proven to be the most suitable vehicles meeting NABDs requirements in the medical healthcare framework, limiting their toxicity, and ensuring their delivery and expression into the target tissues. In this review, after a description of the several kinds of liposomes structures and formulations used for in vitro or in vivo NABDs delivery, the broad range of siRNA-liposomes production techniques are discussed in the light of the latest technological progresses. Then, the current status of siRNA-lipid delivery systems in clinical trials is addressed, offering an updated overview on the clinical goals and the next challenges of this new class of therapeutics which will soon replace traditional drugs
Dalmoro, Annalisa; Bochicchio, Sabrina; Lamberti, Gaetano; Bertoncin, Paolo; Janssens, Barbara; Barba, Anna Angela
Micronutrients encapsulation in enhanced nanoliposomal carriers by a novel preparative technology Journal Article
In: RSC Advances, vol. 9, pp. 19800-19812, 2019.
Abstract | Links | BibTeX | Tags: Drug Delivery Systems, Liposome bioadhesive, Liposomi bioadesivi, Micro and Nano Vectors
@article{Dalmoro2019,
title = {Micronutrients encapsulation in enhanced nanoliposomal carriers by a novel preparative technology },
author = {Annalisa Dalmoro and Sabrina Bochicchio and Gaetano Lamberti and Paolo Bertoncin and Barbara Janssens and Anna Angela Barba},
url = {https://pubs.rsc.org/en/content/articlelanding/2019/ra/c9ra03022k},
doi = {10.1039/C9RA03022K},
year = {2019},
date = {2019-06-25},
journal = {RSC Advances},
volume = {9},
pages = {19800-19812},
abstract = {Micronutrients administration by fortification of staple and complementary foods is a followed strategy to fight malnutrition and micronutrient deficiencies and related pathologies. There is a great industrial interest in preparation of formulations for joint administration of vitamin D3 and vitamin K2 for providing bone support, promoting heart health and helping boost immunity. To respond to this topic, in this work, uncoated nanoliposomes loaded with vitamin D3 and K2 were successfully prepared, by using a novel, high-yield and semi continuous technique based on simil-microfluidic principles. By the same technique, to promote and to enhance mucoadhesiveness and stability of the produced liposomal structures, chitosan was tested as covering material. By this way polymer–lipid hybrid nanoparticles, encapsulating vitamin D3 and vitamin K2, with improved features in terms of stability, loading and mucoadhesiveness were produced for potential nutraceutical and pharmaceutical applications.},
keywords = {Drug Delivery Systems, Liposome bioadhesive, Liposomi bioadesivi, Micro and Nano Vectors},
pubstate = {published},
tppubtype = {article}
}
Micronutrients administration by fortification of staple and complementary foods is a followed strategy to fight malnutrition and micronutrient deficiencies and related pathologies. There is a great industrial interest in preparation of formulations for joint administration of vitamin D3 and vitamin K2 for providing bone support, promoting heart health and helping boost immunity. To respond to this topic, in this work, uncoated nanoliposomes loaded with vitamin D3 and K2 were successfully prepared, by using a novel, high-yield and semi continuous technique based on simil-microfluidic principles. By the same technique, to promote and to enhance mucoadhesiveness and stability of the produced liposomal structures, chitosan was tested as covering material. By this way polymer–lipid hybrid nanoparticles, encapsulating vitamin D3 and vitamin K2, with improved features in terms of stability, loading and mucoadhesiveness were produced for potential nutraceutical and pharmaceutical applications.
2018
Bochicchio, Sabrina; Dalmoro, Annalisa; Bertoncin, Paolo; Lamberti, Gaetano; Moustafine, Rouslan I.; Barba, Anna Angela
Design and production of hybrid nanoparticles with polymeric-lipid shell–core structures: conventional and next-generation approaches Journal Article
In: RSC Advances, vol. 8, pp. 34614–34624, 2018.
Abstract | Links | BibTeX | Tags: Liposome bioadhesive, Liposomi bioadesivi
@article{Bochicchio2018,
title = {Design and production of hybrid nanoparticles with polymeric-lipid shell–core structures: conventional and next-generation approaches },
author = {Sabrina Bochicchio and Annalisa Dalmoro and Paolo Bertoncin and Gaetano Lamberti and Rouslan I. Moustafine and Anna Angela Barba },
url = {https://pubs.rsc.org/en/Content/ArticleLanding/2018/RA/C8RA07069E#!divAbstract},
doi = {10.1039/c8ra07069e},
year = {2018},
date = {2018-09-27},
journal = {RSC Advances},
volume = {8},
pages = {34614–34624},
abstract = {Liposomes constitute a class of prominent drug delivery systems due their cell-mimetic behaviour. Despite
their high biocompatibility, biodegradability and low intrinsic toxicity, their poor stability in biological fluids
as well as in stock conditions (high tendency to degrade or aggregate) have led to new approaches for
liposome stabilization (e.g., surface covering with polymers). Here, liposomes were enwrapped by the
natural biocompatible polymer chitosan to achieve stable shell–core nanostructures. Covered
nanoliposomes were produced using an innovative continuous method based on microfluidic principles.
The produced hybrid polymeric-lipid nanoparticles were characterized in terms of structural properties,
size and stability. Moreover, phenomenological aspects in formation of nanoliposomal vesicles and
chitosan layering, product quality (structure, size) and manufacturing yield related to this novel method
were compared with those of the conventional dropwise method and the obtained products. The
proposed simil-microfluidic method led to the production of stable and completely chitosan-covered
liposomes with a shell–core nanostructure that avoided the disadvantages inherent in the conventional
method (which are time-consuming and/or require bulky and more expensive equipment).},
keywords = {Liposome bioadhesive, Liposomi bioadesivi},
pubstate = {published},
tppubtype = {article}
}
Liposomes constitute a class of prominent drug delivery systems due their cell-mimetic behaviour. Despite
their high biocompatibility, biodegradability and low intrinsic toxicity, their poor stability in biological fluids
as well as in stock conditions (high tendency to degrade or aggregate) have led to new approaches for
liposome stabilization (e.g., surface covering with polymers). Here, liposomes were enwrapped by the
natural biocompatible polymer chitosan to achieve stable shell–core nanostructures. Covered
nanoliposomes were produced using an innovative continuous method based on microfluidic principles.
The produced hybrid polymeric-lipid nanoparticles were characterized in terms of structural properties,
size and stability. Moreover, phenomenological aspects in formation of nanoliposomal vesicles and
chitosan layering, product quality (structure, size) and manufacturing yield related to this novel method
were compared with those of the conventional dropwise method and the obtained products. The
proposed simil-microfluidic method led to the production of stable and completely chitosan-covered
liposomes with a shell–core nanostructure that avoided the disadvantages inherent in the conventional
method (which are time-consuming and/or require bulky and more expensive equipment).
their high biocompatibility, biodegradability and low intrinsic toxicity, their poor stability in biological fluids
as well as in stock conditions (high tendency to degrade or aggregate) have led to new approaches for
liposome stabilization (e.g., surface covering with polymers). Here, liposomes were enwrapped by the
natural biocompatible polymer chitosan to achieve stable shell–core nanostructures. Covered
nanoliposomes were produced using an innovative continuous method based on microfluidic principles.
The produced hybrid polymeric-lipid nanoparticles were characterized in terms of structural properties,
size and stability. Moreover, phenomenological aspects in formation of nanoliposomal vesicles and
chitosan layering, product quality (structure, size) and manufacturing yield related to this novel method
were compared with those of the conventional dropwise method and the obtained products. The
proposed simil-microfluidic method led to the production of stable and completely chitosan-covered
liposomes with a shell–core nanostructure that avoided the disadvantages inherent in the conventional
method (which are time-consuming and/or require bulky and more expensive equipment).
Dalmoro, Annalisa; Bochicchio, Sabrina; Nasibullin, Shamil F.; Bertoncin, Paolo; Lamberti, Gaetano; Barba, Anna Angela; Moustafine, Rouslan I.
Polymer-lipid hybrid nanoparticles as enhanced indomethacin delivery systems Journal Article
In: European Journal of Pharmaceutical Sciences, vol. 121, pp. 16-28, 2018.
Abstract | Links | BibTeX | Tags: Liposome bioadhesive, Liposomi bioadesivi, Micro and Nano Vectors
@article{Dalmoro2018,
title = {Polymer-lipid hybrid nanoparticles as enhanced indomethacin delivery systems},
author = {Annalisa Dalmoro and Sabrina Bochicchio and Shamil F. Nasibullin and Paolo Bertoncin and Gaetano Lamberti and Anna Angela Barba and Rouslan I. Moustafine},
url = {https://www.sciencedirect.com/science/article/pii/S0928098718302331},
doi = {10.1016/j.ejps.2018.05.014},
year = {2018},
date = {2018-08-30},
journal = {European Journal of Pharmaceutical Sciences},
volume = {121},
pages = {16-28},
abstract = {Non-steroidal anti-inflammatory drugs (NSAIDs), i.e. indomethacin used for rheumatoid arthritis and non-rheumatoid inflammatory diseases, are known for their injurious actions on the gastrointestinal (GI) tract. Mucosal damage can be avoided by using nanoscale systems composed by a combination of liposomes and biodegradable natural polymer, i.e. chitosan, for enhancing drug activity.
Aim of this study was to prepare chitosan-lipid hybrid delivery systems for indomethacin dosage through a novel continuous method based on microfluidic principles. The drop-wise conventional method was also applied in order to investigate the effect of the two polymeric coverage processes on the nanostructures features and their interactions with indomethacin. Thermal-physical properties, mucoadhesiveness, drug entrapment efficiency, in vitro release behavior in simulated GI fluids and stability in stocking conditions were assayed and compared, respectively, for the uncoated and chitosan-coated nanoliposomes prepared by the two introduced methods.
The prepared chitosan-lipid hybrid structures, with nanometric size, have shown high indomethacin loading (about 10%) and drug encapsulation efficiency up to 99%. TEM investigation has highlighted that the developed novel simil-microfluidic method is able to put a polymeric layer, surrounding indomethacin loaded nanoliposomes, thicker and smoother than that achievable by the drop-wise method, improving their storage stability. Finally, double pH tests have confirmed that the chitosan-lipid hybrid nanostructures have a gastro retentive behavior in simulated gastric and intestinal fluids thus can be used as delivery systems for the oral-controlled release of indomethacin.
Based on the present results, the simil-microfluidic method, working with large volumes, in a rapid manner, without the use of drastic conditions and with a precise control over the covering process, seems to be the most promising method for the production of suitable indomethacin delivery system, with a great potential in industrial manufacturing.},
keywords = {Liposome bioadhesive, Liposomi bioadesivi, Micro and Nano Vectors},
pubstate = {published},
tppubtype = {article}
}
Non-steroidal anti-inflammatory drugs (NSAIDs), i.e. indomethacin used for rheumatoid arthritis and non-rheumatoid inflammatory diseases, are known for their injurious actions on the gastrointestinal (GI) tract. Mucosal damage can be avoided by using nanoscale systems composed by a combination of liposomes and biodegradable natural polymer, i.e. chitosan, for enhancing drug activity.
Aim of this study was to prepare chitosan-lipid hybrid delivery systems for indomethacin dosage through a novel continuous method based on microfluidic principles. The drop-wise conventional method was also applied in order to investigate the effect of the two polymeric coverage processes on the nanostructures features and their interactions with indomethacin. Thermal-physical properties, mucoadhesiveness, drug entrapment efficiency, in vitro release behavior in simulated GI fluids and stability in stocking conditions were assayed and compared, respectively, for the uncoated and chitosan-coated nanoliposomes prepared by the two introduced methods.
The prepared chitosan-lipid hybrid structures, with nanometric size, have shown high indomethacin loading (about 10%) and drug encapsulation efficiency up to 99%. TEM investigation has highlighted that the developed novel simil-microfluidic method is able to put a polymeric layer, surrounding indomethacin loaded nanoliposomes, thicker and smoother than that achievable by the drop-wise method, improving their storage stability. Finally, double pH tests have confirmed that the chitosan-lipid hybrid nanostructures have a gastro retentive behavior in simulated gastric and intestinal fluids thus can be used as delivery systems for the oral-controlled release of indomethacin.
Based on the present results, the simil-microfluidic method, working with large volumes, in a rapid manner, without the use of drastic conditions and with a precise control over the covering process, seems to be the most promising method for the production of suitable indomethacin delivery system, with a great potential in industrial manufacturing.
Aim of this study was to prepare chitosan-lipid hybrid delivery systems for indomethacin dosage through a novel continuous method based on microfluidic principles. The drop-wise conventional method was also applied in order to investigate the effect of the two polymeric coverage processes on the nanostructures features and their interactions with indomethacin. Thermal-physical properties, mucoadhesiveness, drug entrapment efficiency, in vitro release behavior in simulated GI fluids and stability in stocking conditions were assayed and compared, respectively, for the uncoated and chitosan-coated nanoliposomes prepared by the two introduced methods.
The prepared chitosan-lipid hybrid structures, with nanometric size, have shown high indomethacin loading (about 10%) and drug encapsulation efficiency up to 99%. TEM investigation has highlighted that the developed novel simil-microfluidic method is able to put a polymeric layer, surrounding indomethacin loaded nanoliposomes, thicker and smoother than that achievable by the drop-wise method, improving their storage stability. Finally, double pH tests have confirmed that the chitosan-lipid hybrid nanostructures have a gastro retentive behavior in simulated gastric and intestinal fluids thus can be used as delivery systems for the oral-controlled release of indomethacin.
Based on the present results, the simil-microfluidic method, working with large volumes, in a rapid manner, without the use of drastic conditions and with a precise control over the covering process, seems to be the most promising method for the production of suitable indomethacin delivery system, with a great potential in industrial manufacturing.
2017
Bochicchio, Sabrina; Lamberti, Gaetano; Barba, Anna Angela
Phenomenological and Formulation Aspects in Tailored Nanoliposome Production Book Chapter
In: Liposomes, Chapter 2, IntechOpen, 2017.
Abstract | Links | BibTeX | Tags: cell‐mimetic system, gene therapy, liposomal iron, Liposome, nanoliposomes delivery systems, nutraceuticals, personalized carriers, simil‐microfluidic approach, ultrasonic size reduction
@inbook{Bochicchio2017b,
title = {Phenomenological and Formulation Aspects in Tailored Nanoliposome Production},
author = {Sabrina Bochicchio and Gaetano Lamberti and Anna Angela Barba},
url = {https://www.intechopen.com/books/liposomes/phenomenological-and-formulation-aspects-in-tailored-nanoliposome-production},
doi = {10.5772/intechopen.68157},
year = {2017},
date = {2017-10-25},
booktitle = {Liposomes},
publisher = {IntechOpen},
chapter = {2},
abstract = {Liposomes as cell‐mimetic system have attracted wide attention of researchers in various branches of the drug delivery topic as they can be highly functionalized and personalized, thus solving the major drawbacks of bioactive molecules linked to their low stability, limited membrane permeability, short half‐life and low bioavailability. The development of sustainable processes able to produce ad hoc liposomes in a rapid manner through the use of not‐laboured techniques, avoiding drastic conditions, is of great relevance for the industrial sector. In this chapter, two novel liposome production processes, the ultrasound‐assisted thin‐film hydration and the simil‐microfluidic techniques sharing the same size reduction/homogenization preparative step, are presented. The phenomenological aspects involved in vectors constitution through the duty cycle sonication process (bilayer rupture/vesicles formation mechanisms) and through the simil‐microfluidic approach (intubated flows interdiffusion mechanisms) are described. Finally, two applications as case histories involving the use of the developed techniques for relevant classes of active molecule delivery are described. In particular, a pharmaceutical application concerns the encapsulation of short‐interfering RNA (siRNA) molecule, used for gene therapy, inside cationic nanoliposomes, and a nutraceutical application consists in the production of ferrous sulphate anionic liposomal formulations with improved features compared to those already present on the market.},
keywords = {cell‐mimetic system, gene therapy, liposomal iron, Liposome, nanoliposomes delivery systems, nutraceuticals, personalized carriers, simil‐microfluidic approach, ultrasonic size reduction},
pubstate = {published},
tppubtype = {inbook}
}
Liposomes as cell‐mimetic system have attracted wide attention of researchers in various branches of the drug delivery topic as they can be highly functionalized and personalized, thus solving the major drawbacks of bioactive molecules linked to their low stability, limited membrane permeability, short half‐life and low bioavailability. The development of sustainable processes able to produce ad hoc liposomes in a rapid manner through the use of not‐laboured techniques, avoiding drastic conditions, is of great relevance for the industrial sector. In this chapter, two novel liposome production processes, the ultrasound‐assisted thin‐film hydration and the simil‐microfluidic techniques sharing the same size reduction/homogenization preparative step, are presented. The phenomenological aspects involved in vectors constitution through the duty cycle sonication process (bilayer rupture/vesicles formation mechanisms) and through the simil‐microfluidic approach (intubated flows interdiffusion mechanisms) are described. Finally, two applications as case histories involving the use of the developed techniques for relevant classes of active molecule delivery are described. In particular, a pharmaceutical application concerns the encapsulation of short‐interfering RNA (siRNA) molecule, used for gene therapy, inside cationic nanoliposomes, and a nutraceutical application consists in the production of ferrous sulphate anionic liposomal formulations with improved features compared to those already present on the market.
Bochicchio, Sabrina; Dalmoro, Annalisa; Recupido, Federica; Lamberti, Gaetano; Barba, Anna Angela
Nanoliposomes Production by a Protocol Based on a Simil-Microfluidic Approach Journal Article
In: Springer International Publishing AG 2018, no. 3-10, 2017, ISBN: 978-3-319-62026-8.
Abstract | Links | BibTeX | Tags: liposomal iron, Liposome, Liposomi
@article{Bochicchio2017,
title = {Nanoliposomes Production by a Protocol Based on a Simil-Microfluidic Approach},
author = {Sabrina Bochicchio and Annalisa Dalmoro and Federica Recupido and Gaetano Lamberti and Anna Angela Barba},
editor = {Stefano Piotto and Federico Rossi and Simona Concilio and Ernesto Reverchon and Giuseppe Cattaneo},
url = {https://link.springer.com/chapter/10.1007/978-3-319-62027-5_1},
doi = {10.1007/978-3-319-62027-5_1},
isbn = {978-3-319-62026-8},
year = {2017},
date = {2017-08-01},
journal = {Springer International Publishing AG 2018},
number = {3-10},
abstract = {In this work a protocol based on the microfluidic principles has been developed and applied to produce nanoliposomes. The protocol basically consists in the realization of a contact between two flows, lipids/ethanol and water solutions, inside a tubular device where interdiffusion phenomena allow the formation of lipid vesicles. Effects of solutions flow rates and lipids concentrations on size and size distribution have been investigated. Moreover, ultrasonic energy was used to enhance homogenization of the hydroalcoholic final solutions and to promote the vesicles size reduction. By this protocol a massive output has been achieved; increasing the ratio between the water volumetric flow rate to the lipids-ethanol volumetric flow rate the liposomes dimension decreases; at equal flow rates, when the lipids concentration increases also the liposomes size has been observed increasing. },
keywords = {liposomal iron, Liposome, Liposomi},
pubstate = {published},
tppubtype = {article}
}
In this work a protocol based on the microfluidic principles has been developed and applied to produce nanoliposomes. The protocol basically consists in the realization of a contact between two flows, lipids/ethanol and water solutions, inside a tubular device where interdiffusion phenomena allow the formation of lipid vesicles. Effects of solutions flow rates and lipids concentrations on size and size distribution have been investigated. Moreover, ultrasonic energy was used to enhance homogenization of the hydroalcoholic final solutions and to promote the vesicles size reduction. By this protocol a massive output has been achieved; increasing the ratio between the water volumetric flow rate to the lipids-ethanol volumetric flow rate the liposomes dimension decreases; at equal flow rates, when the lipids concentration increases also the liposomes size has been observed increasing.