High-throughput fabrication of hepatic cell clusteroids with enhanced growth and functionality for tissue engineering applications

Wang, Anheng; Madden, Leigh A.; Paunov, Vesselin N.

doi:10.1039/d0ma00635a

Cited by 6 publications

(4 citation statements)

References 32 publications

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“…We used the w/w Pickering emulsion template to produce the HaCaT cell clusteroids with high yield; prior studies have noted the growth of the clusteroid layer of different cells using the w/w Pickering emulsion template including Hep-G2, HEK 293, and HaCaT cells. , The clusteroids were collected by breaking down the w/w emulsion system using dilution with DMEM/FBS complete medium. As shown in Figure S1, the clusteroids were successfully obtained from the emulsion droplets with relatively uniform sizes and shapes.…”

Section: Resultsmentioning

confidence: 99%

“…No previous study has seriously investigated a 3D cell culture platform as an alternative of in vivo biofilm testing due to the complex preparation and low yield rate . Recently, we developed a novel technique for production of tissue clusteroids (cell clusters) from human keratinocytes and hepatocytes (HaCaT and Hep-G2) by trapping them in water-in-water (w/w) Pickering emulsion droplets. − This technique is based on the formation of w/w Pickering emulsions, in which cells are efficiently encapsulated in emulsion droplets. This was followed by rapid osmotic shrinking of the droplets to compress the cells against each other and form a large amount of nearly spherical clusteroids in a very short time.…”

Section: Introductionmentioning

confidence: 99%

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Biofilm-Infected Human Clusteroid Three-Dimensional Coculture Platform to Replace Animal Models in Testing Antimicrobial Nanotechnologies

Wang

Weldrick

Madden

et al. 2021

ACS Appl. Mater. Interfaces

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Microbial biofilms are a major concern in wound care, implant devices, and organ infections. Biofilms allow higher tolerance to antimicrobial drugs, can impair wound healing, and potentially lead to sepsis. There has been a recent focus on developing novel nanocarrier-based delivery vehicles to enhance the biofilm penetration of traditional antibacterial drugs. However, a feasible in vitro human skin model to mimic the biofilm formation and its treatment for clearance have not yet been reported. This study describes the benefits of using an innovative bacterial biofilm-infected keratinocyte clusteroid model for the first time. It paves a new way for testing innovative nanomedicine delivery systems in a rapid and reproducible way on a realistic human cell-based platform, free of any animal testing. Herein, we have developed a novel composite 3D biofilm/human keratinocyte clusteroid coculture platform, which was used to measure biofilm clearance efficiency of nanoparticle (NP)-based therapeutics. We tested this model by treating the biofilm-infected 3D coculture layers by a ciprofloxacin-loaded Carbopol nanogel particles, surface-functionalized by the cationic protease Alcalase. We measured the antibacterial efficiency of the NP treatment on clearing Staphylococcus aureus and Pseudomonas aeruginosa biofilms on the 3D keratinocyte clusteroid/biofilm coculture model. Our experiments showed that these bacteria can infect the 3D layer of keratinocyte clusteroids and produce a stable biofilm. The biofilms were efficiently cleared by treatment with a formulation of 0.0032 wt % ciprofloxacin-loaded in 0.2 wt % Carbopol NPs surface-functionalized with 0.2 wt % Alcalase. Taken together, these promising results demonstrate that our coculture model can be exploited as a novel platform for testing the biofilm-eliminating efficiency of various NP formulations emulating skin and wound infections and could have wider applicability to replace animal models in similar experiments. This 3D cell culture-based platform could help in developing and testing of more effective antibacterial agents for clinical applications of antiplaque dental treatments, implants, infection control, and wound dressings.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Biofilm-Infected Human Clusteroid Three-Dimensional Coculture Platform to Replace Animal Models in Testing Antimicrobial Nanotechnologies

Wang

Weldrick

Madden

et al. 2021

ACS Appl. Mater. Interfaces

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“…In the emulsions with internal phase fractions of 75% are all surfactant-free o/w Pickering HIPEs. In contrast, HIPEs stabilized by surfactants are usually w/o [ 26 ] and extensive surfactants (5–50%) are frequently demanded to effectively produce stable HIPEs [ 20 ]. Our emulsions did not undergo creaming of dispersed phase fractions at pH around the isoelectric point (3, 3.5, and 4) after 15 days’ storage.…”

Section: Resultsmentioning

confidence: 99%

“…Therefore, utilizing olegels to secure bioavailable substitutes for PHOs is seriously restricted in the food industry. Another possible routine for developing substitutes for PHOs is the water-in-water emulsion template [ 24 , 25 , 26 ]. The shortcoming of such a system is the flavor and commercial acceptance.…”

Section: Introductionmentioning

confidence: 99%

Novel Pickering High Internal Phase Emulsion Stabilized by Food Waste-Hen Egg Chalaza

Wang

2021

Foods

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A massive amount of chalaza with nearly 400 metric tons is produced annually as waste in the liquid-egg industry. The present study aimed to look for ways to utilize chalaza as a natural emulsifier for high internal phase emulsions (HIPEs) at the optimal production conditions to expand the utilization of such abundant material. To the author’s knowledge, for the first time, we report the usage of hen egg chalaza particles as particulate emulsifiers for Pickering (HIPEs) development. The chalaza particles with partial wettability were fabricated at different pH or ionic strengths by freeze-drying. The surface electricity of the chalaza particles was neutralized when the pH was adjusted to 4, where the chalaza contained a particle size around 1500 nm and held the best capability to stabilize the emulsions. Similarly, the chalaza reaches proper electrical charging (−6 mv) and size (700 nm) after the ionic strength was modified to 0.6 M. Following the characterization of chalaza particles, we successfully generated stable Pickering HIPEs with up to 86% internal phase at proper particle concentrations (0.5–2%). The emulsion contained significant stability against coalescence and flocculation during long term storage due to the electrical hindrance raised by the chalaza particles which absorbed on the oil–water interfaces. Different rheological models were tested on the formed HIPEs, indicating the outstanding stability of such emulsions. Concomitantly, a percolating 3D-network was formed in the Pickering HIPES stabilized by chalaza which provided the emulsions with viscoelastic and self-standing features. Moreover, the current study provides an attractive strategy to convert liquid oils to viscoelastic soft solids without artificial trans fats.

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Albumin

Ebhodaghe

2023

Handbook of Biopolymers

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High-throughput fabrication of hepatic cell clusteroids with enhanced growth and functionality for tissue engineering applications

Abstract: We report a novel inexpensive and up-scalable fabrication technique for viable hepatocyte clusteroids and demonstrate that they grow faster than individual cells in tissue engineering applications.

Cited by 6 publications

References 32 publications

Biofilm-Infected Human Clusteroid Three-Dimensional Coculture Platform to Replace Animal Models in Testing Antimicrobial Nanotechnologies

Biofilm-Infected Human Clusteroid Three-Dimensional Coculture Platform to Replace Animal Models in Testing Antimicrobial Nanotechnologies

Novel Pickering High Internal Phase Emulsion Stabilized by Food Waste-Hen Egg Chalaza

Albumin

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