Surfactant-Coated Ultrafine Bubbles for Safe Cell Transfection

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Solution Overview

Problem

Current methods for delivering nucleic acids and proteins into cells, such as those using microbubble liposomes, face challenges including antigenicity concerns and safety issues due to high ultrasound intensity requirements, while conventional ultrafine bubbles were thought to severely damage cell membranes, making them unsuitable for cell transfection.

Innovation Solution

The use of ultrafine bubble water containing bubbles with an average diameter of not more than 200 nm, devoid of phospholipids, in combination with low-intensity ultrasound to enhance the delivery of nucleic acids and proteins into cells, specifically skeletal muscle and nerve cells, by creating a safer and more efficient transfection system.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If microbubble liposomes are used for delivering nucleic acids and proteins into cells, then delivery efficiency is improved, but antigenicity and safety concerns arise due to phospholipid content and high ultrasound intensity requirements

Engineering Contradiction:

Improvedelivery efficiencyVSAvoidantigenicity and safety concerns

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The invention extracts and removes the phospholipid component from the bubble structure, using only surfactant-based ultrafine bubbles without phospholipid encapsulation. This eliminates the antigenicity problem associated with phospholipid while maintaining the ability to enhance nucleic acid and protein delivery into cells through ultrasound exposure.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention changes the physical parameters of the bubble system by using ultrafine bubbles with average diameter of 10-200 nm instead of conventional microbubbles, and operates at lower ultrasound intensities (50-500 mW/cm²) compared to traditional methods (1.5-2.5 W/cm²). This parameter optimization maintains delivery efficiency while reducing harmful effects.

Inventive Principle:

Principle #35Parameter changes

2Length of moving object

If conventional ultrafine bubbles are used for cell transfection, then bubble size is reduced, but cell membrane damage occurs making them unsuitable for transfection

Engineering Contradiction:

Improvebubble sizeVSAvoidcell membrane damage

Core Design Contradiction:

Length of moving objectVSObject-affected harmful factors

Solution Approach 1:

The invention introduces surfactants as intermediary substances that form a protective interface around the ultrafine bubbles. These surfactant-coated bubbles act as gentle mediators that can enter cells through ultrasound-induced endocytosis without causing direct membrane damage, enabling successful transfection of nucleic acids and proteins.

Inventive Principle:

Principle #24Intermediary (Mediator)

Solution Approach 2:

The invention replaces the mechanical rupture mechanism of conventional ultrafine bubbles with a biochemical uptake mechanism. Instead of bubbles physically bursting cell membranes, the surfactant-coated ultrafine bubbles are internalized by cells through ultrasound-enhanced endocytosis, avoiding mechanical damage while achieving effective delivery.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

3Productivity

If high ultrasound intensity is used for bubble liposome transfection, then delivery efficiency is improved, but safety concerns arise

Engineering Contradiction:

Improvedelivery efficiencyVSAvoidsafety concerns

Core Design Contradiction:

ProductivityVSObject-affected harmful factors

Solution Approach 1:

The invention optimizes the ultrasound intensity parameter to a safe range of 50-500 mW/cm², significantly lower than conventional methods. Combined with the use of surfactant-based ultrafine bubbles, this parameter change achieves effective nucleic acid and protein delivery without the safety concerns associated with high-intensity ultrasound.

Inventive Principle:

Principle #35Parameter changes

Applied Scientific Principles

This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.

Function Achieved in This Case

This approach significantly increases the introduction efficiency of nucleic acids and proteins into cells compared to conventional methods, providing a non-invasive and safe delivery system that avoids the limitations of liposome use and high ultrasound intensity, while maintaining long-term stability of ultrafine bubbles.

Implementation Method 1

a method for increasing the delivery of a nucleic acid, a protein or a low-molecular-weight compound into a cell (excluding immune cells) by using the ultrafine bubble water or aqueous solution, and ultrasound

Methodology Applied

Scientific EffectUltrasound: Ultrasound

Implementation Method 2

Ultrafine bubbles more severely damage cell membrane than microbubbles when crushed

Methodology Applied

Scientific EffectAcoustic cavitation: Acoustic Cavitation

Data Source

PatentUS20220249665A1Transfection method

Publication Date: 2022.08.11 TAKEDA PHARMA CO LTD

AI summary

A novel means for safely and efficiently introducing a target substance such as nucleic acid, protein, or the like into cells (excluding immune cells) is provided by the present invention. Specifically, a system for delivering a target substance into a cell (excluding immune cells), including ultrafine bubble water or ultrafine bubble aqueous solution containing ultrafine bubbles with an average diameter of not more than 200 nm and not containing phospholipid, and an ultrasound generator in combination; a method for increasing the delivery of a nucleic acid, a protein or a low-molecular-weight compound into a cell (excluding immune cells) by using the ultrafine bubble water, etc.; and the like are provided.