Name: magnetic-, acoustic-, and optical-triple-responsive microbubbles for magnetic hyperthermia and pothotothermal combination cancer therapy
Uploaded: 2020-05-22T14:30:00
Description: Watch this Scientific Journal Video about Magnetic-, Acoustic-, and Optical-Triple-Responsive Microbubbles for Magnetic Hyperthermia and Pothotothermal Combination Cancer Therapy at JoVE.com

This work was supported by the National Natural Science Foundation of China (81601608) and NUPTSF (NY216024).

All animal experiments were performed in accordance with the protocols approved by the OG Pharmaceutical guidelines for Animal Care and Use of Laboratory Animals. The protocols followed the guidelines of Ethics Committee for laboratory animals of OG Pharmaceutical.
1. Nanoparticle shelled microbubbles (NSMs) synthesis
<ol>
	<li>Disperse magnetic nanoparticles (Fe3O4, iron oxide) in deionized water to form a 10 mg/mL stock solution.</li>
	<li>Place the tube containing th...

<ol>
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Here, we presented a protocol of fabricating iron oxide nanoparticle shelled microbubbles (NSMs) through self-assembly, synergizing magnetic, acoustic, and optical responsiveness in one nanotherapeutic platform. The IONPs were densely packed around the air core to form a magnetic shell, which can be controlled by the external magnetic field for targeting. Once delivered, the release of IONPs can be achieved by ultrasound trigger. The released IONPs can be activated by both NIR light and AFM for PTT and MHT, or the combin...

Cancer is one of the deadliest diseases, causing millions of deaths every year worldwide and huge economic losses1. In clinics, conventional anticancer therapies, such as surgical resection, radiotherapy, and chemotherapy still cannot provide a satisfactory therapeutic efficacy2. Limitations of these therapies are high toxic side effects, high recurrence rate and high metastasis rate3. For example, chemotherapy is suffered from the low delivery efficiency of chemo drugs precisely to the tumor site4. The inability of drugs to penetrate deep into the tumor tissue across the biological barriers, including extracellular matrix and high tumor interstitial fluid pressure, is also responsible for the low therapeutic efficacy5. Besides, the tumor resistance usually happens in the patients who received treatment by single chemotherapy6. Therefore, techniques where thermal ablation of tumor occurs, such as photothermal therapy (PTT) and magnetic hyperthermia therapy (MHT), have shown promising results to reduce tumor resistance and have been emerging in clinical trials7,8,9.
PTT triggers thermal ablation of cancer cells by the action of photothermal conversion agents under the irradiation of the laser energy. The generated high temperature (above 50 &#730;C) induces complete cell necrosis10. Very recently, iron oxide nanoparticles (IONPs) were demonstrated to be a photothermal conversion agent that can be activated by near-infrared (NIR) light11. &#160;Despite the low molar absorption coefficient in the near infrared region, IONPs are candidates for low-temperature (43&#160; &#730;C) photothermal therapy, a modified therapy to reduce the damage caused by heat exposure to normal tissues and to initiate antitumor immunity against tumor metastasis12. One of the limitations of PTT is the low penetration depth of the laser. For deep seated tumors, alternating magnetic field (AFM) induced heating of iron oxide nanoparticles, also called magnetic hyperthermia, is an alternative therapy for PTT13,14. The main advantage of MHT is the high penetration of magnetic field15. However, the required relatively high concentration of IONPs remains a major disadvantage for its clinical application. The delivery efficiency of nanomedicine (or nanoparticles) to solid tumors in animals has been 1-10% due to a series of obstacles including circulation, accumulation, and penetration16,17. Therefore, a controlled and targeted IONPs delivery strategy with the ability to achieve high tissue penetration is of great interest in cancer treatment.
Ultrasound mediated nanoparticle delivery has shown its ability to facilitate the penetration of nanoparticles deep into the tumor tissue, due to the phenomenon called microbubble cavitation18,19. In the present study, we fabricate IONPs shelled microbubbles (NSMs) through self-assembly, synergizing magnetic, acoustic, and optical responsiveness in one nanotherapeutic platform. The NSM contains an air core and a shell of iron oxide nanoparticles, with a diameter of approximately 5.4 &#181;m. The NSMs can be magnetically guided to the tumor site. Then the release of IONPs is triggered by ultrasound, accompanied by microbubble cavitation and microstreaming. The momentum received from the microstreaming facilitates the penetration of IONPs into the tumor tissue. The PTT and MHT can be achieved by NIR laser irradiation or AFM application, or with the combination of both.

<table border="0" cellpadding="0" cellspacing="0" style="width:1331px;" width="1330">
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	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:337px;">808 nm laser power</td>
			<td style="width:315px;">Changchun New Industries Optoelectronics Tech</td>
			<td style="width:263px;">MDL-F-808-5W-18017023</td>
			<td style="width:416px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Calcein-AM</td>
			<td>Thermo Fisher SCIENTIFIC</td>
			<td>C3099</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Fetal bovine serum</td>
			<td>Invitrogen</td>
			<td>16000-044</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Fluorescence Microscope</td>
			<td>Olympus</td>
			<td>IX71</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;">Function generator</td>
			<td>Keysight</td>
			<td>33500B series</td>
			<td style="width:416px;">20 MHz, 2 channels with arbitrary waveform generation capability</td>
		</tr>
		<tr height="48">
			<td height="48" style="height:48px;">Gelatin gel</td>
			<td>Sigma</td>
			<td>9000-70-8</td>
			<td style="width:416px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Heating machine</td>
			<td>Shuangping</td>
			<td>SPG-06- II</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Homemade focused transducer</td>
			<td></td>
			<td></td>
			<td>Frequency=855, R-X=36.2W+5.8W, |Z|-&#952;=37W+8&#176;</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Homogenizer</td>
			<td>SCILOGEX</td>
			<td>D-160</td>
			<td>8000-30000 rpm</td>
		</tr>
		<tr height="41">
			<td height="41" style="height:41px;">Hydrophone</td>
			<td>T&#38;C</td>
			<td>NH1000</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">ICR male mice</td>
			<td>OG Pharmaceutical. Co. Ltd</td>
			<td></td>
			<td>8-week-old</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Inductively coupled plasma optical emission spectrometry</td>
			<td>PerkinElmer</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Infrared thermal imaging camera.</td>
			<td>FLIR</td>
			<td>E50</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Iron(II,III) oxide</td>
			<td>Alfa Aesar</td>
			<td>1317-61-9</td>
			<td>50-100nm APS Powder</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Laser power meter</td>
			<td colspan="2">Changchun New Industries Optoelectronics Tech</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Oscilloscope</td>
			<td>Keysight</td>
			<td>DSOX3054T</td>
			<td>Bandwidth 500 MHz, Sampling Rate 5 GS/S, 4 channels</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">RF Power Amplifier</td>
			<td>T&#38;C</td>
			<td>AG1020</td>
			<td style="width:416px;">The signal source can also be connected to an external signal source. The gain can be adjusted from 0 to 100%. It has multiple functions such as frequency sweep, pulse, and triangle.</td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Roswell Park Memorial Institute-1640</td>
			<td>KeyGEN BioTECH</td>
			<td>KGM31800</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Sodium dodecyl sulfate</td>
			<td>Sigma</td>
			<td>151-21-3</td>
			<td></td>
		</tr>
	</tbody>
</table>

magnetic-, acoustic-, and optical-triple-responsive microbubbles for magnetic hyperthermia and pothotothermal combination cancer therapy

The precision delivery of anti-cancer agents which aim for targeted and deep-penetrated delivery as well as a controlled release at the tumor site has been challenged. Here, we fabricate iron oxide nanoparticle shelled microbubbles (NSMs) through self-assembly, synergizing magnetic, acoustic, and optical responsiveness in one nanotherapeutic platform. Iron oxide nanoparticles serve as both magnetic and photothermal agents. Once intravenously injected, NSMs can be magnetically guided to the tumor site. Ultrasound triggers the release of iron oxide nanoparticles, facilitating the penetration of nanoparticles deep into the tumor due to the cavitation effect of microbubbles. Thereafter, magnetic hyperthermia and photothermal therapy can be performed on the tumor for combinational cancer therapy, a solution for cancer resistance due to the tumor heterogeneity. In this protocol, the synthesis and characterization of NSMs including structural, chemical, magnetic and acoustic properties were performed. In addition, the anti-cancer efficacy by thermal therapy was investigated using in vitro cell cultures. The proposed delivery strategy and combination therapy holds great promise in cancer treatment to improve both delivery and anticancer efficacies.

The triple-responsive nanoparticle-shelled microbubbles (NSMs) used in this study were prepared by agitating the mixture of the surfactant and IONPs. The IONPs (50 nm) self-assembled at the interface of liquid and gas core, to form a densely packed magnetic shell. The morphology of NSMs is shown in Figure. 1A. The resulted NSMs presented a spherical shape and with an average diameter of 5.41 &#177; 1.78 &#956;m (Figure 1B). The results indicated the NSMs were prepared successfully. When ...

Watch this Scientific Journal Video about Magnetic-, Acoustic-, and Optical-Triple-Responsive Microbubbles for Magnetic Hyperthermia and Pothotothermal Combination Cancer Therapy at JoVE.com

Magnetic-, Acoustic-, and Optical-Triple-Responsive Microbubbles for Magnetic Hyperthermia and Pothotothermal Combination Cancer Therapy

Presented here is a protocol for the fabrication of iron oxide nanoparticle-shelled microbubbles (NSMs) through self-assembly, synergizing magnetic, acoustic, and optical responsiveness in one nanotherapeutic platform for magnetic hyperthermia and photothermal combination cancer therapy.

The precision delivery of anti-cancer agents which aim for targeted and deep-penetrated delivery as well as a controlled release at the tumor site has ...

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