Pulsed laser assisted high-throughput intracellular delivery in hanging drop based three dimensional cancer spheroids
Targeted intracellular delivery of biomolecules and therapeutic cargo enables the controlled manipulation of cellular processes. Laser-based optoporation has emerged as a versatile, non-invasive technique that employs light-based transient physical disruption of the cell membrane and achieves high t...
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description | Targeted intracellular delivery of biomolecules and therapeutic cargo enables the controlled manipulation of cellular processes. Laser-based optoporation has emerged as a versatile, non-invasive technique that employs light-based transient physical disruption of the cell membrane and achieves high transfection efficiency with low cell damage. Testing of the delivery efficiency of optoporation-based techniques has been conducted on single cells in monolayers, but its applicability in three-dimensional (3D) cell clusters/spheroids has not been explored. Cancer cells grown as 3D tumor spheroids are widely used in anti-cancer drug screening and can be potentially employed for testing delivery efficiency. Towards this goal, we demonstrated the optoporation-based high-throughput intracellular delivery of a model fluorescent cargo (propidium iodide, PI) within 3D SiHa human cervical cancer spheroids. To enable this technique, nano-spiked core-shell gold-coated polystyrene nanoparticles (ns-AuNPs) with a high surface-to-volume ratio were fabricated. ns-AuNPs exhibited high electric field enhancement and highly localized heating at an excitation wavelength of 680 nm. ns-AuNPs were co-incubated with cancer cells within hanging droplets to enable the rapid aggregation and assembly of spheroids. Nanosecond pulsed-laser excitation at the optimized values of laser fluence (45 mJ cm
−2
), pulse frequency (10 Hz), laser exposure time (30 s), and ns-AuNP concentration (5 × 10
10
particles per ml) resulted in the successful delivery of PI dye into cancer cells. This technique ensured high delivery efficiency (89.6 ± 2.8%) while maintaining high cellular viability (97.4 ± 0.4%), thereby validating the applicability of this technique for intracellular delivery. The optoporation-based strategy can enable high-throughput single cell manipulation, is scalable towards larger 3D tissue constructs, and may provide translational benefits for the delivery of anti-cancer therapeutics to tumors.
This is the first study to report laser mediated optoporation-based intracellular delivery in 3D cellular constructs grown in hanging drop cultures. |
doi_str_mv | 10.1039/d0an02432e |
format | Article |
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−2
), pulse frequency (10 Hz), laser exposure time (30 s), and ns-AuNP concentration (5 × 10
10
particles per ml) resulted in the successful delivery of PI dye into cancer cells. This technique ensured high delivery efficiency (89.6 ± 2.8%) while maintaining high cellular viability (97.4 ± 0.4%), thereby validating the applicability of this technique for intracellular delivery. The optoporation-based strategy can enable high-throughput single cell manipulation, is scalable towards larger 3D tissue constructs, and may provide translational benefits for the delivery of anti-cancer therapeutics to tumors.
This is the first study to report laser mediated optoporation-based intracellular delivery in 3D cellular constructs grown in hanging drop cultures.</description><identifier>ISSN: 0003-2654</identifier><identifier>EISSN: 1364-5528</identifier><identifier>DOI: 10.1039/d0an02432e</identifier><identifier>PMID: 34240729</identifier><language>eng</language><publisher>England: Royal Society of Chemistry</publisher><subject>Biomolecules ; Cancer ; Cargo ; Cell membranes ; Cell Survival ; Efficiency ; Electric fields ; Excitation ; Fluence ; Fluorescence ; Gold ; Gold coatings ; Humans ; Laser applications ; Lasers ; Metal Nanoparticles ; Nanoparticles ; Nanosecond pulses ; Neoplasms - drug therapy ; Polystyrene resins ; Pulsed lasers ; Spheroids ; Spheroids, Cellular ; Tumors</subject><ispartof>Analyst (London), 2021-08, Vol.146 (15), p.4756-4766</ispartof><rights>Copyright Royal Society of Chemistry 2021</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c428t-264894045926e859bea086e943e2ff76ebdaa5fad954de9860f01c7512d982813</citedby><cites>FETCH-LOGICAL-c428t-264894045926e859bea086e943e2ff76ebdaa5fad954de9860f01c7512d982813</cites><orcidid>0000-0001-8648-383X ; 0000-0002-4073-9980 ; 0000-0001-7654-6905 ; 0000-0002-9403-2155</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,776,780,881,2817,2818,27903,27904</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/34240729$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Gupta, Pallavi</creatorcontrib><creatorcontrib>Kar, Srabani</creatorcontrib><creatorcontrib>Kumar, Ashish</creatorcontrib><creatorcontrib>Tseng, Fan-Gang</creatorcontrib><creatorcontrib>Pradhan, Shantanu</creatorcontrib><creatorcontrib>Mahapatra, Pallab Sinha</creatorcontrib><creatorcontrib>Santra, Tuhin Subhra</creatorcontrib><title>Pulsed laser assisted high-throughput intracellular delivery in hanging drop based three dimensional cancer spheroids</title><title>Analyst (London)</title><addtitle>Analyst</addtitle><description>Targeted intracellular delivery of biomolecules and therapeutic cargo enables the controlled manipulation of cellular processes. Laser-based optoporation has emerged as a versatile, non-invasive technique that employs light-based transient physical disruption of the cell membrane and achieves high transfection efficiency with low cell damage. Testing of the delivery efficiency of optoporation-based techniques has been conducted on single cells in monolayers, but its applicability in three-dimensional (3D) cell clusters/spheroids has not been explored. Cancer cells grown as 3D tumor spheroids are widely used in anti-cancer drug screening and can be potentially employed for testing delivery efficiency. Towards this goal, we demonstrated the optoporation-based high-throughput intracellular delivery of a model fluorescent cargo (propidium iodide, PI) within 3D SiHa human cervical cancer spheroids. To enable this technique, nano-spiked core-shell gold-coated polystyrene nanoparticles (ns-AuNPs) with a high surface-to-volume ratio were fabricated. ns-AuNPs exhibited high electric field enhancement and highly localized heating at an excitation wavelength of 680 nm. ns-AuNPs were co-incubated with cancer cells within hanging droplets to enable the rapid aggregation and assembly of spheroids. Nanosecond pulsed-laser excitation at the optimized values of laser fluence (45 mJ cm
−2
), pulse frequency (10 Hz), laser exposure time (30 s), and ns-AuNP concentration (5 × 10
10
particles per ml) resulted in the successful delivery of PI dye into cancer cells. This technique ensured high delivery efficiency (89.6 ± 2.8%) while maintaining high cellular viability (97.4 ± 0.4%), thereby validating the applicability of this technique for intracellular delivery. The optoporation-based strategy can enable high-throughput single cell manipulation, is scalable towards larger 3D tissue constructs, and may provide translational benefits for the delivery of anti-cancer therapeutics to tumors.
This is the first study to report laser mediated optoporation-based intracellular delivery in 3D cellular constructs grown in hanging drop cultures.</description><subject>Biomolecules</subject><subject>Cancer</subject><subject>Cargo</subject><subject>Cell membranes</subject><subject>Cell Survival</subject><subject>Efficiency</subject><subject>Electric fields</subject><subject>Excitation</subject><subject>Fluence</subject><subject>Fluorescence</subject><subject>Gold</subject><subject>Gold coatings</subject><subject>Humans</subject><subject>Laser applications</subject><subject>Lasers</subject><subject>Metal Nanoparticles</subject><subject>Nanoparticles</subject><subject>Nanosecond pulses</subject><subject>Neoplasms - drug therapy</subject><subject>Polystyrene resins</subject><subject>Pulsed lasers</subject><subject>Spheroids</subject><subject>Spheroids, Cellular</subject><subject>Tumors</subject><issn>0003-2654</issn><issn>1364-5528</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2021</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkU1v2zAMhoViQ5tmu_TeQcBuBdzq07YuA4KsX0Cw7rCdBcWibQWO5Ul2gP77KkubrSeC5MOXJF6ELii5poSrG0tMT5jgDE7QjPJcZFKy8gOaEUJ4xnIpztB5jJuUUiLJKTrjgglSMDVD08-pi2BxZyIEbGJ0cUxp65o2G9vgp6YdphG7fgymgq6bOhOwhc7tIDynMm5N37i-wTb4Aa_NXivNAWDrttBH53vT4cr0VZKPQwvBOxs_oY-1SXs_v8Y5-n13-2v5kK2e7h-Xi1VWCVaO6XRRKkGEVCyHUqo1GFLmoAQHVtdFDmtrjKyNVVJYUGVOakKrQlJmVclKyufo20F3mNZbsBXs3-j0ENzWhGftjdPvO71rdeN3usgp5apIAl9fBYL_M0Ec9cZPIb0UNZNSKC6k4Im6OlBV8DEGqI8bKNF7i_R3svjx16LbBH_5_6Yj-uZJAi4PQIjVsfvPY_4CTMGY8A</recordid><startdate>20210807</startdate><enddate>20210807</enddate><creator>Gupta, Pallavi</creator><creator>Kar, Srabani</creator><creator>Kumar, Ashish</creator><creator>Tseng, Fan-Gang</creator><creator>Pradhan, Shantanu</creator><creator>Mahapatra, Pallab Sinha</creator><creator>Santra, Tuhin Subhra</creator><general>Royal Society of Chemistry</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7SR</scope><scope>7U5</scope><scope>8BQ</scope><scope>8FD</scope><scope>JG9</scope><scope>L7M</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0001-8648-383X</orcidid><orcidid>https://orcid.org/0000-0002-4073-9980</orcidid><orcidid>https://orcid.org/0000-0001-7654-6905</orcidid><orcidid>https://orcid.org/0000-0002-9403-2155</orcidid></search><sort><creationdate>20210807</creationdate><title>Pulsed laser assisted high-throughput intracellular delivery in hanging drop based three dimensional cancer spheroids</title><author>Gupta, Pallavi ; Kar, Srabani ; Kumar, Ashish ; Tseng, Fan-Gang ; Pradhan, Shantanu ; Mahapatra, Pallab Sinha ; Santra, Tuhin Subhra</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c428t-264894045926e859bea086e943e2ff76ebdaa5fad954de9860f01c7512d982813</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2021</creationdate><topic>Biomolecules</topic><topic>Cancer</topic><topic>Cargo</topic><topic>Cell membranes</topic><topic>Cell Survival</topic><topic>Efficiency</topic><topic>Electric fields</topic><topic>Excitation</topic><topic>Fluence</topic><topic>Fluorescence</topic><topic>Gold</topic><topic>Gold coatings</topic><topic>Humans</topic><topic>Laser applications</topic><topic>Lasers</topic><topic>Metal Nanoparticles</topic><topic>Nanoparticles</topic><topic>Nanosecond pulses</topic><topic>Neoplasms - drug therapy</topic><topic>Polystyrene resins</topic><topic>Pulsed lasers</topic><topic>Spheroids</topic><topic>Spheroids, Cellular</topic><topic>Tumors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Gupta, Pallavi</creatorcontrib><creatorcontrib>Kar, Srabani</creatorcontrib><creatorcontrib>Kumar, Ashish</creatorcontrib><creatorcontrib>Tseng, Fan-Gang</creatorcontrib><creatorcontrib>Pradhan, Shantanu</creatorcontrib><creatorcontrib>Mahapatra, Pallab Sinha</creatorcontrib><creatorcontrib>Santra, Tuhin Subhra</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Engineered Materials Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Materials Research Database</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Analyst (London)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Gupta, Pallavi</au><au>Kar, Srabani</au><au>Kumar, Ashish</au><au>Tseng, Fan-Gang</au><au>Pradhan, Shantanu</au><au>Mahapatra, Pallab Sinha</au><au>Santra, Tuhin Subhra</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Pulsed laser assisted high-throughput intracellular delivery in hanging drop based three dimensional cancer spheroids</atitle><jtitle>Analyst (London)</jtitle><addtitle>Analyst</addtitle><date>2021-08-07</date><risdate>2021</risdate><volume>146</volume><issue>15</issue><spage>4756</spage><epage>4766</epage><pages>4756-4766</pages><issn>0003-2654</issn><eissn>1364-5528</eissn><abstract>Targeted intracellular delivery of biomolecules and therapeutic cargo enables the controlled manipulation of cellular processes. Laser-based optoporation has emerged as a versatile, non-invasive technique that employs light-based transient physical disruption of the cell membrane and achieves high transfection efficiency with low cell damage. Testing of the delivery efficiency of optoporation-based techniques has been conducted on single cells in monolayers, but its applicability in three-dimensional (3D) cell clusters/spheroids has not been explored. Cancer cells grown as 3D tumor spheroids are widely used in anti-cancer drug screening and can be potentially employed for testing delivery efficiency. Towards this goal, we demonstrated the optoporation-based high-throughput intracellular delivery of a model fluorescent cargo (propidium iodide, PI) within 3D SiHa human cervical cancer spheroids. To enable this technique, nano-spiked core-shell gold-coated polystyrene nanoparticles (ns-AuNPs) with a high surface-to-volume ratio were fabricated. ns-AuNPs exhibited high electric field enhancement and highly localized heating at an excitation wavelength of 680 nm. ns-AuNPs were co-incubated with cancer cells within hanging droplets to enable the rapid aggregation and assembly of spheroids. Nanosecond pulsed-laser excitation at the optimized values of laser fluence (45 mJ cm
−2
), pulse frequency (10 Hz), laser exposure time (30 s), and ns-AuNP concentration (5 × 10
10
particles per ml) resulted in the successful delivery of PI dye into cancer cells. This technique ensured high delivery efficiency (89.6 ± 2.8%) while maintaining high cellular viability (97.4 ± 0.4%), thereby validating the applicability of this technique for intracellular delivery. The optoporation-based strategy can enable high-throughput single cell manipulation, is scalable towards larger 3D tissue constructs, and may provide translational benefits for the delivery of anti-cancer therapeutics to tumors.
This is the first study to report laser mediated optoporation-based intracellular delivery in 3D cellular constructs grown in hanging drop cultures.</abstract><cop>England</cop><pub>Royal Society of Chemistry</pub><pmid>34240729</pmid><doi>10.1039/d0an02432e</doi><tpages>11</tpages><orcidid>https://orcid.org/0000-0001-8648-383X</orcidid><orcidid>https://orcid.org/0000-0002-4073-9980</orcidid><orcidid>https://orcid.org/0000-0001-7654-6905</orcidid><orcidid>https://orcid.org/0000-0002-9403-2155</orcidid><oa>free_for_read</oa></addata></record> |
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source | MEDLINE; Royal Society of Chemistry Journals Archive (1841-2007); Royal Society Of Chemistry Journals 2008-; Alma/SFX Local Collection |
subjects | Biomolecules Cancer Cargo Cell membranes Cell Survival Efficiency Electric fields Excitation Fluence Fluorescence Gold Gold coatings Humans Laser applications Lasers Metal Nanoparticles Nanoparticles Nanosecond pulses Neoplasms - drug therapy Polystyrene resins Pulsed lasers Spheroids Spheroids, Cellular Tumors |
title | Pulsed laser assisted high-throughput intracellular delivery in hanging drop based three dimensional cancer spheroids |
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