title: Development of a custom biological scaffold for investigating ultrasound-mediated intracellular delivery.
abstract: In vitro investigations of ultrasound mediated, intracellular drug and gene delivery (i.e. sonoporation) are typically carried out in cells cultured in standard plastic well plates. This creates conditions that poorly resemble in vivo conditions, as well as generating unwanted ultrasound phenomena that may confound the interpretation of results. Here, we present our results in the development of a biological scaffold for sonoporation studies. The scaffolds were comprised of cellulose fibers coated with chitosan and gelatin. Scaffold formulation was optimized for adherence and proliferation of mouse fibroblasts in terms of the ratio and relative concentration of the two constituents. The scaffolds were also shown to significantly reduce ultrasound reflections compared to the plastic well plates. A custom treatment chamber was designed and built, and the occurrence of acoustic cavitation in the chamber during the ultrasound treatments was detected; a requirement for the process of sonoporation. Finally, experiments were carried out to optimize the ultrasound exposures to minimize cellular damage. Ultrasound exposure was then shown to enable the uptake of 100nm fluorescently labeled polystyrene nanoparticles in suspension into the cells seeded on scaffolds, compared to incubation of cell-seeded scaffolds with nanoparticles alone. These preliminary results set the basis for further development of this platform. They also provide motivation for the development of similar platforms for the controlled investigation of other ultrasound mediated cell and tissue therapies.
authors: Department of Bioengineering, University of Texas at Arlington, Arlington, TX 76010, United States.
authors: Loan
authors: L
authors: Bui
authors: Department of Biomedical Technology, King Saud University, Riyadh 12372, Saudi Arabia.
authors: Adham
authors: A
authors: Aleid
authors: Department of Biomedical Engineering, University of Miami, Coral Gables, FL 33146, United States; Department of Medical Equipment Technology, Majmaah University, Majmaah City 11952, Saudi Arabia.
authors: Ahmad
authors: A
authors: Alassaf
authors: Department of Biomedical Engineering, Catholic University of America, Washington, DC 20064, United States.
authors: Otto C
authors: OC
authors: Wilson
authors: Department of Biomedical Engineering, Catholic University of America, Washington, DC 20064, United States.
authors: Christopher B
authors: CB
authors: Raub
authors: Department of Diagnostic Radiology and Nuclear Medicine, University of Maryland School of Medicine, Baltimore, MD 21201, United States; Marlene and Stewart Greenebaum Cancer Center, University of Maryland School of Medicine, Baltimore, MD 21201, United States. Electronic address: vfrenkel@som.umaryland.edu.
authors: Victor
authors: V
authors: Frenkel
mesh_terms: D000162:Acoustics; D000818:Animals; D002460:Cell Line; D048271:Chitosan / Q000737:chemistry; D016503:Drug Delivery Systems; D018274:Electroporation; D004705:Endocytosis; D005780:Gelatin / Q000737:chemistry; D018014:Gene Transfer Techniques; D007091:Image Processing, Computer-Assisted; D042541:Intracellular Space / Q000378:metabolism*; D049449:Luminescence; D051379:Mice; D008856:Microscopy, Fluorescence; D053758:Nanoparticles; D015203:Reproducibility of Results; D054457:Tissue Scaffolds / Q000737:chemistry*; D014465:Ultrasonics / Q000379:methods*
chemical_list: D005780:Gelatin; D048271:Chitosan
keywords: Biological Scaffolds; Inertial cavitation; Intracellular drug delivery; Reflections; Sonoporation; Ultrasound