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Showing content from https://pubmed.ncbi.nlm.nih.gov/25200612/ below:

Cellular level robotic surgery: Nanodissection of intermediate filaments in live keratinocytes

Affiliations

• ¹ Department of Electrical and Computer Engineering, Michigan State University, East Lansing, MI, 48824, USA.
• ² Department of Mechanical and Biomedical Engineering, City University of Hong Kong, Hong Kong.
• ³ Hong Kong Productivity Council, Hong Kong.
• ⁴ College of Human Medicine, Michigan State University, East Lansing, MI, 48824, USA.
• ⁵ Department of Dermatology, University at Buffalo, Buffalo, NY, 14203, USA.
• ⁶ Department of Dermatology, University at Buffalo, Buffalo, NY, 14203, USA. Electronic address: aasinha@buffalo.edu.
• ⁷ Department of Electrical and Computer Engineering, Michigan State University, East Lansing, MI, 48824, USA. Electronic address: xin@egr.msu.edu.

• PMID: 25200612
• PMCID: PMC4280342
• DOI: 10.1016/j.nano.2014.08.008

Item in Clipboard

Ruiguo Yang et al. Nanomedicine. 2015 Jan.

• ¹ Department of Electrical and Computer Engineering, Michigan State University, East Lansing, MI, 48824, USA.
• ² Department of Mechanical and Biomedical Engineering, City University of Hong Kong, Hong Kong.
• ³ Hong Kong Productivity Council, Hong Kong.
• ⁴ College of Human Medicine, Michigan State University, East Lansing, MI, 48824, USA.
• ⁵ Department of Dermatology, University at Buffalo, Buffalo, NY, 14203, USA.
• ⁶ Department of Dermatology, University at Buffalo, Buffalo, NY, 14203, USA. Electronic address: aasinha@buffalo.edu.
• ⁷ Department of Electrical and Computer Engineering, Michigan State University, East Lansing, MI, 48824, USA. Electronic address: xin@egr.msu.edu.

• PMID: 25200612
• PMCID: PMC4280342
• DOI: 10.1016/j.nano.2014.08.008

Item in Clipboard

We present the nanosurgery on the cytoskeleton of live cells using AFM based nanorobotics to achieve adhesiolysis and mimic the effect of pathophysiological modulation of intercellular adhesion. Nanosurgery successfully severs the intermediate filament bundles and disrupts cell-cell adhesion similar to the desmosomal protein disassembly in autoimmune disease, or the cationic modulation of desmosome formation. Our nanomechanical analysis revealed that adhesion loss results in a decrease in cellular stiffness in both cases of biochemical modulation of the desmosome junctions and mechanical disruption of intercellular adhesion, supporting the notion that intercellular adhesion through intermediate filaments anchors the cell structure as focal adhesion does and that intermediate filaments are integral components in cell mechanical integrity. The surgical process could potentially help reveal the mechanism of autoimmune pathology-induced cell-cell adhesion loss as well as its related pathways that lead to cell apoptosis.

Keywords: Atomic Force Microscopy; Cell–cell adhesion; Desmosome; Intermediate filament; Mechanical property; Nanosurgery.

Copyright © 2015 Elsevier Inc. All rights reserved.

PubMed Disclaimer

The authors have no conflict of interest to declare.

Intermediate filament bundles imaged with…

Intermediate filament bundles imaged with AFM show stranded filamentous structures at the peripheral…

Intermediate filament bundles imaged with AFM show stranded filamentous structures at the peripheral of the cells (arrows). A: Topography image, B: Deflection error image, C: Three dimensional rendering. Scan size: 30 μm.

The tensegrity structure with 6…

The tensegrity structure with 6 compressional (black) and 24 tensional (blue) elements was…

The tensegrity structure with 6 compressional (black) and 24 tensional (blue) elements was used to model the cell stiffness with (A) and without (B) the intermediate filament which is considered tensional elements (red) projecting from the nucleus to the cell peripheral.

(A) Illustration of composite cell…

(A) Illustration of composite cell structure during cell indentation and dissection; (B) Force-displacement…

(A) Illustration of composite cell structure during cell indentation and dissection; (B) Force-displacement curve without penetration of the cell membrane; (C) Force displacement curve with penetration of the cell membrane with the kink, indicating the force drop, zoomed-in in the inset, a 1.9 nN penetration force was observed.

( A ) The lateral…

( A ) The lateral force was the laser position shift in the…

(A) The lateral force was the laser position shift in the horizontal direction on the detector; (B) The tangent direction of the trajectory defines the angle (θ) between the cutting force F and the lateral force F_n; (C) The trajectory of the nanosurgery on the cell periphery between neighboring cells with the black triangle indicating the tip position, the 3D structure was recreated from the AFM image obtained before nanosurgery using custom-made nanomanipulation software; (D) the X, Y, Z piezo position and the cutting force F in the process of the nanosurgery along the path in (C). Three repetitions of dissection operation were recorded with a cutting depth of 100 nm. The detailed force profile is magnified with the insect showing the stretching distance Δl=148 nm and the distance between two strands of filaments d=360 nm.

( A ), ( B…

( A ), ( B ) The AFM image of keratinocyte cells before…

(A), (B) The AFM image of keratinocyte cells before and after three repeating nanosurgeries (scan size: 24.5 μm); (C), (D) the height information of the line section indicated in (A)(B) before and after nanosurgery along the trajectory in Figure 4 shows the detailed structural change. The severing was successfully completed indicated by the structural rearrangement of the cytoskeleton, since the pre-defined cutting depth, the structures at the lower topography area was not affected.

(A) The stiffness change of…

(A) The stiffness change of individual cells before ten repetitions of nanosurgery with…

(A) The stiffness change of individual cells before ten repetitions of nanosurgery with cutting depth of 100 nm each and after a 3 hour continuous observation; (B) the stiffness drop was assessed statistically before and after nanosurgery from 36.1± 3.5 kPa to 18.4 ± 1.9 kPa (mean ± SEM, n=200). (C) The stiffness drop was observed for anti-Dsg3 antibody treated samples and the Ca²⁺ depleted samples; (D) the normal keratinocyte cell structure and (E) the cell structure after non-formation of desmosomes. (F) The stiffness comparison with and without intermediate filament over strain; a significant drop of stiffness was observed without intermediate filament connection.

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  Shakoor A, Gao W, Zhao L, Jiang Z, Sun D. Shakoor A, et al. Microsyst Nanoeng. 2022 Apr 29;8:47. doi: 10.1038/s41378-022-00376-0. eCollection 2022. Microsyst Nanoeng. 2022. PMID: 35502330 Free PMC article. Review.

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