Development of a culture chamber for mechanical loading of adherent cells with large uniform strain_Journal of Biomedical Engineering

Authors：

WANG Ziqi ^1,2 , GAO Lilan ^1,2 ,  LYU Linwei ^1,2 , WANG Xin ^1,2 , ZHANG Chunqiu ^1,2

1. Tianjin Key Laboratory of Advanced Mechatronic System Design and Intelligent Control, Tianjin University of Technology, Tianjin 300384, P. R. China;
2. National Experimental Teaching Demonstration Center of Mechatronics Engineering, Tianjin University of Technology, Tianjin 300384, P. R. China;

Corresponding author：

LYU Linwei, Email: lvlw@tjut.edu.cn

Keywords：

Loading device; Adherent cells; Strain uniformity; New culture chamber; Optimization simulation

DOI：

10.7507/1001-5515.202204008

Video：

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Based on the current study of the influence of mechanical factors on cell behavior which relies heavily on experiments in vivo, a culture chamber with a large uniform strain area containing a linear motor-powered, up-to-20-Hz cell stretch loading device was developed to exert mechanical effects on cells. In this paper, using the strain uniformity as the target and the substrate thickness as the variable, the substrate bottom of the conventional incubation chamber is optimized by using finite element technique, and finally a new three-dimensional model of the incubation chamber with “M” type structure in the section is constructed, and the distribution of strain and displacement fields are detected by 3D-DIC to verify the numerical simulation results. The experimental results showed that the new cell culture chamber increased the accuracy and homogeneous area of strain loading by 49.13% to 52.45% compared with that before optimization. In addition, the morphological changes of tongue squamous carcinoma cells under the same strain and different loading times were initially studied using this novel culture chamber. In conclusion, the novel cell culture chamber constructed in this paper combines the advantages of previous techniques to deliver uniform and accurate strains for a wide range of cell mechanobiology studies.

Citation： WANG Ziqi, GAO Lilan, LYU Linwei, WANG Xin, ZHANG Chunqiu. Development of a culture chamber for mechanical loading of adherent cells with large uniform strain. Journal of Biomedical Engineering, 2022, 39(5): 997-1004. doi: 10.7507/1001-5515.202204008 Copy

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2.	Hamill O P, Martinac B. Molecular basis of mechanotransduction in living cells. Physiol Rev, 2001, 81(2): 685-740.
3.	Seelbinder B, Scott A K, Nelson I, et al. TENSCell: imaging of stretch-activated cells reveals divergent nuclear behavior and tension. Biophys J, 2020, 118(11): 2627-2640.
4.	Cheng J, Zou Q, Xue Y, et al. Mechanical stretch promotes antioxidant responses and cardiomyogenic differentiation in P19 cells. J Tissue Eng Regen Med, 2021, 15(5): 453-462.
5.	Duncan R L, Turner C H. Mechanotransduction and the functional response of bone to mechanical strain. Calcif Tissue Int, 1995, 57(5): 344-358.
6.	Gao J, Fu S, Zeng Z, et al. Cyclic stretch promotes osteogenesis-related gene expression in osteoblast-like cells through a cofilin-associated mechanism. Mol Med Rep, 2016, 14(1): 218-224.
7.	Beloussov L V, Saveliev S V, Naumidi I I, et al. Mechanical stresses in embryonic tissues: patterns, morphogenetic role, and involvement in regulatory feedback. Int Rev Cytol, 1994, 150: 1-34.
8.	Simpson D G, Carver W, Borg T K, et al. Role of mechanical stimulation in the establishment and maintenance of muscle cell differentiation. Int Rev Cytol, 1994, 150: 69-94.
9.	Pfister B J, Grasman J M, Loverde J R. Exploiting biomechanics to direct the formation of nervous tissue. Curr Opin Biomed Eng, 2020, 14(1): 59-66.
10.	Brown T D. Techniques for mechanical stimulation of cells in vitro: a review. J Biomech, 2000, 33(1): 3-14.
11.	Chen J H, Liu C, You L, et al. Boning up on Wolff's law: mechanical regulation of the cells that make and maintain bone. J Biomech, 2010, 43(1): 108-118.
12.	Sears C, Kaunas R. The many ways adherent cells respond to applied stretch. J Biomech, 2016, 49(8): 1347-1354.
13.	Meng L, Xue G, Liu Q, et al. In-situ electromechanical testing and loading system for dynamic cell-biomaterial interaction study. Biomed Microdevices, 2020, 22(3): 56.
14.	Dai Z X, Shih P J, Yen J Y, et al. Functional assistance for stress distribution in cell culture membrane under periodically stretching. J Biomech, 2021, 125: 110564.
15.	Zhang Chunqiu, Qiu Lulu, Gao Lilan, et al. A novel dual-frequency loading system for studying mechanobiology of load-bearing tissue. Mater Sci Eng C, 2016, 69: 262-267.
16.	Van Dyke W S, Sun X, Richard A B, et al. Novel mechanical bioreactor for concomitant fluid shear stress and substrate strain. J Biomech, 2012, 45(7): 1323-1327.
17.	Bianchi F, George J H, Malboubi M, et al. Engineering a uniaxial substrate-stretching device for simultaneous electrophysiological measurements and imaging of strained peripheral neurons. Med Eng Phys, 2019, 67(1): 1-10.
18.	Apa L, Carraro S, Pisu S, et al. Development and validation of a device for in vitro uniaxial cell substrate deformation with real-time strain control. Meas Sci Technol, 2020, 31: 125702.
19.	Lei Y, Masjedi S, Ferdous Z. A study of extracellular matrix remodeling in aortic heart valves using a novel biaxial stretch bioreactor. J Mech Behav Biomed Mater, 2017, 75: 351-358.
20.	Dermenoudis S, Missirlis Y. Design of a novel rotating wall bioreactor for the in vitro simulation of the mechanical environment of the endothelial function. J Biomech, 2010, 43(7): 1426-1431.
21.	Toume S, Gefen A, Weihs D. Printable low-cost, sustained and dynamic cell stretching apparatus. J Biomech, 2016, 49(8): 1336-1339.
22.	Tsukamoto S, Asakawa T, Kimura S, et al. Intranuclear strain in living cells subjected to substrate stretching: A combined experimental and computational study. J Biomech, 2021, 119: 110292.
23.	Morita Y, Watanabe S, Ju Y, et al. In vitro experimental study for the determination of cellular axial strain threshold and preferential axial strain from cell orientation behavior in a non-uniform deformation field. Cell Biochem Biophys, 2013, 67(3): 1249-1259.
24.	Bieler F H, Ott C E, Thompson M S, et al. Biaxial cell stimulation: A mechanical validation. J Biomech, 2009, 42(11): 1692-1696.
25.	Gu Xiaojun, Cao Yinfeng, Zhu Jihong, et al. Shape optimization of SMA structures with respect to fatigue. Mater Design, 2020, 189: 108456.
26.	Upadhyay B D, Sonigra S S, Daxini S D. Numerical analysis perspective in structural shape optimization: A review post 2000. Adv Eng Softw, 2021, 155: 102992.
27.	郭新路, 刘蓉, 王永轩. 仿骨小梁力学性能的多孔结构拓扑优化设计. 医用生物力学, 2018, 33(5): 402-409.
28.	邱璐璐, 宋阳, 李可, 等. 数字图像相关技术在生物力学方面的研究进展. 生物医学工程与临床, 2017, 21(6): 676-681.
29.	Gilbert J A, Weinhold P S, Banes A J, et al. Strain profiles for circular cell culture plates containing flexible surfaces employed to mechanically deform cells in vitro. J Biomech, 1994, 27(9): 1169-1177.
30.	Albiges-Rizo C, Destaing O, Fourcade B, et al. Actin machinery and mechanosensitivity in invadopodia, podosomes and focal adhesions. J Cell Sci, 2009, 122(17): 3037-3049.
31.	Gasparski A N, Ozarkar S, Beningo K A. Transient mechanical strain promotes the maturation of invadopodia and enhances cancer cell invasion in vitro. J Cell Sci, 2017, 130(11): 1965-1978.

1. Gladilin E, Gonzalez P, Eils R. Dissecting the contribution of actin and vimentin intermediate filaments to mechanical phenotype of suspended cells using high-throughput deformability measurements and computational modeling. J Biomech, 2014, 47(11): 2598-2605.
2. Hamill O P, Martinac B. Molecular basis of mechanotransduction in living cells. Physiol Rev, 2001, 81(2): 685-740.
3. Seelbinder B, Scott A K, Nelson I, et al. TENSCell: imaging of stretch-activated cells reveals divergent nuclear behavior and tension. Biophys J, 2020, 118(11): 2627-2640.
4. Cheng J, Zou Q, Xue Y, et al. Mechanical stretch promotes antioxidant responses and cardiomyogenic differentiation in P19 cells. J Tissue Eng Regen Med, 2021, 15(5): 453-462.
5. Duncan R L, Turner C H. Mechanotransduction and the functional response of bone to mechanical strain. Calcif Tissue Int, 1995, 57(5): 344-358.
6. Gao J, Fu S, Zeng Z, et al. Cyclic stretch promotes osteogenesis-related gene expression in osteoblast-like cells through a cofilin-associated mechanism. Mol Med Rep, 2016, 14(1): 218-224.
7. Beloussov L V, Saveliev S V, Naumidi I I, et al. Mechanical stresses in embryonic tissues: patterns, morphogenetic role, and involvement in regulatory feedback. Int Rev Cytol, 1994, 150: 1-34.
8. Simpson D G, Carver W, Borg T K, et al. Role of mechanical stimulation in the establishment and maintenance of muscle cell differentiation. Int Rev Cytol, 1994, 150: 69-94.
9. Pfister B J, Grasman J M, Loverde J R. Exploiting biomechanics to direct the formation of nervous tissue. Curr Opin Biomed Eng, 2020, 14(1): 59-66.
10. Brown T D. Techniques for mechanical stimulation of cells in vitro: a review. J Biomech, 2000, 33(1): 3-14.
11. Chen J H, Liu C, You L, et al. Boning up on Wolff's law: mechanical regulation of the cells that make and maintain bone. J Biomech, 2010, 43(1): 108-118.
12. Sears C, Kaunas R. The many ways adherent cells respond to applied stretch. J Biomech, 2016, 49(8): 1347-1354.
13. Meng L, Xue G, Liu Q, et al. In-situ electromechanical testing and loading system for dynamic cell-biomaterial interaction study. Biomed Microdevices, 2020, 22(3): 56.
14. Dai Z X, Shih P J, Yen J Y, et al. Functional assistance for stress distribution in cell culture membrane under periodically stretching. J Biomech, 2021, 125: 110564.
15. Zhang Chunqiu, Qiu Lulu, Gao Lilan, et al. A novel dual-frequency loading system for studying mechanobiology of load-bearing tissue. Mater Sci Eng C, 2016, 69: 262-267.
16. Van Dyke W S, Sun X, Richard A B, et al. Novel mechanical bioreactor for concomitant fluid shear stress and substrate strain. J Biomech, 2012, 45(7): 1323-1327.
17. Bianchi F, George J H, Malboubi M, et al. Engineering a uniaxial substrate-stretching device for simultaneous electrophysiological measurements and imaging of strained peripheral neurons. Med Eng Phys, 2019, 67(1): 1-10.
18. Apa L, Carraro S, Pisu S, et al. Development and validation of a device for in vitro uniaxial cell substrate deformation with real-time strain control. Meas Sci Technol, 2020, 31: 125702.
19. Lei Y, Masjedi S, Ferdous Z. A study of extracellular matrix remodeling in aortic heart valves using a novel biaxial stretch bioreactor. J Mech Behav Biomed Mater, 2017, 75: 351-358.
20. Dermenoudis S, Missirlis Y. Design of a novel rotating wall bioreactor for the in vitro simulation of the mechanical environment of the endothelial function. J Biomech, 2010, 43(7): 1426-1431.
21. Toume S, Gefen A, Weihs D. Printable low-cost, sustained and dynamic cell stretching apparatus. J Biomech, 2016, 49(8): 1336-1339.
22. Tsukamoto S, Asakawa T, Kimura S, et al. Intranuclear strain in living cells subjected to substrate stretching: A combined experimental and computational study. J Biomech, 2021, 119: 110292.
23. Morita Y, Watanabe S, Ju Y, et al. In vitro experimental study for the determination of cellular axial strain threshold and preferential axial strain from cell orientation behavior in a non-uniform deformation field. Cell Biochem Biophys, 2013, 67(3): 1249-1259.
24. Bieler F H, Ott C E, Thompson M S, et al. Biaxial cell stimulation: A mechanical validation. J Biomech, 2009, 42(11): 1692-1696.
25. Gu Xiaojun, Cao Yinfeng, Zhu Jihong, et al. Shape optimization of SMA structures with respect to fatigue. Mater Design, 2020, 189: 108456.
26. Upadhyay B D, Sonigra S S, Daxini S D. Numerical analysis perspective in structural shape optimization: A review post 2000. Adv Eng Softw, 2021, 155: 102992.
27. 郭新路, 刘蓉, 王永轩. 仿骨小梁力学性能的多孔结构拓扑优化设计. 医用生物力学, 2018, 33(5): 402-409.
28. 邱璐璐, 宋阳, 李可, 等. 数字图像相关技术在生物力学方面的研究进展. 生物医学工程与临床, 2017, 21(6): 676-681.
29. Gilbert J A, Weinhold P S, Banes A J, et al. Strain profiles for circular cell culture plates containing flexible surfaces employed to mechanically deform cells in vitro. J Biomech, 1994, 27(9): 1169-1177.
30. Albiges-Rizo C, Destaing O, Fourcade B, et al. Actin machinery and mechanosensitivity in invadopodia, podosomes and focal adhesions. J Cell Sci, 2009, 122(17): 3037-3049.
31. Gasparski A N, Ozarkar S, Beningo K A. Transient mechanical strain promotes the maturation of invadopodia and enhances cancer cell invasion in vitro. J Cell Sci, 2017, 130(11): 1965-1978.

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