title: Biomimetic stochastic topography and electric fields synergistically enhance directional migration of corneal epithelial cells in a MMP-3-dependent manner.
abstract: Directed migration of corneal epithelial cells (CECs) is critical for maintenance of corneal homeostasis as well as wound healing. Soluble cytoactive factors and the intrinsic chemical attributes of the underlying extracellular matrix (ECM) participate in stimulating and directing migration. The central importance of the intrinsic biophysical attributes of the microenvironment of the cell in modulating an array of fundamental epithelial behaviors including migration has been widely documented. Among the best measures of these attributes are the intrinsic topography and stiffness of the ECM and electric fields (EFs). How cells integrate these multiple simultaneous inputs is not well understood. Here, we present a method that combines the use of (i) topographically patterned substrates (mean pore diameter 800nm) possessing features that approximate those found in the native corneal basement membrane; and (ii) EFs (0-150mVmm(-1)) mimicking those at corneal epithelial wounds that the cells experience in vivo. We found that topographic cues and EFs synergistically regulated directional migration of human CECs and that this was associated with upregulation of matrix metalloproteinase-3 (MMP3). MMP3 expression and activity were significantly elevated with 150mVmm(-1) applied-EF while MMP2/9 remained unaltered. MMP3 expression was elevated in cells cultured on patterned surfaces against planar surfaces. The highest single-cell migration rate was observed with 150mVmm(-1) applied EF on patterned and planar surfaces. When cultured as a confluent sheet, EFs induced collective cell migration on stochastically patterned surfaces compared with dissociated single-cell migration on planar surfaces. These results suggest significant interaction of biophysical cues in regulating cell behaviors and will help define design parameters for corneal prosthetics and help to better understand corneal wound healing.
authors.affiliation: Department of Dermatology, School of Medicine, University of California Davis, Sacramento, CA 95817, USA; School of Life Science, Yunnan Normal University, Kunming, Yunnan 650500, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Gao
authors.affiliation: Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California Davis, Davis, CA 95616, USA.
authors.forename: Vijay Krishna
authors.initials: VK
authors.lastname: Raghunathan
authors.affiliation: Department of Dermatology, School of Medicine, University of California Davis, Sacramento, CA 95817, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Reid
authors.affiliation: Department of Otolaryngology, School of Medicine, University of California Davis, Sacramento, CA 95817, USA.
authors.forename: Dongguang
authors.initials: D
authors.lastname: Wei
authors.affiliation: Department of Otolaryngology, School of Medicine, University of California Davis, Sacramento, CA 95817, USA.
authors.forename: Rodney C
authors.initials: RC
authors.lastname: Diaz
authors.affiliation: Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California Davis, Davis, CA 95616, USA.
authors.forename: Paul
authors.initials: P
authors.lastname: Russell
authors.affiliation: Department of Surgical and Radiological Sciences, School of Veterinary Medicine, University of California Davis, Davis, CA 95616, USA; Department of Ophthalmology & Vision Science, School of Medicine, University of California Davis, Sacramento, CA 95817, USA. Electronic address: cjmurphy@ucdavis.edu.
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Murphy
authors.affiliation: Department of Dermatology, School of Medicine, University of California Davis, Sacramento, CA 95817, USA; School of Life Science, Yunnan Normal University, Kunming, Yunnan 650500, People's Republic of China; Department of Ophthalmology & Vision Science, School of Medicine, University of California Davis, Sacramento, CA 95817, USA. Electronic address: minzhao@ucdavis.edu.
authors.forename: Min
authors.initials: M
authors.lastname: Zhao
mesh_terms: D032701:Biomimetics; D002465:Cell Movement; D019573:Epithelium, Corneal / Q000166:cytology*; D019278:Matrix Metalloproteinase 3 / Q000235:genetics / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D013269:Stochastic Processes
chemical_list: D019278:Matrix Metalloproteinase 3
keywords: Basement membrane; Cornea; Corneal epithelial cells; Electric field; Topography