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Article: Development of a micromanipulator-based loading device for mechanoregulation study of human mesenchymal stem cells in three-dimensional collagen constructs

TitleDevelopment of a micromanipulator-based loading device for mechanoregulation study of human mesenchymal stem cells in three-dimensional collagen constructs
Authors
Issue Date2010
PublisherMary Ann Liebert, Inc. Publishers. The Journal's web site is located at http://www.liebertpub.com/publication.aspx?pub_id=261
Citation
Tissue Engineering - Part C: Methods, 2010, v. 16 n. 1, p. 93-107 How to Cite?
AbstractMechanical signal is important for regulating cellular activities, including proliferation, metabolism, matrix production, and orientation. Bioreactors with loading functions can be used to precondition cells in three-dimensional (3D) constructs so as to study the cellular responses to mechanical stimulation. However, full-scale bioreactor is not always an affordable option considering the high cost of equipments and the liter-sized medium with serum and growth factor supplements. In this study, a custom-built loading system was developed by coupling a conventional camera-equipped inverted research microscope with two micromanipulators. The system was programmed to deliver either cyclic compressive loading with different frequencies or static compressive loading for 1 week to investigate the cellular responses of human mesenchymal stem cells (hMSCs) entrapped in a 3D construct consists of reconstituted collagen fibers. Cellular properties, including their alignment, cytoskeleton, and cell metabolism, and properties of matrix molecules, such as collagen fiber alignment and glycosaminoglycan deposition, were evaluated. Using a MatLab-based image analysis program, reorientation of the entrapped cells from a random distribution to a preferred alignment along the loading direction in constructs with both static and cyclic compression has been demonstrated, but no such alignment was found in the free-floating controls. Fluorescent staining on filamentous actin cytoskeleton also confirmed the finding. Nevertheless, the collagen fiber meshwork entrapping the hMSCs remained randomly distributed, and no change in cellular metabolism and glycosaminoglycans production was noted. The current study provides a simple and affordable option toward setting up a mechanoregulation facility based on existing laboratory equipments and sheds new insights on the effect of mechanical loading on the alignment of hMSCs in 3D collagen constructs. Copyright © 2010, Mary Ann Liebert, Inc.
Persistent Identifierhttp://hdl.handle.net/10722/68271
ISSN
2015 SCImago Journal Rankings: 1.141
ISI Accession Number ID
Funding AgencyGrant Number
AOSpineAOSBRC-07-06
Innovation and Technology Commission
Hong Kong GovernmentITS/133/08
University Research Committee
University of Hong Kong11159180
University Strategic Research Theme on Nanobiotechnology
Funding Information:

This work was supported by grants from AOSpine (AOSBRC-07-06), Innovation and Technology Commission, the Hong Kong Government (ITS/133/08), University Research Committee, the University of Hong Kong (11159180), and the University Strategic Research Theme on Nanobiotechnology. We thank Ms. Chow Mei Kwan for her help in the development of the program for FFT image analysis.

References
Grants

 

DC FieldValueLanguage
dc.contributor.authorAuYeung, KLen_HK
dc.contributor.authorSze, KYen_HK
dc.contributor.authorSham, MHen_HK
dc.contributor.authorChan, BPen_HK
dc.date.accessioned2010-09-06T06:02:59Z-
dc.date.available2010-09-06T06:02:59Z-
dc.date.issued2010en_HK
dc.identifier.citationTissue Engineering - Part C: Methods, 2010, v. 16 n. 1, p. 93-107en_HK
dc.identifier.issn1937-3384en_HK
dc.identifier.urihttp://hdl.handle.net/10722/68271-
dc.description.abstractMechanical signal is important for regulating cellular activities, including proliferation, metabolism, matrix production, and orientation. Bioreactors with loading functions can be used to precondition cells in three-dimensional (3D) constructs so as to study the cellular responses to mechanical stimulation. However, full-scale bioreactor is not always an affordable option considering the high cost of equipments and the liter-sized medium with serum and growth factor supplements. In this study, a custom-built loading system was developed by coupling a conventional camera-equipped inverted research microscope with two micromanipulators. The system was programmed to deliver either cyclic compressive loading with different frequencies or static compressive loading for 1 week to investigate the cellular responses of human mesenchymal stem cells (hMSCs) entrapped in a 3D construct consists of reconstituted collagen fibers. Cellular properties, including their alignment, cytoskeleton, and cell metabolism, and properties of matrix molecules, such as collagen fiber alignment and glycosaminoglycan deposition, were evaluated. Using a MatLab-based image analysis program, reorientation of the entrapped cells from a random distribution to a preferred alignment along the loading direction in constructs with both static and cyclic compression has been demonstrated, but no such alignment was found in the free-floating controls. Fluorescent staining on filamentous actin cytoskeleton also confirmed the finding. Nevertheless, the collagen fiber meshwork entrapping the hMSCs remained randomly distributed, and no change in cellular metabolism and glycosaminoglycans production was noted. The current study provides a simple and affordable option toward setting up a mechanoregulation facility based on existing laboratory equipments and sheds new insights on the effect of mechanical loading on the alignment of hMSCs in 3D collagen constructs. Copyright © 2010, Mary Ann Liebert, Inc.en_HK
dc.languageengen_HK
dc.publisherMary Ann Liebert, Inc. Publishers. The Journal's web site is located at http://www.liebertpub.com/publication.aspx?pub_id=261en_HK
dc.relation.ispartofTissue Engineering - Part C: Methodsen_HK
dc.rightsThis is a copy of an article published in the Tissue Engineering Part C: Methods © 2010 [copyright Mary Ann Liebert, Inc.]; Tissue Engineering Part C: Methods is available online at: http://www.liebertonline.com.-
dc.rightsCreative Commons: Attribution 3.0 Hong Kong License-
dc.subject.meshBone Marrow Cells - cytology-
dc.subject.meshCell Differentiation-
dc.subject.meshCollagen - chemistry-
dc.subject.meshMesenchymal Stem Cells - cytology-
dc.subject.meshMicromanipulation - instrumentation - methods-
dc.titleDevelopment of a micromanipulator-based loading device for mechanoregulation study of human mesenchymal stem cells in three-dimensional collagen constructsen_HK
dc.typeArticleen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1937-3384&volume=16&issue=1&spage=93&epage=107&date=2010&atitle=Development+of+a+micromanipulator-based+loading+device+for+mechanoregulation+study+of+human+mesenchymal+stem+cells+in+three-dimensional+collagen+constructsen_HK
dc.identifier.emailSze, KY:szeky@graduate.hku.hken_HK
dc.identifier.emailSham, MH:mhsham@hkucc.hku.hken_HK
dc.identifier.emailChan, BP:bpchan@hkucc.hku.hken_HK
dc.identifier.authoritySze, KY=rp00171en_HK
dc.identifier.authoritySham, MH=rp00380en_HK
dc.identifier.authorityChan, BP=rp00087en_HK
dc.description.naturepublished_or_final_versionen_US
dc.identifier.doi10.1089/ten.tec.2008.0707en_HK
dc.identifier.pmid19368498-
dc.identifier.scopuseid_2-s2.0-77952364810en_HK
dc.identifier.hkuros169432en_HK
dc.relation.referenceshttp://www.scopus.com/mlt/select.url?eid=2-s2.0-77952364810&selection=ref&src=s&origin=recordpageen_HK
dc.identifier.volume16en_HK
dc.identifier.issue1en_HK
dc.identifier.spage93en_HK
dc.identifier.epage107en_HK
dc.identifier.isiWOS:000274125800010-
dc.publisher.placeUnited Statesen_HK
dc.relation.projectCreating Stable Tissue Interfaces for Bioengineered Intervertebral Disc Segment-
dc.identifier.scopusauthoridAuYeung, KL=24166171100en_HK
dc.identifier.scopusauthoridSze, KY=7006735060en_HK
dc.identifier.scopusauthoridSham, MH=7003729109en_HK
dc.identifier.scopusauthoridChan, BP=7201530390en_HK

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