ASTM F2664-11
(Guide)Standard Guide for Assessing the Attachment of Cells to Biomaterial Surfaces by Physical Methods
Standard Guide for Assessing the Attachment of Cells to Biomaterial Surfaces by Physical Methods
SIGNIFICANCE AND USE
Cell attachment or, lack of it, to biomaterials is a critical factor affecting the performance of a device or implant. Cell attachment is a complicated, time-dependent, process involving significant morphological changes of the cell and deposition of a bed of extracellular matrix. Details of the adhesive bond that is formed have been reviewed by, for example, Pierres et al (2002) (4), Lukas and Dvorak (2004) (5), and Garcia and Gallant (2003) (6). The strength of this coupling can be determined either by monitoring the force of attachment between a cell and a substrate over time or by measuring the force required to detach the cell once it has adhered.
Cell adhesion to a surface depends on a range of biological and physical factors that include the culture history, the age of the cell, the cell type, and both the chemistry and morphology of the underlying surface and time. These elements that need to be considered in developing a test protocol.
Devising robust methods for measuring the propensity of cells to attach to different substrates is further complicated since either cell adhesion or detachment can be assessed. These processes that are not always similar or complementary.
Most studies of cell attachment focus on obtaining some measure of the time-dependent force required to detach, or de-adhere, cells that have already adhered to a surface (James et al, 2005) (7). More recently investigators have begun to measure the adhesive forces that develop between cells and the underlying surface during attachment (Lukas and Dvorak, 2004) (5). From a practical point of view, it is much easier to measure the force required to detach or de-adhere cells from a surface than to measure those that develop during attachment. However, in both cases, the experimental data should be interpreted with a degree of caution that depends on the intended use of the measurements. The methods of measuring cell adhesion described herein are measures of the force required to detach an...
SCOPE
1.1 This guide describes protocols that can be used to measure the strength of the adhesive bond that develops between a cell and a surface as well as the force required to detach cells that have adhered to a substrate. Controlling the interactions of mammalian cells with surfaces is fundamental to the development of safe and effective medical products. This guide does not cover methods for characterizing surfaces. The information generated by these methods can be used to obtain quantitative measures of the susceptibility of surfaces to cell attachment as well as measures of the adhesion of cells to a surface. This guide also highlights the importance of cell culture history and influences of cell type.
1.2 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.
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Designation: F2664 − 11
Standard Guide for
Assessing the Attachment of Cells to Biomaterial Surfaces
1
by Physical Methods
This standard is issued under the fixed designation F2664; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope Surface Texture Parameters
ISO 13565-1 Geometrical Product Specifications (GPS)—
1.1 This guide describes protocols that can be used to
Surface Texture: Profile Method; Surfaces Having Strati-
measure the strength of the adhesive bond that develops
fied Functional Properties—Part 1: Filtering and General
between a cell and a surface as well as the force required to
Measurement Conditions
detach cells that have adhered to a substrate. Controlling the
interactions of mammalian cells with surfaces is fundamental
3. Terminology
tothedevelopmentofsafeandeffectivemedicalproducts.This
guide does not cover methods for characterizing surfaces. The
3.1 Definitions:
information generated by these methods can be used to obtain 3.1.1 adhesion, n—a physiochemical state by which a cell is
quantitative measures of the susceptibility of surfaces to cell
coupled to a non-cell surface by interfacial forces, which may
attachment as well as measures of the adhesion of cells to a consist of covalent or ionic forces.
surface. This guide also highlights the importance of cell
3.1.2 biocompatibility, n—a material may be considered
culture history and influences of cell type.
biocompatibleifthematerialsperformwithanappropriatehost
1.2 This standard does not purport to address all of the
response in a specific application. F2312
safety concerns, if any, associated with its use. It is the
3.1.3 biomarker, n—biochemical feature or facet that can be
responsibility of the user of this standard to establish appro-
used to measure the progress of disease or the effects of
priate safety and health practices and determine the applica-
treatment.
bility of regulatory limitations prior to use.
3.1.4 biomaterial, n—any substance (other than a drug),
synthetic or natural, that can be used as a system or part of a
2. Referenced Documents
system that treats, augments, or replaces any tissue, organ, or
2
2.1 ASTM Standards:
function of the body. F2312
D4410 Terminology for Fluvial Sediment
3.1.5 detachment, n—process whereby an adhered cell or
F22 Test Method for Hydrophobic Surface Films by the
group of cells is actively detached from a surface.
Water-Break Test
3.1.6 hydrophilic, adj—having a strong affinity for water,
F2312 Terminology Relating to Tissue Engineered Medical
Products wettable. F22
F2603 Guide for Interpreting Images of Polymeric Tissue
3.1.7 implant,n—inmedicine,anobject,structure,ordevice
Scaffolds
intended to reside within the body for diagnostic, prosthetic, or
3
2.2 ISO Standards:
other therapeutic purposes.
ISO 4287 Geometrical Product Specifications (GPS)—
3.1.8 laminar flow, n—well-ordered, patterned flow of fluid
Surface Texture: Profile Method—Terms, Definitions and
4
layers assumed to slide over one another. (See Ref (1).)
3.1.9 lay, n—direction of the predominant surface pattern.
1
This guide is under the jurisdiction of ASTM Committee F04 on Medical and
ISO 13565-1
Surgical Materials and Devices and is the direct responsibility of Subcommittee
3.1.10 passage, n—the transfer or transplantation of cells,
F04.43 on Cells and Tissue Engineered Constructs for TEMPs.
Current edition approved April 1, 2011. Published May 2011. Originally
with or without dilution, from one culture vessel to another. It
approved in 2007. Last previous edition approved in 2007 as F2664 – 07. DOI:
is understood that any time cells are transferred from one
10.1520/F2664-11.
vessel to another, a certain portion of the cells may be lost and,
2
For referenced ASTM standards, visit the ASTM website, www.astm.org, or
contact ASTM Customer Service at service@astm.org. For Annual Book of ASTM
Standards volume information, refer to the standard’s Document Summary page on
the ASTM website.
3 4
Available fromAmerican National Standards Institute (ANSI), 25 W. 43rd St., The boldface numbers in parentheses refer to the list of references at the end of
4th Floor, New York, NY 10036, http://www.ansi.org. this standard.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959. United States
1
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F2664 − 11
therefore, dilution of cells, whether deliberate or not, may de-adhere, cells that have already adhered to a surface (James
occur. This term is synonymous with the term subculture. (See et al, 2005) (7). More recently investigators
...
This document is not anASTM standard and is intended only to provide the user of anASTM standard an indication of what changes have been made to the previous version. Because
it may not be technically possible to adequately depict all changes accurately, ASTM recommends that users consult prior editions as appropriate. In all cases only the current version
of the standard as published by ASTM is to be considered the official document.
Designation:F2664–07 Designation:F2664–11
Standard Guide for
Assessing the Attachment of Cells to Biomaterial Surfaces
1
by Physical Methods
This standard is issued under the fixed designation F2664; the number immediately following the designation indicates the year of
original adoption or, in the case of revision, the year of last revision. A number in parentheses indicates the year of last reapproval. A
superscript epsilon (´) indicates an editorial change since the last revision or reapproval.
1. Scope
1.1 This guide describes protocols that can be used to measure the strength of the adhesive bond that develops between a cell
and a surface as well as the force required to detach cells that have adhered to a substrate. Controlling the interactions of
mammaliancellswithsurfacesisfundamentaltothedevelopmentofsafeandeffectivemedicalproducts.Thisguidedoesnotcover
methods for characterizing surfaces. The information generated by these methods can be used to obtain quantitative measures of
the susceptibility of surfaces to cell attachment as well as measures of the adhesion of cells to a surface.This guide also highlights
the importance of cell culture history and influences of cell type.
1.2 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility
of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory
limitations prior to use.
2. Referenced Documents
2
2.1 ASTM Standards:
D4410 Terminology for Fluvial Sediment
F22 Test Method for Hydrophobic Surface Films by the Water-Break Test
F2312 Terminology Relating to Tissue Engineered Medical Products
F2603 Guide for Interpreting Images of Polymeric Tissue Scaffolds
3
2.2 ISO Standards:
ISO 4287 Geometrical Product Specifications (GPS)—Surface Texture: Profile Method—Terms, Definitions and Surface
Texture Parameters
ISO 13565-1 Geometrical Product Specifications (GPS)—Surface Texture: Profile Method; Surfaces Having Stratified
Functional Properties—Part 1: Filtering and General Measurement Conditions
3. Terminology
3.1 Definitions:
3.1.1 adhesion, n—a physiochemical state by which a cell is coupled to a non-cell surface by interfacial forces, which may
consist of covalent or ionic forces.
3.1.2 biocompatibility, n—a material may be considered biocompatible if the materials perform with an appropriate host
response in a specific application. F2312
3.1.3 biomarker, n—biochemical feature or facet that can be used to measure the progress of disease or the effects of treatment.
3.1.4 biomaterial, n—any substance (other than a drug), synthetic or natural, that can be used as a system or part of a system
that treats, augments, or replaces any tissue, organ, or function of the body. F2312
3.1.5 detachment, n—process whereby an adhered cell or group of cells is actively detached from a surface.
3.1.6 hydrophilic, adj—having a strong affinity for water, wettable. F22
3.1.7 implant, n—a substance or object that is put in the body as a prosthesis, or for treatment or diagnosis. —in medicine,an
object, structure, or device intended to reside within the body for diagnostic, prosthetic, or other therapeutic purposes.
1
This guide is under the jurisdiction of ASTM Committee F04 on Medical and Surgical Materials and Devices and is the direct responsibility of Subcommittee F04.43
on Cells and Tissue Engineered Constructs for TEMPs.
Current edition approved June 1, 2007. Published June 2007. DOI: 10.1520/F2664-07.
CurrenteditionapprovedApril1,2011.PublishedMay2011.Originallyapprovedin2007.Lastpreviouseditionapprovedin2007asF2664 – 07.DOI:10.1520/F2664-11.
2
For referencedASTM standards, visit theASTM website, www.astm.org, or contactASTM Customer Service at service@astm.org. For Annual Book of ASTM Standards
volume information, refer to the standard’s Document Summary page on the ASTM website.
3
Available from American National Standards Institute (ANSI), 25 W. 43rd St., 4th Floor, New York, NY 10036, http://www.ansi.org.
Copyright © ASTM International, 100 Barr Harbor Drive, PO Box C700, West Conshohocken, PA 19428-2959, United States.
1
---------------------- Page: 1 ----------------------
F2664–11
4
3.1.8 laminar flow, n—well-ordered, patterned flow of fluid layers assumed to slide over one another. (See Ref (1).)
3.1.9 lay, n—direction of the predominant surface pattern. ISO 13565-1
3.1.10 passage, n—the transfer or transplantation of cells, with or without dilution, from one culture vessel to another. It is
understood
...
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