A Novel Method for Single Sample Multi-Axial Nanoindentation of Hydrated Heterogeneous Tissues Based on Testing Great White Shark Jaws
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{"title"=>"A novel method for single sample multi-axial nanoindentation of hydrated heterogeneous tissues based on testing great white shark jaws", "type"=>"journal", "authors"=>[{"first_name"=>"Toni L.", "last_name"=>"Ferrara", "scopus_author_id"=>"25721911100"}, {"first_name"=>"Philip", "last_name"=>"Boughton", "scopus_author_id"=>"23488091800"}, {"first_name"=>"Eve", "last_name"=>"Slavich", "scopus_author_id"=>"56038601800"}, {"first_name"=>"Stephen", "last_name"=>"Wroe", "scopus_author_id"=>"35726136500"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"sgr"=>"84894170758", "doi"=>"10.1371/journal.pone.0081196", "pui"=>"372407397", "pmid"=>"24260558", "scopus"=>"2-s2.0-84894170758", "issn"=>"19326203"}, "id"=>"76e5786b-9677-30a5-ab79-1ac85d123004", "abstract"=>"Nanomechanical testing methods that are suitable for a range of hydrated tissues are crucial for understanding biological systems. Nanoindentation of tissues can provide valuable insights into biology, tissue engineering and biomimetic design. However, testing hydrated biological samples still remains a significant challenge. Shark jaw cartilage is an ideal substrate for developing a method to test hydrated tissues because it is a unique heterogeneous composite of both mineralized (hard) and non-mineralized (soft) layers and possesses a jaw geometry that is challenging to test mechanically. The aim of this study is to develop a novel method for obtaining multidirectional nanomechanical properties for both layers of jaw cartilage from a single sample, taken from the great white shark (Carcharodon carcharias). A method for obtaining multidirectional data from a single sample is necessary for examining tissue mechanics in this shark because it is a protected species and hence samples may be difficult to obtain. Results show that this method maintains hydration of samples that would otherwise rapidly dehydrate. Our study is the first analysis of nanomechanical properties of great white shark jaw cartilage. Variation in nanomechanical properties were detected in different orthogonal directions for both layers of jaw cartilage in this species. The data further suggest that the mineralized layer of shark jaw cartilage is less stiff than previously posited. Our method allows multidirectional nanomechanical properties to be obtained from a single, small, hydrated heterogeneous sample. Our technique is therefore suitable for use when specimens are rare, valuable or limited in quantity, such as samples obtained from endangered species or pathological tissues. We also outline a method for tip-to-optic calibration that facilitates nanoindentation of soft biological tissues. Our technique may help address the critical need for a nanomechanical testing method that is applicable to a variety of hydrated biological materials whether soft or hard.", "link"=>"http://www.mendeley.com/research/novel-method-single-sample-multiaxial-nanoindentation-hydrated-heterogeneous-tissues-based-testing-g", "reader_count"=>9, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>2, "Student > Ph. D. Student"=>3, "Student > Master"=>1, "Student > Bachelor"=>1, "Lecturer"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>2, "Student > Ph. D. Student"=>3, "Student > Master"=>1, "Student > Bachelor"=>1, "Lecturer"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>3, "Materials Science"=>1, "Agricultural and Biological Sciences"=>5}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>3}, "Materials Science"=>{"Materials Science"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>5}}, "reader_count_by_country"=>{"Mexico"=>1, "Germany"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1288158"], "description"=>"<p>Note: Numbers that are in bold or underlined are significant at α  =  0.05 and 0.1, respectively; “Not significant” indicates contrasts were not performed since no directional effect was observed.</p>", "links"=>[], "tags"=>["adjusted", "contrasts", "directional"], "article_id"=>856576, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.t003", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Bonferroni_adjusted_p_values_for_results_of_contrasts_testing_directional_differences_/856576", "title"=>"Bonferroni adjusted <i>p</i>-values for results of contrasts testing directional differences.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-11-19 03:53:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1288156"], "description"=>"<p>Optical Images from the Hysitron Triboindenter of mineralized (A, B) and non-mineralized (C, D) great white shark jaw cartilage. The image of the anterior surface of mineralized cartilage (A) shows the optical cross hairs (blue) on a block of mineralized cartilage. The image of the ventral surface (B) shows numerous nodules (e.g. white arrow) present in the jaw cartilage. Images C and D are both from the ventral surface of the same sample of cartilage and show differences observed in topology in the non-mineralized layer.</p>", "links"=>[], "tags"=>["images", "hysitron", "triboindenter", "mineralized", "non-mineralized", "anterior", "cartilage", "shows", "optical", "hairs", "ventral", "nodules", "differences", "observed", "topology"], "article_id"=>856574, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.g008", "stats"=>{"downloads"=>1, "page_views"=>24, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Optical_Images_from_the_Hysitron_Triboindenter_of_mineralized_A_B_and_non_mineralized_C_D_great_white_shark_jaw_cartilage_The_image_of_the_anterior_surface_of_mineralized_cartilage_A_shows_the_optical_cross_hairs_blue_on_a_block_of_mineralized_cartilage_/856574", "title"=>"Optical Images from the Hysitron Triboindenter of mineralized (A, B) and non-mineralized (C, D) great white shark jaw cartilage. The image of the anterior surface of mineralized cartilage (A) shows the optical cross hairs (blue) on a block of mineralized cartilage. The image of the ventral surface (B) shows numerous nodules (e.g. white arrow) present in the jaw cartilage. Images C and D are both from the ventral surface of the same sample of cartilage and show differences observed in topology in the non-mineralized layer.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-11-19 03:53:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1288153"], "description"=>"<p>Clips were moulded using thermoplastic aliphatic polyester encompassing a ferromagnetic steel stub which is visible in Clip B. The steel stub allowed the clips to adhere to the magnetic stage of the nanoindenter. In Clip A, the sample was held in place with a clasp (A) and a textured surface was moulded into the base of the clip to further minimize movement of the sample during testing. Samples were loaded into the groove of Clip B by depressing the thumb rest. The width of the groove in the flexible clip was chosen to accommodate samples between 1–3 mm in thickness and held the cartilage firmly in place without compressing the sample. The anterior surface of the sample was tested using Clip A, while the buccal and ventral surfaces were tested by rotating the sample in Clip B. Only one sample was required to complete multi-axial testing on both layers of cartilage because the sample was not bonded to the stub and could be easily rotated into different positions using the clips.</p>", "links"=>[], "tags"=>["built", "nanomechanical"], "article_id"=>856571, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.g005", "stats"=>{"downloads"=>0, "page_views"=>7, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Clips_purpose_built_for_nanomechanical_testing_/856571", "title"=>"Clips purpose built for nanomechanical testing.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-11-19 03:53:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1288152"], "description"=>"<p>The sample is shown <i>in situ</i> (grey outline) and removed, rotated and enlarged to show both mineralized (M) and non-mineralized (NM) layers of cartilage. Arrows indicate the anterior (A), buccal (B) and ventral (V) surfaces of the sample relative to the jaw. Surfaces are defined as follows: anterior surface of the sample corresponded to the frontal plane (anteroposterior axis) of the jaws; the buccal surface of the sample corresponded to the sagittal plane (mediolateral axis) of the jaws; the ventral surface of the sample corresponded to the transverse plane (longitudinal axis) of the jaws. Dashed arrows indicate directions perpendicular to the page.</p>", "links"=>[], "tags"=>["cartilage", "removed", "nanomechanical"], "article_id"=>856570, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.g004", "stats"=>{"downloads"=>7, "page_views"=>61, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Diagram_of_the_cartilage_sample_removed_from_the_great_white_shark_jaw_for_nanomechanical_testing_/856570", "title"=>"Diagram of the cartilage sample removed from the great white shark jaw for nanomechanical testing.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-11-19 03:53:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1288155"], "description"=>"<p>Boxplots show variation in Young’s modulus (A) and hardness (B) for mineralized and non-mineralized great white shark jaw cartilage in different directions. Axes are in gigapascals (GPa). Whiskers on boxplots represent minimum and maximum values.</p>", "links"=>[], "tags"=>[], "article_id"=>856573, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.g007", "stats"=>{"downloads"=>0, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Boxplots_/856573", "title"=>"Boxplots.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-11-19 03:53:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1288154"], "description"=>"<p>In panel 1A, the sample of cartilage with mineralized (light grey) and non-mineralized (dark grey) cartilage is placed flat in Clip A (black circle) under a microscope (orange box). The testing position (on mineralized cartilage) is defined by the optical cross hairs (orange circle, A) of the microscope and this position is coupled to the indenter position (orange circle, B). Indentation occurs at the position defined by the optical cross hairs (C, D). Each indentation is a repeat of steps A-D and numerous indentations are performed on the sample. Upon completion of testing mineralized cartilage, the optical cross hairs are moved to a location on non-mineralized cartilage (yellow circle, 1A) and steps in Panel 1A-D are repeated to test the anterior surface of this layer (not shown). In Panel 2, the cartilage sample is placed in Clip B (black box) and the indentation sequence (A-D) is repeated numerous times to test the buccal surface (orange) of mineralized cartilage. The upper portion of the sample is removed (dashed line, Panel 2A) and the sequence is subsequently repeated to test the buccal surface of non-mineralized cartilage (not shown). In Panel 3 the sample is rotated in Clip B to test the ventral surface (orange) of mineralized cartilage. The upper portion of the sample was then removed to test the ventral surface (orange) of non-mineralized cartilage (Panel 4A-D). All tests were conducted in a hydrating chamber (not shown).</p>", "links"=>[], "tags"=>["multi-axial", "nanomechanical"], "article_id"=>856572, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.g006", "stats"=>{"downloads"=>0, "page_views"=>14, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Schematic_of_multi_axial_nanomechanical_testing_/856572", "title"=>"Schematic of multi-axial nanomechanical testing.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-11-19 03:53:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1288151"], "description"=>"<p>The image shows the effect of water loss if no attempt is made to maintain hydration during nanoindentation. Due to significant moisture loss in the sample, the non-mineralized layer (1) is noticeably shrivelled and paper thin. The mineralized blocks (tesserae) present in the outer layer are also visible (2).</p>", "links"=>[], "tags"=>["cartilage"], "article_id"=>856569, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.g003", "stats"=>{"downloads"=>0, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Desiccation_of_cartilage_in_the_8220_dry_8221_specimen_/856569", "title"=>"Desiccation of cartilage in the “dry” specimen.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-11-19 03:53:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1288149"], "description"=>"<p>An arch-shaped wedge of cartilage (A) possessing both mineralized (1) and non-mineralized layers (2) was held in place with the clasp of a clip (B). The “V” shaped notch cut into the clasp (B) maintained equal pressure on both sides of the sample during nanomechanical testing and assisted with positioning the sample under the microscope.</p>", "links"=>[], "tags"=>[], "article_id"=>856567, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.g001", "stats"=>{"downloads"=>0, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Preparation_of_the_8220_dry_8221_bull_shark_sample_/856567", "title"=>"Preparation of the “dry” bull shark sample.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-11-19 03:53:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1288150"], "description"=>"<p>A plastic bag was heat sealed along 3 sides (1A) to form a pouch. A section of cartilage (A) was inserted into the pouch. A syringe filled with a hydrating medium was placed in the pouch with the opening of the needle positioned beneath the sample (A). The pouch was then heat-shrunk with a hair dryer allowing the pouch to conform to the sample and the needle, forming a channel (1B) along the length of the needle. The hydrating medium (1C) was injected into the channel as the needle was carefully withdrawn. The end of the channel was heat sealed and cut forming a tail (2C). A small rectangular window (not shown) was cut in the pouch to expose the surface for testing and the sample and pouch were then placed in a clip (D). The sample was then placed in a hydrating chamber (E) with the tail of the pouch tucked beneath the rectangular opening of the chamber (1E) to facilitate nanomechanical testing.</p>", "links"=>[], "tags"=>["enveloping"], "article_id"=>856568, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.g002", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Sample_preparation_in_an_enveloping_pouch_/856568", "title"=>"Sample preparation in an enveloping pouch.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-11-19 03:53:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1288159"], "description"=>"<p>Note: Numbers represent the mean (in MPa) followed by the number of indentations.</p>", "links"=>[], "tags"=>["modulus", "hardness", "cartilage"], "article_id"=>856577, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.t002", "stats"=>{"downloads"=>1, "page_views"=>15, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Young_8217_s_modulus_and_hardness_for_great_white_shark_jaw_cartilage_in_three_directions_/856577", "title"=>"Young’s modulus and hardness for great white shark jaw cartilage in three directions.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-11-19 03:53:25"}
  • {"files"=>["https://ndownloader.figshare.com/files/1288157"], "description"=>"<p>Young’s modulus (<i>E</i>), hardness (H) and contact depth (h<sub>c</sub>) for bull shark jaw cartilage.</p>", "links"=>[], "tags"=>["modulus", "hardness"], "article_id"=>856575, "categories"=>["Biological Sciences"], "users"=>["Toni L. Ferrara", "Philip Boughton", "Eve Slavich", "Stephen Wroe"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0081196.t001", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Young_s_modulus_E_hardness_H_and_contact_depth_h_c_for_bull_shark_jaw_cartilage_/856575", "title"=>"Young’s modulus (<i>E</i>), hardness (H) and contact depth (h<sub>c</sub>) for bull shark jaw cartilage.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-11-19 03:53:25"}

PMC Usage Stats | Further Information

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Relative Metric

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