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Characterization of topographical effects on macrophage behavior in a foreign body response model
Open accessAbstract
Current strategies to limit macrophage adhesion, fusion and fibrous capsule formation in the foreign body response have focused on modulating material surface properties. We hypothesize that topography close to biological scale, in the micron and nanometric range, provides a passive approach without bioactive agents to modulate macrophage behavior. In our study, topography-induced changes in macrophage behavior was examined using parallel gratings (250 nm-2 mum line width) imprinted on poly(epsilon-caprolactone) (PCL), poly(lactic acid) (PLA) and poly(dimethyl siloxane) (PDMS). RAW 264.7 cell adhesion and elongation occurred maximally on 500 nm gratings compared to planar controls over 48 h. TNF-alpha and VEGF secretion levels by RAW 264.7 cells showed greatest sensitivity to topographical effects, with reduced levels observed on larger grating sizes at 48 h. In vivo studies at 21 days showed reduced macrophage adhesion density and degree of high cell fusion on 2 mum gratings compared to planar controls. It was concluded that topography affects macrophage behavior in the foreign body response on all polymer surfaces examined. Topography-induced changes, independent of surface chemistry, did not reveal distinctive patterns but do affect cell morphology and cytokine secretion in vitro, and cell adhesion in vivo particularly on larger size topography compared to planar controls.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The observed increase in R-ratio correlated with an increase in sub-population of cells attaining the elongated morphology, defined as having R ≥ 2.5 and aligned within 30° of the parallel gratings (Figure 4 B).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.2 Cell Adhesion and Morphology Changes on NIL gratings
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In the paper
“RAW 264.7 cell adhesion and elongation occurred maximally on 500 nm gratings compared to planar controls over 48 hr.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > Abstract
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In the paper
“Similarly, the MCP-1 secretion profile shared the same trend as TNF-α secretion (Figure 6 C).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.4 Cytokine Secretion Profile
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In the paper
“While the fraction of elongated cells on each topography was small (less than 25% of total cell population), statistical significance was observed on 250 nm and 2 μm PCL gratings compared to planar control (*p<0.05), and on 500 nm gratings (**p<0.01).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.2 Cell Adhesion and Morphology Changes on NIL gratings
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In the paper
“The low average R-ratio of all adherent cells (R<2, Figure 5 I) was due to the round cells predominating in the sub-population (1.2<R<1.6, Figure 5 I, inset).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.3 RAW 246.7 Response on Constant Height Topography
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In the paper
“However, the elongated cells had a higher R-ratio (R>3, Figure 5 J).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.3 RAW 246.7 Response on Constant Height Topography
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In the paper
“Macrophage adhesion and foreign body giant cell (FBGC) fusion were present on all grating samples at Day 7 (Figure 7 A, C, E) and Day 21 (Figure 7 B, D, F).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.5 In vivo Characterization in the Rat Cage Implant System
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In the paper
“The average R-ratio of adherent cells on PCL gratings imprinted on glass was comparable to that of PCL gratings on Mylar, except for 250 nm gratings (#p<0.01) (Figure 4 C).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.2 Cell Adhesion and Morphology Changes on NIL gratings
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In the paper
“Similarly, macrophages elongated in the direction of gratings on three different widths of PCL gratings imprinted on glass: 2 μm (Figure 3 E), 500 nm (Figure 3 F), and 250 nm (Figure 3 G).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.2 Cell Adhesion and Morphology Changes on NIL gratings
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In the paper
“Only on the 2 μm PLA gratings did the cells clearly elongate in the direction of gratings (left portion), as illustrated by the immunostained F-actin (green) and nuclei (blue) of the cells.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.2 Cell Adhesion and Morphology Changes on NIL gratings
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In the paper
“Approximately 25-55% of fields on surfaces registered high levels of fusion (>90%) while only 28-37% of fields supported low fusion (<10%).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.5 In vivo Characterization in the Rat Cage Implant System
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In the paper
“The average R-ratios of cells on planar PCL (0 nm) imprinted on glass and on Mylar were significantly different (#p<0.01), and marginally higher than that on planar TCPS and glass controls.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.2 Cell Adhesion and Morphology Changes on NIL gratings
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In the paper
“with comparable nuclei density observed on all grating samples, except for increased cell density on 2 μm gratings compared to planar control at Day 21 (*p<0.05).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.5 In vivo Characterization in the Rat Cage Implant System
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In the paper
“Topography-induced cellular fusion showed that low fusion decreased with increasing grating sizes at Day 7, but the trend reversed at Day 21 (Figure 7 H).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.5 In vivo Characterization in the Rat Cage Implant System
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In the paper
“The topographical effect on MIP-1α secretion was difficult to discern (Figure 6 B) but significance was observed at 6 hr for 300 nm samples compared to planar control (*p<0.05), at 24 hr for 1 μm gratings compared to 300 nm and 500 nm samples (*p<0.01), and at 48 hr for 300 nm, 500 nm and 1 μm gratings compared to planar samples (*p<0.05).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.4 Cytokine Secretion Profile
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In the paper
“Patterns in topography-induced cytokine secretion were difficult to discern but greater statistical significance was observed for 24 hr and 48 hr readings.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.4 Cytokine Secretion Profile
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In the paper
“While most adherent cells and FBGCs displayed a thinly spread morphology on the PCL/Mylar surfaces, some cells displayed elongated morphologies and the extension of pseudopodia in the direction of gratings, confirmed by SEM imaging (Supplementary Figure 4).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.5 In vivo Characterization in the Rat Cage Implant System
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In the paper
“The pro-inflammatory and pro-wound healing cytokine IL-1β, and the anti-inflammatory and pro-wound healing cytokines IL-6, IL-10, and IL-12 were not detected in the supernatant (detection limit: 3.2 pg/mL).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.4 Cytokine Secretion Profile
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In the paper
“At Day 7, fusion was minimal on all surfaces with 58-74% of fields having low (<10%) or no fusion (0%), and only 0-5% of fields with high fusion (>90%).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.5 In vivo Characterization in the Rat Cage Implant System
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“In contrast, VEGF was low at 6 hr to 24 hr (<0.5 pg/mL per 1000 cells, Figure 6 D), but increased at 48 hr.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.4 Cytokine Secretion Profile
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In the paper
“A comparison of the observed values (O) to the expected value (E) of the topographical gratings showed fidelity to be within 8% of expected feature size (O/E value of 1.08).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.1 Substrate Fabrication and Characterization
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In the paper
“The effect of topography on fractional distribution of elongated cells and average R-ratio was not statistically significant.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.3 RAW 246.7 Response on Constant Height Topography
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In the paper
“Similar to macrophage adhesion and elongation on PCL gratings, the majority of adherent cells retained their native round shape (Figure 5 G) while elongated cells formed a small fraction, less than 15%, of total cells (Figure 5 F).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.3 RAW 246.7 Response on Constant Height Topography
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In the paper
“At 24 hr, increasing grating width increased TNF-α secretion (*p<0.001), but the trend was reversed at 48 hr (*p<0.05), with statistical significance noted for 1 μm gratings compared to 300 nm gratings.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.4 Cytokine Secretion Profile
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In the paper
“In contrast, high fusion was not prevalent at Day 7 but showed a decreasing trend with larger grating sizes.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.5 In vivo Characterization in the Rat Cage Implant System
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In the paper
“Cell spreading of adherent macrophage was comparable across all grating sizes at 6 hr and 24 hr, but increased significantly on 500 nm and 1 μm gratings compared to planar PDMS control (Figure 5 F).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.3 RAW 246.7 Response on Constant Height Topography
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In the paper
“TNF-α (tumor necrosis factor-α), MIP-1α (macrophage inflammatory protein-1α), MCP-1 (monocyte chemoattractant protein-1) and VEGF (vascular endothelial growth factor) cytokines were detected in the supernatant.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.4 Cytokine Secretion Profile
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In the paper
“Adhesion, elongation and alignment of RAW 264.7 to its underlying topography was clearly observed on PCL and PLA gratings (width:height:period = 1:1:2) and on PDMS substrates of constant height (width:period = 1:2, height = 350 nm), independent of surface chemistry.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
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In the paper
“In vivo studies at 21 days showed reduced macrophage adhesion density and degree of high cell fusion on 2 μm gratings compared to planar controls.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > Abstract
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- Rests on an assumption: The rat cage implant model reflects the foreign body response relevant to clinical implants.
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In the paper
“The fraction of elongated cells increased from 0.10 on planar controls to 0.17 (66% increase), 0.23 (123% increase) and 0.20 (85% increase) on 250 nm, 500 nm and 2 μm gratings, respectively.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.2 Cell Adhesion and Morphology Changes on NIL gratings
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In the paper
“The average R-ratio of macrophages on planar control was 1.78, but increased to 2.18 on 250 nm gratings and 2.53 on 500 nm gratings, and then decreased slightly to 2.24 on 2 μm gratings.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.2 Cell Adhesion and Morphology Changes on NIL gratings
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In the paper
“Between 24 hr to 48 hr, the levels of pro-inflammatory cytokines TNF-α, MIP-1α and MCP-1 were maintained or reduced, while VEGF, a pro-inflammation/pro-wound healing cytokine, increased.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.4 Cytokine Secretion Profile
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In the paper
“While the fraction of elongated cells on each topography was small (less than 25% of total cell population), statistical significance was observed on 250 nm and 2 μm PCL gratings compared to planar control (*p<0.05), and on 500 nm gratings (**p<0.01).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.2 Cell Adhesion and Morphology Changes on NIL gratings
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In the paper
“Morphometric analysis of RAW 264.7 response to topography showed that cell adhesion density on 6 hr samples was low (around 1 × 100 cells/mm2) and did not increase above the cell seeding density (at 2 × 100 cells/mm2) until between 24 hr and 48 hr (Figure 5 E), when proliferation occurred.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.3 RAW 246.7 Response on Constant Height Topography
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In the paper
“Topography-induced MCP-1 secretion was most significant at 24 hr (*p<0.001) with increasing MCP-1 levels on increasing grating sizes: planar (*p<0.01), 300 nm (*p<0.05) and 500 nm (*p<0.05) compared to 1 μm gratings, but was difficult to discern at 48 hr.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.4 Cytokine Secretion Profile
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In the paper
“Macrophage cells studies in our system showed that elongation increased with decreasing topography, but only to a limit of 500 nm before the trend diverged.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
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In the paper
“In the long term response (21 days), high cellular fusion (>90%) was reduced as grating sizes increased: represented as high fusion on planar > 500 nm > 2 μm (Figure 7 G).”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
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In the paper
“Triplicates (n=3) of each grating size were used for each time point.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 2. Materials and Methods > 2.2 Cell Culture and Immunohistochemistry Staining
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In the paper
“but do affect cell morphology and cytokine secretion in vitro”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > Abstract
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- Rests on an assumption: The RAW 264.7 murine macrophage cell line is a valid model for macrophage behavior in the foreign body response.
- Rests on an assumption: Differences in surface chemistry among PCL, PLA, and PDMS do not confound the observed effects of topography.
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In the paper
“The Bio-Plex reader was calibrated and validated prior to every reading, strictly following the manufacturer's instructions.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 2. Materials and Methods > 2.3 Cytokine Secretion Profile Immunoassays
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In the paper
“All protocols were approved by the Institutional Animal Care and Use Committee of Case Western Reserve University.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 2. Materials and Methods > 2.4 In vivo Cage Implant System
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In the paper
“The lack of distinctive trends in cell morphology and orientation changes of both RAW 264.7 and in vivo macrophage cells to the topography differed from our previous studies involving other cell types, such as mesenchymal stem cells [26] and smooth muscle cells [45], that showed over 80% cell elongation and alignment.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
What the sources say
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In the paper
“Morphology measurements were made from at least 150 cells in each triplicate sample (n=3) per grating size.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 2. Materials and Methods > 2.2 Cell Culture and Immunohistochemistry Staining
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In the paper
“using the cage implant model, an established system for studying host tissue-implant response [44].”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.5 In vivo Characterization in the Rat Cage Implant System
What the sources say
- Not checked yetcites [44] Miller, 1989, Generation of IL-1-like activity in response to biomedical polymer implants: a comparison of in vitro and in vivo models, J Biomed Mater Res source
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In the paper
“confirmed by SEM imaging (Supplementary Figure 4)”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 3. Results > 3.5 In vivo Characterization in the Rat Cage Implant System
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In the paper
“Our results suggest that larger size topography (500 nm-2 μm) limits macrophage activation and inflammation-related processes but topography-induced sensitivity reaches a limit around 500 nm.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
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“MCP-1 (macrophage chemoattractant protein-1), a chemokine that recruits macrophage and promotes migration, also stimulates murine FBGC fusion in vivo [54].”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
What the sources say
- Not checked yetcites [54] Kyriakides, 2004, The CC chemokine ligand, CCL2/MCP1, participates in macrophage fusion and foreign body giant cell formation, Am J Pathol source
Strength:plain· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✗ Load-bearing: fails✓ Based: passesProbably not a substantial claim. The paper’s conclusions may not depend on it.
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In the paper
“Although the response to topography in the form of gratings from micro- to submicron scale has not revealed a distinctive pattern, topography indeed affects cell morphology, cytokine secretion in vitro, and cell adhesion in vivo particularly for larger size patterns compared to planar controls.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 5. Conclusions
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
The paper states this without a source or data behind it here, so there is nothing to check it against. The quote shows it is in the paper.
In the paper
“while MIP-1α is involved in chemotaxis and recruitment of monocytes and leukocytes during the inflammatory response.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
Strength:plain· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✗ Load-bearing: fails✓ Based: passesProbably not a substantial claim. The paper’s conclusions may not depend on it.
None of the sources cited for this claim have been checked yet.
In the paper
“The phenomenon appears to mirror that of phagocytosis, where macrophages preferentially internalize particulates in the micron range over the nanometric range, with peak phagocytosis observed with particles of approximately 2 μm in diameter [46-48].”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
What the sources say
- Not checked yetcites [46] Ahsan, 2002, Targeting to macrophages: role of physicochemical properties of particulate carriers–liposomes and microspheres – on the phagocytosis by macrophages, J Control Release source
- Not checked yetcites [47] Schafer, 1992, Phagocytosis of nanoparticles by human immunodeficiency virus (HIV)-infected macrophages: a possibility for antiviral drug targeting, Pharm Res source
- Not checked yetcites [48] Tabata, 1988, Effect of the size and surface charge of polymer microspheres on their phagocytosis by macrophage, Biomaterials source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The larger topographical cues (2 μm gratings) could be presenting a physical interruption or constraint to the fibrous scar tissue formation process, reducing cell adhesion density and limiting FBGC fusion on larger gratings compared to planar controls.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
None of the sources cited for this claim have been checked yet.
In the paper
“While the infinitely long parallel gratings, compared to a cell dimension, provide a non-phagocytosable surface, the limit of macrophage sensitivity to nanotopographical cues could be compared to a surface area dependence on pro-inflammation response, analogous to surface area dependence of nanoparticles in determining inflammatory response [50, 51], and the influence of particle shape and size on”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
What the sources say
- Not checked yetcites [50] Limbach, 2005, Oxide nanoparticle uptake in human lung fibroblasts: effects of particle size, agglomeration, and diffusion at low concentrations, Environ Sci Technol source
- Not checked yetcites [51] Hussain, 2009, Oxidative stress and proinflammatory effects of carbon black and titanium dioxide nanoparticles: role of particle surface area and internalized amount, Toxicology source
- Not checked yetcites [52] Gratton, 2008, The effect of particle design on cellular internalization pathways, Proc Natl Acad Sci U S A source
Strength:hedged· not yet calibratedCausal:correlational· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✗ Load-bearing: fails✓ Based: passesProbably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Macrophage cells appeared to be relatively insensitive to nanotopography (less than 500 nm), preferring to respond to 500 nm, 1 μm and 2 μm gratings.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
Load-bearingStrength:hedged· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesIt may state two separate claims as one.
The paper states this without a source or data behind it here, so there is nothing to check it against. The quote shows it is in the paper.
In the paper
“VEGF promotes blood vessel dilation and angiogenesis in the event of inflammation to promote wound healing in damaged tissue.”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
Strength:plain· not yet calibratedCausal:direct· not yet calibratedTests✓ Asserted: passes✓ Checkable: passes✗ Load-bearing: fails✗ Based: failsProbably not a substantial claim. The paper’s conclusions may not depend on it. The paper may give no reason why it holds.
None of the sources cited for this claim have been checked yet.
In the paper
“Cellular responses to topography are known to affect cell function in terms of adhesion strength, spreading, intracellular signaling and differentiation potential [59-61].”
§ Characterization of Topographical Effects on Macrophage Behavior in a Foreign Body Response Model > 4. Discussion
What the sources say
- Not checked yetcites [59] Kulangara, 2009, Substrate topography shapes cell function, Soft Matter source
- Not checked yetcites [60] Vogel, 2006, Local force and geometry sensing regulate cell functions, Nat Rev Mol Cell Biol source
- Not checked yetcites [61] Yim, 2005, Significance of synthetic nanostructures in dictating cellular response, Nanomedicine source
Tests✓ Asserted: passes✓ Checkable: passes✓ Load-bearing: passes✓ Based: passesProbably not a substantial claim.
The paper relies on this without support.
The RAW 264.7 murine macrophage cell line is a valid model for macrophage behavior in the foreign body response.
“Adherent murine macrophage cell line, RAW 264.7 (ATCC, Manassas, VA), was grown in Dulbecco's Modified Eagle's Medium (DMEM) cell culture media supplemented with 10% fetal bovine serum, penicillin (100 μg/mL) and streptomycin (100 μg/mL).”
The paper relies on this without support.
The rat cage implant model reflects the foreign body response relevant to clinical implants.
“Samples of PCL gratings nanoimprinted on Mylar were evaluated in rats using the cage implant model to determine the effect of topography on foreign body reaction.”
The paper relies on this without support.
Differences in surface chemistry among PCL, PLA, and PDMS do not confound the observed effects of topography.
“RAW 264.7 was cultured on 3 different types of polymers, PCL, PLA and PDMS gratings, to examine macrophage response to topography independent of surface chemistry.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“Upregulation of TNF‐α and VEGF provided a mixture of M1 and M2 phenotype in smaller gratings (higher surface roughness).”
From Unraveling and Harnessing the Immune Response at the Cell–Biomaterial Interface for Tissue Engineering Purposes (2024) · cites this paper as [147]
Experimental — accuracy not yet measuredThe source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“Chen et al. confirmed this topography‐induced polarization on PCL, poly(lactic acid) (PLA), and poly(dimethyl siloxane) imprinted with 250–2000 nm grooves.[ 147 ]”
From Unraveling and Harnessing the Immune Response at the Cell–Biomaterial Interface for Tissue Engineering Purposes (2024) · cites this paper as [147]
Experimental — accuracy not yet measuredThe cited source says what the claim attributes to it.
In the citing paper
“Seeded RAW 264.7 established its longest elongation in 500 nm grooves.”
From Unraveling and Harnessing the Immune Response at the Cell–Biomaterial Interface for Tissue Engineering Purposes (2024) · cites this paper as [147]
In the citing paper
“irregular topographies with similar dimensions produce circularly spreading macrophages with M1 characteristics”
From Modeling mechanical activation of macrophages during pulmonary fibrogenesis for targeted anti-fibrosis therapy (2024) · cites this paper as [39]
In the citing paper
“Aligned patterns with feature sizes in the micron level were shown to promote M2 macrophage differentiation through forced alignment”
From Modeling mechanical activation of macrophages during pulmonary fibrogenesis for targeted anti-fibrosis therapy (2024) · cites this paper as [39]
In the citing paper
“its severity depends on numerous factors, such as material chemistry18 and topography.19”
From Student award winner in the undergraduate category for the society of biomaterials 9th World Biomaterials Congress, Chengdu, China, June 1–5, 2012: The effects of substrate stiffness on the in vitro activation of macrophages and in vivo host response to poly(ethylene glycol)‐based hydrogels (2012) · cites this paper as [19]
In the citing paper
“bone marrow derived macrophages aligned along the grooved surface topographies, generally in the direction parallel to the grooves”
From Micro- and Nanopatterned Topographical Cues for Regulating Macrophage Cell Shape and Phenotype (2015) · cites this paper as [12]
In the citing paper
“Lithographically generated gratings as well as different post geometries have also been shown to modulate macrophage TNF-α and vascular endothelial growth factor (VEGF) secretion.12,13”
From Micro- and Nanopatterned Topographical Cues for Regulating Macrophage Cell Shape and Phenotype (2015) · cites this paper as [12]
In the citing paper
“Interestingly, upon in vivo implantation, materials with 2 mm gratings exhibited the least number of FBGC (1 mm grating materials were not analyzed).”
From Biomaterial based modulation of macrophage polarization: a review and suggested design principles (2015) · cites this paper as [46]
In the citing paper
“This study by Chen et al. [46] also showed that gratings that were 1 mm wide elicited the lowest inflammatory response in vitro (reduced levels of TNF-a and VEGF secretion) compared to nano gratings and planar controls.”
From Biomaterial based modulation of macrophage polarization: a review and suggested design principles (2015) · cites this paper as [46]
In the citing paper
“It was recently found that topographical cues could override the effects of surface chemistry in certain materials, especially in the first 6-48 hours after initial contact.”
From Biomaterial based modulation of macrophage polarization: a review and suggested design principles (2015) · cites this paper as [46]
In the citing paper
“RAW 264.7 cells seeded on substrates with three distinctive surface chemistries ((Poly (s-caprolactone) (PCL)), (Poly (Lactic acid)(PLA)), (Poly (Dimethyl Siloxane) (PDMS))) but different width parallel gratings (Fig. 4f,g) (with a range of 250 nm to 2 mm) showed increased elongation with decreasing topography irrespective of the underlying surface chemistry; however, cells were largely”
From Biomaterial based modulation of macrophage polarization: a review and suggested design principles (2015) · cites this paper as [46]
In the citing paper
“studies suggest around 1 mm thick strips or fibers are optimum [46]”
From Biomaterial based modulation of macrophage polarization: a review and suggested design principles (2015) · cites this paper as [46]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Day-21 adhesion trend is reversed in the abstract
MajorThe abstract says macrophage adhesion density was lower on 2 μm gratings than on planar controls at 21 days. The results report a statistically significant increase on those gratings at Day 21, reversing the stated conclusion.
“In vivo studies at 21 days showed reduced macrophage adhesion density and degree of high cell fusion on 2 μm gratings compared to planar controls.”
“except for increased cell density on 2 μm gratings compared to planar control at Day 21 (*p<0.05).”
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Metadata
- Authors
- Chen, S., Jones, J. A., Xu, Y., Low, H.-Y., Anderson, J. M., & Leong, K. W.
- Affiliations
- Johns Hopkins University (US)
- Case Western Reserve University (US)
- Institute of Materials Research and Engineering (SG)
- Duke University (US)
- Corresponding author
- Leong, Kam W.
- Venue
- Biomaterials
- Publisher
- Elsevier BV
- Published
- 2010-05
- Volume
- 31
- Issue
- 13
- Pages
- 3479-3491
- ISSN
- 0142-9612 (ISSN-L)
- PMC
- PMC2837101
- OpenAlex
- W2038846800
- Cited by
- 389
- References
- 73
- Citations by year
- 2026: 13 · 2025: 21 · 2024: 21 · 2023: 24 · 2022: 30 · 2021: 38 · 2020: 28 · 2019: 31 · 2018: 23 · 2017: 32 · 2016: 24 · 2015: 24 · 2014: 30 · 2013: 16 · 2012: 15
- FWCI
- 7.00
- Type
- journal article
- Language
- en
- Open access
- green
- europepmc.org
- Licence
- Locations
- Biomaterials (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- DukeSpace (Duke University) (submittedVersion, cc-by-nc-nd)
- Funders
- National Institutes of Health (EB000282, HL83008)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Physical Sciences › Engineering › Biomedical Engineering › Bone Tissue Engineering Materials
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- topography, foreign body reaction, cell adhesion, cell fusion, fibrous capsule formation, TNF-α, vascular endothelial growth factor, RAW 264.7 macrophages, polycaprolactone, polylactic acid, polydimethylsiloxane
- MeSH terms
- Animals, Cell Line, Foreign-Body Reaction, Macrophages, Microscopy, Electron, Scanning, Models, Biological, Polymers, Surface Properties, Tumor Necrosis Factor-alpha, Vascular Endothelial Growth Factor A, Mice
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
A language model drew the claims from the paper on 2026-10-05, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).
A second model labelled each claim on 2026-10-05 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
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