Abstract:BackgroundCell migration is a vital process for growth and repair. In vitro migration assays, utilized to study cell migration, often rely on physical scraping of a cell monolayer to induce cell migration. The physical act of scrape injury results in numerous factors stimulating cell migration – some injury-related, some solely due to gap creation and loss of contact inhibition. Eliminating the effects of cell injury would be useful to examine the relative contribution of injury versus other mechanisms to cell… Show more
“…It was reported that smooth muscle cells migrated faster on the gelatin-coated surface than on the uncoated polystyrene surface [33]. An increase of 63% in the migration rate from 24 to 48 h was observed in a barrier wound-healing assay [33]. Under the same conditions, for human umbilical vein endothelial cells, the migration rate was even six times faster on the gelatin-coated surface than on the uncoated polystyrene one [33].…”
Section: Resultsmentioning
confidence: 99%
“…Gelatin, an irreversibly hydrolyzed form of collagen, is widely used as a coating material in cell culture for improving cell attachment [32]. It was reported that smooth muscle cells migrated faster on the gelatin-coated surface than on the uncoated polystyrene surface [33]. An increase of 63% in the migration rate from 24 to 48 h was observed in a barrier wound-healing assay [33].…”
The wound-healing assay is commonly and widely used for investigating collective cell migration under various physical and chemical stimuli. Substrate-coating materials are shown to affect the wound-healing process in a cell-type dependent manner. However, experiment-to-experiment variations make it difficult to compare results from different assays. In this paper, a modified barrier wound-healing assay was reported for studying the wound-healing process on different substrates in one single petri dish. In short, half of a dish was covered with the tape, and coating materials, poly-l-lysine and gelatin, were applied to the surface. After peeling off the tape, half of the surface was coated with the desired material. Then a customized barrier was placed inside the dish to create the wound. The results indicated that surface coating did not affect cell proliferation/viability, and the wound-healing rate increased in coated surfaces compared to uncoated ones. The present study provides a platform for further understanding the mechanisms of substrate coating-dependent wound-healing processes.
“…It was reported that smooth muscle cells migrated faster on the gelatin-coated surface than on the uncoated polystyrene surface [33]. An increase of 63% in the migration rate from 24 to 48 h was observed in a barrier wound-healing assay [33]. Under the same conditions, for human umbilical vein endothelial cells, the migration rate was even six times faster on the gelatin-coated surface than on the uncoated polystyrene one [33].…”
Section: Resultsmentioning
confidence: 99%
“…Gelatin, an irreversibly hydrolyzed form of collagen, is widely used as a coating material in cell culture for improving cell attachment [32]. It was reported that smooth muscle cells migrated faster on the gelatin-coated surface than on the uncoated polystyrene surface [33]. An increase of 63% in the migration rate from 24 to 48 h was observed in a barrier wound-healing assay [33].…”
The wound-healing assay is commonly and widely used for investigating collective cell migration under various physical and chemical stimuli. Substrate-coating materials are shown to affect the wound-healing process in a cell-type dependent manner. However, experiment-to-experiment variations make it difficult to compare results from different assays. In this paper, a modified barrier wound-healing assay was reported for studying the wound-healing process on different substrates in one single petri dish. In short, half of a dish was covered with the tape, and coating materials, poly-l-lysine and gelatin, were applied to the surface. After peeling off the tape, half of the surface was coated with the desired material. Then a customized barrier was placed inside the dish to create the wound. The results indicated that surface coating did not affect cell proliferation/viability, and the wound-healing rate increased in coated surfaces compared to uncoated ones. The present study provides a platform for further understanding the mechanisms of substrate coating-dependent wound-healing processes.
“…4). While the injury free release of contact inhibition is sufficient to induce cell migration in cell exclusion assays 2,6,9,14 , the injury inflicted upon the cells in scratch wound healing models with the associated disruption of cell adhesions and release of intracellular contents and signalling molecules has been shown to alter the microenvironment in the cell culture and to provide a critical input required for cell-cell coordination in the spreading of the wounded monolayer 1,3,4,6,7,9 . Thus, while cell exclusion assays provide repeatability and geometric control, they fail to induce an injury and the insert used in this study correspondingly led to slower gap closure kinetics compared to the three assays that inflicted injury upon the cell layer.Figure 4Definition of regions of interest (ROI) along the scratch margins.…”
Section: Resultsmentioning
confidence: 99%
“…Subsequently serial high-resolution images are captured and analysed to quantify the dynamics of cell migration into the gap 3 . However, as the scratch is most commonly created manually using a pipette tip, the extent of the cellular injury and the geometry of the scratch is influenced not only by the scraping tool utilized, but also the level of manual control, the pressure exerted, the angle of the instrument and the velocity of scraping 3,4,6,7 . As a result, repeatability and reproducibility of the assays is limited and dependent on the manual dexterity of the researchers.…”
Section: Introductionmentioning
confidence: 99%
“…As a result, repeatability and reproducibility of the assays is limited and dependent on the manual dexterity of the researchers. Variation during wounding may confound interpretation and quantification of the wound healing process and impede comparison of results amongst different researchers and laboratories 1,3,4,6,7 .…”
A novel magnetic scratch method achieves repeatability, reproducibility and geometric control greater than pipette scratch assays and closely approximating the precision of cell exclusion assays while inducing the cell injury inherently necessary for wound healing assays. The magnetic scratch is affordable, easily implemented and standardisable and thus may contribute toward better comparability of data generated in different studies and laboratories.
Diaphragmatic wall defects caused by congenital disorders or disease remain a major challenge for physicians worldwide. Polymeric patches have been extensively explored within research laboratories and the clinic for soft tissue and diaphragm reconstruction. However, patch usage may be associated with allergic reaction, infection, granulation, and recurrence of the hernia. In this study, we designed and fabricated a porous scaffold using a combination of 3D printing and freeze‐drying techniques. A 3D printed polycaprolactone (PCL) mesh was used to reinforcegelatin scaffolds, representing an advantage over previously reported examples since it provides mechanical strength and flexibility. In vitro studies showed that adherent cells were anchorage‐dependent and grew as a monolayer attached to the scaffolds. Microscopic observations indicated better cell attachments for the scaffolds with higher gelatin content as compared with the PCL control samples. Tensile testing demonstrated the mechanical strength of samples was significantly greater than adult diaphragm tissue. The biocompatibility of the specimens was investigated in vivo using a subcutaneous implantation method in Bagg albino adult mice for 20 days, with the results indicating superior cellular behavior and attachment on scaffolds containing gelatin in comparison to pure PCL scaffolds, suggesting that the porous PCL/gelatin scaffolds have potential as biodegradable and flexible constructs for diaphragm reconstruction.
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