Wound healing assay (2024)

FAQs

What is the assay for wound healing? ›

The wound healing (or scratch) assay is a method to measure two-dimensional cell migration. An artificial gap is generated on a confluent cell monolayer, and movement tracked via microscopy or other imaging. U2OS cells were seeded in 24-well plates at 2 × 10⁵ cells/mL and incubated for 24–48 h.

The basic steps involve creating a "wound" in a cell monolayer, capturing the images at the beginning and at regular intervals during cell migration to close the wound, and comparing the images to quantify the migration rate of the cells.

Scratch wound assay creates a gap in confluent monolayer of keratinocytes to mimic a wound. The protocol of scratch wound is based on few steps: cell culture preparation, scratch wound assay, data acquisition, and data analysis. Keywords: Cell migration; Data analysis; Healing assay; Scratch wound assay; Wound repair.

Place in incubator, and image on phase-contrast microscope every 4-8 hours until cells migrate to meet in the middle (24-48 hours). If resources are available, cells can be imaged ideally using automatic time-lapse photography on an incubating microscope.

Measure the length "head-to-toe" at the longest point (A). Measure the width side-to-side at the widest point (B) that is perpendicular to the length, forming a "+". Measure the depth (C) at the deepest point of the wound. All measures should be in centimeters.

Removal of a piece of tissue using a scalpel or punch biopsy is critical to achieve a quantitative culture of wound tissue, which is the gold standard for identification of wound infection.

Scratch assay on non-transfected cells

Scrape the cell monolayer in a straight line to create a “scratch” with a p200 pipet tip. Remove the debris and smooth the edge of the scratch by washing the cells once with 1 ml of the growth medium and then replace with 5 ml of medium specific for the in vitro scratch assay.

Conducting a wound healing and migration assay is an easy procedure: Create a physical gap within a cell monolayer . Monitor the process of cell migration into the gap with live cell imaging or by taking photos at different time points .

They usually heal within 3 to 7 days. A large, deep scrape may take 1 to 2 weeks or longer to heal. A scab may form on some scrapes. Follow-up care is a key part of your treatment and safety.

What are the advantages of scratch wound assay? ›

Advantages of this assay are that it does not require the use of specific chemoattractants or gradient chambers and it generates a strong directional migratory response, even in cell types that do not show robust responses in "single cell" migration assays.

Minor scrapes may be uncomfortable, but they usually heal within 3 to 7 days. The larger and deeper the scrape, the longer it will take to heal. A large, deep scrape may take up to 1 to 2 weeks or longer to heal. It's common to have small amounts of fluid drain or ooze from a scrape.

Inflammation that doesn't go away

If your wound is still red and raw after a couple weeks, it's a good idea to talk to a doctor – even if you don't think you have an infection.

Cuts and scratches are areas of damage on the surface of the skin. A cut is a line of damage that can go through the skin and into the muscle tissues below, whereas a scratch is surface damage that does not penetrate the lower tissues. Cuts and scratches may bleed or turn red, become infected, and leave scars.

In medicine, an assay is an analysis used to determine the presence of a particular substance and its concentration. Thus, for example, an assay may be conducted on a vaccine, to determine its effectiveness or potency in preventing illness.

The transwell invasion assay involves the addition of an extracellular matrix on top of the porous membrane which only permits chemotaxis of cells which possess invasive properties such as tumor cells.

Laboratory Studies

Determination of serum protein, albumin, prealbumin, and transferrin levels to assess the patient's nutritional status. Coagulation studies to evaluate for abnormalities, especially if deep wound excision is required. Tissue cultures of wound to determine appropriate antibiotic therapy.

The Boyden chamber assay, originally introduced by Boyden for the analysis of leukocyte chemotaxis, is based on a chamber of two medium-filled compartments separated by a microporous membrane.