Two In Vitro Experiments to Assess a Tissue Engineered Scaffold

What are two experiments that can be performed in vitro to assess if a tissue engineered scaffold is eluting degradation products causing inflammation?

1. Analysis of the degradation products of the scaffolds

2. Assessment of the inflammatory response

Analysis of the degradation products of the scaffolds

One experiment that can be performed in vitro to assess if a tissue engineered scaffold is eluting degradation products causing inflammation is the analysis of the degradation products of the scaffolds.

For the first experiment, you can create a set of replica tissue-engineered scaffolds that are similar to the ones that will be implanted. Then, set up separate culture mediums for the scaffolds that replicate physiological circumstances, such as simulated bodily fluid or cell culture media. Incubate the scaffolds under carefully monitored settings and regularly replace the media to mimic the deterioration process. Gather media samples at predetermined intervals that correspond to important in vivo time points for inflammation. Perform an analysis of the collected media samples using methods like enzyme-linked immunosorbent assay (ELISA) or liquid chromatography-mass spectrometry (LC-MS) to find and measure the presence of degradation products that are known to cause an inflammatory reaction.

Assessment of the inflammatory response

Another experiment that can be performed in vitro to assess if a tissue engineered scaffold is eluting degradation products causing inflammation is the assessment of the inflammatory response.

For the second experiment, in vitro cultivation of immune cells like macrophages or monocytes can be done. Replicating tissue-engineered scaffolds are exposed to pertinent culture media under controlled degradation conditions to create conditioned media. Gather the conditioned medium at predetermined intervals that coincide with the in vivo inflammation. Use the conditioned medium produced by the scaffold degradation to treat the cultured immune cells. Measure pertinent indicators, such as cytokine production or the activation of inflammatory signaling pathways, to evaluate the immune cells' inflammatory response. Evaluate the inflammatory response of immune cells exposed to conditioned media from scaffold degradation in comparison to control groups. If the immune cells' inflammatory reaction to the conditioned media from the scaffold degradation is significantly greater than that of control samples, it may confirm that the degradation products produced by the scaffold are capable of causing inflammation.