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A Multi-well Format Polyacrylamide-based Assay for Studying the Effect of Extracellular Matrix Stiffness on the Bacterial Infection of Adherent Cells
Published on: July 5, 2018
High-throughput assay for bacterial adhesion on acellular dermal matrices and synthetic surgical materials
Theodore T Nyame1, Katherine P Lemon, Roberto Kolter
1Boston, Mass. From the Division of Plastic and Reconstructive Surgery, Massachusetts General Hospital, Harvard Medical School; the Center for Regenerative Medicine, Harvard Stem Cell Institute; the Division of Infectious Diseases, Children's Hospital Boston, Harvard Medical School; and the Department of Microbiology and Molecular Genetics, Harvard Medical School.
Background:
There has been increasing use of synthetic and acellular dermal matrix materials in surgery, ranging from breast reconstruction to hernia repairs. There is a paucity of data on how acellular dermal matrix compares with other surgical materials as a substrate for bacterial adhesion, the first step in formation biofilm, which occurs in prosthetic wound infections. The authors have designed a high-throughput assay to evaluate Staphylococcus aureus adherence on various synthetic and biologically derived materials.
Methods:
Clinical isolates of S. aureus (strains SC-1 and UAMS-1) were cultured with different materials, and bacterial adherence was measured using a resazurin cell vitality assay. Four materials that are commonly used in surgery were evaluated: Prolene mesh, Vicryl mesh, and two different acellular dermal matrix preparations (AlloDerm and FlexHD). The authors were able to develop a high-throughput and reliable assay for quantifying bacterial adhesion on synthetic and biologically derived materials.
Results:
The resazurin vitality assay can be reliably used to quantify bacterial adherence to acellular dermal matrix material and synthetic material. S. aureus strains SC-1 and UAMS-1 both adhered better to acellular dermal matrix materials (AlloDerm versus FlexHD) than to the synthetic material Prolene. S. aureus also adhered better to Vicryl than to Prolene. Strain UAMS-1 adhered better to Vicryl and acellular dermal matrix materials than did strain SC-1.
Conclusions:
The results show that S. aureus adheres more readily to acellular dermal matrix material than to synthetic material. The resazurin assay provides a standard method for evaluating surgical materials with regard to bacterial adherence and potential propensity for biofilm development.
