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Laboratory Research
Despite broadening usage of human placental tissues in the management of wounds, the placental disc remains an underutilized material. The placental disc is a vascular tissue rich in extracellular matrix components, but transplantation is complicated by the maternal cells. The risk of adverse immunological responses to maternal elements necessitates complete decellularization prior to use. A proprietary processing technique has been developed to decellularize the placental disc yet maintain the native components of the matrix, yielding a product amenable to cellular infiltration. This study characterizes the impact of processing on the makeup and quality of a placental extracellular matrix (PECM*) particulate.
Methods:
PECM was manufactured using a proprietary process. Decellularization was visualized with H&E staining. Immunohistochemical analysis was performed for collagen types I and IV. Collagen content was quantified by hydroxyproline assay. Molecular integrity of collagen in PECM was assessed by differential scanning calorimetry (DSC). Sterilized PECM was evaluated using a shotgun proteomics approach, performed by Creative Proteomics. High pressure liquid chromatography and tandem mass spectrometry assessed the presence of a wider range of extracellular matrix and matrix-associated proteins in PECM. A 3D in vitro culture system containing primary human dermal fibroblasts was used to assess cellular infiltration into the final product.
Results:
H&E staining confirmed decellularization of the placental disc. Presence of collagen types I and IV was confirmed with immunostaining of PECM, and quantified by hydroxyproline assay to be approximately ~60% collagen. DSC analysis demonstrated that the molecular integrity of collagen in PECM was conserved throughout the decellularization process. Post-sterilization, proteomic analysis identified a diverse range of matrix-associated proteins preserved in PECM. In vitro, PECM provided a scaffold supportive of cellular interaction.
Discussion:
PECM has been optimized to effectively decellularize the tissue while retaining the inherent composition of the placental extracellular matrix. Characterization reveals not only an intact collagenous extracellular matrix but also a diverse array of proteins. Cellular compatibility of sterilized PECM is further demonstrated through the cellular interaction with the matrix in an in vitro model. This initial characterization of the decellularized disc matrix, PECM particulate, supports its continued development for a variety of wound management applications.
Trademarked Items: *AXIOFILL ECM Particulate, MIMEDX Group Inc., Marietta, GA
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