Assessing the compatibility of primary human hepatocyte culture within porous silk sponges

Abstract

Porous silk scaffolds hybridized with extracellular matrix proteins are useful for culture of primary human hepatocytes ± supportive non-parenchymal cells.

Impact of bioactive molecule inclusion in lyophilized silk scaffolds varies betweenin vivoandin vitroassessments

“…After general anesthesia of oxygen and isoflurane, the back of the rat was shaved and the skin was prepped with betadine disinfectant, followed by alcohol wipe, and repeated three times. Up to three small longitudinal incisions were made through the skin of the rat and sterilized acellular silk scaffolds 27-28 (3 mm radius x 3 mm height) were subcutaneously implanted in lateral pockets of each rat. The incisions were closed with surgical clips that were removed after 5-7 days.…”

“…These biological signaling modalities include the ability to easily enhance the silk biomaterial with relevant ECM components. 27-30 This is important as the composition of the ECM can modulate the time scales over which growth factors can bind and remain sequestered in the local environment. 31-33 In silk-based biomaterials with a subcutaneous rodent implant model, aligned lyophilized silk sponges with the addition of decellularized cardiac ECM led to cell infiltration and promoted vascularization when compared to aligned lyophilized silk sponges alone.…”

Bioengineered Liver Models for Investigating Disease Pathogenesis and Regenerative Medicine

“…5b). 243 Compared with conventional 2D monolayers, PHHs in silk scaffolds modified with ECM proteins had significantly higher hepatic functions, which were enhanced up to fivefold with the incorporation of supportive NPCs. Decellularized liver matrix has also been used for tissue engineering because of its ability to preserve the diversity of the liver ECM.…”

“…A different approach formed multilayered heparin microwells using micromolding and microcontact printing to spatially arrange rat hepatocytes and fibroblasts. 253 In another recent study, an HA methacrylate and PEGDA microporous cylindrical shell was seeded with supportive HUVECs, human 243 (c) Hepatic aggregates containing PHHs (magenta) and normal human dermal fibroblasts were created using pyramidal microwells and were then combined with geometrically patterned human endothelial cords composed of HUVEC cells (yellow) in a fibrin hydrogel to create free-standing liver tissue seeds (top). Albumin promoter activity was enhanced in cocultures compared with hepatocyte-only constructs after implantation into rodents (bottom).…”

“…Bottom left to right: coculture with 3T3-J2 fibroblast elevated albumin production compared with monocultures in silk/collagen scaffolds. Hybridization of silk scaffolds with Matrigel improves hepatic functions as compared with silk-and Matrigel-only control cultures 243. (c) Hepatic aggregates containing PHHs (magenta) and normal human dermal fibroblasts were created using pyramidal microwells and were then combined with geometrically patterned human endothelial cords composed of HUVEC cells (yellow) in a fibrin hydrogel to create free-standing liver tissue seeds (top).…”