Decellularized porcine liver scaffold for maintaining the viability and capacity of pancreatic islets

Bioengineered pancreatic constructs based on scaffolds made from decellularized tissues and pancreatic islets (PIs) may be used to extend the functional activity of transplanted PIs in patients with type I diabetes. Objective: to investigate in vitro the effect of decellularized porcine liver scaffold (DPLS) on the viability and insulin-producing capacity of isolated human PIs. Materials and methods. The resulting DPLS was subjected to histological examination, DNA quantification, and cytotoxic effect testing. The PIs were isolated from human pancreas fragments using the collagenase technique. Under standard conditions, PIs were cultured in three different environments: monoculture (control group), with DPLS present (experimental group 1) or with decellularized human pancreas scaffold (DHPS) present (experimental group 2). Vital fluorescent dyes were used to evaluate the viability of PIs. Basal and glucose-loaded insulin concentrations were determined by enzyme immunoassay. Results. The basic composition and structure of the extracellular matrix of liver tissue in DPLS samples were preserved thanks to the selected decellularization procedure. The samples had no cytotoxic effect, and the residual amount of DNA in the scaffold did not exceed 1.0%. PIs from the experimental groups showed no significant signs of degradation and fragmentation during the 10-day incubation period compared to PIs from the control group. On day 10, the viability of PIs from experimental group 1 was 64%, that of experimental group 2 was 72%, and that of the control group was less than 20%. After the first day of culturing, insulin concentration were 29.0% higher in experimental group 1 and 39.1% higher in experimental group 2 compared to the control group. On day 10 of the experiment, insulin levels in experimental groups 1 and 2 differed by 124.8% and 150.9%, respectively, from the control group. Under a glucose load, the insulin level in experimental group 1 was 1.7 times higher than in the control group, whereas that of experimental group 2 was 2.2 times higher. Conclusion. The resulting DPLS has a positive effect on the viability and insulin-producing capacity of PIs. When creating a bioengineered construct of PIs, DPLS can be used as a component obtained in sufficient quantity from an available source.

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