AI Article Synopsis
- Adult porcine islets (APIs) show promise for treating type 1 diabetes but face challenges like IBMIR and poor reestablishment of extracellular connections when transplanted.
- Intrahepatic transplantation risks destroying about two-thirds of islets, while intraperitoneal transplantation offers advantages such as immunoprotection and larger graft accommodations.
- Experiments demonstrated that hypoxia significantly harmed free islet viability, causing necrosis, while encapsulated islets retained better integrity; both types exhibited reduced metabolic activity and insulin secretion under low oxygen conditions.
Article Abstract
Background: Adult porcine islets (APIs) constitute a promising alternative to human islets in treating type 1 diabetes. The intrahepatic site has been used in preclinical primate studies of API xenografts; however, an estimated two-thirds of donor islets are destroyed after intraportal infusion due to a number of factors, including the instant blood-mediated inflammatory reaction (IBMIR), immunosuppressant toxicity, and poor reestablishment of extracellular matrix connections. Intraperitoneal (ip) transplantation of non-vascularized encapsulated islets offers several advantages over intrahepatic transplantation of free islets, including avoidance of IBMIR, immunoprotection, accommodation of a larger graft volume, and reduced risk of hemorrhage. However, there exists evidence that the peritoneal site is hypoxic, which likely impedes islet function.
Methods: We tested the effect of hypoxia (2%-5% oxygen or pO : 15.2-38.0 mm Hg) on free and encapsulated APIs over a period of 6 days in culture. Free and encapsulated APIs under normoxia served as controls. Islet viability was evaluated with a viability/cytotoxicity assay using calcein AM and ethidium bromide on days 1, 3, and 6 of culture. Alamar blue assay was used to measure the metabolic activity on days 1 and 6. Insulin in spent medium was assayed by ELISA on days 1 and 6.
Results: Viability staining indicated that free islet clusters lost their integrity and underwent severe necrosis under hypoxia; encapsulated islets remained intact, even when they began to undergo necrosis. Under hypoxia, the metabolic activity and insulin secretion (normalized to metabolic activity) of both free and encapsulated islets decreased relative to islets cultured under normoxic conditions.
Conclusions: Hypoxia (2%-5% oxygen or pO : 15.2-38.0 mm Hg) affects the viability, metabolic activity, and insulin secretion of both free and encapsulated APIs over a six-day culture period. Encapsulation augments islet integrity under hypoxia, but it does not prevent loss of viability, metabolic activity, or insulin secretion.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5334784 | PMC |
| http://dx.doi.org/10.1111/xen.12275 | DOI Listing |
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