Toward a bioartificial pancreas: diffusion of insulin and IgG across immunoprotective membranes with controlled hydrophilic channel diameters.

Research continued toward a bioartificial pancreas (BAP). Our BAPs consist of a perforated nitinol scaffold coated with reinforced amphiphilic conetwork membranes and contain live pancreatic islets. The membranes are assemblages of cocontinuous hydrophobic domains and hydrophilic channels whose diameters were varied by the MW of hydrophilic segments between crosslinks (M(c,HI) = 32, 44, and 74 kg x mol(-1)). We studied the diffusion rate of insulin, BSA, and IgG across the membrane of the BAP in the absence of islets. Membranes of M(c,HI) = 74 kg x mol(-1) showed rapid insulin and BSA transport and negligible IgG diffusion. BAPs containing approximately 300 mouse islets showed appropriate response upon glucose challenge in vitro. The BAP implanted into diabetic mice reduced hyperglycemia and maintained islet viability for at least 4 d.

A new bioartificial pancreas utilizing amphiphilic membranes for the immunoisolation of porcine islets: a pilot study in the canine.

We have developed a replaceable bioartificial pancreas to treat diabetes utilizing a unique cocontinous amphiphilic conetwork membrane created for macroencapsulation and immunoisolation of porcine islet cells (PICs). The membrane is assembled from hydrophilic poly(N,N-dimethyl acrylamide) and hydrophobic/oxyphilic polydimethylsiloxane chains cross-linked with hydrophobic/oxyphilic polymethylhydrosiloxane chains. Our hypothesis is that this membrane allows the survival of xenotransplanted PICs in the absence of prevascularization or immunosuppression because of its extraordinarily high-oxygen permeability and small hydrophilic channel dimensions (3-4 nm). The key components are a 5-10 microm thick semipermeable amphiphilic conetwork membrane reinforced with an electrospun nanomat of polydimethylsiloxane-containing polyurethane, and a laser-perforated nitinol scaffold to provide geometric stability. Devices were loaded with PICs and tested for their ability to maintain islet viability without prevascularization, prevent rejection, and reverse hyperglycemia in three pancreatectomized dogs without immunosuppression. Tissue tolerance was good and there was no systemic toxicity. The bioartificial pancreas protected PICs from toxic environments in vitro and in vivo. Islets remained viable for up to 3 weeks without signs of rejection. Neovascularization was observed. Hyperglycemia was not reversed, most likely because of insufficient islet mass. Further studies to determine long-term islet viability and correction of hyperglycemia are warranted.

A novel macroencapsulating immunoisolatory device: the preparation and properties of nanomat-reinforced amphiphilic co-networks deposited on perforated metal scaffold.

This paper describes the design and preparation of the non-biological components (the "hardware") of a conceptually novel bioartificial pancreas (BAP) to correct diabetes. The key components of the hardware are (1) a thin (5-10 microm) semipermeable amphiphilic co-network (APCN) membrane [i.e., a membrane of cocontinuous poly(dimethyl acryl amide) (PDMAAm)/polydimethylsiloxane (PDMS) domains cross-linked by polymethylhydrosiloxane (PMHS)] expressly created for macroencapsulation and immunoisolation of a tissue graft; (2) an electrospun nanomat of PDMS-containing polyurethane to reinforce the water-swollen APCN membrane; and (3) a perforated hollow-ribbon nitinol scaffold to stiffen and provide geometric stability to the construct. The reinforcement of water-swollen hydrogels with an electrospun nanomat is a generally applicable new method for hydrogel reinforcement. Details of device design and preparation are discussed. The advantages and disadvantages of micro- and macro-immunoisolation are analyzed, and the requirements for the ideal immunoisolatory membrane are presented. Burst pressure, and glucose and insulin permeabilities of representative devices have been determined and the effect of device composition and wall thickness on these properties is discussed.

Direct probe-atmospheric pressure chemical ionization mass spectrometry of cross-linked copolymers and copolymer blends.

Complex copolymers are heated to slowly increasing temperatures on a direct probe (DP) inside the plasma of the atmospheric pressure chemical ionization (APCI) source of a quadrupole ion trap. Slow heating allows for temporal separation of the thermal degradation products according to the stabilities of the bonds being cleaved. The products released from the DP are identified in situ by APCI mass spectrometry and tandem mass spectrometry. DP-APCI experiments on amphiphilic copolymers provide conclusive information about the nature of the hydrophobic and hydrophilic components present and can readily distinguish between copolymers with different comonomer compositions as well as between cross-linked copolymers and copolymer blends with similar physical properties. The dependence of DP-APCI mass spectra on temperature additionally reveals information about the thermal stability of the different domains within a copolymer.

Novel biostable and biocompatible amphiphilic membranes.

We determined the biostability and biocompatibility of two types of amphiphilic conetworks (APCNs): (1) hydrophilic poly(N,N-dimethyl acrylamide) (PDMAAm) and hydrophobic polydimethylsiloxane (PDMS) microdomains co-crosslinked with polymethylhydrosiloxane (PMHS) clusters (PDMAAm/PMHS/PDMS), and (2) poly(ethylene glycol) (PEG) and PDMS microdomains co-crosslinked with two specially designed small-molecule crosslinking agents SiC(6)H(5)(SiH)(2)OEt (Y) and polypentamethylhydrocyclosiloxane (PD(5)) (PEG/Y or PD(5)/PDMS). Negative standards for comparing biocompatibility and biostability were crosslinked PDMS. Biostability was assessed by quantitatively determining extractables, equilibrium water swelling, mechanical properties (stress-strain response) of polymer samples before and after implantation in rats for up to 8 weeks, and oxidative accelerated degradation test. Biocompatibility was assessed by determining body weight, fibrous tissue encapsulation, fluid accumulation, and by histological evaluation of lymphocyte infiltration, fibrous tissue accumulation and collagen deposition. According to these stringent metrics PDMAAm/PMHS/PDMS is both biostable and biocompatible, whereas PEG/Y or PD(5)/PDMS degrades in living tissue but is biocompatible. Surprisingly, the overall biocompatibility scores of these APCNs were superior to those of the PDMS negative standard.