Adam D. Mendelsohn

Long-Term Small Molecule and Protein Elution from TiO2 Nanotubes

In this study, TiO(2) nanotubes of various dimensions were used to elute albumin, a large protein molecule, as well as sirolimus and paclitaxel, common small molecule drugs. The nanotubes controlled small molecule diffusion for weeks and large molecule diffusion for a month. Drug eluted from the nanotubes was bioactive and decreased cell proliferation in vitro. Elution kinetics was most profoundly affected by tube height. This study demonstrates that TiO(2) nanotubes may be a promising candidate for a drug-eluting implant coating.

Differentiation of human embryonic stem cells into pancreatic endoderm in patterned size-controlled clusters.

Pancreatic β-cells function optimally when clustered in islet-like structures. However, nutrient and oxygen deprivation limits the viability of cells at the core of excessively large clusters. Hence, production of functional β-cells from human embryonic stem cells (hESCs) for patients with diabetes would benefit from the growth and differentiation of these cells in size-controlled aggregates. In this study, we controlled cluster size by seeding hESCs onto glass cover slips patterned by the covalent microcontact-printing of laminin in circular patches of 120 μm in diameter. These were used as substrates to grow and differentiate hESCs first into SOX17-positive/SOX7-negative definitive endoderm, after which many clusters released and formed uniformly sized three-dimensional clusters. Both released clusters and those that remained attached differentiated into HNF1β-positive primitive gut tube-like cells with high efficiency. Further differentiation yielded pancreatic endoderm-like cells that co-expressed PDX1 and NKX6.1. Controlling aggregate size allows efficient production of uniformly-clustered pancreatic endocrine precursors for in vivo engraftment or further in vitro maturation.

Prevention of pulmonary metastasis from subcutaneous tumors by binary system-based sustained delivery of catalase

Catalase delivery can be effective in inhibiting reactive oxygen species (ROS)-mediated acceleration of tumor metastasis. Our previous studies have demonstrated that increasing the plasma half-life of catalase by pegylation (PEG-catalase) significantly increases its potency of inhibiting experimental pulmonary metastasis in mice. In the present study, a biodegradable gelatin hydrogel formulation was used to further increase the circulation time of PEG-catalase. Implantation of (111)In-PEG-catalase/hydrogel into subcutaneous tissues maintained the radioactivity in plasma for more than 14 days. Then, the effect of the PEG-catalase/hydrogel on spontaneous pulmonary metastasis of tumor cells was evaluated in mice with subcutaneous tumor of B16-BL6/Luc cells, a murine melanoma cell line stably expressing luciferase. Measuring luciferase activity in the lung revealed that the PEG-catalase/hydrogel significantly (P<0.05) inhibited the pulmonary metastasis compared with PEG-catalase solution. These findings indicate that sustaining catalase activity in the blood circulation achieved by the use of pegylation and gelatin hydrogel can reduce the incidence of tumor cell metastasis.

Patterning of mono- and multilayered pancreatic beta-cell clusters.

Cluster-size dependent behavior of pancreatic beta-cells has direct implications in islet transplantation therapy for type I diabetes treatment. Control over the cluster size enables evaluation of cluster-size-dependent function, ultimately leading to the production of beta-cell clusters with improved transplant efficacy. This work for the first time demonstrates the use of microcontact-printing-based cell patterning of discrete two- and three-dimensional clusters of pancreatic beta-cells. Both single and multiple cell layers are confined to a 2D area by attaching to patterns of covalently linked laminin and not adhering to surrounding polyethylene glycol. Cell clusters were successfully formed within 24 h for printed patterns in the range 40-120 microm, and simple modulation of the initial cell seeding density leads to the formation of multiple cell layers. Semiquantitative fluorescence microscopy, X-ray photoelectron spectroscopy, and Fourier transform infrared spectroscopy were used to extensively characterize the surface chemistry. This technique offers exceptional control over cell cluster shape and size, and not only provides an effective tool to study the cluster-size-dependent behavior of pancreatic beta-cells but also has potential applicability to numerous other cell lines.

Inorganic Nanoporous Membranes for Immunoisolated Cell-Based Drug Delivery

Materials advances enabled by nanotecbnology have brought about promising approaches to improve the encapsulation mechanism for immunoisolated cell-based drug delivery. Cell-based drug delivery is a promising treatment for many diseases but has thus far achieved only limited clinical success. Treatment of insulin dependent diabetes mellitus (IDDM) by transplantation of pancreatic beta-cells represents the most anticipated application ofcell-based drug delivery technology. This review outlines the challenges involved with maintaining transplanted cell viability and discusses how inorganic nanoporous membranes may be useful in achieving clinical success.

Size-controlled insulin-secreting cell clusters

The search for an effective cure for type I diabetes from the transplantation of encapsulated pancreatic β-cell clusters has so far produced sub-optimal clinical outcomes. Previous efforts have not controlled the size of transplanted clusters, a parameter implicated in affecting long-term viability and the secretion of therapeutically sufficient insulin. Here we demonstrate a method based on covalent attachment of patterned laminin for fabricating uniformly size-controlled insulin-secreting cell clusters. We show that cluster size within the range 40-120μm in diameter affects a variety of therapeutically relevant cellular responses including insulin expression, content and secretion. Our studies elucidate two size-dependent phenomena: (1) as the cluster size increases from 40μm to 60μm, glucose stimulation results in a greater amount of insulin produced per cell; and (2) as the cluster size increases beyond 60μm, sustained glucose stimulation results in a greater amount of insulin secreted per cell. Our study describes a method for producing uniformly sized insulin-secreting cell clusters, and since larger cluster sizes risk nutrient availability limitations, our data suggest that 100-120μm clusters may provide optimal viability and efficacy for encapsulated β-cell transplants as a treatment for type I diabetes and that further in vivo evaluation is warranted.