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Non-invasive evaluation of alginate/poly-l-lysine/alginate microcapsules by magnetic resonance microscopy
Ioannis Constantinidis1, Samuel C Grant, Susanne Celper
1Division of Endocrinology, Department of Medicine, University of Florida, PO Box 100226, Gainesville, FL 32610-0226, USA. consti@medicine.ufl.edu
Abstract:
In this report, we present data to demonstrate the utility of (1)H MR microscopy to non-invasively examine alginate/poly-l-lysine/alginate (APA) microcapsules. Specifically, high-resolution images were used to visualize and quantify the poly-l-lysine (PLL) layer, and monitor temporal changes in the alginate gel microstructure during a month long in vitro culture. The thickness of the alginate/PLL layer was quantified to be 40.6+/-6.2 microm regardless of the alginate composition used to generate the beads or the time of alginate/PLL interaction (2, 6, or 20 min). However, there was a notable difference in the contrast of the PLL layer that depended upon the guluronic content of the alginate and the alginate/PLL interaction time. The T(2) relaxation time and the apparent diffusion coefficient (ADC) of the alginate matrix were measured periodically throughout the month long culture period. Alginate beads generated with a high guluronic content alginate demonstrated a temporal decrease in T(2) over the duration of the experiment, while ADC was unaffected. This decrease in T(2) is attributed to a reorganization of the alginate microstructure due to periodic media exchanges that mimicked a regular feeding regiment for cultured cells. In beads coated with a PLL layer, this temporal decrease in T(2) was less pronounced suggesting that the PLL layer helped maintain the integrity of the initial alginate microstructure. Conversely, alginate beads generated with a high mannuronic content alginate (with or without a PLL layer) did not display temporal changes in either T(2) or ADC. This observation suggests that the microstructure of high mannuronic content alginate beads is less susceptible to culture conditions.
Insights
Proton MR microscopy non-invasively examined alginate/poly-l-lysine/alginate microcapsules. The poly-l-lysine layer maintained alginate microstructure integrity during in vitro culture, unlike high-mannuronic alginate beads.
Area of Science:
- Biomaterials Science
- Medical Imaging
- Cell Encapsulation Technology
Background:
- Alginate microcapsules are widely used for cell encapsulation.
- Non-invasive methods are needed to assess microcapsule integrity during culture.
- The poly-l-lysine (PLL) layer is crucial for stabilizing alginate microcapsules.
Purpose of the Study:
- To demonstrate the utility of (1)H MR microscopy for examining alginate/poly-l-lysine/alginate (APA) microcapsules.
- To visualize and quantify the PLL layer and monitor changes in alginate microstructure in vitro.
- To investigate the effect of alginate composition and PLL coating on microcapsule stability.
Main Methods:
- (1)H MR microscopy was employed for high-resolution imaging of APA microcapsules.
- The thickness of the alginate/PLL layer was quantified.
- T(2) relaxation time and apparent diffusion coefficient (ADC) of the alginate matrix were measured over a month of in vitro culture.
Main Results:
- The alginate/PLL layer thickness was consistently ~40.6 µm.
- High-guluronic alginate beads showed decreased T(2) over time, indicating microstructural reorganization, which was less pronounced with a PLL layer.
- High-mannuronic alginate beads (with or without PLL) exhibited stable T(2) and ADC, suggesting resistance to culture-induced changes.
Conclusions:
- (1)H MR microscopy is a valuable tool for non-invasively assessing APA microcapsule structure and stability.
- The poly-l-lysine layer enhances the structural integrity of alginate microcapsules, particularly those made from high-guluronic alginate, during in vitro culture.
- High-mannuronic alginate microcapsules demonstrate inherent stability under the tested culture conditions.

