Hollow multilayered capsules have shown massive potential for being used in the\nbiomedical and biotechnology fields, in applications such as cellular internalization, intracellular\ntrafficking, drug delivery, or tissue engineering. In particular, hollow microcapsules, developed\nby resorting to porous calcium carbonate sacrificial templates, natural-origin building blocks and\nthe prominent Layer-by-Layer (LbL) technology, have attracted increasing attention owing to\ntheir key features. However, these microcapsules revealed a great tendency to aggregate, which\nrepresents a major hurdle when aiming for cellular internalization and intracellular therapeutics\ndelivery. Herein, we report the preparation of well-dispersed polysaccharide-based hollow multilayered\nmicrocapsules by combining the LbL technique with an optimized purification process. Cationic\nchitosan (CHT) and anionic alginate (ALG) were chosen as the marine origin polysaccharides\ndue to their biocompatibility and structural similarity to the extracellular matrices of living\ntissues. Moreover, the inexpensive and highly versatile LbL technology was used to fabricate\ncore-shell microparticles and hollow multilayered microcapsules, with precise control over their\ncomposition and physicochemical properties, by repeating the alternate deposition of both materials.\nThe microcapsules� synthesis procedure was optimized to extensively reduce their natural aggregation\ntendency, as shown by the morphological analysis monitored by advanced microscopy techniques.\nThe well-dispersed microcapsules showed an enhanced uptake by fibroblasts, opening new perspectives\nfor cellular internalization.
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