A novel gellan gel-based microcarrier for anchorage-dependent cell delivery

Competent vehicles are highly sought after as a means to transplant cells for tissue regeneration. In this study, novel hydrogel-based microspherical cell carriers are designed and developed with an FDA-approved natural polysaccharide, gellan gum. The bulk fabrication of these microspheres is performed via a water-in-oil (W/O) emulsion process followed by a series of redox (oxidation-reduction) crosslinking treatments; this enables the microspherical dimensions to be precisely manipulated in terms of injectability, and simultaneously ensures the structural stability. To acquire adhesion affinity with anchorage-dependent cells (ADCs), a covalent coating of gelatin is further applied on the microspherical surfaces. The final product is constructed as a variety of gelatin-grafted-gellan microspherical cell carriers, abbreviated as "TriG" microcarriers. The cell-loading tests are conducted, respectively, with human dermal fibroblasts (HDFs) and human fetal osteoblasts (hFOBs). Morphological observation from optical microscopy and field emission scanning electron microscopy indicates that the HDFs spread well and populate rapidly on surfaces of TriG microcarriers. Immunofluorescent staining reveals the activation of focal adhesion and subsequent organization of F-actin from the attached cell surfaces, which suggests the TriG microspherical substrate is favorable to ADC adhesion and therefore capable of promoting HDF proliferation to achieve confluence by turning over three times within 10 days. The hFOBs are also cultivated on the TriG carriers, where ideal viability and clear potentials for osteogenesis are demonstrated by fluorescent "Live/Dead" screening and specific histobiochemical indications. All these findings suggest that the TriG microcarriers are suitable to provide open platforms for therapeutic ADC proliferation and differentiation.