Microsphere‐Incorporated Hybrid Thermogel for Neuronal Differentiation of Tonsil Derived Mesenchymal Stem Cells

Abstract

Neuronal differentiation of tonsil-derived mesenchymal stem cells (TMSCs) is investigated in a 3D hybrid system. The hybrid system is prepared by increasing the temperature of poly(ethylene glycol)-poly(l-alanine) aqueous solution to 37 °C through the heat-induced sol-to-gel transition, in which TMSCs and growth factor releasing microspheres are suspended. The in situ formed gel exhibits a modulus of 800 Pa at 37 °C, similar to that of brain tissue, and it is robust enough to hold the microspheres and cells during the 3D culture of TMSCs. The neuronal growth factors are released over 12-18 d, and the TMSCs in a spherical shape initially undergo multipolar elongation during the 3D culture. Significantly higher expressions of the neuronal biomarkers such as nuclear receptor related protein (Nurr-1), neuron specific enolase, microtubule associated protein-2, neurofilament-M, and glial fibrillary acidic protein are observed in both mRNA level and protein level in the hybrid systems than in the control experiments. This study proves the significance of a controlled drug delivery concept in tissue engineering or regenerative medicine, and a 3D hybrid system with controlled release of growth factors from microspheres in a thermogel can be a very promising tool.

Application of Tonsil-Derived Mesenchymal Stem Cells in Tissue Regeneration: Concise Review

“…In addition to their mesodermal differentiation potential (bone, cartilage, and fat), it was verified that TMSCs have the capacity to differentiate into endodermal lineage, expressing markers of parathyroid cells . Subsequently, other studies also identified differentiation capacities of TMSCs into various cell types, including myocytes , insulin‐releasing cells , neuron‐like or glial cells , tenocytes , and hepatocytes (Fig. ).…”

“…Similar to BM‐MSCs, TMSCs have a potential to differentiate into either neuron‐like or glial cells . In a 3D hybrid thermogel that fabricates growth factor‐releasing microspheres, TMSCs expressed phenotypes similar to immature neurons, exhibiting multipolar elongation with weak expression of neuronal markers such as neuronal growth factor, nuclear receptor‐related protein, and neuron‐specific enolase .…”

Gelatin‐Based Capsules through Interfacial Polymerization: Batch and Continuous Flow Synthesis

“…Polymeric micro-and nanoparticles gained a lot of research interest over last decades as carriers for the controlled delivery of therapeutic peptides, vaccines, antibiotics [1][2][3], and for the purpose of cellular differentiation [4]. Several nanomaterials are already approved for medical applications, e.g., liposomes DoxilTM and polymeric micelles NanoxelTM for cancer treatment [5][6][7].…”

“…The amino acids along the gelatin chain present numerous functional amino and hydroxyl groups as targets for surface modification or crosslinking reactions [26]. Glutaraldehyde [4,[27][28][29][30][31][32][33][34], genipin [35], and a water-soluble carbodiimide [36] are the widely used crosslinking agents for gelatin.…”

Polymeric microspheres: a delivery system for osteogenic differentiation

“…Polymeric MSs derived from synthetic sources are widely used in clinical applications because of their safety and biocompatibility . In recent times, MSs are used not only for drug delivery applications but also for the purpose of cellular differentiation . MSs coated with antibodies and peptides have been widely explored in the diagnosis of diseases as well …”