3D Porous Chitosan Scaffolds Suit Survival and Neural Differentiation of Dental Pulp Stem Cells

Abstract

A key aspect of cell replacement therapy in brain injury treatment is construction of a suitable biomaterial scaffold that can effectively carry and transport the therapeutic cells to the target area. In the present study, we created small 3D porous chitosan scaffolds through freeze-drying, and showed that these can support and enhance the differentiation of dental pulp stem cells (DPSCs) to nerve cells in vitro. The DPSCs were collected from the dental pulp of adult human third molars. At a swelling rate of ~84.33 ± 10.92 %, the scaffold displayed high porosity and interconnectivity of pores, as revealed by SEM. Cell counting kit-8 assay established the biocompatibility of the chitosan scaffold, supporting the growth and survival of DPSCs. The successful neural differentiation of DPSCs was assayed by RT-PCR, western blotting, and immunofluorescence. We found that the scaffold-attached DPSCs showed high expression of Nestin that decreased sharply following induction of differentiation. Exposure to the differentiation media also increased the expression of neural molecular markers Microtubule-associated protein 2, glial fibrillary acidic protein, and 2',3'-cyclic nucleotide phosphodiesterase. This study demonstrates that the granular 3D chitosan scaffolds are non-cytotoxic, biocompatible, and provide a conducive and favorable micro-environment for attachment, survival, and neural differentiation of DPSCs. These scaffolds have enormous potential to facilitate future advances in treatment of brain injury.

In vitro comparative survey of cell adhesion and proliferation of human induced pluripotent stem cells on surfaces of polymeric electrospun nanofibrous and solution‐cast film scaffolds

“…A synthetic 3D system to replace the natural ECM is one of the most important goals in tissue engineering. 10,11 According to previous reports the pores, ridges, and fibers present in the natural ECM are in the nanometer range. It is known that cells attach and organize well around fibers with diameters smaller than the diameter of the cells (in the nanometer range).…”

“…In tissue engineering scaffolds Correspondence to: E. Hoveizi; e-mail: e.hoveizi@scu.ac.ir and e.hoveizi@yahoo.com also serve as a three dimensional (3D) architectures for cell adhesion, proliferation, differentiation and provide structural support for the tissue regeneration. 11 Generally, the ideal scaffold for tissue regeneration should be biocompatible, biodegradable and allow for adequate cell loading, facilitate cell proliferation and differentiation, and possess mechanical and physical properties that are suitable for the target application. 12,13 In this study we made PCL scaffold by two methods and then we investigated the relationships between properties of two different scaffolds and the activity of hiPSCs on them.…”

Application of stem cells and chitosan in the repair of spinal cord injury

“…On the one hand, chitosan has the potential to repair SCI when co‐cultured with stem cells in vitro . Particularly, 3D porous scaffolds created by chitosan could provide a conductive and favorable microenvironment for attachment, survival and neural differentiation of dental pulp stem cells (DPSCs) (Feng et al, 2014), human adipose‐derived stem cells (hADSCs) (Gao et al, 2014) and human endometrial stem cells (hEnSCs)(Ebrahimi‐Barough et al, 2015). Also, purine‐crosslink chitosan sponges entrapped with NT‐3 promote oligodendrocyte progenitor cells (OPCs) to differentiate into mature oligodendrocytes and yield promising therapies for application in the CNS repair (Mekhail et al, 2015).…”

“…Stem cell type TrKc-MSC MAP-2: Microtubule-associated protein 2; V2a: spinal interneurons; BDNF: brain derived neurotrophic factor; MNTS1: a multineurotrophin that binds TrkA; RA: retinoic acid; SHH: Sonic hedgehog; BMSC: bone mesenchymal stem cell; NT-3: neurotrophic factor-3; TrKc: tropomyosin receptor kinase C. on this, good biocompatibility is significantly demonstrated between silk fibroin-chitosan scaffolds and grafted stem cells (Ji et al, 2013).On the one hand, chitosan has the potential to repair SCI when co-cultured with stem cells in vitro. Particularly, 3D porous scaffolds created by chitosan could provide a conductive and favorable microenvironment for attachment, survival and neural differentiation of dental pulp stem cells (DPSCs) (Feng et al, 2014), human adipose-derived stem cells (hADSCs) (Gao et al, 2014) and human endometrial stem cells (hEnSCs) (Ebrahimi-Barough et al, 2015). Also, purine-crosslink chitosan sponges entrapped with NT-3 promote oligodendrocyte progenitor cells (OPCs) to differentiate into mature oligodendrocytes and yield promising therapies for application in the CNS repair (Mekhail et al, 2015).…”

3D Microfibrous Scaffolds Selectively Promotes Proliferation and Glial Differentiation of Adult Neural Stem Cells: A Platform to Tune Cellular Behavior in Neural Tissue Engineering

“…The ability of these cells to differentiate into neurons, astrocytes, and oligodendrocytes, the principal neural cell types of the CNS, provides an advantage for studying directed cell differentiation . Combined with biomaterials, scaffolds can also influence cell differentiation thus promoting stem cells into neural lineages for therapeutic strategies for nervous system rescue and repair . However, little is known about the extent to which PCL microfibers may impact cell proliferation and directed cell differentiation.…”