Research Models of the Nanoparticle-Mediated Drug Delivery across the Blood–Brain Barrier

Brain diseases and damages come in many forms such as neurodegenerative diseases, tumors, and stroke. Millions of people currently suffer from neurological diseases worldwide. While Challenges of current diagnosis and treatment for neurological diseases are the drug delivery to the central nervous system. The Blood–Brain Barrier (BBB) limits the drug from reaching the targeted site thus showing poor effects. Nanoparticles that have advantage of the assembly at the nanoscale of available biomaterials can provide a delivery platform with potential to raising brain levels of either imaging therapeutic drugs or imaging. Therefore, successful modeling of the BBB is another crucial factor for the development of nanodrugs. In this review, we analyze the in vitro and in vivo findings achieved in various models, and outlook future development of nanodrugs for the successful treatment of brain diseases and damages.

Chondrogenic Potential of Dedifferentiated Rat Chondrocytes Reevaluated in Two- and Three-Dimensional Culture Conditions

For the cartilage repair, the cell sources currently adopted are primarily chondrocytes or mesenchymal stem cells (MSCs). Due to the fact that chondrocytes dedifferentiate during 2-dimensional (2D) expansion, MSCs are generally more studied and considered to have higher potential for cartilage repair purposes. Here we question if the dedifferentiated chondrocytes can regain the chondrogenic potential, to find potential applications in cartilage repair. For this we chose chondrocytes at passage 12 (considered to have sufficiently dedifferentiated) and the expression of chondrogenic phenotypes and matrix syntheses were examined over 14 days. In particular, the chondrogenic potential of MSCs was also compared. Results showed that the dedifferentiated chondrocytes proliferated actively over 14 days with almost 2.5-fold increase relative to MSCs. Moreover, the chondrogenic ability of chondrocytes was significantly higher than that of MSCs, as confirmed by the expression of a series of mRNA levels and the production of cartilage extracellular matrix molecules in 2D-monolayer and 3-dimensional (3D)-spheroid cultures. Of note, the significance was higher in 3D-culture than in 2D-culture. Although more studies are needed such as the use of different cell passages and human cell source, and the chondrogenic confirmation under in vivo conditions, this study showing that the dedifferentiated chondrocytes can also be a suitable cell source for the cell-based cartilage repair, as a counterpart of MSCs, will encourage further studies regarding this issue.

Effects of Type I Collagen Concentration in Hydrogel on the Growth and Phenotypic Expression of Rat Chondrocytes

It is controversial whether type I collagen itself can maintain and improve chondrogenic phenotype of chondrocytes in a three-dimensional (3D) environment. In this study, we examined the effect of type I collagen concentration in hydrogel (0.5, 1, and 2 mg/ml) on the growth and phenotype expression of rat chondrocytes in vitro. All collagen hydrogels showed substantial contractions during culture, in a concentration-dependent manner, which was due to the cell proliferation. The cell viability was shown to be the highest in 2 mg/ml collagen gel. The mRNA expression of chondrogenic phenotypes, including SOX9, type II collagen, and aggrecan, was significantly up-regulated, particularly in 1 mg/ml collagen gel. Furthermore, the production of type II collagen and glycosaminoglycan (GAG) content was also enhanced. The results suggest that type I collagen hydrogel is not detrimental to, but may be useful for, the chondrocyte culture for cartilage tissue engineering.

Co-culture of Human Dental Pulp Stem Cells and Endothelial Cells Using Porous Biopolymer Microcarriers: A Feasibility Study for Bone Tissue Engineering

Delivery of stem cells with osteogenesis while enabling angiogenesis is important for vascularized bone tissue engineering. Here a three-dimensional (3D) co-culture system of dental pulp stem cells (DPSCs) and endothelial cells (ECs) was designed using porous microcarriers, and the feasibility of applying to bone tissue engineering was investigated in vitro. Highly porous spherical microcarriers made of degradable biopolymers were prepared with sizes of hundreds of micrometers. The microcarriers loaded with DPSCs were co-cultured with ECs embedded in a hydrogel of type I collagen. An optimal coculture medium that preserves the viability of ECs while stimulating the osteogenic differentiation of DPSCs was found to be a 10:1 of osteogenic medium:endothelial medium. The co-cultured constructs of DPSCs/ECs showed significantly higher level of alkaline phosphatase activity than the mono-cultured cells. Moreover, the expressions of genes related with osteogenesis and angiogenesis were significantly up-regulated by the co-cultures with respect to the mono-cultures. Results imply the interplay between ECs and DPSCs through the designed 3D co-culture models. The microcarrier-enabled co-cultured cell system is considered to be useful as an alternative tool for future vascularized bone tissue engineering.

Alginate-Hyaluronic Acid-Collagen Composite Hydrogel Favorable for the Culture of Chondrocytes and Their Phenotype Maintenance

Articular cartilage has limited regeneration capacity, thus significant challenge has been made to restore the functions. The development of hydrogels that can encapsulate and multiply cells, and then effectively maintain the chondrocyte phenotype is a meaningful strategy to this cartilage repair. In this study, we prepared alginate-hyaluronic acid based hydrogel with type I collagen being incorporated, namely Alg-HA-Col composite hydrogel. The incorporation of Col enhanced the chemical interaction of molecules, and the thermal stability and dynamic mechanical properties of the resultant hydrogels. The primary chondrocytes isolated from rat cartilage were cultured within the composite hydrogel and the cell viability recorded revealed active proliferation over a period of 21 days. The mRNA levels of chondrocyte phenotypes, including SOX9, collagen type II, and aggrecan, were significantly up-regulated when the cells were cultured within the Alg-HA-Col gel than those cultured within the Alg-HA. Furthermore, the secretion of sulphated glycosaminoglycan, a cartilage-specific matrix molecule, was recorded higher in the collagen-added composite hydrogel. Although more in-depth studies are required such as the in vivo functions, the currently-prepared Alg-HA-Col composite hydrogel is considered to provide favorable 3-dimensional matrix conditions for the cultivation of chondrocytes. Moreover, the cell-cultured constructs may be useful for the cartilage repair and tissue engineering.

Method-Isolation and Culture of Primary Rat Adipose Derived Stem Cells Using Porous Biopolymer Microcarriers

Adipose-derived stem cells (ADSCs) are an attractive source of material for mesenchymal stem cell research due to the abundance of adipose and relative ease of access compared with bone marrow. A key consideration for research is whether cell isolation methods can be improved, to reduce the process steps needed to isolate and expand cell material. In the current study, we used macroporous biopolymer microcarriers to isolate primary ADSCs. We found that the method was capable of isolating ADSCs that were subsequently capable of being transferred to culture dishes and expanded in vitro. Moreover, flow cytometry revealed that they expressed typical stem cell markers and were capable of undergoing tri-lineage differentiation. In summary, it is feasible to use biopolymer microcarriers for retrieval of viable ADSCs that retain identity markers of stem cell function.

Porous Microcarrier-Enabled Three-Dimensional Culture of Chondrocytes for Cartilage Engineering: A Feasibility Study

Cartilage repair is substantially intractable due to poor self-healing ability. Porous microspheres can be a fascinating three-dimensional matrix for cell culture and injectable carrier in cartilage engineering. In this study, we assessed the feasible use of porous biopolymer microspheres for chondrocyte carriers. When seeded onto the blended biopolymer microspheres and followed by a dynamic spinner flask culture, the chondrocytes showed robust growth behaviors during the culture period. The gene expressions of SOX9, type II collagen, and aggrecan were significantly upregulated after 2-week of culture. Furthermore, immunolocalization of type II collagen and secretion of glycosaminolglycan became prominent. The results suggest the feasible usefulness of the porous microspheres as the cell culture matrix and the subsequent delivery into cartilage defects.

Enhanced tyrosine hydroxylase expression in PC12 cells co-cultured with feline mesenchymal stem cells

Mesenchymal stem cells (MSCs) secrete a variety of neuroregulatory molecules, such as nerve growth factor, brain-derived neurotrophic factor, and glial cell-derived neurotrophic factor, which upregulate tyrosine hydroxylase (TH) gene expression in PC12 cells. Enhancing TH gene expression is a critical step for treatment of Parkinson's disease (PD). The objective of this study was to assess the effects of co-culturing PC12 cells with MSCs from feline bone marrow on TH protein expression. We divided the study into three groups: an MSC group, a PC12 cell group, and the combined MSC + PC12 cell group (the co-culture group). All cells were cultured in DMEM-HG medium supplemented with 10% fetal bovine serum for three days. Thereafter, the cells were examined using western blot analysis and immunocytochemistry. In western blots, the co-culture group demonstrated a stronger signal at 60 kDa than the PC12 cell group (p < 0.001). TH was not expressed in the MSC group, either in western blot or immunocytochemistry. Thus, the MSCs of feline bone marrow can up-regulate TH expression in PC12 cells. This implies a new role for MSCs in the neurodegenerative disease process.