Osteoblastic differentiation potential of human amniotic fluid-derived mesenchymal stem cells in different culture conditions.

Osteoporosis is a bone degenerative disease characterized by a decrease in bone strength and an alteration in the osseous micro-architecture causing an increase in the risk of fractures. These diseases usually happen in post-menopausal women and elderly men. The most common treatment involves anti-resorptive agent drugs. However, the inhibition of bone resorption alone is not adequate for recovery in patients at the severe stage of osteoporosis who already have a fracture. Therefore, the combination of utilizing osteoblast micro mimetic scaffold in cultivation with the stimulation of osteoblastic differentiations to regain bone formation is a treatment strategy of considerable interest. The aims of this current study are to investigate the osteoblastic differentiation potential of mesenchymal stem cells derived from human amniotic fluid and to compare the monolayer culture and scaffold culture conditions. The results showed the morphology of cells in human amniotic fluid as f-type, which is a typical cell shape of mesenchymal stem cells. In addition, the proliferation rate of cells in human amniotic fluid reached the highest peak after 14 days of culturing. After which time, the growth rate slowly decreased. Moreover, the positive expression of specific mesenchymal cell surface markers including CD44, CD73, CD90, and also HLA-ABC (MHC class I) were recorded. On the other hand, the negative expressions of the endothelial stem cells markers (CD31), the hematopoietic stem cells markers (CD34, 45), the amniotic stem cells markers (CD117), and also the HLA-DR (MHC class II) were also recorded. The expressions of osteoblastogenic related genes including OCN, COL1A1, and ALP were higher in the osteogenic-induced group when compared to the control group. Interestingly, the osteoblastogenic related gene expressions that occurred under scaffold culture conditions were superior to the monolayer culture conditions. Additionally, higher ALP activity and greater calcium deposition were recorded in the extracellular matrix in the osteogenic-induced group than in the culture in the scaffold group. In summary, the mesenchymal stem cells derived from human amniotic fluid can be induced to be differentiated into osteoblastic-like cells and can promote osteoblastic differentiation using the applied scaffold.

Sesamin encouraging effects on chondrogenic differentiation of human amniotic fluid-derived mesenchymal stem cells.

Worldwide, the most recognized musculoskeletal degenerative disease is osteoarthritis (OA). Sesamin, a major abundant lignan compound present in Sesamun Indicum Linn, has been described for its various pharmacological effects and health benefits. However, the promoting effects of sesamin on chondrogenic differentiation have not yet been observed. Herein, the aim of this study was to investigate the effects of sesamin on cell cytotoxicity and the potent supporting effects on chondrogenic differentiation of human amniotic fluid-derived mesenchymal stem cells (hAF-MSCs). The results indicated that sesamin was not toxic to hAF-MSCs after sesamin treatment. When treating the cells with a combination of sesamin and inducing factors, sesamin was able to up-regulate the expression level of specific genes which play an essential role during the cartilage development process, including SOX9, AGC, COL2A1, COL11A1, and COMP and also simultaneously promote the cartilage extracellular protein synthesis, aggrecan and type II collagen. Additionally, histological analysis revealed a high amount of accumulated sGAG staining inside the porous scaffold in the sesamin co-treating group. In conclusion, the results of this study have indicated that sesamin can be considered a chondrogenic inducing factor and a beneficial dietary supplement for cartilage repair.

Mesenchymal stem cells differentiated into chondrocyte-Like cells.

Among the stem cells contained in human amniotic fluid (hAF), the human amniotic fluid derived-mesenchymal stem cells (hAF-MSCs) are derived from fetal membranes and tissues that are produced during fetal development. The aim of this study was to characterize the 'stem-ness' properties of hAF-MSCs and their potency with regard to the chondrogenic differentiations using the scaffold cultivation method. This study revealed that the easily accessed and isolated MSCs were highly cell prolific and there were fewer ethical concerns regarding their usage. The MSCs were studied through the use of the alamar blue technique. In addition, after cell isolation, hAF-MSCs displayed typical MSCs morphologies including MSCs biomarker characteristics and immune privilege properties (CD44, CD73, CD90, CD105 and HLA-ABC) through immunofluorescence and flow cytometry. Interestingly, this result indicated a negative expression when using the C-Kit (CD117, tyrosine kinase receptor type III ligand for cytokine stem cell factor). This expression can be found at the cell's surface of the amniotic fluid-derived stem cells (AFSCs). This study found evidence that hAF-MSCs had the ability to differentiate the cells into the chondrogenic lineage by exhibiting chondrogenic related genes and proteins (SOX9, AGC, COL2A1 and COMP) through RT-qPCR, immunoenzymatic assays and immunofluorescence analysis. Furthermore, MSCs presented sGAGs accumulation, which was confirmed by histological analysis and SEM. Therefore, this study showed that the MSCs characteristics are contained in AF and are of significant value for further research. It appears that MSCs possess the potential for use in treatments that would necessitate the use of regenerative cell therapy.