OBJECTIVE: Adipose tissue-derived stem cells (ASCs) are a promising source of cells for articular cartilage regeneration. However, ASCs isolated from different adipose tissue depots have heterogeneous cell characterizations and differentiation potential when cultured in 3-dimensional (3D) niches.
METHODS: We compared the chondrogenicity of ASCs isolated from infrapatellar fat pads (IPFPs) and subcutaneous fat pads (SCFPs) in 3D gelatin-based biomimetic matrix.
RESULTS: The IPFP-ASC-differentiated chondrocytes had higher ACAN, COL2A1, COL10, SOX6, SOX9, ChM-1, and MIA-3 mRNA levels and lower COL1A1 and VEGF levels than the SCFP-ASCs in 3D matrix. The difference in mRNA profile may have contributed to activation of the Akt, p38, RhoA, and JNK signaling pathways in the IPFP-ASCs. The chondrocytes differentiated from IPFP-ASCs had pronounced glycosaminoglycan and collagen type II production and a high chondroitin-6-sulfate/chondroitin-4-sulfate ratio with less polymerization of β-actin filaments. In an ex vivo mice model, magnetic resonance imaging revealed a shorter T2 relaxation time, indicating that more abundant extracellular matrix was secreted in the IPFP-ASC-matrix group. Histological examinations revealed that the IPFP-ASC matrix had higher chondrogenic efficacy of new cartilaginous tissue generation as evident in collagen type II and S-100 staining. Conclusion. ASCs isolated from IPFPs may be better candidates for cartilage regeneration, highlighting the translational potential of cartilage tissue engineering using the IPFP-ASC matrix technique.

Bovine pericardium has been proposed as an available material for tissue engineering and bioprosthetic reconstruction. In this study, bovine pericardium was fabricated into a scaffold for culturing and chondrogenic differentiation of human adipose-derived stem cells (hADSCs). Bovine pericardium was treated in 10 mM Tris-HCl and 0.15% SDS, followed by crosslinking in 0.1% glutaraldehyde. Treated bovine pericardium (tBP) was characterized as a slight yellowish thin membrane with enhanced tensile strength and strain property. The membrane maintained stability under enzymatic conditions for up to 16 days of incubation. The results confirmed tBP as a cell-friendly scaffold for hADSCs due to low cytotoxicity and its ability to support an appropriate attachment and proliferation of hADSCs. Moreover, there was an accumulation of the extracellular matrix proteoglycan in tBP seeded with hADSCs after 7 and 14 days of chondrogenic induction. COMP as a specific marker of chondrogenesis was detected after 7 days, whereas type X-a1 collagen (Col10a1) expression was stable up to day 14. However, minor expression of aggrecan was found. Taken together, these results indicate that tBP is a potential scaffold for hADSCs for cartilage tissue engineering.Key words: Bovine pericardium, scaffold, adipose-derived stem cells, chondrogenic differentiation, cartilage regeneration, augmentation rhinoplasty.

Microfluidic devices are beneficial in miniaturizing and multiplexing various cellular assays in a single platform. Chondrogenesis is known to pertain to chemical, topographical, and mechanical cues in the microenvironment. Mechanical cues themselves have numerous parameters such as strain magnitude, frequency, and stimulation time. Effects of different strain magnitudes on the chondrogenic differentiation of adult stem cells have not been explored thoroughly. Here, a new multilayer microdevice is presented for the unidirectional compressive stimulation of cells in a three-dimensional cell culture. Numerical simulations were performed to evaluate and optimize the design. Results showed a favorable highly uniform axial strain distribution and negligible radial and circumferential strain for the optimized design. Moreover, an experimental study was performed on rabbit adipose-derived stem cells encapsulated in-situ in alginate hydrogel. Strain levels of 20%, 15%, 10%, 5%, and 0% were studied simultaneously on a microfluidic platform. Dynamic mechanical compression positively influenced cellular viability and upregulated collagen II, Sox-9, and aggrecan expression in the absence of exogenous growth factors. The expression of collagen type II as specific marker for articular chondrocytes was further confirmed by immunofluorescence staining of collagen type II. Taking together, 10% strain can be considered as optimal stimulation factor for chondrogenic differentiation of adipose derived stem cells.

Fetal-derived mesenchymal stem cells especially human umbilical cord matrix mesenchymal stem cells (hUCMSCs), with their ease of availability, pluripotency, and high expansion potential have emerged as an alternative solution for stem cell based cartilage therapies. An attempt to elucidate the effect of dynamic mechanical compression in modulating the chondrogenic differentiation of hUCMSCs is done in this study to add on to the knowledge of optimizing chondrogenic signals necessary for the effective differentiation of these stem cells and subsequent integration to the surrounding tissues. hUCMSCs were seeded in porous poly (vinyl alcohol)-poly (caprolactone) (PVA-PCL) scaffolds and cultured in chondrogenic medium with/without TGF-β3 and were subjected to a dynamic compression of 10% strain, 1 Hz for 1/4 h for 7 days. The results on various analysis shows that the extent of dynamic compression is an important factor affecting cell viability. Mechanical stimulation in the form of dynamic compression stimulates expression of chondrogenic genes even in the absence of chondrogenic growth factors and also augments growth factor induced chondrogenic potential of hUCMSC. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 2554-2566, 2016.

Adipose-derived mesenchymal stromal cells are promising as a regenerative therapy tool for defective tissues in mesenchymal lineage, including fat, bone, cartilage, and blood vessels. In potential future clinical applications, adipose-derived stem cell cryopreservation is an essential fundamental technology. The aim of this study is to define an adequate protocol for the cryopreservation of adipose-derived mesenchymal stromal cells, by comparing various protocols so as to determine the effects of cryopreservation on viability and chondrogenic differentiation potential of adipose-derived stem cells upon freeze-thawing of AT-MSCs colonies cryopreserved with standard and modified protocols, using flow cytometry and confocal microscopy. The study concludes that adipose-derived mesenchymal stromal cells could be long-term cryopreserved without any loss of their proliferative or differentiation potential.

The study results of in vitro formation of tissue-engineered cartilage construct on the basis of cell-engineered construct composed of biopolymer hydrogel matrix and human adipose tissue-derived mesenchymal stromal cells (hADSCs) are presented. It was revealed that hADSCs in biopolymer hydrogel matrix Sphero®GEL under chondrogenic conditions generate three-dimensional structures and produce cartilaginous extracellular matrix components: collagen type II and glycosaminoglycans.