Spermatogenesis is a process in which animals generate spermatozoa from spermatogonial stem cells (SSCs). Successful in vitro differentiation of SSCs towards spermatids holds a significant promise for regeneration of impaired spermatogenesis. The present study aims to evaluate the efficiency of a 3D culture containing naringenin on proliferation and differentiation potentials of mouse SSCs. In this study, multiwalled carbon nanotubes (MWCNTs) were incorporated into poly(L-lactic acid) (PLLA) fibers via electrospinning technique. The fibrous PLLA/MWCNTs were studied by Fourier-transform infrared spectroscopy (FTIR), transmission electron microscope (TEM), water contact angle measurements, electrical conductivity, and mechanical properties. Next, the SSCs were seeded into the PLLA/MWCNTs scaffolds and exhibited preferable survival and differentiation efficiency to subsequent cell lines. To shed more light on this matter, the immunocytochemistry, reverse-transcription polymerase chain reaction (RT-PCR), and qRT-PCR results showed that the aforementioned cells on the 3D fabrics overexpressed the C-kit and SYCP3 proteins. In addition, the reactive oxygen species (ROS) measurement data demonstrated that naringenin, an effective antioxidant, plays an important role in in vitro spermatogenesis. Taken together, the results of this study revealed the synergistic effects of 3D scaffolds and naringenin for efficient spermatogenesis in laboratories. KEYWORDS antioxidants, in vitro spermatogenesis, naringenin, PLLA/MWCNTs scaffolds, spermatogonial stem cells (SSCs) Abbreviations: bp, base pairs; C-Kit, proto-oncogene c-Kit or tyrosine-protein kinase Kit; DMEM, Dulbecco's modified eagle medium; DMF, N, N-dimethyl formamide; ECM, natural extra cellular matrix; FTIR, Fourier transform-infrared spectroscopy; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; ICSI, intra-cytoplasmic sperm injection; Id4, inhibitor of differentiation 4; IVM, in vitro oocyte maturation; m-TESE, testicular sperm extraction; PLLA/MWCNTs, poly (L-lactic acid)/multi-walled carbon nanotube; PLZF, promyelocytic leukemia zinc finger; ROS, reactive oxygen species; ROSI, round spermatid injection; RT-PCR, reverse-transcription polymerase chain reaction; SEM, scanning electron microscopy; SSCs, spermatogonial stem cells; SYCP3, synaptonemal complex protein 3

Background: Synthetic tissue engineering scaffolds has poor biocompatiblity with very low angiogenic properties. Conditioning the scaffolds with functional groups, coating with biological components, especially extracellular matrix (ECM), is an excellent strategy for improving their biomechanical and biological properties.Methods: In the current study, a composite of polycaprolactone and gelatin (PCL/ Gel) was electrospun in the ratio of 70/30 and surface modified with 1% gelatincoating (G-PCL/Gel) or plasma treatment (P-PCL/Gel). The surface modification was determined by SEM and ATR-FTIR spectroscopy, respectively. The scaffolds were cultured with fibroblast 3T3, then decellularized during freeze-thawing process to fabricate a fibroblast ECM-conditioned PCL/Gel scaffold (FC-PCL/Gel).The swelling and degaradtion as well as in vitro and in vivo biocompatibility and angiogenic properties of the scaffolds were evaluated.

Background Several factors like three‐dimensional microstructure, growth factors, cytokines, cell–cell communication, and coculture with functional cells can affect the stem cells behavior and differentiation. The purpose of this study was to investigate the potential of decellularized placental sponge as adipose‐derived mesenchymal stem cells (AD‐MSCs) and macrophage coculture systems, and guiding the osteogenic differentiation of stem cells. Methods The decellularized placental sponge (DPS) was fabricated, and its mechanical characteristics were evaluated using degradation assay, swelling rate, and pore size determination. Its structure was also investigated using hematoxylin and eosin staining and scanning electron microscopy. Mouse peritoneal macrophages and AD‐MSCs were isolated and characterized. The differentiation potential of AD‐MSCs co‐cultured with macrophages was evaluated by RT‐qPCR of osteogenic genes on the surface of DPS. The in vivo biocompatibility of DPS was determined by subcutaneous implantation of scaffold and histological evaluations of the implanted site. Results The DPS had 67% porosity with an average pore size of 238 μm. The in vitro degradation assay showed around 25% weight loss during 30 days in PBS. The swelling rate was around 50% during 72 h. The coculture of AD‐MSCs/macrophages on the DPS showed a significant upregulation of four differentiation osteogenic lineage genes in AD‐MSCs on days 14 and 21 and a significantly higher mineralization rate than the groups without DPS. Subcutaneous implantation of DPS showed in vivo biocompatibility of scaffold during 28 days follow‐up. Conclusions Our findings suggest the decellularized placental sponge as an excellent bone substitute providing a naturally derived matrix substrate with biostructure close to the natural bone that guided differentiation of stem cells toward bone cells and a promising coculture substrate for crosstalk of macrophage and mesenchymal stem cells in vitro.

Attempts are being made to develop an ideal wound dressing with excellent biomechanical and biological properties. Here, a thermos-responsive hydrogel is fabricated using chitosan (CTS) with various concentrations (1%, 2.5%, and 5% w/v) of solubilized placental extracellular matrix (ECM) and 20% 𝜷-glycerophosphate to optimize a smart wound dressing hydrogel with improved biological behavior. The thermo-responsive CTS (TCTS) alone or loaded with ECMs (ECM-TCTS) demonstrate uniform morphology using SEM. TCTS and ECM1%-TCTS and ECM2.5%-TCTS show a gelation time of 5 min at 37 °C, while no gel formation is observed at 4 and 25 °C. ECM5%-TCTS forms gel at both 25 and 37 °C. The degradation and swelling ratios increase as the ECM content of the hydrogel increase. All the constructs show excellent biocompatibility in vitro and in vivo, however, the hydrogels with a higher concentration of ECM demonstrate better cell adhesion for fibroblast cells and induce expression of angiogenic factors (VEGF and VEGFR) from HUVEC. Only the ECM5%-TCTS has antibacterial activity against Acinetobacter baumannii ATCC 19606. The data obtained from the current study suggest the ECM2.5%-TCTS as an optimized smart biomimetic wound dressing with improved angiogenic properties now promises to proceed with pre-clinical and clinical investigations.

Recent studies have demonstrated the influential role of microbial stimulus in characteristics and immunomodulatory effects of mesenchymal stem cells (MSCs). Due to the migration of MSCs to infection site, it is of importance to understand the interaction of microbial ligands with MSCs in order to clarify the positive or negative role of MSCs in the control of infection. In this research, we assess leishmanial soluble antigen (LSA)-primed MSCs on macrophage immune responses to lipopolysaccharide (LPS). For this purpose, the effects of both conditioned media (CM) and cell-cell contact of LSA primed MSCs were determined on macrophage responses to LPS. According to the obtained results, MSC-treated macrophages demonstrated an alternatively activated macrophages with higher levels of interleukin-10 (IL-10) and transforming growth factor-alpha (TNF-α) and lower levels of IL-6 and nitric oxide (NO) production as compared to the controls. In addition, phagocytosis of apoptotic thymocytes was induced in MSC-treated macrophages. In conclusion, it seems that MSCs trigger an anti-inflammatory phenotype in macrophages at Leishmania infected sites in order to enhance the induction of immune regulatory cells and clearance of apoptotic cells. J. Cell. Biochem. 118: 2764-2779, 2017. © 2017 Wiley Periodicals, Inc.