TRITC-Loaded PLGA Nanoparticles as Drug Delivery Carriers in Mouse Oocytes and Embryos.

The structural layers around oocytes make it difficult to deliver drugs aimed at treating infertility. In this study, we sought to identify nanoparticles (NPs) that could easily pass through zona pellucida (ZP), a special layer around oocytes, for use as a drug delivery carrier. Three types of NPs were tested: quantum dot NPs, PE-polyethylene glycol (PEG)-loaded poly(lactic-co-glycolic acid) (PLGA) NPs (PEG/PL), and tetramethylrhodamine-loaded PLGA NPs (TRNPs). When mouse oocytes were treated with NPs, only TRNPs could fully pass through the ZP and cell membrane. To assess the effects of TRNPs on fertility and potential nanotoxicity, we performed mRNA sequencing analysis to confirm their genetic safety. We established a system to successfully internalize TRNPs into oocytes. The genetic stability and normal development of TRNP-treated oocytes and embryos were confirmed. These results imply that TRNPs can be used as a drug delivery carrier applicable to germ cells.

In situ pocket-type microcarrier (PMc) as a therapeutic composite: Regeneration of cartilage with stem cells, genes, and drugs.

We prepared pocket-type micro-carriers (PMc) with pores larger than 30 µm for use in cell delivery by adding 40 mg pluronic F-127 copolymers (F-127) to biodegradable PLGA dissolved in dichloromethane solution. The controlling the size of the pockets in this way facilitates the adhesion of cells by regulating the size of the pockets according to the cells having various sizes. The size of PMc pores could be controlled within a range of 2 to 30 µm by varying the F-127 content. The ratio of F-127 to DOPA-bPEI was most appropriate at 1: 1, and the pocket size at 10 mg/ml of F-127 was appropriate for adhering 20-30 µm stem cells. F-127 containing SOX9 pDNA, in combination with DOPA-polyethylene-coated gold nanoparticles and dexamethasone loaded in PMcs, promoted cartilage differentiation. Gold nanoparticles complex and dexamethasone (DEX) loaded in PMcs were identified by micro-CT imaging and fluorescence imaging, respectively. By captured in pore generated on/in microspheres, the stem cells were safe and stable for use in delivery, both in vitro and in an animal model. Thus, microsphere pores can safely capture stem cells, and at the same time provide a microenvironment in which the captured stem cells can differentiate into chondrocytes.

Fabrication of Nanocomposites Complexed with Gold Nanoparticles on Polyaniline and Application to Their Nerve Regeneration.

Electrically conductive materials can stimulate stem cells through electric shock and thereby contribute to the regulation of cell proliferation and differentiation. Recently, polymer-metal complexes composed of polyaniline and gold nanoparticles have emerged as novel candidates for use in regenerative medicine. By mixing two different materials, such composites maximize the benefits while alleviating the disadvantages of using either material alone. Based on their excellent conductivity, these complexes can be applied to nerve regeneration using stem cells. In this study, we investigated a method for producing hybrid nanocomposites by complexing gold nanoparticles to polyaniline and tested the resultant composites in a model of nerve regeneration. We manipulated the shape, size, and electrical conductivity of the hybrid composites by compounding the component materials at various ratios. The most efficient nanocomposite was named conductive reinforced nanocomposites (CRNc's). When the CRNc was delivered directly to cells, no cytotoxicity was observed. After the intracellular delivery of the CRNc, the stem cells were electrically stimulated using an electroporator. As a result of performing mRNA-sequencing (Seq) analysis after electrical stimulation (ES) of the CRNc-internalized cells, it was confirmed that the CRNc-internalized cells have a pattern similar to that of the positive group-induced neuron cells. In particular, microtubule-associated protein 2 is more than twice that of the control group (negative control), and the nerve fiber protein is strongly expressed as in the positive control group. In addition, we verified that neural differentiation progressed by monitoring the growth of neurites from stem cells. Together, these findings show that the CRNc can be used to induce the formation of neuron-like cells by applying ES to stem cells.

Primary retroperitoneal dysgerminoma presenting as an adrenal tumor: a case report and literature review.

Primary extragonadal germ cell tumors are rare and mostly occur in young men with predominance of nonseminomatous histology. We report an undescribed case of primary retroperitoneal dysgerminoma presenting as an adrenal tumor in a 17-year-old girl. Surgery was performed on a 10 × 9.5 cm sized adrenal gland tumor and the resected tumor showed unequivocal histological features of dysgerminoma. The diagnosis was confirmed by the tumor's germ cell immunophenotype. Postoperative ultrasonography, CT and PET over a 6-month period revealed no evidence of ovarian lesion. The patient is stable, but with a suspicious residual tumor after adjuvant chemotherapy.