Effects of Oxysterols on Chondrogenesis of Human Adipose-derived Stem Cells.

Human adipose-derived stem cells (hADSCs) have been implicated as a high potency source of chondrocytes in cell therapy for cartilage defects. However, appropriate stimulators for the chondrogenesis of hADSCs are needed. Oxysterols have the potential to act as a stimulator. This study aims to investigate the effect of oxysterols on the chondrogenesis of hADSCs. hADSCs were collected from the abdominal subcutaneous tissue samples of patients undergoing caesarean section, and were cultured to passage 5. Mesenchymal stem cell markers were examined by flow cytometry. After the cells were subjected to adipogenic, osteogenic, and chondrogenic induction, the differentiation of each cell lineage was evaluated by RT-PCR and specific staining (Oil red O, Alizarin red S, and Alcian blue, respectively). The cell pellets of hADSCs were cultured in chondrogenic induced media containing 2µM 22(R)-hydroxycholesterol, 22(S)-hydroxycholesterol, or 25-hydroxycholesterol for 4 weeks. At 3 and 4 weeks of culture, the size and wet weight of the pellets were measured. The expressions of chondrogenesis-related genes and glycosaminoglycans production were examined by quantitative real-time PCR and Alcian blue staining. hADSCs were positive for the mesenchymal markers CD75, CD90, and CD105, while being negative for the hematopoietic markers CD31, CD34 and CD45. The multilineage potential of hADSCs was confirmed by the expression of adipogenic-, osteogenic-, and chondrogenic-specific genes, along with specific staining. 22(R)-hydroxycholesterol treatment significantly increased the size and wet weight of the pellet, glycosaminoglycans production, and expression of chondrogenic-related genes compared to the control group and other oxysterols (P<0.05). These results indicate that 22(R)-hydroxycholesterol can be effective as a stimulator for the chondrogenesis of hADSCs.

Serum estradiol levels during controlled ovarian hyperstimulation influence the pregnancy outcome of in vitro fertilization in a concentration-dependent manner.

OBJECTIVE: To determine an optimal serum E(2) level on the day of hCG administration in controlled ovarian hyperstimulation (COH) during IVF-ET without compromising pregnancy outcome. DESIGN: Retrospective study. SETTING: Large urban medical center. PATIENT(S): Data of 455 cycles of fresh IVF-ET with COH. INTERVENTION(S): Serum E(2) levels on the day of hCG administration were categorized into five groups: group A (<1000 pg/mL), group B (1000-2000 pg/mL), group C (2000-3000 pg/mL), group D (3000-4000 pg/mL), and group E (>4000 pg/mL). MAIN OUTCOME MEASURE(S): Serum E(2) levels, number of oocytes retrieved, pregnancy outcomes. RESULT(S): Of 455 cycles, 148 (32.5%) cycles resulted in clinical pregnancy. The implantation rate was 12.2%, and the delivery rate was 18.7%. The number of oocytes obtained increased with increasing serum E(2) levels. The pregnancy rate gradually increased from group A to D as E(2) levels increased but decreased in group E. In women <38 years, the IVF-ET outcomes were similar to those of total patients. However, in women >/=38 years old, pregnancy and delivery rates were higher in group C than in other groups. CONCLUSION(S): These results show that serum E(2) levels have a concentration-dependent effect on the pregnancy outcome, suggesting an optimal range of E(2) level for achieving a successful pregnancy. This optimal range of serum E(2) level in women is age dependent: 3000-4000 pg/mL for women <38 years and 2000-3000 pg/mL for women >/=38 years.

Decreased amniotic fluid leptin levels in the second trimester in Down's syndrome.

OBJECTIVE: To investigate a possible role of leptin contributing to the pathophysiology of Down's syndrome by assessing amniotic fluid leptin levels in the 2nd trimester of normal pregnancy and pregnancy complicated by Down's syndrome. METHODS: We retrospectively assessed leptin levels of 2nd trimester amniotic fluid from 15 pregnancies diagnosed with Down's syndrome and 15 matched unaffected singleton pregnancies (controls). All amniotic fluid samples were obtained from women undergoing genetic amniocentesis at 16-20 weeks of gestation. Leptin levels were measured by enzyme-linked immunosorbent assay. RESULTS: Amniotic fluid leptin levels were significantly decreased in the Down's syndrome group (3.26 +/- 2.36 ng/ml) compared to the controls (8.26 +/- 9.11 ng/ml, p < 0.05). CONCLUSION: This study showed significantly decreased leptin levels in the 2nd trimester amniotic fluid of fetuses with Down's syndrome. This result suggests a possible role of leptin in the pathophysiology of Down's syndrome during the fetal period.

Correlation of synovial fluid leptin concentrations with the severity of osteoarthritis.

Leptin is known to play an important role in the pathophysiology of osteoarthritis (OA). This study investigated whether synovial fluid (SF) leptin level is related to the radiographic severity of OA and its role as a quantitative marker for the detection of OA. SF was obtained from 42 OA patients who underwent knee surgery and 10 who had no abnormality of articular cartilage during arthroscopic examination. The progression of OA was classified by Kellgren-Lawrence grading scale. The concentrations of leptin were measured with commercial enzyme-linked-immunosorbent serologic assay kits. Median leptin concentrations in SF were significantly higher in OA patients (median 4.40 ng/ml; range 0.5-15.8) compared to controls (median 2.05 ng/ml; range 1.0-4.6; P = 0.006). SF leptin levels showed significant difference according to the severity of OA (P = 0.0125). Median SF leptin level was highest in stage IV patients (11.1 ng/ml), which was significantly higher compared to all other groups including controls (P < 0.05). Age showed a significant positive correlation with leptin concentrations in OA patients (P < 0.05), but not in controls. These results demonstrate that SF leptin concentrations were closely related to the radiographic severity of OA, suggesting that SF leptin levels could be used as an effective marker for quantitative detection of OA.