Objective To explore the possibility of constructing tissue engineered cartilage complex three-dimensional nano-scaffold with collagen type II and hyaluronic acid (HA) by electrospinning. Methods The three-dimensional porous nano-scaffolds were prepared by electrospinning techniques with collagen type II and HA (8 ∶ 1, W ∶ W), which was dissolved in mixed solvent of 3-trifluoroethanol and water (1 ∶ 1, V ∶ V). The morphology were observed by light microscope and scanning electron microscope (SEM). And the porosity, water absorption rate, contact angle, and degradation rate were detected. Chondrocytes were harvested from 1-week-old Japanese white rabbit, which was disgested by 0.25% trypsin 30 minutes and 1% collagenase overlight. The passage 2 chondrocytes were seeded on the nano-scaffold. The cell adhesion and proliferation were evaluated by cell counting kit 8 (CCK-8). The cell-scaffold composites were cultured for 2 weeks in vitro, and the biological morphology and extracelluar matrix (ECM) secretion were observed by histological analysis. Results The optimal electrospinning condition of nano-scaffold was 10% electrospinning solution concentration, 10 cm receiver distance, 5 mL/ h spinning injection speed. The scaffold had uniform diameter and good porosity through the light microscope and SEM. The diameter was 300-600 nm, and the porosity was 89.5% ± 25.0%. The contact angle was (35.6 ± 3.4)°, and the water absorption was 1 120% ± 34% at 24 hours, which indicated excellent hydrophilicity. The degradation rate was 42.24% ± 1.51% at 48 days. CCK-8 results showed that the adhesive rate of cells with scaffold was 169.14% ± 11.26% at 12 hours, and the cell survival rate was 126.03% ± 4.54% at 7 days. The histological and immunohistochemical staining results showed that the chondrocytes could grow well on the scaffold and secreted ECM. And the similar cartilage lacuma structure could be found at 2 weeks after co-culture, which suggested that hyaline cartilage formed. Conclusion The collage type II and HA complex three-dimensional nano-scaffold has good physicochemical properties and excellent biocompatibility, so it can be used as a tissue engineered cartilage scaffold.
ObjectiveTo explore the effect of laparoscopic hepatectomy in patients with complex hepatolithiasis.MethodsThe clinical data of 31 patients with complex hepatolithiasis treated by laparoscopic hepatectomy in our hospital from January 2015 to September 2019 were retrospectively analyzed, and the effect was followed up.ResultsTwo cases were converted to open surgery, and the remaining 29 cases successfully completed laparoscopic surgery. The operative time of 31 patients was 185–490 min (260±106) min; the intraoperative bleeding volume was 200–1 300 mL (491±225) mL; the time of hepatic blood flow occlusion was 20–45 min (29±18) min; the time of choledochoscopy was 10–50 min (28±15) min. The scope of hepatectomy includes: Ⅱ, Ⅲ, Ⅵ, and Ⅶ in 14 cases, Ⅰ, Ⅱ, Ⅲ,Ⅵ, and Ⅶ in 8 cases, Ⅱ, Ⅲ, Ⅳ, Ⅵ, and Ⅶ in 3 cases, Ⅳ, Ⅴ, and Ⅷ in 3 cases, Ⅱ, Ⅲ, Ⅴ, Ⅵ, Ⅶ, and Ⅷ in 2 cases, Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅵ, and Ⅶ in 1 case. The postoperative anal exhaust time was 24–73 h (41.8±15.2) h; postoperative feeding time was 14–23 h, median feeding time was 19 h; postoperative ambulation time was 15–46 h, median ambulation time was 27 h; postoperative drainage tube extraction time was 3–14 d, median drainage tube extraction time was 5 d; postoperative hospitalization time was 6–15 d, median postoperative hospitalization time was 9 d. Twenty-seven patients were followed up for 7 to 63 months, with a median follow-up time of 25 months. The incidence of complications was 19.4% (6/31), according to Claviein classification, there were 2 cases in grade Ⅰ (6.5%), 1 case in grade Ⅱ (3.2%), 3 cases in grade Ⅲa (9.7%).ConclusionLaparoscopic hepatectomy is safe and feasible for the treatment of complex hepatolithiasis, which is performed by a skilled hepatobiliary surgeon.