1.Modulation of osteogenic differentiation in mouse osteoprogenitor cells by curved microgrooves with varied roughness on polycaprolactone surfaces
Weixin WANG ; Yixuan LEI ; Yingzhen LAI ; Haitao YE ; Menglin CAO ; Yuqi HE
Chinese Journal of Stomatology 2025;60(10):1152-1161
Objective:To construct curved microgrooves with gradient surface roughness on polycaprolactone (PCL) members by regulating alkali etching time and to investigate the synergistic effect of surface roughness and curved microgrooves on the in vitro osteogenic differentiation of mouse pre-osteoblasts (MC3T3-E1), aiming to determine the optimal PCL surface modification strategy. Methods:Soft lithography and melt-casting techniques were used to fabricate PCL membranes with regularly arranged curved microgrooves (CMP). Alkali etching was performed for 24, 48, and 72 h. Groups: smooth PCL (control), CMP (curved microgrooves only), CMP-24 h, CMP-48 h, CMP-72 h (CMP etched for 24, 48, 72 h, respectively). Surface physicochemical properties were characterized: surface morphology was observed by scanning electron microscopy (SEM), surface roughness was measured by atomic force microscopy (AFM), and surface hydrophilicity was evaluated by contact angle measurement. MC3T3-E1 cells were cultured in vitro. Cell adhesion, proliferation, and osteogenic differentiation were assessed using cell counting (CCK-8), immunofluorescence staining, alkaline phosphatase (ALP) and Alizarin red staining with quantification. The mRNA expression levels of osteogenesis-related genes [ALP, collagen type Ⅰ (COL-1), Runt-related transcription factor 2 (RUNX-2), osteocalcin (OCN), osteopontin (OPN)] were detected by real-time fluorescence quantitative PCR (RT-qPCR). Results:Curved microgrooves were successfully fabricated on PCL membranes. Alkali treatment improved surface hydrophilicity and increased roughness. The CMP-72 h group exhibited the best hydrophilicity (contact angle: 33.2°±5.5°), with significant differences compared to all other groups (all P<0.05). The CMP-72 h group showed the highest roughness [(59.966±4.729) nm], which was significantly different from all other groups (all P<0.05). CCK-8 results on day 5 showed that both curved microgrooves and roughness promoted cell proliferation: CMP-24 h (0.292±0.003) and CMP-72 h (0.383±0.004) were significantly higher than those in the smooth group (0.270±0.005) (all P<0.05). Immunofluorescence staining revealed that curved microgrooves induced significant contact guidance of cells; this effect weakened with increasing etching time. ALP and Alizarin red staining indicated the deepest osteogenic staining in the CMP-48 h group. Both ALP activity (0.013 021±0.000 032) and Alizarin red quantification (0.290±0.003) were highest in the CMP-48 h group, significantly different from all other groups (all P<0.05). RUNX-2 expression in CMP-24 h and CMP-48 h groups (1.845±0.087 and 1.837±0.027, respectively) was significantly higher than in other groups (all P<0.05), with no significant difference between these two groups ( P>0.05). CMP-48 h group exhibited the highest mRNA expression of all osteogenic genes tested, specifically ALP (2.194±0.028), COL-1 (1.983±0.024), OCN (7.644±0.156), and OPN (2.648±0.031), all significantly greater than other groups (all P<0.05). Conclusions:Both curved microgrooves and surface roughness modification enhance the in vitro osteogenic differentiation of cells on PCL membranes. Among the tested strategies, alkali etching of curved microgrooves for 48 hours (CMP-48h) provided the optimal enhancement of osteogenic capability for MC3T3-E1 cells and represented a promising surface modification strategy for future PCL membranes.
2.Modulation of osteogenic differentiation in mouse osteoprogenitor cells by curved microgrooves with varied roughness on polycaprolactone surfaces
Weixin WANG ; Yixuan LEI ; Yingzhen LAI ; Haitao YE ; Menglin CAO ; Yuqi HE
Chinese Journal of Stomatology 2025;60(10):1152-1161
Objective:To construct curved microgrooves with gradient surface roughness on polycaprolactone (PCL) members by regulating alkali etching time and to investigate the synergistic effect of surface roughness and curved microgrooves on the in vitro osteogenic differentiation of mouse pre-osteoblasts (MC3T3-E1), aiming to determine the optimal PCL surface modification strategy. Methods:Soft lithography and melt-casting techniques were used to fabricate PCL membranes with regularly arranged curved microgrooves (CMP). Alkali etching was performed for 24, 48, and 72 h. Groups: smooth PCL (control), CMP (curved microgrooves only), CMP-24 h, CMP-48 h, CMP-72 h (CMP etched for 24, 48, 72 h, respectively). Surface physicochemical properties were characterized: surface morphology was observed by scanning electron microscopy (SEM), surface roughness was measured by atomic force microscopy (AFM), and surface hydrophilicity was evaluated by contact angle measurement. MC3T3-E1 cells were cultured in vitro. Cell adhesion, proliferation, and osteogenic differentiation were assessed using cell counting (CCK-8), immunofluorescence staining, alkaline phosphatase (ALP) and Alizarin red staining with quantification. The mRNA expression levels of osteogenesis-related genes [ALP, collagen type Ⅰ (COL-1), Runt-related transcription factor 2 (RUNX-2), osteocalcin (OCN), osteopontin (OPN)] were detected by real-time fluorescence quantitative PCR (RT-qPCR). Results:Curved microgrooves were successfully fabricated on PCL membranes. Alkali treatment improved surface hydrophilicity and increased roughness. The CMP-72 h group exhibited the best hydrophilicity (contact angle: 33.2°±5.5°), with significant differences compared to all other groups (all P<0.05). The CMP-72 h group showed the highest roughness [(59.966±4.729) nm], which was significantly different from all other groups (all P<0.05). CCK-8 results on day 5 showed that both curved microgrooves and roughness promoted cell proliferation: CMP-24 h (0.292±0.003) and CMP-72 h (0.383±0.004) were significantly higher than those in the smooth group (0.270±0.005) (all P<0.05). Immunofluorescence staining revealed that curved microgrooves induced significant contact guidance of cells; this effect weakened with increasing etching time. ALP and Alizarin red staining indicated the deepest osteogenic staining in the CMP-48 h group. Both ALP activity (0.013 021±0.000 032) and Alizarin red quantification (0.290±0.003) were highest in the CMP-48 h group, significantly different from all other groups (all P<0.05). RUNX-2 expression in CMP-24 h and CMP-48 h groups (1.845±0.087 and 1.837±0.027, respectively) was significantly higher than in other groups (all P<0.05), with no significant difference between these two groups ( P>0.05). CMP-48 h group exhibited the highest mRNA expression of all osteogenic genes tested, specifically ALP (2.194±0.028), COL-1 (1.983±0.024), OCN (7.644±0.156), and OPN (2.648±0.031), all significantly greater than other groups (all P<0.05). Conclusions:Both curved microgrooves and surface roughness modification enhance the in vitro osteogenic differentiation of cells on PCL membranes. Among the tested strategies, alkali etching of curved microgrooves for 48 hours (CMP-48h) provided the optimal enhancement of osteogenic capability for MC3T3-E1 cells and represented a promising surface modification strategy for future PCL membranes.
3.Clinical effectiveness of endoscopic versus surgical treatment of chronic pancreatitis: a meta-analysis
Chuyue ZHANG ; Guangjin LIANG ; Yingzhen SU ; Chunfeng WANG ; Wang ZENG ; Fan CAO ; Bo HUANG
Chinese Journal of Hepatobiliary Surgery 2023;29(1):54-59
Objective:To compare the efficacy of endoscopy and surgery in chronic pancreatitis.Methods:CNKI, CBM, Wanfang, PubMed, Cochrane Library, Embase and Web of Science were searched to compared endoscopy and surgery for the clinical efficacy of chronic pancreatitis. Literatures were searched from the establishment of the database to August 14, 2022. Compared pain relief, clinical response to initial treatment, complications, endocrine/exocrine insufficiency, length of hospital stay and mean number of procedures between the two groups. Manager 5.4.1 software was used for data analysis. Odds ratio ( OR) or weighted mean difference ( WMD) was calculated with 95% confidence interval (95% CI). Results:A total of seven studies were included, including three randomized controlled trials and four retrospective studies with 708 patients. There were 513 males and 195 females. Endoscopic interventions were performed in 364 patients and 344 patients underwent surgery. The results of meta-analysis showed that the total pain relief rate ( OR=0.38, 95% CI: 0.24-0.59) and the complete pain relief rate ( OR=0.47, 95% CI: 0.29-0.77), short-term (1-1.5 years) pain relief rate ( OR=0.42, 95% CI: 0.24-0.74), clinical relief rate ( OR=0.23, 95% CI: 0.10-0.55) were better than the endoscopic group, and could significantly reduce the number of reoperation ( WMD=1.64, 95% CI: 0.89-2.40), and the difference was statistically significant (all P<0.05). There were no significant differences in complications, new-onset endocrine insufficiency, new-onset exocrine insufficiency and length of hospital stay between the endoscopy group and the surgical group (all P>0.05). Conclusion:Surgical intervention is superior to endoscopic treatment in controlling pain associated with chronic pancreatitis and in clinical relief after the first treatment, and can effectively reduce the number of reoperations.
4.Application of circulating free DNA in the diagnosis and treatment of hepatocellular carcinoma
Yijian YANG ; Bimang FU ; Nan XIE ; Fan CAO ; Yingzhen SU
Journal of Clinical Hepatology 2020;36(12):2835-2838
Hepatocellular carcinoma has a low early diagnostic rate, and there is a lack of highly sensitive and specific tumor markers. In recent years, fluid biopsy technique, represented by circulating free DNA (cfDNA), has become an auxiliary method for the diagnosis of cancer and has attracted more and more attention due to its advantages of noninvasiveness, convenience, and repeatability. With reference to the recent studies in China and foreign countries, this article summarizes and analyzes the advances in cfDNA in the diagnosis and treatment of hepatocellular carcinoma from the aspects of biological characteristics, detection techniques, and clinical application, so as to provide a basis for clinical diagnosis and treatment.
5.Identification of Fengliaoxing Fengshi Dieda Medicinal Wines in Fake by GC and TLC
Zhen TAN ; Yingzhen CAO ; Yongjian WANG
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(09):-
Objective To establish a method for the fake indentification of Fengliaoxing Fengshi Dieda medicinal wines by TLC and GC. Methods The column was Supelco SE-30 (30 m?0.25 mm?0.25 ?m). The flow rate was 1.4 mL/min, and gas was N2. The inlets temperature was 210 ℃;the detectors temperature was 300 ℃;the column temperature use program rising temperature. Results TLC can identify whether ephedra and erycibe were contained in the medicinal wines. GC can identify whether diclofenac sodium was contained in the medicinal wines. Conclusion The method is accurate and reproducible, and can be used for the identification of Fengliaoxing Fengshi Dieda medicinal wines.
6.Content Determination of Waishang Ruyi Ointments by GC
Chinese Journal of Information on Traditional Chinese Medicine 2006;0(09):-
Objective To establish a method of GC for determination of Waishang Ruyi Ointments.Methods WAX PEG-20M capillary column was used.The column temperature was 140 ℃,the injection port temperature was 230 ℃,the detector temperature was 250 ℃,the flow rate was 1.0 mL/min.Results For Waishang Ruyi Ointments,the linear range was 0.513 8~5.138 mg,and the average recovery rate was 96.2% with RSD=1.0%.Conclusion The method is simple,easy-to-use,and can effectively control the quality of the ointments.

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