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.Combined liver-kidney transplantation for giant polycystic liver and polycystic kidney involving iliac fossa: one case report
Longshan LIU ; Wenbin ZHANG ; Weiqiang JU ; Maogen CHEN ; Yongcheng WEI ; Yingzhen HE ; Jun LI ; Changxi WANG ; Xiaoshun HE
Chinese Journal of Organ Transplantation 2024;45(10):728-729
This report described one patient of giant polycystic liver and polycystic kidney involving iliac fossa. Preoperative computed tomography (CT) revealed a large polycystic kidney occupying partially iliac fossa space. A decompression of lower pole of original kidney was planned for placing transplanted kidney. During total liver resection plus orthotopic liver transplantation, right polycystic kidney could move up on its own and iliac fossa space was released for placing transplanted kidney smoothly. Polycystic kidney shrunk markedly post-operation. It provided references for surgical planning of combined liver-kidney transplantation for this type of disease.
4.Comparative study on clinical characteristics of pattern of phlegm heat and bowel excess in acute stage of stroke involving meridians-collaterals or involving zang-fu organs
Tingting CHEN ; Jinting REN ; Lina REN ; Zhongyan HE ; Chang SUN ; Donghui WANG ; Mingguang SUN ; Fang WANG ; Yingzhen XIE
Journal of Beijing University of Traditional Chinese Medicine 2018;41(1):83-88
Objective To explore the clinical characteristics of pattern of phlegm heat and bowel excess in acute stage of stroke involving meridian-collateral or involving zang-fu organs from the aspects of severity of neurological impairment, pattern distribution at different time points and duration of bowel excess.Methods The basic information,scores of National Institute of Health Stroke Scale(NIHSS), whether bowel qi being unobstructed or not,duration of bowel excess,and pattern distribution at different time points(onset for 1 d to 3 d,onset for 4 d to 7 d and onset for 8 d to 14 d)were inputted based on structured data by using retrospective analysis of medical records in 204 cases of pattern of phlegm heat and bowel excess of stroke involving meridian-collateral(meridian-collateral group)and 217 cases of pattern of phlegm heat and bowel excess of stroke involving zang-fu organs(zang-fu organ group).The clinical characteristics from the data were compared, analyzed and studied.Results The severity of neurological impairment except of conscious state was significantly higher in zang-fu organ group than that in meridian-collateral group(P<0.01).The average duration of unobstructed bowel qi was significantly longer in zang-fu organ group than that in meridian-collateral group(P<0.01), and the percentage of patients with throughout obstructed bowel qi was significantly higher in zang -fu organ group than that in meridian-collateral group in acute stage of stroke.The pattern of phlegm heat and bowel excess showed a decline trend after stroke onset for 1 d to 14 d in meridian-collateral group, and showed an ascending trend after stroke onset for 1 d to 7 d and a decling trend after stroke onset for 8 d to 14 d in zang-fu organ group.The cases of pattern of qi deficiency with blood stasis or pattern of collaterals blocked by wind -phlegm and static blood were significantly higher in meridian-collateral group at 4-14 d than those at 1-3 d,and cases of pattern of wind stirring due to yin deficiency,pattern of phlegm heat and blood stasis or pattern of wind phlegm and blood stasis were significantly higher in zang-fu organ group at 8 -14 d than those at time point 1.Conclusion The duration of pattern of phlegm heat and bowel excess is posi-tively correlated to the severity of stroke,that is,neurological impairment is more serious and duration is longer in patients with pattern of phlegm heat and bowel excess of stroke involving zang -fu organs.

Result Analysis
Print
Save
E-mail