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PURPOSE: This study aimed to explore the effects and mechanisms of long non-coding RNA (lncRNA) anti-differentiation non-coding RNA (ANCR) on the osteogenesis of osteoblast cells in postmenopausal osteoporosis (PMOP). MATERIALS AND METHODS: Mice models of PMOP were established. ANCR expression and intracellular calcium ions were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and laser confocal microscopy, respectively. ANCR was silenced in osteoblast cells from PMOP mice by the transfection of siRNA-ANCR (si-ANCR). The proliferation and apoptosis of osteoblast cells was analyzed by MTT and flow cytometry, respectively. Alkaline phosphatase (ALP) activity and calcium nodules were examined by ALP and alizarin red staining assay, respectively. The expression of enhancer of zeste homolog 2 (EZH2), runt related transcription factor 2 (RUNX2), and OSTERIX was detected by qRT-PCR and Western blot. Furthermore, an osteogenesis model was constructed in mice, and osteoid formation was observed by hematoxylin-eosin (HE) staining. The interaction between lncRNA-ANCR and EZH2 was further identified by RNA pull-down assay. RESULTS: ANCR expression and intracellular calcium ions were increased in PMOP mice. Si-ANCR significantly increased the proliferation, ALP activity, calcium deposition of osteoblast cells and decreased apoptosis. ANCR and EZH2 were down-regulated by si-ANCR, while RUNX2 and OSTERIX were upregulated. Si-ANCR also promoted osteoid formation in mice treated with hydroxyapatite-tricalcium phosphate. In addition, ANCR specifically bound to EZH2. CONCLUSION: Silencing ANCR promotes the osteogenesis of PMOP osteoblast cells. The specific binding of ANCR with EZH2 suppressed RUNX2, thereby inhibiting osteogenesis.
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Animals , Female , Humans , Mice , Alkaline Phosphatase , Apoptosis , Blotting, Western , Calcium , Flow Cytometry , Ions , Microscopy, Confocal , Osteoblasts , Osteogenesis , Osteoporosis, Postmenopausal , Real-Time Polymerase Chain Reaction , RNA , RNA, Long Noncoding , RNA, Small Interfering , RNA, Untranslated , Transcription Factors , TransfectionABSTRACT
Objective: To investigate the heterotopic osteogenesis of tissue engineered bone using the co-culture system of vascular endothelial cells (VECs) and adipose-derived stem cells (ADSCs) as seed cells. Methods: The partially deproteinized biological bone (PDPBB) was prepared by fibronectin combined with partially deproteinized bone (PDPB). The ADSCs of 18-week-old Sprague Dawley (SD) rats and VECs of cord blood of full-term pregnant SD rats were isolated and cultured. Three kinds of tissue engineered bone were constructed in vitro: PDPBB+VECs (group A), PDPBB+ADSCs (group B), PDPBB+co-cultured cells (VECs∶ADSCs was 1∶1, group C), and PDPBB was used as control group (group D). Scanning electron microscopy was performed at 10 days after cell transplantation to observe cell adhesion on scaffolds. Forty-eight 18-week-old SD rats were randomly divided into groups A, B, C, and D, with 12 rats in each group. Four kinds of scaffolds, A, B, C, and D, were implanted into the femoral muscle bags of rats in corresponding groups. The animals were killed at 2, 4, 8, and 12 weeks after operation for gross observation, HE staining and Masson staining histological observation, and the amount of bone collagen was measured quantitatively by Masson staining section. Results: Scanning electron microscopy showed that the pores were interconnected in PDPB materials, and a large number of lamellar protein crystals on the surface of PDPBB modified by fibronection were loosely attached to the surface of the scaffold. After 10 days of co-culture PDPBB and cells, a large number of cells attached to PDPBB and piled up with each other to form cell clusters in group C. Polygonal cells and spindle cells were mixed and distributed, and some cells grew along bone trabeculae to form cell layers. Gross observation showed that the granulation tissue began to grow into the material pore at 2 weeks after operation. In group C, a large number of white cartilage-like substances were gradually produced on the surface of the material after 4 weeks, and the surface of the material was uneven. At 12 weeks, the amount of blood vessels on the surface of group A increased, and the material showed consolidation; there was a little white cartilage-like material on the surface of group B, but the pore size of the material did not decrease significantly; in group D, the pore size of the material did not decrease significantly. Histological observation showed that there was no significant difference in the amount of bone collagen between groups at 2 weeks after operation ( F=2.551, P=0.088); at 4, 8, and 12 weeks after operation, the amount of bone collagen in group C was significantly higher than that in other 3 groups, and that in group B was higher than that in group D ( P0.05). Conclusion: The ability of heterotopic osteogenesis of tissue engineered bone constructed by co-culture VECs and ADSCs was the strongest.
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Objective@#To evaluate the accuracy of a 3D-printed template used to assist the preparation of tibial tunnel in reconstruction of anterior cruciate ligament (ACL).@*Methods@#Twenty healthy adult cadaveric knees were scanned by computed tomography(CT) and magnetic resonance imaging (MRI). The knees were from 11 males and 9 females who had died at an average age of 36 years (range, from 27 to 68 years) and from 8 left and 12 right sides. Individualized 3D reconstruction models of the knee joint were established based on their imaging data. According to the anatomic footprints of the virtual tibial tunnel, 20 individualized navigation templates were designed and printed by 3D printing. The templates were used to assisst preparation of tibial tunnels in the ACL reconstruction for the 20 cadaveric knees. After operation, CT scanning was conducted again to compare the corresponding postitions between the preoperative virtual tunnel and the postoperative actual tunnel. The positions of the tibial tunnel were described by the Tsukada method.@*Results@#The ratio of the distance between the tunnel outlet center and the medial edge of the tibia to the distance between the median and lateral edges of the tibial plateau was 49.7%±2.1% for the preoperative virtual tibial tunnel and 48.8%±2.8% for the postoperative actural tunnel, showing no significant difference between them (t=1.971, P=0.063). The ratio of the distance between the tunnel outlet center and the anterior edge of the tibia to the distance between the anerior and posterior edges of the tibial plateau was 41.3%±1.3% for the preoperative virtual tibial tunnel and 40.3%±3.7% for the postoperative actural tunnel, showing no significant difference between them(t=1.494, P=0.152).@*Conclusion@#3D printing can be used to design and manufacture a navigation template which can accurately assist preparation of tibial bone tunnel in ACL reconstruction.
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Objective To evaluate the accuracy of a 3D-pfinted template used to assist the preparation of tibial tunnel in reconstruction of anterior cruciate ligament (ACL).Methods Twenty healthy adult cadaveric knees were scanned by computed tomography (CT) and magnetic resonance imaging (MRI).The knees were from 11 males and 9 females who had died at an average age of 36 years (range,from 27 to 68 years) and from 8 left and 12 fight sides.Individualized 3D reconstruction models of the knee joint were established based on their imaging data.According to the anatomic footprints of the virtual tibial tunnel,20 individualized navigation templates were designed and printed by 3D printing.The templates were used to assisst preparation of tibial tunnels in the ACL reconstruction for the 20 cadaveric knees.After operation,CT scanning was conducted again to compare the corresponding postitions between the preoperative virtual tunnel and the postoperative actual tunnel.The positions of the tibial tunnel were described by the Tsukada method.Results The ratio of the distance between the tunnel outlet center and the medial edge of the tibia to the distance between the median and lateral edges of the tibial plateau was 49.7％ ±2.1％ for the preoperative virtual tibial tunnel and 48.8％ ± 2.8％ for the postoperative actural tunnel,showing no significant difference between them (t =1.971,P =0.063).The ratio of the distance between the tunnel outlet center and the anterior edge of the tibia to the distance between the anerior and posterior edges of the tibial plateau was 41.3％ ± 1.3％ for the preoperative virtual tibial tunnel and 40.3％ ± 3.7％ for the postoperative actural tunnel,showing no significant difference between them (t =1.494,P =0.152).Conclusion 3D printing can be used to design and manufacture a.navigation template which can accurately assist preparation of tibia] bone tunnel in ACL reconstruction.
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ObjectiveTo investigate the detection results of WBC,CRP,ESR and GR% in patients with knee infection after operation. Methods The study was conducted from March 2012 to January 2015. Twenty patients were collected in the study. The values of WBC,CRP,ESR and GR% were compared and analyzed between pre-operation and post-operation of 1,3 and 7 days. Results The values of WBC,CRP,ESR and GR% in the post-operation of 1 day were higher than those in the pre-operation,and there were statistically significances (P 0.05). Conclusion WBC,CRP,ESR and GR% have good reference value for early diagnosis and treatment,which can be used as screen testing indexes in the early knee infection.
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We used ultra-high performance liquid chromatography-mass spectrometry (UPLC/MS) for analyzing and identifying the active components of newborn calf serum (NCS) and fetal bovine serum (FBS). The results demonstrated significant differences in the components between NCS and FBS. FBS appeared to have more complex components than NCS, with mass to ratios (m/z) of the substances of 498, 273 and 448. These substances in FBS may be the main active components to support the proliferation and differentiation of cells.
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Animals , Cattle , Animals, Newborn , Blood , Chromatography, High Pressure Liquid , Fetal Blood , Chemistry , Mass Spectrometry , Serum , ChemistryABSTRACT
Objective To investigate the effects of BMP2 in the co-culture system of hBMSCs and hUVECs.Methods Based on the co-culture system of cells,a simple culture group (hBMSCs group),a combined culture group (BMSCs + hUVECs group) and a BMP2 silenced group (BMSCs + BMP2 silenced hUVECs) were established.On days 4,6,8,10,cells were counted for drawing cell growth curves.In addition,ALP and OC expression in BMSCs was detected in each group.Results At each time point,cell number in the BMSCs + hUVECs group were significantly greater than in the BMSCs group,while the BMP2 silenced group were in the median.ALP and OC expression in the BMSCs + hUVECs group were significantly greater than in the BMSCs group,while the BMP2 silenced group were in the median.Conclusion BMP2 plays an important role in the co-culture system,which can promote osteogenic differentiation and the growth of BMSCs.
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OBJECTIVE@#To compare the results of supracricoid partial laryngectomy-cricohyoidopexy (SCPL-CHP) and horizontal-vertical hemilaryngectomy in the treatment of mid and late laryngeal carcinoma.@*METHOD@#Retrospective analysis on the types of mid and late stage of laryngeal carcinoma clinical material, 22 patients supracricoid partial laryngectomy-cricohyoidopexy, 20 patients horizontal-vertical hemilaryngectomy, each with the added radiotherapy. The long term results of operation and glottic reconstruction were evaluated by postoperative visiting, semi-quantitative speech intelligibility analysis, electroglottograph (EGG) and so on.@*RESULT@#Forty-two cases of laryngeal cancer patients were decannulated, the decannulation rate was 100%. Postoperative decannulation time: surgical CHP for (44.0 +/- 4.6) d, 3/4 throat operation for (39.0 +/- 2.7) d, two groups of postoperative decannulation time difference was statistically significant (t = 4.2395, P 0.05). GRBAS in the evaluation of G rating, the difference between the two groups was statistically significant (P 0.05), and the jitter, shimmer and NNE compared CHP group to 3/4 laryngectomy group were significantly increased (P 0.05).@*CONCLUSION@#According to the laryngeal of middle-late carcinoma, the region and the involvement of the scope were considered to choose appropriate surgical treatments, and both can complete resection of the tumor, and can retain good laryngeal functions,and CHP has a wider range of operation indications and clinical application prospect, is worthy to be popularized.
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Adult , Aged , Humans , Male , Middle Aged , Carcinoma, Squamous Cell , General Surgery , Cricoid Cartilage , General Surgery , Deglutition , Hyoid Bone , General Surgery , Laryngeal Neoplasms , General Surgery , Laryngectomy , Methods , Pharynx , General Surgery , Retrospective Studies , Treatment OutcomeABSTRACT
BACKGROUND: Nerve growth factor is secreted and synthetized by a variety of cells, such as inflammatory calls and repairing calls, its biological effects are diverse and closely related to the process of wound repair, but its mechanism is not yet clear.OBJECTIVE: To observe the influence of nerve growth factor on the biological characteristics of scar fibroblasts.METHODS: Eight clinical surgical resection specimens, including 5 face and neck hyperplastic scar or keloid specimens, did not receive any treatment; three were prepuce specimens following circumcision (normal tissue). By use of tissue block method, the scar and normal skin fibroblasts were cultured, followed by digestion passage. The scar tissue and normal tissue flbroblasts at 3-6passages in the logarithmic phase were seeded in 96-well plate and divided into the experimental group (scar flbroblest group) and the control group (normal skin fibroblasts group), with two parallel holes in each group were added with 3,33, 0.33 mg/L nerve growth factor, 50 μL. Inverted microscope was used to observe fibroblast morphology. At 24, 48, 72 hours after culture, the absorbanca value was measured using MTT. Fibroblast DNA content and cell apoptosis were determined by flow cytometry.RESULTS AND CONCLUSION: The fibroblasts were adherent cells, the scar and normal skin tissues were shown to cell free out of tissue block and gradual expansion at 4-6 days after incubation. Compared with normal skin fibroblasts, the pathological scar fibroblasts became larger, irregular shape and arrangement. MTT results showed that nerve growth factor could promote the normal and hypertrophic scar fibroblasts growth, which becomes more apparent. Flow cytometry results showed that by adding nerve growth factor, the percentage of scar fibroblasts at proliferating S-G_2-M phase was higher than that in the control;group; with a Iower level of apoptosis. It is indicated that nerve growth factor plays an obviously promoting role on normal and scar skin fibroblasts growth and proliferation, especially on the scar skin.
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Objective To explore the mechanism of cytokines for the scars,and to study the effect of platelet derived growth factor(PDGF)on the biological behavior of fibroblasts in scars.Methods Fibroblasts of scars and normal skins were cultured in vitro.The results were observed and analyzed by light inverted microscopy(LM),and 3-(4,5-dimethyl-thiazol-2-yl)-2,5 ciphenyl tetrazolium bromide (MTT)assay.The effects of PDGF on the biological behaviors of fibroblasts of scars were also determined. Results In vitro study,using LM,FCM and MTT assay,showed that proliferation of fibroblasts were inereased significantly when PDGF was added to the cultures,as compared to the control groups.Conclusions PDGF can increase fibroblast proliferation.These results demonstrate that PDGF is beneficial for wound healing at early stage.
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The research in bone tissue engineering is abundant and its development is rapid,however,there has been no ideal scaffold materials.We review the recent articles on bone tissue engineering,including ceramics materials,polymerized materials,materials deriver from natural biological organism and their compound materials