ABSTRACT
Objective To detect the influence of miRNA-34a (miR-34a) on the proliferation of osteosarcoma and the mechanisms responsible for miR-34a regulation.Methods The osteoblastic cell line MG-63 and Saos-2,human osteoblastic cell line hFOB 1.19,10 osteosarcoma tissues and 10 normal bone tissues were selected.The expression of miRNA-34a in osteosarcoma cells and tissues was detected by quantitative real-time polymerase chain reaction (qPCR).Next,a eukaryotic expression vector named pcDNA/miR-34a was constructed.Then,osteosarcoma cells were transfected with this eukaryotic expression vector and the effects of miR-34a overexpression on the proliferation and growth of osteosarcoma were measured using CCK-8,colony formation and xenograft model of nude mice.Finally,Western blot analysis was used to detect the expression of ether-à-go-go 1 (Eag1) gene in osteosarcoma cells after transfected with pcDNA/miR-34a or a miR-34a inhibitor miR-34a-2'-O-Methyl antisense oligoribonucleotide (miR-34a-2'-O-Me).Results Compared with normal bone tissues and osteoblastic cell line,miR-34a was down-regulated in osteosarcoma cell lines and tissues.Compared with the blank group and the control group,the cell survival rates of miR-34a group of the two cell lines were significantly lower [MG-63 72 h:blank group (40.05±4.82) ％,control group (36.88± 4.66) ％,miRNA-34a group (26.24±6.22) ％;MG-63 96 h:blank group (83.55±5.95) ％,control group (80.13± 4.48) ％,miRNA-34a group (30.21±7.26) ％;Saos-2 72 h:blank group (46.45±8.15) ％,control group (43.33± 6.89) ％,miRNA-34a group (26.81±3.17) ％;Saos-2 96 h:blank group (84.79±4.10) ％,control group (80.14± 3.11) ％,miRNA-34a group (31.77±5.17) ％].The similar results were obtained from colony formation assay (MG-63:blank group 83.40±3.29,control group 80.00±3.06,miR-34a group 24.40±2.71;Saos-2:blank group 85.00±3.32,control group 80.60±3.29,miR-34a group 30.40±4.94).The tumor volumes of osteosarcoma xenograft in the miR-34a group was significantly smaller than that in the blank group and control group after 21 days treatment (all P ＜ 0.001).Overexpression of miR-34a could decrease Eag1 expression in osteosarcoma cell lines while inhibition of miR-34a induced the of expression Eag1 (P ＜ 0.001).Conclusion MiR-34a plays a tumor suppressor role in osteosarcoma and could suppress the proliferation and growth of osteosarcoma through the regulation of Eag1.Moreover,it may be a novel target for osteosarcoma therapy.
ABSTRACT
<p><b>UNLABELLED</b>OBJECTIVE This study aims to construct a chitosan (CS)-polycaprolactone (PCL)-hydroxyapatite (HA) composite biomimetic scaffold to replace condyle and to explore the tissue engineering applications of condylar.</p><p><b>METHODS</b>A resin mold of the mandibular condyle was prepared by using rapid prototyping techniques. A mandibular condylar integrated biomimetic scaffold model was prepared by solution casting-ice Lek. PCL and CS were mixed at a ratio of 4:1. HA at quality ratios of 40%, 50%, 60%, and 70% was added to groups a, b, c, and d, respectively. The microscopic morphology, porosity, infrared spectra, X-ray diffraction pattern, and mechanical properties of the scaffold were observed.</p><p><b>RESULTS</b>The scaffold that includes both upper and lower parts displayed the same features (i.e., shape, yellow-white appearance, and hard texture) as the mandibular condyle. Scanning electron microscopy showed that the composite scaffold had a 3D network spatial structure, 70%-85% porosity, and 10-200 µm pore size. Infrared spectra showed that the peak intensity reduced with decreasing HA content. X-ray diffraction showed that the diffraction peak decreased with increasing HA content. Suitable tensile and compressive and flexural strength were discovered in the presence of 50% HA.</p><p><b>CONCLUSION</b>The scaffold prepared by solution casting-ice Lek shows favorable comprehensive features and is expected to replace human condylar.</p>
Subject(s)
Humans , Chitosan , Durapatite , Hardness , Mandible , Microscopy, Electron, Scanning , Polyesters , Porosity , Tissue Engineering , Tissue Scaffolds , X-Ray DiffractionABSTRACT
BACKGROUND:Swel ing after knee arthroplasty can produce tension bul ae, worsen pain, and even block venous return. Thus, intraventricular pressure of osseous fascia increases, which can block arterial blood circulation, even induce osteofascial compartment syndrome. OBJECTIVE:To evaluate efficacy of Cryf/cuff Systems and intermittent cold compress with hypertonic saline fol owing total knee arthroplasty. METHODS:A total of 60 patients with unilateral total knee arthroplasty were randomly assigned into two groups. Persistent freezing group received treatment with Cryo/cuff Systems after arthroplasty, and intermittent cold group received intermittent cold compress with hypertonic saline after arthroplasty. RESULTS AND CONCLUSION:Significant differences in circumference differences in superior patel ar pole, patel ar midpoint, and thickest point of gastrocnemius muscle were detected between persistent freezing group and intermittent cold group at 1 and 2 days after total knee arthroplasty (P<0.05), but no significant difference was detectable at day 3. Visual analogue scale scores at rest and during activity were significantly lower in the persistent freezing group than those in the intermittent cold group at 1 and 2 days after total knee arthroplasty (P<0.01), but no significant difference was visible at day 3. Range of motion was better in the persistent freezing group than that in the intermittent cold group at 1, 2 and 3 days (P<0.01), but no significant difference was observed at 1 and 2 weeks. Mean skin temperature was higher in the persistent freezing group than that in the intermittent cold group at 3 days (P<0.05). Results suggested that Cryf/cuff Systems could lessen tissue swel ing and pain, increased range of motion compared with intermittent cold compress with hypertonic saline at 1 and 2 days after total knee arthroplasty, but no significant difference was detected at day 3. That is, intermittent cold compress with hypertonic saline can reach the same effect as Cryf/cuff Systems at day 3.