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16 December 2016, Volume 20 Issue 52 Previous Issue    Next Issue

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In vitro bioactivity and degradability of injectable poly(propylene fumarate)/ beta-tricalcium phosphate bone cement Ma Zheng-yu, Yang Feng, Wang Jing, Liu Chang-sheng 2016, 20 (52):  7757-7764.  doi: 10.3969/j.issn.2095-4344.2016.52.001 Abstract ( 319 )   PDF (2500KB) ( 235 )   Save BACKGROUND: Poly(propylene fumarate) (PPF) can crosslink at room temperature, and β-tricalcium phosphate (β-TCP) has good biocompatibility, but PPF/β-TCP composite bone cement has not yet been systematically studied. OBJECTIVE: To prepare PPF/β-TCP composite bone cement and to explore its in vitro bioactivity and degradability. METHODS: β-TCP and PPF were respectively synthesized by liquid-phase precipitation and a two-step method, and PPF/β-TCP composite bone cement was prepared through mixing PPF with β-TCP. The    in vitro bioactivity of PPF/β-TCP was compared with the commercial poly(methyl methacrylate) (PMMA) through the ability of forming hydroxyapatite after immersed in simulated body fluid for 7 days. The in vitro degradability of PPF/β-TCP was studied via investigating the transformation of pH values, water uptake and mass loss, compressive strength and morphology at each time point. RESULTS AND CONCLUSION: There were hydroxyapatites formed on the PPF/β-TCP material, but none on the commercial PMMA material. The pH values of the PPF/β-TCP were stable in PBS for 63 days, indicating its degradation is moderate; the mass loss was up to 13.5% after 84 days. Scanning electron microscope displayed the degraded PPF/β-TCP surface, and its compressive strength was decreased gradually, which good for the integrity and sustainability of mechanical properties during degradation. These results suggest that PPF/β-TCP bone cement holds mineralization and degradability in vitro. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Physiochemical properties and cytotoxicity of silk fibroin based calcium phosphate cement incorporated with N-acetylcysteine Feng Tao, Pi Bin, Jiang Lei, Li Bin, Lu Ying-jie, Yang Hui-lin, Zhu Xue-song 2016, 20 (52):  7765-7772.  doi: 10.3969/j.issn.2095-4344.2016.52.002 Abstract ( 252 )   PDF (9896KB) ( 144 )   Save BACKGROUND: Calcium phosphate cements (CPCs) possess the bio-degradation and osteoconduction, and its final hydration product, hydroxyapatite, is the main inorganic constituent of bones. However, its poor mechanical property makes it unable to be used for repairing weight-bearing bone defects. OBJECTIVE: To develop a kind of bioactive bone cements with decent biomechanical property and biocompatibility. METHODS: 6% silk fibroin aqueous solutions containing different concentrations of N-acetylcysteine (0, 10 and 25 mmol/L) were prepared. Each cement sample was prepared by mixing the curing liquid and α-tricalcium phosphate powder with the ratio of 0.4 mL: 1 g; α-tricalcium phosphate powder mixed with ddH2O as control group. The compressive strength, setting time of the cements were measured. The crystal components of the cements were characterized using X-ray diffraction and the microstructure was observed using scanning electron microscope. MC3T3-E1 cells were seeded onto the material in each group, and cell morphology was observed under scanning electron microscope at 24 hours. MC3T3-E1 cells were cultured in the extract of each material, cell proliferation was detected at 1, 3, 5 and 7 days, and the lactate dehydrogenase level was detected at 1 and 3 days. RESULTS AND CONCLUSION: X-ray diffraction and scanning electron microscope showed that the final hydration products of α-tricalcium phosphate in all specimens were hydroxyapatite. When the concentration of N-acetylcysteine was 25 mmol/L, the compressive strength of the material reached (49.39±1.68) MPa, with the initial setting time of (21.77±1.07) minutes and the final setting time of (31.88±1.69) minutes. There was no significant difference in cell morphology among cements. These results suggest that the cement containing N-acetylcysteine exhibites good biocompatibility and high mechanical strength. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Construction of a polycaprolactone/bone extracellular matrix scaffold with three-dimensional printing technology and its osteoinductivity in vitro Yang Peng, Li Chun-de 2016, 20 (52):  7773-7780.  doi: 10.3969/j.issn.2095-4344.2016.52.003 Abstract ( 281 )   PDF (11694KB) ( 221 )   Save BACKGROUND: Scholars are still looking for ideal bone tissue-engineered scaffolds, and  three-dimensional (3D) printing technology is a novel construction method. In the meanwhile, bone extracellular matrix is becoming a hotspot in osteogenic induction. OBJECTIVE: To construct the polycaprolactone/bone extracellular matrix scaffold using 3D printing technology and co-culture method, and to detect its osteogenic property. METHODS: 216 3D-printed polycaprolactone scaffolds were divided into group A (96 pores, n=72) and group B(48 pores, n=144). Passage 5 bone marrow mesenchymal stem cells from Sprague-Dawley rats were seeded onto the two kinds of polycaprolactone scaffolds, and the group A was used for alizarin red staining and Masson staining, while the group B for collagen and glycosaminoglycan detection at 1, 2 and 3 weeks of incubation. Afterwards, the scaffolds at 1, 2 and 3 weeks of culture were decellularized and labeled as groups AE1, AE2, AE3, BE1, BE2 and BE3. Then passage 5 bone marrow mesenchymal stem cells from Sprague-Dawley rats were seeded onto each scaffold again, and the former three groups underwent alizarin red staining, and the latter three were used for calcium, alkaline phosphatase activity and DNA quantitative analysis at 1, 2 and 3 weeks of culture. RESULTS AND CONCLUSION: Masson staining, glycosaminoglycan and hydroxyproline quantitative analysis showed that the extracellular matrix on the composite scaffold increased with time. Alkaline phosphatase activity revealed that the composite scaffold had a significantly stronger osteogenic differentiation than the normal polycaprolactone scaffold (P < 0.05). Alizarin red staining and calcium quantitative analysis showed that the mineralization of the composite scaffold was more obvious than that of the normal polycaprolactone scaffold (P < 0.05), but the total DNA analysis did not differ significantly between scaffolds. These results suggest that the composite scaffold with extracellular matrix is constructed successfully using the 3D technology and co-culture method and exhibits a better osteoinductivity. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of titanium ions on the proliferation and activation of T lymphocytes in vitro Chen Fu-jun, Chen Dong-hui, Yang Qian, Li Chang, Tang Li 2016, 20 (52):  7781-7787.  doi: 10.3969/j.issn.2095-4344.2016.52.004 Abstract ( 210 )   PDF (6088KB) ( 477 )   Save BACKGROUND: Titanium ions have been proved to stimulate the secretion of bone remodeling-related factors from T lymphocytes; however, the effects of titanium ions on the early activation, intermediate activation, and cell cycle of T lymphocytes remain unclear. OBJECTIVE: To investigate the effects of titanium ions on the proliferation and activation of T lymphocytes in vitro. METHODS: Cell proliferation and cycle test: Jurkat E6-1 T lymphocytes in logarithmic phase were collected and cultured in the medium containing 0 (control), 25 (low concentration), 50 (middle concentration), and 100 μmol/L (high concentration) titanium ions for 24 hours to detect the cell relative proliferation rate and cell cycle. Cell activation trial: Jurkat E6-1 T lymphocytes were divided into two groups that were subdivided into four groups containing 0, 25, 50, and 100 μmol/L titanium ions, respectively with or without phytohemagglutinin (PHA) pre-stimulation. The expressions of CD69 and CD25 were measured after cultured for 24 hours. RESULTS AND CONCLUSION: Titanium ions enhanced T lymphocytes proliferation in a concentration-dependent manner (P < 0.05). Compared with the control group, the percentages of G0/G1 phase decreased and the proportions of cells in S and G2/M phase increased significantly in the low, middle and high concentration groups (P < 0.05). The proportion of G0/G1-phase cells in the high concentration group was less and the proportion of G2/M phase cells was higher than those in the middle and low concentration groups (P < 0.05). With PHA pre-stimulation, the expression of CD69 in the high concentration group was higher than that in the middle and low concentration groups (P < 0.05); whereas the difference of CD25 expression was not significant among four subgroups. Titanium ions promoted the expression of CD69 in a concentration-dependent manner (P < 0.05), but there was no CD25 expression in each subgroup without PHA pre-stimulation. To conclude, titanium ions can significantly promote T lymphocyte proliferation and early activation in vitro, and moreover, induce S and G2/M phase arrest in T lymphocytes. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Osteoinductivity and performance of silk fibroin solution Yi Bing-cheng, Zhang Hui-lan, Yu Zhe-pao, Yuan Hui-hua, Wang Xian-liu, Shen Yan-bing, Bao Jia-yu, Lou Xiang-xin, Zhang Yan-zhong 2016, 20 (52):  7788-7795.  doi: 10.3969/j.issn.2095-4344.2016.52.005 Abstract ( 460 )   PDF (5937KB) ( 209 )   Save BACKGROUND: Silk fibroin, as a kind of high-performance biomaterial, has been widely used to construct scaffolds in bone tissue engineering. However, whether silk fibroin itself holds osteoinductive ability has not been reported yet. OBJECTIVE: To investigate the impact of different concentrations of silk fibroin solution on the proliferation and differentiation of rat bone marrow mesenchymal stem cells (BMSCs) in vitro. METHODS: Silk fibroin and BMSCs were respectively isolated from silkworm cocoon and rat tibia, and were identified. Then, BMSCs were cultured in different concentrations of silk fibroin solution (0.01%, 0.05% and 0.1%), and the cell proliferation and the alkaline phosphatase activity were detected at different time points. RESULTS AND CONCLUSION: FTIR spectra of the sample extracted from silkworm cocoon showed distinct absorption peaks at 1 653 (amide I), 1 530.5 (amide II) and 1 212.3 cm-1 (amide III), which could be confirmed to be silk fibroin. Thus generated BMSCs showed long fusiform or astral morphology, positive for representative markers (CD29, CD44 and CD90) relating to mesenchymal stem cells, and could differentiate into osteocytes, chondrocytes and adipocytes under specific induction conditions, which further confirmed the extracted cells were BMSCs. Compared with the control group (without silk fibroin), 0.05% silk fibroin not only significantly promoted the cell adhesion, migration and proliferation, but also enhanced the alkaline phosphatase activity (P < 0.01). With the increasing of the silk fibroin concentrations, the osteodifferentiation capacity of the BMSCs was progressively improved within the range of 0-0.05% and then declined at 0.01% of silk fibroin solutions. These results suggest that silk fibroin can promote osteogenesis, thus providing scientific evidence for developing silk fibroin-based tissue-engineered scaffolds. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Magnetic resonance imaging of canine oral epithelial cells labeled with ultrasmall superparamagnetic iron oxide Zhou Shu-kui, Yao Ting-ting, Zhang Kai-le, Zou Qing-song, Fu Qiang 2016, 20 (52):  7796-7802.  doi: 10.3969/j.issn.2095-4344.2016.52.006 Abstract ( 269 )   PDF (4706KB) ( 220 )   Save BACKGROUND: Epithelial cells are commonly used as the seed cell in tissue engineering; however, there is still a lack of an effective in vivo noninvasive trace technology. OBJECTIVE: To investigate the feasibility of labeling canine oral epithelial cells with ultrasmall superparamagnetie iron oxide (USPIO) and magnetic resonance imaging (MRI) in vitro.  METHODS: Oral epithelial cells from beagles were primary cultured, and then labeled by 0.75 mg/L poly-L-lysine combined with USPIO (0, 5, 10, 25, 50 and 100 mg/L), respectively. To determine the optimal dosage, the intracellular iron expression was identified by Prussian blue staining, and the cell viability in different groups was detected by cell counting kit-8. Finally, 2×105 labeled cells were suspended with 1 mL PBS buffer, and were screened using 3.0 T MR on T2*WI sequences in vitro.  RESULTS AND CONCLUSION: USPIO prepared with 0.75 mg/L poly-L-lysine could successfully label dog oral epithelial cells. Prussian blue staining showed intracellular blue spots, and the intracellular blue spots became more with the concentration increasing and saturated at the concentration of 25 mg/L. Cell counting kit-8 indicated that the cell viability did not change when the concentration < 25 mg/L. Among the T2*WI sequences, the MRI signal intensity decreased with the concentration increasing. In conclusion, canine oral epithelial cells can be effectively labeled with USPIO making no impact on cell viability when the concentration < 25 mg/L, and MRI can be used to track these labeled cells in vitro． 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effect of sodium hyaluronate/chitosan nanoparticles on the neovascularization in burned cornea Lu Jing, Wu Shi-ke, Chen Guang, Zhao Yue, Li Dan 2016, 20 (52):  7803-7808.  doi: 10.3969/j.issn.2095-4344.2016.52.007 Abstract ( 350 )   PDF (4212KB) ( 167 )   Save BACKGROUND: Chitosan nanoparticles-encapsuled sodium hyaluronate is an effective drug for the burned cornea. OBJECTIVE: To verify the effect of sodium hyaluronate/chitosan nanoparticles on the neovascularization in burned cornea. METHODS: Thirty Sprague-Dawley rats were randomly divided into three groups, and the model of burned cornea caused by base was established in the rats of model and experimental groups, followed by respectively treated with 10 μL sodium hyaluronate/chitosan nanoparticle suspension and normal saline, once daily, for consecutive 4 weeks. Rats only given normal saline were used as controls. Four weeks later, the dynamic growth of newly formed blood vessels in the cornea was observed using silt lamp. The levels of tumor necrosis factor-α and interleukin-6 were detected by ELISA, histological changes of the cornea were observed by hematoxylin-eosin staining, and the mRNA expression levels of vascular endothelial growth factor and cyclooxygenase 2 were detected by real-time PCR. RESULTS AND CONCLUSION: Compared with the control group, the area of the newly formed blood vessel and the levels of tumor necrosis factor-α, vascular endothelial growth factor and cyclooxygenase 2 were significantly increased in the model group (P < 0.05, P < 0.01). In the experimental group, all above indicators were significantly lower than those in the model group (P < 0.05). There were a large number of inflammatory cells and neovascularization in the model group, but only few inflammatory cells in the experimental group. These results show that sodium hyaluronate/chitosan nanoparticles can inhibit the neovascularization in the burned cornea. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Decellularized whole liver bioscaffold repairs liver injury Zhang Qing-feng, Li Zi-rong 2016, 20 (52):  7809-7814.  doi: 10.3969/j.issn.2095-4344.2016.52.008 Abstract ( 289 )   PDF (3631KB) ( 220 )   Save BACKGROUND: Decellularized scaffolds are special for retaining the tubular structure used for nutrition transport, and providing a similar inner environment for cell growth. OBJECTIVE: To study the preparation of the decellularized whole liver bioscaffold and to explore its repair outcomes for liver injury. METHODS: Livers from 12 Sprague-Dawley rats were used for preparing the decellularized whole liver bioscaffold by chemical detergent-enzymes decellularized technology. Models of liver injury were established in another 24 Sprague-Dawley rats and randomized into two groups: the decellularized whole liver bioscaffold was implanted into the rat liver lesions in experimental group, and controls were given the injection of normal saline. Thirty days later, the serum levels of alanine aminotransferase and glutamic-oxaloacetic transaminase were detected, and liver tissues were removed for hematoxylin-eosin staining. RESULTS AND CONCLUSION: Hematoxylin-eosin staining showed that extracellular matrix-like structures existed in the decellularized bioscaffold; cell components were completely removed from the liver, the collagen fibers in the scaffold arranged regularly and were not dissolved under electron microscope. The serum levels of alanine aminotransferase and glutamic-oxaloacetic transaminase in the experimental group were significantly lower than those in the control group (P < 0.05). Hematoxylin-eosin staining showed a large number of blue-stained and dense distributed nuclei, and pink distribution of collagen fibers that had no overt breakages in the control group, while pink and dense structures in the experimental group. These results suggest that the decellularized whole liver bioscaffold is easy to obtain, and can promote the injured liver repair. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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In vitro study on a degradable poly-L-lactic acid biliary-enteric composite stent Yao Jin-feng, Han Jin-li, Guo Yu, Cui Dong-lai 2016, 20 (52):  7815-7820.  doi: 10.3969/j.issn.2095-4344.2016.52.009 Abstract ( 316 )   PDF (2077KB) ( 186 )   Save BACKGROUND: The degradable poly-L-lactic acid (PLLA) biliary-enteric composite stent has been developed. OBJECTIVE: To analyze the solubility and biocompatibility of the degradable PLLA biliary-enteric composite stent. METHODS: Solubility: the PLLA composite stent was implanted into artificial gastric acid to detect the dissolution rate within 12 weeks. Pyrogen test: the PLLA composite stent extracts were injected into the rabbits via ear vein to detect the changes of body temperature. Hemolysis test: the PLLA composite stent extracts, normal saline and distilled water were added into the rabbit anticoagulant, respectively, to detect the hemolysis ratio. Cytocompatibility test: Caco-2 cells were respectively cultured in the DMEM medium containing 10% fetal bovine serum, rubber material extracts and the PLLA composite stent extracts, and the cell proliferation was detected at 12, 24, 36 and 48 hours; the lactic dehydrogenase release was detected at 2 days. RESULTS AND CONCLUSION: The PLLA composite stent showed a long stability in vivo, and approximately 80% was dissolved at about 20 weeks. No reactions of pyrogen and henolysis were observed in the pyrogen and hemolysis tests. The PLLA composite stent made no effects on the Caco-2 cell proliferation and lactic dehydrogenase release. In conclusion, the PLLA composite material holds a good solubility and cytocompatibility. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of three-dimensional printed porous titanium scaffolds on bone ingrowth Wang Zhong-han, Wang Chen-yu, Liu He, Li Chen, Qin Yan-guo, Cai Xiao-yu, Wang Jin-cheng 2016, 20 (52):  7821-7828.  doi: 10.3969/j.issn.2095-4344.2016.52.010 Abstract ( 423 )   PDF (1273KB) ( 526 )   Save BACKGROUND: With the development of three-dimensional (3D) printing technology, 3D printed porous titanium scaffolds as bone substitutes have become a research hotspot. OBJECTIVE: To introduce and discuss the effects of each parameter of 3D printed porous titanium scaffolds on bone ingrowth, and to sum out the optimal parameters for bone ingrowth. METHODS: The first author retrieved PubMed, Springerlink and Medline databases with “three-dimensional (3D) printing, scaffold, titanium, bone ingrowth” as keywords for relevant articles published from 2006 to 2016. 125 articles were retrieved initially, and finally 42 eligible articles were included for analysis. RESULTS AND CONCLUSION: Pore size, porosity, pore structures and surface modifications of 3D printed porous titanium scaffolds all make effects on bone ingrowth or osteoblasts in scaffolds. Scaffolds with appropriate pore size and porosity can promote the vascularization and provide adequate nutrition and oxygen supplement, to ensure high cell viability. Regulations of cell performances, such as cell attachment, proliferation and differentiation, are also affected by pore structures and nano-scale surface modification. Herein, a detailed combination of the parameters, as mentioned above, can create a better porous scaffold for better bone ingrowth. Hence, the high-stability interface between bone and scaffolds may be obtained through the parameter adjustment. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Biocompatibility of calcium phosphate cement/fibrin glue in filling radius defects Huang Tian1, Zheng Nan-sheng2, Zhang Yu-zhuan1, Wu Yong-le1, Wang Gang1 2016, 20 (52):  7829-7835.  doi: 10.3969/j.issn.2095-4344.2016.52.011 Abstract ( 312 )   PDF (5629KB) ( 459 )   Save BACKGROUND: The chemical compositions and structure of calcium phosphate bone cement are similar to those of human bone, which can fill the bone collapse caused by fracture and induce osteogenesis, but its degradation rate is slow. OBJECTIVE: To evaluate the biocompatibility of the calcium phosphate cement/fibrin glue and the feasibility of repairing radius defects. METHODS: In vitro cytotoxicity experiment: Mouse fibroblasts were cultured in the calcium phosphate bone cement/fibrin glue extracts, phenol solution, and RPMI 1640 culture medium containing 10% fetal bovine serum, respectively, to detect the cytotoxicity grade. Hemocompatibility experiment: Calcium phosphate bone cement/fibrin glue extracts, normal saline and distilled water were respectively added into the rabbit anticoagulation, to detect the hemolytic rate. Forty-five New Zealand white rabbits were enrolled and modeled into bilateral radius defects, followed by randomly allotted into three groups: blank control group without any intervention, experimental and control groups were given the implantation with calcium phosphate bone cement/fibrin glue and autologous radius, respectively. X-ray, histology, bone mineral density and biomechanical test were performed at postoperative 4, 8 and 16 weeks. RESULTS AND CONCLUSION: The toxicity grade of the calcium phosphate cement/fibrin glue was 0 to 1. The hemolytic rate of the calcium phosphate cement/fibrin glue was 3.15%. At 16 weeks postoperatively, X-ray showed that in the experimental and control groups, the fracture line disappeared completely, pulp cavity was recanalized, and in plastic completely. Histology showed that the reconstructed bone trabecular was obvious, plate layer of bone was mature, and medullary cavity recanalization appeared in the control group; there were a large number of new grid-shaped woven bone tissues growing into the material in the experimental group, with overt degradation, and degradation rate was in parallel to bone ingrowth. The bone density, the maximum load, maximum stress and failure energy in the experimental and control groups were significantly higher than those in the blank control group (P < 0.05), and all above indicators showed no significant differences between the experimental and control groups. These results manifest that the calcium phosphate bone cement/fiber protein glue composite material holding a good biocompatibility can promote bone tissue regeneration for bone defect repair, achieving similar curative effect with autologous bone transplantation. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Ethyl linoleate inhibits inflammatory reaction induced by titanium particles and its mechanism Liu Guo-dong, Xin Bing, Huang Dong, Zheng Bai, Sun Bai-han, Guo Kai-jin 2016, 20 (52):  7836-7843.  doi: 10.3969/j.issn.2095-4344.2016.52.012 Abstract ( 324 )   PDF (5491KB) ( 206 )   Save BACKGROUND: Ethyl linoleate has been proved to attenuate the inflammatory-cytokines release induced by lipopolysaccharide, but whether it can inhibit titanium-induced osteolysis and the underlying mechanism remain unclear. OBJECTIVE: To observe the effect of ethyl linoleate on the expression of inflammatory-related factors induced by titanium particles and explore its mechanism. METHODS: Forty-eight Kunming mice were randomly divided into blank control, titanium, dimethylsulfoxide (DMSO) and experimental groups. The back air pouch Inflammatory models were established in the mice of the titanium, DMSO and experimental groups, in which the 200 µL menstruum of DMSO (0.5%) and 200 µL ethyl linoleate (0.5%) were respectively administered into the pouch of the mice at 12 hours. Mice in the blank control group received no intervention. Fourteen days later, the inflammatory cell infiltration in the skin was examined through hematoxylin-eosin staining; the expression levels of inhibitor κB-α, nuclear factor-κB, inducible nitric oxide synthase, cyclooxygenase-2, tumor necrosis factor-α and interleukin-6 as well as ERK, p-ERK, JNK, p-JNK, p38 and p-p38 in MAPK signaling pathways were evaluated by western blot assay. RESULTS AND CONCLUSION: In the titanium group and DMSO group, there were numerous inflammatory cells and vacuole-like necrotic tissues in the hair follicle lacuna of dermis and loose connective tissues of hypodermis. The experiment group showed significant reduction in inflammatory cell infiltration and vacuole-like necrosis. Compared with the blank control group, the expression levels of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-κB, tumor necrosis factor-α, interleukin-6, ERK, JNK and p38 in the DMSO and titanium groups were significantly increased, while inhibitor κB-α significantly decreased (P < 0.05). Compared with the DMSO and titanium groups, there were significantly down-regulated levels of inducible nitric oxide synthase, cyclooxygenase-2, nuclear factor-κB, tumor necrosis factor-α, interleukin-6, ERK, JNK and p38, and up-regulated inhibitor κB-α level in the experimental group (P < 0.05). In conclusion, ethyl linoleate can remarkably suppress the expressions of titanium-induced inflammatory factors associated with the inhibition of nuclear factor-κB and MAPK signaling pathway activation. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Effects of diameters and spacing of silicon micro pillars on the differentiation of neural stem cells Yan Dong-dong, Gao Xing-hua, Xiong Yi 2016, 20 (52):  7844-7850.  doi: 10.3969/j.issn.2095-4344.2016.52.013 Abstract ( 273 )   PDF (4397KB) ( 218 )   Save BACKGROUND: Different structures of matrix models, such as grating, holes and pillars make different effects on the differentiation of neural stem cells. OBJECTIVE: To explore the effects of the diameter and spacing, known as physical signals of micro pillars on neural stem cell differentiation. METHODS: Micro pillars with different diameters and spacing, both of which had four dimensions of 2.5, 5, 10 and 20 μm, were fabricated on silicon substrates by photolithographic method. Purified primary neural stem cells were incubated on the each micro pillar for 7 days in vitro. Then the differentiation of neural stem cells into neuron-like cells was observed using immunofluorescence staining and quantitative real-time PCR. RESULTS AND CONCLUSION: When the diameters of the micro pillars were constant and the spacing of micro pillars varied in the range of 2.5-10 μm, the differentiation rate of neural stem cells increased with the spacing increase. When the spacing was invariable and the diameters changed in the range of 2.5-20 μm, the differentiation rate of neural stem cells declined with the diameter increase. Especially, the micro pillars with 2.5 μm diameter and 10 μm spacing significantly promoted the differentiation of neural stem cells into neuron-like cells. These results show that specific micro pillars with small diameters and large spacing facilitate the differentiation of neural stem cells, thus providing guidance for developing tissue-engineered scaffolds.  中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Performance comparison of 3D printing sheep vertebral bone meal/polyvinyl alcohol scaffold, nano-hydroxyapatite/polyvinyl alcohol scaffold and sheep vertebral bone meal/polyvinyl alcohol nonporous bone plate Zhou Qi-Qi, Han Xiang-zhen, Song Yan-yan, Lv Ming-fan, Hu Yang, He Hui-yu 2016, 20 (52):  7851-7857.  doi: 10.3969/j.issn.2095-4344.2016.52.014 Abstract ( 326 )   PDF (1916KB) ( 229 )   Save BACKGROUND: With the promotion of 3D printing technology, 3D printing scaffolds for bone tissue engineering have become the new ideas for jaw bone repair. OBJECTIVE: To compare the physical and biological properties of sheep vertebral bone meal/polyvinyl alcohol (PVA) scaffold, nano-hydroxyapatite (nHA)/PVA scaffold, and sheep vertebral bone meal/PVA nonporous bone plate. METHODS: 3D printing technology was used to print sheep vertebral bone meal/PVA scaffold, nHA/PVA scaffold, and sheep vertebral bone meal/PVA nonporous bone plate. Porosity, morphology, water absorption rate and mechanical properties of different scaffolds were detected. Three kinds of scaffolds were all used to culture bone marrow mesenchymal stem cells, and cell proliferation ability was detected using cell counting kit-8 at 1, 4, 7 days of culture. RESULTS AND CONCLUSION: Under scanning electron microscope, the sheep vertebral bone meal/PVA scaffold and nHA/PVA scaffold exhibited regular and interconnected pores with good continuity and clear network structure; the sheep vertebral bone meal/PVA nonporous bone plate had no obvious pores; however, it had dense and evenly distributed micropores with different sizes on its surface. The porosity of nHA/PVA scaffold was lower than that of the sheep vertebral bone meal/PVA scaffold (P < 0.05). The water absorption rate was highest for the nHA/PVA scaffold followed by the sheep vertebral bone meal/PVA scaffold and the sheep vertebral bone meal/PVA nonporous bone plate (P < 0.05). In contrast, the scaffold toughness was highest for the sheep vertebral bone meal/PVA nonporous bone plate, followed by the sheep vertebral bone meal/PVA scaffold and nHA/PVA scaffold. In addition, the cell proliferation activity of cells cultured on the sheep vertebral bone meal/PVA scaffold was significantly higher than that cultured on the other two kinds of scaffolds. Taken together, the 3D printing sheep vertebral bone/PVA scaffold has good physical and chemical performance. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Bone filling mesh container versus balloon percutaneous kyphoplasty repairs osteoporotic vertebral compressive fracture Fu Guo-yong, Geng Xiao-peng, Wang Xia, Li Xue-cheng, Gao Sheng, Niu Chong, Dou Yong-feng, Zhu Kai 2016, 20 (52):  7858-7864.  doi: 10.3969/j.issn.2095-4344.2016.52.015 Abstract ( 323 )   PDF (1212KB) ( 397 )   Save BACKGROUND: It remains controversial about the clinical outcomes of bone filling mesh containers (BFMCs) and percutaneous kyphoplasty (PKP) in pain relief, kyphosis correction, vertebral height restoration and reduction of cement leakage. OBJECTIVE: To compare the clinical outcomes of BFMCs and PKP for osteoporotic vertebral compressive fracture. METHODS: A total of 90 patients with osteoporotic vertebral compressive fracture were equivalently randomized into two groups, followed by treated with BFMCs or PKP, respectively. During a more than 3-month follow-up, pain relief, kyphotic angle, the vertebral height and cement leakage were observed in the two groups to assess the therapeutic effects. RESULTS AND CONCLUSION: Pain in all patients was relieved at 24 hours after operation. There was no significant difference in pain relief between two groups (P > 0.05). PKP was more effective to restore the vertebral height (P < 0.05), while BMCFs significantly reduced the leakage rate of bone cement (P < 0.05). These results suggest that BFMCs and PKP have their own advantages in the treatment of osteoporotic vertebral compressive fracture, but both exert analgesic effects. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Inhalation of upconversion nanoparticles does transient damage to the mouse liver and lung Yang Run-xiu, Cai Li-yun, Zheng Ai-hua, Guan Ji-tao 2016, 20 (52):  7865-7871.  doi: 10.3969/j.issn.2095-4344.2016.52.016 Abstract ( 256 )   PDF (1613KB) ( 395 )   Save BACKGROUND: Lots of in vitro experiments have explored the toxicity of upconversion nanoparticles, but its toxicity in vivo is little reported. OBJECTIVE: To investigate the toxicity of upconversion nanoparticles in mouse organs. METHODS: After tracheotomy, 36 Balb/c mice were randomly divided into three groups, followed by instilled with 28 mg/kg upconversion nanoparticle (experimental group), the same volume of normal saline (control group), and nothing (sham operation group), respectively. The functional changes of the lung, liver and kidney were detected at 1 day, 1 and 2 weeks postoperatively, and meanwhile, the morphological changes of the lung, liver, kidney, and heart were observed. RESULTS AND CONCLUSION: At 1 day postoperatively, the pH values in the experimental group were lower than those in the control and sham operation groups (P < 0.05), while the glutamic-pyruvic transaminase level was higher than that in the control and sham operation groups (P < 0.05). The oxygen partial pressure in the sham operation group was higher than that in the other two groups at 1 day postoperatively (P < 0.05). The oxygen partial pressure and glutamic-pyruvic transaminase level did not significantly differ among groups at 1 and 2 weeks postoperatively. The carbon dioxide differential pressure and kidney function showed no significant differences among groups at different time points after surgery. At postoperative 1 day, in the experimental group, hyperplasia and inflammation were most obvious, distorted alveolar cavity and congestion of blood vessels were visible. In the control group, obvious hyperplasia and inflammation were found, the alveolar cavity was crimped and the gap between alveoli was broadened. The sham operation group had normal alveoli with no inflammations. Lung lesions in the experimental and control groups became mild with time at postoperative 1 and 2 weeks. One day postoperatively, hepatocyte swelling and vacuolar degeneration were severer in the experimental group. Moderate hepatocyte swelling and vacuolar degeneration occurred in the control group. The sham operation group showed mild hepatocyte swelling and vacuolar degeneration. The morphology of the liver in each group returned to normal at 2 weeks postoperatively. Fortunately, the heart and kidney structure showed no overt changes in each group. These findings suggest that upconversion nanoparticles cause transient damage to the mouse lung and liver. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Histocompatibility and distribution of ferroferric oxide nanoparticles   Gong Jun-zuo1, Tu Zhong-qi2, Duan Hong2, Zhou Shao-bing3 2016, 20 (52):  7872-7877.  doi: 10.3969/j.issn.2095-4344.2016.52.017 Abstract ( 296 )   PDF (2171KB) ( 172 )   Save BACKGROUND: Ferroferric oxide (Fe3O4) nanoparticles are a research hotspot in drug delivery system, which can transport antineoplastic drugs to the lesion under external magnetic field. Additionally, its submicrons even can reach the tumor site several centimeters far away from the magnetic source. OBJECTIVE: To investigate the histocompatibility and in vivo distribution of Fe3O4 nanoparticles and to explore its application prospect and limitations as a drug carrier in the chemotherapy of osteosarcoma. METHODS: 10.0 mg/kg Fe3O4 nanoparticles were administrated into Wistar rats via tail vein, then the rats were executed at 15, 60 and 120 minutes, respectively, and the rat lung, brain, heart, liver, kidney, hind limb and skeletal muscle were removed. The ferric ion content in each tissue was determined by atomic absorption spectrometer, and the morphological changes of different tissues were observed by hematoxylin-eosin staining at each time point. RESULTS AND CONCLUSION: After administrated for 15 minutes, the concentration of Fe3O4 nanoparticles in the liver and kidney reached peak, followed by a decrease at 60 and 120 minutes, but still remained a high level. The concentration of Fe3O4 nanoparticles at three time points showed significant difference compared with the control group (P < 0.05), demonstrating that the nanoparticles can be quickly enriched and long-term persistent in the liver and kidney. After administrated for 15 minutes, the concentration of Fe3O4 nanoparticles in the heart, lung, skeletal muscle and bone reached peak, which had significant difference compared with the control group (P < 0.05), and significantly decreased subsequently except that in the bone. This significant difference still displayed at 60 minutes between groups (P < 0.05), indicating that the nanoparticle can reach a high concentration but persist short time in the high blood perfused tissues. Compared with the control group, the concentration of Fe3O4 nanoparticles in the brain tissue showed no significant difference at each time point (P > 0.05), suggesting that the blood-brain barrier can inhibit the nanoparticle penetration. No overt morphological changes were found in each tissue after hematoxylin-eosin staining. In conclusion, the distribution of Fe3O4 nanoparticles conjugate sodium oleate in organism is influenced by the blood perfusion and mononuclear phagocyte system, and they cannot penetrate the blood-brain barrier and make no significant effect on tissues, but maintain a high level in the liver kidney and bone for a long-term, thus providing a theoretical basis for the drug delivery system in the magenetic hyperthermia therapy of malignant tumors. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Alkylated-chitosan preparation and its hemostatic effect Huang Yu-fen, Zou Li-hong, Gao Jie, Liu Lu 2016, 20 (52):  7878-7884.  doi: 10.3969/j.issn.2095-4344.2016.52.018 Abstract ( 415 )   PDF (1483KB) ( 206 )   Save BACKGROUND: 18-Alkylated-chitosan has been proved to exhibit a good coagulation capacity through hydrophobic modification. OBJECTIVE: To improve the adhesion between the material and the wound through the introduction of alkylated-chitosan molecules. METHODS: The alkyl group was introduced in the chitosan by potentiodynamic method to prepare the alkylated-chitosan with different substitution degrees (7%,16%, 26% and 40%), and then their structure, viscosity, porosity, water absorption, contact angle and in vitro coagulation capacity were detected. Thirty New Zealand rabbits were enrolled, modeled into the left femoral artery hemorrhage, and randomized into six groups, including four experimental groups, positive control and control groups, followed by treated with alkylated-chitosan sponges with different degrees of substitution, absorbable hemostatic gauze and chitosan sponge, respectively. Afterwards, the blood loss and clotting time were observed. RESULTS AND CONCLUISON: With the degree of substitution increased, the viscosity of alkylated-chitosan increased firstly and then decreased, the porosity and water absorption decreased gradually, and the contact angle on a rise. Compared with chitosan, different alkylated-chitosans showed a better henostatic effect in vitro, especially the alkylated-chitosan with 16% substitution degree. The blood loss and clotting time in the experimental groups except that the alkylated-chitosan with 7% substitution degree were significantly lower than those in the control group (P < 0.05); the bold loss in the alkylated-chitosan with 40% substitution degree group was significantly lower than that in the positive control group (P < 0.05). These findings suggest that the alkylated-chitosan with 40% substitution degree exhibits the best hemostatic effect in vivo. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Research status of skull repair materials and the prospect of three-dimensional printing technology Lu Lei, Chen Xu-yi, Li Yi-peng, Gang Lin, Tu Yue 2016, 20 (52):  7885-7890.  doi: 10.3969/j.issn.2095-4344.2016.52.019 Abstract ( 378 )   PDF (874KB) ( 331 )   Save BACKGROUND: Skull repair materials cannot only restore the normal shape of the skull, but also play an important role in brain functional recovery. OBJECTIVE: To summarize the research status of polyetheretherketone (PEEK), titanium alloy and tissue engineering technique in cranioplasty and the prospect of three-dimensional (3D) printing technology. METHODS: Literatures related to skull repair materials were retrieved in databases of CNKI and PubMed published from 1995 to 2016, using the keywords of “bone regeneration material in calvarial, 3d printing bone scaffold” in Chinese and English, respectively.  RESULTS AND CONCLUSION: Although titanium and PEEK have been used in clinic, titanium holds conductivity, thermal conductivity, while PEEK that may be displaced or lost is not involved in osseointegration. Tissue engineering technology participates in the skull tissue reconstruction, achieving satisfactory repair outcomes, but the problems of scaffold selection and preparation, seed cell obtainment, and growth factor release need to be overcomed. 3D printing technology can print personalized shape, fit the defect precisely, but the raw materials should have good biocompatibility and biomechanical property. Combination of tissue engineering technology with 3D printing technology shows a broad prospect in cranioplasty. 中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Tissue-engineered scaffolds for articular cartilage repair Fang Hong-song, Zhou Jian-lin, Peng Hao, Deng Shuang, Weng Jin-qing, Liu Feng, Chen Sen, Zhou Guan-jin 2016, 20 (52):  7891-7898.  doi: 10.3969/j.issn.2095-4344.2016.52.020 Abstract ( 434 )   PDF (1053KB) ( 255 )   Save BACKGROUND: Articular cartilage repair has been a difficulty in the clinical setting, which is mainly treated with autologous or allogeneic osteochondral grafts, and cartilage periosteum or periosteum grafts. However, the limited source, secondary lesion and immunological rejection force some researchers to search for a novel treatment strategy, cartilage tissue engineering, that is of great significance for cartilage regeneration and repair. OBJECTIVE: To investigate the tissue-engineered scaffolds for the repair of articular cartilage defects. METHODS: The first author searched the PubMed and WanFang databases for the articles addressing tissue-engineered cartilage for articular cartilage defects published between 1991 and 2015 using the keywords “articular cartilage defect, scaffold, tissue engineered cartilage” in English and Chinese, respectively. The irrelative and repetitive literatures were excluded. RESULTS AND CONCLUSION: Finally 48 eligible literatures were enrolled based on the inclusion and exclusion criteria. Cartilage tissue engineering possesses the advantages of controllability, little damage to tissue itself, and biological repair of injured cartilage. Tissue-engineered scaffold material is a critical factor in tissue engineering construction; therefore, it should hold biodegradability and histocompatibility. The commonly used scaffold materials include natural macromolecule materials (collagen, silk fibroin and chitosan), and synthetic polymer materials (polylactic acid and tricalcium phosphate). It is necessary to prepare composite scaffolds with high bioactivity integrate advantages of each material. The tissue engineering is bound to be a hotspot in the field of articular cartilage repair.  中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics

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Research process of the effect of titanium dioxide nanotubes on peri-implant cells Li Hong-cai, Ma Zhuang 2016, 20 (52):  7899-7904.  doi: 10.3969/j.issn.2095-4344.2016.52.021 Abstract ( 225 )   PDF (930KB) ( 234 )   Save BACKGROUND: Compared with the titanium with smooth surface, TiO2 nanotubes are more beneficial for the early cell adhesion. OBJECTIVE: To review the research progress of the effect of TiO2 nanotubes on peri-implant cells. METHODS: A computer-based retrieval of CNKI, CqVip, WanFang and PubMed databases was performed for pertinent literatures published between 1990 to 2016, using the Chinese keywords of “TiO2 nanotubes or titanium dioxide nanotubes and implant”, and English keywords of “TiO2 nanotubes implant, Tio2 nanotubes cell”. RESULTS AND CONCLUSION: The main factors that TiO2 nanotubes affect the peri-implant cell behaviors include the tubular morphology, size, TiO2 crystal structure, as well as surface chemical compositions, roughness and hydrophilicity and free energy. The influence factors of TiO2 nanotubes for cells are not only related to the tube diameter, but also associated with cell types. Different cell types hold different diameters suitable for the cell adhesion, proliferation and differentiation on the material surface. Therefore, it is advisable to select the best nanotube size for different tissues and cells applied in different biomedical fields.  中国组织工程研究杂志出版内容重点：生物材料；骨生物材料; 口腔生物材料; 纳米材料; 缓释材料; 材料相容性；组织工程 Figures and Tables | References | Related Articles | Metrics