20 34 20160819 Chinese Journal of Tissue Engineering Research August 19, 2016 Vol.20, No.34 Cytodext-3/ 12 1 3 1 1 12 1 1 1 1 1 1 1 ( 1 100853 2 300071 3 050017). Cytodext-3 /[J].201620(34):5104-5109. DOI: 10.3969/j.issn.2095-4344.2016.34.014 ORCID: 0000-0002-0033-9501() Cytodext-3 Cytodex t-3 Cytodext-3 / DNA Cytodex-3 Cytodext-3 / Cytodext-3 / 1990 100853 :R318 :A :2095-4344 (2016)34-05104-06 2016-05-29 Cytodext-3 Cytodext-3 / Cytodext-3 DNA Cytodext-3 / (P < 0.05) 1 d 14 d DNA DNA (P < 0.05)Cytodext-3 / 5104 P.O. Box 10002, Shenyang 110180
. Cytodext-3 / Yin He-yong, Master, PLA Institute of Orthopedics, General Hospital of Chinese PLA, Beijing 100853, China; School of Medicine, Nankai University, Tianjin 300071, China (Z141107004414044)(863)(2012AA020502) Constructing injectable tissue-engineered cartilage using cytodex-3 microcarrier and alginate hydrogel Corresponding author: Peng Jiang, M.D., Associate researcher, Master s supervisor, PLA Institute of Orthopedics, General Hospital of Chinese PLA, Beijing 100853, China; Yin He-yong 1, 2, Sun Zhen 1, Li Pan 3, Yu Xiao-ming 1, Xu Yi-chi 1, Sun Xun 1, 2, Xiao Bo 1, Wang Yu 1, Wang Ai-yuan 1, Guo Quan-yi 1, Xu Wen-jing 1, Lu Shi-bi 1, Peng Jiang 1 ( 1 PLA Institute of Orthopedics, General Hospital of Chinese PLA, Beijing 100853, China; 2 School of Medicine, Nankai University, Tianjin 300071, China; 3 Hebei Medical University, Shijiazhuang 050017, Hebei Province, China) Abstract BACKGROUND: Alginate hydrogel and microcarrier both can be used as injectable scaffolds, but their shortcomings such as poor mechanical property and poor plasticity remain unresolved. OBJIECTIVE: To explore the feasibility of constructing an injectable tissue-engineered cartilage with cytodex-3 microcarrier/alginate hydrogel composite. METHODS: Injectable cytodex-3 microcarrier/alginate hydrogel composite scaffold and injectable alginate hydrogel scaffold were established, and the mechanical properties of the two scaffolds were detected. Chondrocytes-seeded cytodex-3 microcarrier was obtained after incubated in the bioreactor, and then composited with alginate hydrogel as experimental group; chondrocytes were co-cultured with alginate hydrogel as control group. Subsequently, cell viability and ability of DNA and glycosaminoglycan synthesis were detected. RESULTS AND CONCLUSION: The Young s modulus of the experimental group was significantly higher than that of the control group (P < 0.05). And in the control group, chondrocytes were in a round shape and evenly distributed in the alginate hydrogel; in the experimental group, chondrocytes adhered on the scaffold surface and evenly distributed in the scaffold. After 1 day of culture, both viable and numerous dead chondrocytes could be found in both two scaffolds; and after 14-day culture, there were no dead chondrocytes in both two scaffolds, abundant proliferating chondrocytes maintained a high cell viability, and the number of chondrocytes in the experimental group was significantly higer than that of the control group. What s more, the contents of DNA and glycosaminoglycans were in a rise with time in both two groups, which were significantly higher in the experimental group than the control group (P < 0.05). These results suggest that the cytodex-3 microcarrier/alginate hydrogel composite is a promising injectable scaffold in cartilage tissue engineering. Subject headings: Chondrocytes; Stents; Tissue Engineering Funding: the Beijing Science and Technology Project, No. Z141107004414044; the National High-tech Research and Development Program of China (863), No. 2012AA020502 Cite this article: Yin HY, Sun Z, Li P, Yu XM, Xu YC, Sun X, Xiao B, Wang Y, Wang AY, Guo QY, Xu WJ, Lu SB, Peng J. Constructing injectable tissue-engineered cartilage using cytodex-3 microcarrier and alginate hydrogel. Zhongguo Zuzhi Gongcheng Yanjiu. 2016;20(34):5104-5109. 0 Introduction [1] [2-3] [4-5] 250 µm [6-8] (phco 2 ) 40 [9] ISSN 2095-4344 CN 21-1581/R CODEN: ZLKHAH 5105
. Cytodext-3 / Cytodext-3 Cytodext-3 Cytodext- [10-11] / 1Materials and methods 1.1 1.2 2014620159 1.3 1 SCXK-()2013-0004 Cytodext-3 Sigma DMEM Gibco DNA Invitrogen Synthecon BX-51DP70 Olympus ElectroForce 3320 BOSS 1.4 1 PBS3 1 cm 1 cm 1 cm0.15% 37 2 h 10%DMEM1 500 r/min 5 min10%dmed 37 5%CO 2 3 d 0.25% [12-13] P24 cm 4 cm Cytodext-3/ Cytodex-3 Cytodex-31 g100 ml PBS( Ca 2+ Mg 2+ )4 hpbs 5 minpbs50 mlpbs (115 103 kpa)15 min 4 1 ml 2.4% 102 mmol/l CaCl 2 Cytodext-3/ 1 ml 2.4% 102 mmol/l CaCl 2 Cytodext-3 2 10 6 P2 20 r/min1 min30 min24 h 50 r/min 37 5%CO 2 1 2.4% 102 mmol/l CaCl 2 6 2 10 6 P2 1 ml 2.4% 102 mmol/l CaCl 2 6 1.5 BOSS [14-16] 13714 d 5 mg/l FDA5 minpbs3 5 µmg/l PI5 minpbs [17] DNA137 1421 d3 4(30 mmol/l 55 mmol/l0.15 mmol/l CaCl 2 )30 min 5106 P.O. Box 10002, Shenyang 110180
. Cytodext-3 / 60 DNADNA [18] 13 71421 d3 19 [19] 1.6 SPSS 13.0 x _ ±s SNKP < 0.05 2Results 2.1 Cytodext-3/ Cytodext-3/ 1 1 Cytodext-3 / Figure 1 Gross observation of the cytodex-3 microcarrier/ alginate hydrogel composite scaffold 2.2 Cytodext-3/ [(24.22±2.32)(0.78±0.063) kpap < 0.05] 2.3 1 d 2AD3 d 7 d 2BE14 d 2CF 2.4 DNA DNA DNA DNA (P < 0.05)3 2.5 (P < 0.05)4 3Discussion 4 cm 2 [20-22] 3 [3] [23] [24] Cytodext-3 / 1 / DNA ISSN 2095-4344 CN 21-1581/R CODEN: ZLKHAH 5107
. Cytodext-3 / A B C D E F DNA(µg) 14 12 10 8 6 4 1 d 3 d 7 d 14 d 21 d 3 DNA Figure 3 DNA contents in the chondrocytes of the two groups at different time points of culture DNA (P < 0.05) (µg) 2 Figure 2 Dead/living chondrocytes stained in the two scaffolds at different time points of culture ABC 1714 d DE F 1714 d 150 100 50 0 1 d 3 d 7 d 14 d 21 d 4 Figure 4 Glycosaminoglycan contents in the chondrocytes of the two groups at different time points of culture (P < 0.05) [25] [26] [14] Cytodext-3 Cytodext-3 / Cytodext-3 / CNKI 5108 P.O. Box 10002, Shenyang 110180
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