38 2010 12 FENXI HUXUE Chinese Journal of nalytical Chemistry 12 1754 ~ 1758 DOI 10. 3724 /SP. J. 1096. 2010. 01754 / * 402160 401331 Nano-H / PL Nano-H /PL MTT BMSCs SEM 4 d 8 d 2 3 4 5 6 8 d MTT 100% 0 26% Nano-H /PL / 1 1-2 ~ 4 PL H 5 Nano-H PL Nano-H /PL Nano-H / PL Nano-H /PL 6 7 8 9 Bone marrow stromal cells BMSCs Nano-H /PL 2 2. 1 BDS200-PH GHP-9050 EDX S3400N Model550 Bio-Rad 2010-04-15 2010-06-28 No KJ081205 No KJ091213 * E-mail zxc_228@ 163. com
12 / 1755 BS-323S-WE1 PL 8 H H GIBCO Invitrogen DMF12 DMSO MTT PBS Con 2. 2 Nano-H /PL g /ml 30 min Nano-H /PL 1 d Nano-H /PL 150 0. 03 MPa 1 d Nano-H /PL SEM 1 1 Nano-H /PL SEM PL H Fig. 1 SEM morphology of nano-hydroxyapatite / 1 polylactic acid H /PL 2. 3 1 ~ 2 d Wistar 4 75% 1 Nano-H /PL 5 min PBS Table 1 Component analysis of nano-h / PL 0. 5 mm 3 particulate from energy spectrum Element Weight tomic C 63. 57 70. 53 O 34. 56 28. 79 20% DMF12 5 ml P 0. 64 0. 27 2 ~ 3 h 37 Ca 1. 23 0. 41 Totals 100. 00 CO 2 5% 3 d 80% 3 3 BMSCs 1 10 4 /ml 2. 4 Nano-H /PL 24 h 24 3 3 BMSCs 1 10 7 /ml 8 h 2. 5 BMSCs Nano-H /PL 4 d 3% PBS 35 BMSCs
1756 38 2. 6 10 ~ MTT 12 BMSCs 2. 4 Nano-H /PL 24 2 10 4 / 100 μl 100 μl Con 0 5 2 3 4 5 6 8 d MTT 5 50 $L 5 g /L MTT 1 ml 37 4 h 400 $ L DMSO 10 min 200 $L 96 570 nm 1 Relative growth rate 2 = - - 100% 3 3. 1 1 2 Table 2 and cytotoxicity stage Relative growth rate Cytotoxicity 100 75 ~ 99 50 ~ 74 25 ~ 49 1 ~ 24 < 1 BMSCs Nano-H /PL 8 d 2 24 h BMSCs 3 Nano-H /PL Nano-H /PL BMSCs SEM BMSCs 4 d Nano-H / PL 3a 3b 0 1 2 3 4 5 2 3. 2 BMSCs Nano-H /PL 8 d Fig. 2 Bone marrow stromal cells MSCs morphology on nano-h /PL microspheres at a culture time of 8 d MTT MTT DMSO 13 3 2 3 4 5 6 8 d Nano-H /PL MTT 100% 0 BMSCs Nano-H / PL 4
12 / 1757 3 Fig. 3 Nano-H /PL 4 d SEM SEM images to show cell morphology on nano-h /PL scaffold at a culture time of 4 d 3 MTT Table 3 Resultes of MTT for cell proliferations 2 d Material extract ID 3 d 4 d 5 d 6 d 8 d 1 0. 165 145. 09 0. 215 126. 80 0. 277 127. 08 0. 371 134. 29 0. 432 110. 09 0. 474 101. 05 2 0. 159 133. 33 0. 200 111. 34 0. 246 105. 55 0. 360 128. 98 0. 430 109. 44 0. 472 100. 52 3 0. 153 121. 56 0. 195 106. 18 0. 242 102. 77 0. 357 127. 53 0. 429 109. 12 0. 470 100. 00 Positive control 0. 091 0. 092 0. 094 0. 093 0. 094 0. 091 Negative control 0. 142 0. 189 0. 238 0. 300 0. 401 0. 470 bsorbance Relative growth rate 2 14% 5 20% 6 8 6 Nano-H /PL BMSCs 4 BMSCs Fig. 4 BMSCs growth curves on materials a. Experimental group on Nano-H /PL scaffold b. Negative control group. References 1 HONG Yi GO Chang-You SHEN Jia-Cong. J. Biomedical Engineering 2007 24 2 463 ~ 465 2 Tan H P Wu J D Lao L H Gao C Y. cta Biomaterialia 2009 5 1 328 ~ 337 3 Lao L H Tan H P Wang Y J Gao C Y. Colloids and Surfaces B Biointerfaces 2008 66 2 218 ~ 225 4 XIONG Xin-Bo LI He-Jun HUNG Jian-Feng M Wei ZHENG Yong GUO Lin-Jun. Rare Metal Material and Engineering 2004 33 3 313 ~ 316 5 Hu X H Zhou J Zhang N Tan H P Gao C Y. J. The Mechanicalbe Havior of Biomedical Materials 2008 1 4 352 ~ 359
1758 38 6 LIN Zong-Qiong XIO Xiu-Feng SHE Hou-De HUNG Li-Zhong LIU Rong- Fang. Polymer Materials Science & Engineering 2008 24 10 155 ~ 158 7 YIN Hai-Rong LIN She-Bao YU Cheng-Long LI Yang. New Chemical Materials 2008 36 12 17 ~ 19 8 Mao Z W Wang B Ma L Gao C Y. Nanomedicine Nanotechnology Biology and Medicine 2007 3 3 215 ~ 223 9 Taboas J M Maddox R D Hollister S J. Biomaterials 2003 24 1 181 ~ 194 10 HUNG Yin-Jiu SONG Bao-n JIN Lin-Hong HU De-Yu. YNG Song LI Ning WU Shou-Wei. J. Biology 2009 26 4 13 ~ 16 11 JING Ze XU Hua LI Sheng-Shui ZHNG Feng-Mei LI Shuang-Biao LIU Yan. China Medical Herald 2010 7 3 28 ~ 29 12 HOU Chun-Mei LI Xin-Ying YE Wei-Liang CO Xi-Yuan XIO He LI Yan. Bull. cad. Mil. Med. Sci. 2009 33 4 400 ~ 401 13 ZHNG Li LU Chun-Tang ZHOU Shu-Xia XIONG Cheng-Dong. J. Practical Stomatology 2000 16 5 363 ~ 365 Evaluation of in Vitro Cell Compatibility for Nano-Hydroxyapatite / Polylactic cid Microspheres and Scaffold XU Wen-Feng * 1 LIO Xiao-Ling 2 1 College of Chemistry and Environment Engineering Chongqing University of rts and Sciences Chongqing 402160 2 College of Metallurgical and Materials Engineering Chongqing University of Science and Technology Chongqing 401331 bstract Nano-H / PL particles have been prepared by using the solution blending and solvent evaporation method and then bonded into the three dimensional porous scaffolds. The cell proliferation and cell morphology were observed and estimated by using inverted phase contrast microscope scanning electron microscope SEM and MTT assay. The cell cytotoxicity were also assessed. The result indicated that the cells mainly adhered and spread on the low-lying place or the hole place of the scaffold on which cultured cells after 4 d and the cell morphology from SEM presented spindle-shaped with many abundant triangle directly attached on the material's surface after 8 d and there were no deformation and necrosis phenomenon. The results of MTT assay indicated that all the relative growth rate of various experimental groups achieves above 100% the level of cytotoxicity is 0 and the experimental group cell vigor is higher 26% than that of the blank group. The results indicated that the Nano-H /PL particle scaffold is non-toxic for cells and has excellent adhesion and proliferation of bone cells. On the basis of these data Nano-H /PL particle scaffold is believed to be an optical bone repairing scaffold for tissue engineering. Keywords Nano-hydroxyapatite /polylactic acid microspheres Scaffold Cell compatibility Cytotoxicity Received 15 pril 2010 accepted 28 Jun 2010