1、骨髓基质干细胞体外复合珊瑚材料的生长和成骨活性2006?10?o?y2?y5?7?i?f?r?o?7?t?D?V?K?M?s?K?7?e?251?Lt?Lm?T?i?(BoneMarrowStromalCells?aBMSCs)?E?a?r?o?f?a?0?r?o?f?L0?(?BMSCs?a?t?w?r?o?a?BMSCs?r?o?E?L0?z?L2?7?L2?Z?L2?V?fAKP)?(0CN)?7?f?L0?AKP?OCN?=?j?E?t?BMSCs?j?BMSCs?L0?r?o?1?w5xlOTem3?a?z?j?M?O?7?E7?d?wBMSCs?i?e?7?7?a?Z?E?BMSC
2、s?r?o?7?d?w?7?a?sOCN?y7?d?L0?BMSCs?r?o?t?D?V?r?o?L0?r?o?1?w5xlOYcm?a?a?f?7?d?0?L0Lt?U?Lm?i?M?V?Lt?h?(?LmQ813?w1+2Lt?U?B?2LmALt?f?Lm1673Ys0364(2006)?05Ys0251Ys05InVitroProliferationandOsteogenesisAbilityofBMSCsontheCoralYUANJie?aLIUGuangpeng,ZHULion?aXUFeng,WENGYulai?aCUILei?aLIUWei?aCA0Yilin?wDepart
3、mentofPlasticandReconstructiveSurgery,Shanghai?PeopleYPSHospital?aShanghaiKeyLaboratoryofTisSueEngineering,Shanghai?Q3?5?hUniversitySchoolofMedicine?aShanghai200011?wChi?LtAbstractLmObjectiveToexplorethepropercellseedingdensityandCOYsculturetimeofthebonemarrowstromalcells(BMSCs)?coralconstruct?wMeth
4、odsBMSCswereisolatedandculturedinosteogenicYsinducedandnon-inducedmedium?wTheinducedcellswerecomplexwithcoraltoco-culture?anon-inducedBMSCs-coralconstructwasservedascontr01?wTheconstructswereexaminedfromadherentrate?agrowthcurve?aScanelectronmicroscopy(SEM)andquantitativehistochemistry?wResultsQuant
5、itativehistochemisuydemonstratedthehigheralkalinephosphatase(AKP)andosteocalcinexpressionofinducedBMSCs-scaffoldthanthoseofnon-inducedBMSCsYsscaffold?wTheadherentratesignificantlydecreasedwhenthecellseedingdensityexceeded1?w5xlOT?Scm3?wThegrowthCHIVedemonstratedtheproliferationofBMSCsoncoralreachedp
6、lateauafter7daysco-culture?wAlsoSEMandquantitativehistoehemistryshowedBMSCsgrewwellandbegantoexpressosteocalcinonthecoralafter7days?wConclusionsTheosteogenicabilitywashigherofinducedBMSCsYsscaffoldthannonYsinducedBMSCsYsscaffold?wTheproperseedingdensityandCOYsculturetimeoftheconstructwere1?w5xlOVcm3
7、and7daysrespectively?wLtKeywordsLmBonemarrowstromalcells?OCoral?OTissueengineering?OAlkalinephosphatase?OOsteocalcin?n?N?a?m?h?i?(BoneMarrowStromalCells?aBMSCs)?a?i?t?M?a?V?n?t?a?J?(?L0?M?F?J?t?J?r?o?r?E?a?n?i?J?aBMSCs?i?w?7?J?N?J?l?(863?)?M?f?J?s?w?(2002AA205011)?O?e?0?Yx?h?YT?L0?5?N200011?e?0?e?N?
8、V?D?z?y?m?D?r?f?O?e?0?M?s?L0?N?5?N?e?M?h?I?2?7?t?D?V?a?0?J?L0?H?n?EBMSCs?J?r?o?n?t?D?V?a?V?N?s?=?i?a?s?n?E?V?a?r?Y?N1YP21L0?q?n?i?f?J?z?a?f?BMSCs?J?r?o?7?t?D?V?L0?E?y?i?r?a?BMSCs?i?J?e?z?a?0?O?a?N?fBMSCs?r?o?7?a?JournalofTissueEngineeringandReconstructiveSurgery?aOctober?w2006Vol2No5?V?(AKP)?(OCN)?a
9、?0?r?o?f?a?G?E?M?r?r?1?E?qL01?w?1?w1?w?2?a?20?a?r?a?e?L0?G?M?s?q?L0?L?L2B?K?2?q?h?Sigma?wAKP?h?Sigma?L0OCN?h?Biomedicaltechnologies?L0?L2?L296?L?h?Falcon?L0?EQuantachrome?L0AG?5KN?7?JShimadzu?L0?h?Dialab?L0?E?h?SonicsMaterials?L0DU?640?b?f?(?h?BeckmanCoulter?L0SN695B?j?G?e?F?s?q?7?L01?w2?q?6?=?5?2?X
10、?a?i?31?a?V?S?N?6?=?5?2?X?a?i?-?a?L?O?(?H?B?t4?lcmxlcmx0?w5cm?a?i?7?M?a?7?0?w5mm?Smin?a?J?E?L01?w3BMSCs?i?e?z?W?Kasten?4?a?(?W?a?i?e?z?L0?n?U?51?(BMSCs?L0?w?(?y2?BMSCs?n5?106cm3(1?w4x105?S?)L01?107?Scm3(2?w7?105?S?)?a1?w5x107?Scm3(4?105?S?)?a2xlOT?Scm3(5?w4x105?S?)?(?b?3mmx3mmx3mm?BYsTCP?J?e?a?g?15?
11、a?40?L0?(?J?r?o?370CL25?5?V?f?h?j?4?S?w?a?a?a?(?h?0?w25?5?0?w02?5EDTA?a?f?H?a?a?k?G?N?r?o?24?S?w?a?(?r?o?h?9?a?a?H?L0?0?w25?5?0?w02?5EDTA?h?f?H?,?L0?w?h?a?g?z?L0?z?=(?k?G?g?z?)?S(?k?G?)L01?w4BMSCs?r?o?f?(?y2?1?3?w7x107cm3flxl04?S?1?3mmx3mmx3mm?V?S?e?w?96?L?h?a?1?a?c?d?E?n?4?r?o?a?4?L?E?,?V?a?L?MTF?2
12、01?e1?a37c6?4?S?w?a?S?F?6?L?e?5?L0?150?1?u?x?a?J?15?(?a?(?a?w?(?96?a?i?e?R(OD490)?a?E?g?L00D490?R?E?H?7?L0?(?y2?BMSCs?H?w?H?t?a?1?w5x107?Scm3?3mm?3mm?3mm?V?S?a?L0?4?S?w?t?a?d?c?a?7?d?w?a?i2?5?e?h?30?(?aXL?30?Z?j?(Philips?a?)?L01?w5BMSCs?r?o?f?wAKP?D?V?(?y2?1?3?w7x107cm3flxl04?S?1?3mmx3mmx3mm?V?S?e?a
13、?96?L?h?a?1?a?w?c?i?y1L23L27L214L221L228?d?d?(?b?1?6?r?o?a?6?E?,?V?a?w?n?M?Nf1)stocksubstrate40mg(2?)?20ml?,?hL0(21?36?J15mL?(?b?B?1?a?1?E?a?1?j?0?w5mlAlkalineBuffersolution?0?w5mlstocksubstratesolution?a37c6?,?10?(?L0(31?B?y?15ml?h?aPBS?A?a?(?B?y?a?3mlTris(pH7?w4)?a?J?(1m30s?a?9s?a4c6)L0f41?0?w1ml?
14、o?B?B?h?a?0?w1ml?,?a?37c6?,?15?(?L0f51?lOml0?w05mol?SLNaOH?a405nm?b?f?(?R?a?6?L0f6)0?w08150D?R=1SigmaU?a1SigmaU=1xmolp-nitrophenol?ShL0?AKP?b?E?a?E?g?a?H?w?AKP?b?L0T?M?w?f?2006?10?o?y2?y5?71?w6BMSCs?r?o?f?wOCN?D?V?(?y2?1?3?w7?105?Scm3f1?104?S?1?3mmx3mmx3mm?V?S?e?w?96?h?a?1?L0?w?c?i?y1L23L27L214L221L
15、228?d?1?6?L?r?o?2001xl?e?5?a?6?L?2001xl?e?5?E?,?V?a?20c6?L0?2?g?B?y?H?w?n?M?B?X?y?h?Y?Nf1)?n?M?B?a?Xl?2?N?D?(TotalCounts)3?4?N?0?V?o5-6?N?B?7?8?N1?wOOng?SmL?B?a501xl9?10?N5?w00ng?SmL?B?a50txl11?12?N10?w00ng?SmL?B?a50?l13?14?N25?w00ng?SmL?B?a501xl15?16?N50?wOOng?SmL?B?a501xl17?18?N?(LowContr01)?a50?l
16、19?20?N?(HighContr01)?a501,d21?68?N?1?a?50txl?e?5?69?92?N?,?V?1?a?501xl?e?5?(2)3?4?h?150pA?C?(Buffer)?a5-6?h?501xl?C?(Buffer)?O(3)?q?n?1?h?=?100I_zl125I?Of4)5?92?h?lOOtxlOCN?q?5?O(5)?J?30?60?O(6)?V?a37c6?,?2?w5?S?Of7)?M1?w2?a?1?h?=?1001xl?IgG?100?1PEG?a?a?d?j?10?(?Nf8)?1L22?f?a?q?n?1?lml?C?(Buffer)?
17、a4L0CL24200rpm?Smin?a?15?(?O(9)?F?6?e?5?a?1?(?a?W?B?Y?L0?OCN?b?Eng?SmlL0?OCN?b?E?a?E?g?a?H?w?OCN?b?L01?w7?=?B?r?s)?E?a?q?i?(?a?iStudent?NewmanYskewls?L0p0?w05?E?n?j?V?b?a?5?ESASVer?w6?w12L02?2?w1?q?L?E150+_501xm?a?E57?w60+1?w50?5?a?E?E6?w46?0?w93MPa(?1)L0?w-YK_YK?m?Q4?N?S?S?N?R?j?_7?a?NYP?N?a?_?4?w7
18、?L0Q4?StrokeStrain(?5)?1?E?2?w2BMSCs?i?e?z?i?5X106?Sem3(1?w4x105?S?1?w?z?E98?w40?0?w72?5?O?1?107?Sem3(2?w7x105?S?)?a?z?E98?w07+0?w41?5?O?1?w5xlOT?Senl3f4x105?S?)?a?z?E98?w08+0?w83?5?O?2?107?Scm3f5?w4?105?S?1?a?z?E89?w77+1?w25?5L0?h?n?g?i?e?z?=?j?V?b(pO?w05)?a?a?L2x107?Sems?w?i?J?e?z?n?g?n?j?M?(p0?w05)?a?E?q?i?J?e?L02L03BMSCs?r?o?f?iMTT?h?BMSCs?i?J?e?7?y?J?a?L0?h?i?L?w?y1L22?d?a?F?R?f?r?w?j?E?S?0?7L0?y3?d?7s?,hi?Ni?a?y7?d?a?y8?d?P?_?M?L0?D?i?e?S?7?E24?S?36?S?a?7?E4?d?6?dL0?w?y7?d?7L0?BMSCs?i?e?7?j?V?b(p0?w05?a?2)L0?Z?E?_?N?a?L?w?L?w?N?t?X?L0BMSCs?r?o?7?d?BMSCs?i?(?F?a?(?f?i?
