中华骨科杂志
中華骨科雜誌
중화골과잡지
CHINESE JOURNAL OF ORTHOPAEDICS
2014年
9期
954-961
,共8页
李琪佳%王茜%甘洪全%刘英杰%王志强
李琪佳%王茜%甘洪全%劉英傑%王誌彊
리기가%왕천%감홍전%류영걸%왕지강
钽%成骨细胞%材料试验%组织工程
鐽%成骨細胞%材料試驗%組織工程
단%성골세포%재료시험%조직공정
Tantalum%Osteoblasts%Materials testing%Tissue engineering
目的 探讨国产多孔钽材料的细胞毒性和生物相容性,并通过兔骨内植入成骨示踪观察其成骨作用.方法 扫描电镜观察并测量多孔钽的形态学特征.来源于新西兰胎兔颅盖骨的成骨细胞以多孔钽浸提液(实验组)及完全培养基(对照组)培养,MTT法检测多孔钽的细胞毒性及其对增殖的影响.成骨细胞与多孔钽体外复合培养,观察成骨细胞的黏附、生长及增殖.雄性新西兰大白兔24只,制备股骨髁上多孔钽棒植入模型;4只动物于术后第5天和第19天分别肌肉注射荧光素钙黄绿素和茜素红,术后10周取材,于488 nm(钙黄绿素激发光)和543 nm(茜素红激发光)波长处,激光扫描共聚焦显微镜观察多孔钽-骨界面成骨;20只动物于术后2、4、8和12周取材行大体及硬组织切片观察.结果 多孔钽表面及断面可见均匀分布的三维立体连通孔隙结构.MTT法检测显示实验组与对照组细胞随培养时间的延长,其OD值的差异均无统计学意义.扫描电镜显示复合培养早期,细胞在多孔支架表面和孔壁上黏附,相互连接;晚期汇合成片并分泌细胞外基质覆盖材料表面.体内成骨实验显示植入的多孔钽棒与宿主骨结合紧密.硬组织切片显示术后2、4周时多孔钽-骨界面已出现新生骨及小血管,并向孔隙内生长;8、12周时多孔钽表面和孔隙内已长满新生骨组织,新生骨小梁已发育成熟并与材料直接接触.激光共聚焦扫描显微镜显示多孔钽-骨界面及孔隙内可见绿色荧光(钙黄绿素)和红色荧光(茜素红)标记的新生骨组织,红色荧光带位于绿色荧光带周围,早期均呈不连续性,晚期则融为一体.结论 国产多孔钽材料无细胞毒性,具有良好的生物相容性,多孔钽-骨界面为接触及传导性成骨并呈时间依赖性.
目的 探討國產多孔鐽材料的細胞毒性和生物相容性,併通過兔骨內植入成骨示蹤觀察其成骨作用.方法 掃描電鏡觀察併測量多孔鐽的形態學特徵.來源于新西蘭胎兔顱蓋骨的成骨細胞以多孔鐽浸提液(實驗組)及完全培養基(對照組)培養,MTT法檢測多孔鐽的細胞毒性及其對增殖的影響.成骨細胞與多孔鐽體外複閤培養,觀察成骨細胞的黏附、生長及增殖.雄性新西蘭大白兔24隻,製備股骨髁上多孔鐽棒植入模型;4隻動物于術後第5天和第19天分彆肌肉註射熒光素鈣黃綠素和茜素紅,術後10週取材,于488 nm(鈣黃綠素激髮光)和543 nm(茜素紅激髮光)波長處,激光掃描共聚焦顯微鏡觀察多孔鐽-骨界麵成骨;20隻動物于術後2、4、8和12週取材行大體及硬組織切片觀察.結果 多孔鐽錶麵及斷麵可見均勻分佈的三維立體連通孔隙結構.MTT法檢測顯示實驗組與對照組細胞隨培養時間的延長,其OD值的差異均無統計學意義.掃描電鏡顯示複閤培養早期,細胞在多孔支架錶麵和孔壁上黏附,相互連接;晚期彙閤成片併分泌細胞外基質覆蓋材料錶麵.體內成骨實驗顯示植入的多孔鐽棒與宿主骨結閤緊密.硬組織切片顯示術後2、4週時多孔鐽-骨界麵已齣現新生骨及小血管,併嚮孔隙內生長;8、12週時多孔鐽錶麵和孔隙內已長滿新生骨組織,新生骨小樑已髮育成熟併與材料直接接觸.激光共聚焦掃描顯微鏡顯示多孔鐽-骨界麵及孔隙內可見綠色熒光(鈣黃綠素)和紅色熒光(茜素紅)標記的新生骨組織,紅色熒光帶位于綠色熒光帶週圍,早期均呈不連續性,晚期則融為一體.結論 國產多孔鐽材料無細胞毒性,具有良好的生物相容性,多孔鐽-骨界麵為接觸及傳導性成骨併呈時間依賴性.
목적 탐토국산다공단재료적세포독성화생물상용성,병통과토골내식입성골시종관찰기성골작용.방법 소묘전경관찰병측량다공단적형태학특정.래원우신서란태토로개골적성골세포이다공단침제액(실험조)급완전배양기(대조조)배양,MTT법검측다공단적세포독성급기대증식적영향.성골세포여다공단체외복합배양,관찰성골세포적점부、생장급증식.웅성신서란대백토24지,제비고골과상다공단봉식입모형;4지동물우술후제5천화제19천분별기육주사형광소개황록소화천소홍,술후10주취재,우488 nm(개황록소격발광)화543 nm(천소홍격발광)파장처,격광소묘공취초현미경관찰다공단-골계면성골;20지동물우술후2、4、8화12주취재행대체급경조직절편관찰.결과 다공단표면급단면가견균균분포적삼유입체련통공극결구.MTT법검측현시실험조여대조조세포수배양시간적연장,기OD치적차이균무통계학의의.소묘전경현시복합배양조기,세포재다공지가표면화공벽상점부,상호련접;만기회합성편병분비세포외기질복개재료표면.체내성골실험현시식입적다공단봉여숙주골결합긴밀.경조직절편현시술후2、4주시다공단-골계면이출현신생골급소혈관,병향공극내생장;8、12주시다공단표면화공극내이장만신생골조직,신생골소량이발육성숙병여재료직접접촉.격광공취초소묘현미경현시다공단-골계면급공극내가견록색형광(개황록소)화홍색형광(천소홍)표기적신생골조직,홍색형광대위우록색형광대주위,조기균정불련속성,만기칙융위일체.결론 국산다공단재료무세포독성,구유량호적생물상용성,다공단-골계면위접촉급전도성성골병정시간의뢰성.
Objective To investigate cytotoxicity,biocompatibility and new bone formation traced of Chinese porous tantalum,and provide experimental strategies for further clinical application.Methods The physical properties of the porous tantalum were observed by the SEM.The osteoblasts were isolated from rabbit embryo.The extract fluid from tantalum was made.The cytotoxicity and proliferation of osteoblasts compounded with porous tantalum in vitro were detected by the MTT assay.The osteoblasts were co-cultured with extract of tantalum in vitro and the morphological changes,proliferation and adhesion were observed under SEM.A total of 24 New Zealand rabbits were used to establish the model of femoral condyles with porous tantalum bars implanted.Among which 4 of them was injected with calcein and alizarin on the 5th day and the 19th day and sacrificed at 10 week postoperatively.The specimens were observed with LSCM at 488 nm and 543 nm wavelength respectively.The remained 20 animals were sacrificed successively at 2,4,8,12 weeks of implantation,then were examined by histological observation.Results The SEM showed that the pore of porous tantalum were three-dimensional connected morphology.MTT assay showed that the osteoblasts grew well in extract and no significant difference between experimental and control groups.The osteoblasts grew and spread extensively on porous tantalum.Early on co-culture,the osteoblasts attached to the surface and inner walls of material,in the later stage,the osteoblasts excreted bone matrix over the surface of porous tantalum.The animal model showed that porous tantalum was bonded closely with host bone.Hard slicing showed that new bone and capillary regenerated on tantalum-bone interface at 2,4 weeks postoperatively.The pores were full with bone tissue at 8,12 weeks.The LSCM indicated that the green and red fluorescence-labeled new bone was displayed on tantalum-bone interface,while the red zone located around the green zones.They appeared to be discontinuous at early stage,but connected with each other at the end.Conclusion The Chinese porous tantalum has good biocompatibility and no cytotoxicity.The contact osteogenesis and bone conduction exist in tantalum-bone interface,and in a time-dependent manner.
