中国组织工程研究与临床康复
中國組織工程研究與臨床康複
중국조직공정연구여림상강복
JOURNAL OF CLINICAL REHABILITATIVE TISSUE ENGINEERING RESEARCH
2009年
21期
4105-4108
,共4页
郑有华%蒋柳宏%张志光%苏凯%匡世军
鄭有華%蔣柳宏%張誌光%囌凱%劻世軍
정유화%장류굉%장지광%소개%광세군
骨髓间充质干细胞%碱性成纤维细胞生长因子%基因转染%珊瑚人工骨%组织工程
骨髓間充質榦細胞%堿性成纖維細胞生長因子%基因轉染%珊瑚人工骨%組織工程
골수간충질간세포%감성성섬유세포생장인자%기인전염%산호인공골%조직공정
背景:颌面骨缺损是临床上常遇到的问题,寻找理想的种子细胞同支架材料复合构建组织工程化人工骨成为该类疾病治疗的发展趋势.目的:将转染碱性成纤维生长因子基因的骨髓间充质干细胞与珊瑚骨的复合培养,观察转染碱性成纤维生长因子基凶的骨髓间充质干细胞在珊瑚支架材料上生长状况.设计、时间及单位:骨组织工程实验,于2006-0312008-06 在中山大学口腔医学研究所完成.材料:选用海南省浅海滩产石头状滨珊瑚为原料,将其制成8mm×8mm×2mm的珊瑚人工骨小块.方法:采用密度梯度离心法分离新西兰大白兔骨髓间充质干细胞,采用贴壁筛选法对分离出的骨髓间充质下细胞进行纯化,利用脂质体转染bFGF-pcDNA3到骨髓间充质干细胞.取生长良好的转染碱性成纤维生长因子基因骨髓间充质干细胞和未转染的骨髓间充质干细胞,分别接种于不同珊瑚表面.主要观察指标:MTT法观察细胞-支架联合培养骨髓间充质干细胞的增殖情况和利用扫描电镜观察珊瑚支架上细胞的生长状况.结果:MTT法检测显示细胞-支架联合培养转染组细胞与联合培养未转染组细胞增殖相比差异有显著性意义(P＜0.05),联合培养转染组细胞生长增殖强于未转染组细胞.而联合培养的转染组细胞同单纯培养转染组细胞增殖相比差异无显著性意义(P＞0.05).扫描电镜观察显示复合骨髓间充质干细胞细胞贴附在珊瑚上,并在材料上完全铺展,形态多样,细胞向孔内长入或跨越微孔表面,部分区域有细胞外基质形成.结论:转染碱性成纤维生长因子基因的骨髓间充质干细胞在珊瑚支架材料上生长状况较未转染组好,珊瑚人工骨不影响骨髓间充质干细胞的增殖,可以作为骨髓间充质干细胞支架材料构建组织工程骨.
揹景:頜麵骨缺損是臨床上常遇到的問題,尋找理想的種子細胞同支架材料複閤構建組織工程化人工骨成為該類疾病治療的髮展趨勢.目的:將轉染堿性成纖維生長因子基因的骨髓間充質榦細胞與珊瑚骨的複閤培養,觀察轉染堿性成纖維生長因子基兇的骨髓間充質榦細胞在珊瑚支架材料上生長狀況.設計、時間及單位:骨組織工程實驗,于2006-0312008-06 在中山大學口腔醫學研究所完成.材料:選用海南省淺海灘產石頭狀濱珊瑚為原料,將其製成8mm×8mm×2mm的珊瑚人工骨小塊.方法:採用密度梯度離心法分離新西蘭大白兔骨髓間充質榦細胞,採用貼壁篩選法對分離齣的骨髓間充質下細胞進行純化,利用脂質體轉染bFGF-pcDNA3到骨髓間充質榦細胞.取生長良好的轉染堿性成纖維生長因子基因骨髓間充質榦細胞和未轉染的骨髓間充質榦細胞,分彆接種于不同珊瑚錶麵.主要觀察指標:MTT法觀察細胞-支架聯閤培養骨髓間充質榦細胞的增殖情況和利用掃描電鏡觀察珊瑚支架上細胞的生長狀況.結果:MTT法檢測顯示細胞-支架聯閤培養轉染組細胞與聯閤培養未轉染組細胞增殖相比差異有顯著性意義(P＜0.05),聯閤培養轉染組細胞生長增殖彊于未轉染組細胞.而聯閤培養的轉染組細胞同單純培養轉染組細胞增殖相比差異無顯著性意義(P＞0.05).掃描電鏡觀察顯示複閤骨髓間充質榦細胞細胞貼附在珊瑚上,併在材料上完全鋪展,形態多樣,細胞嚮孔內長入或跨越微孔錶麵,部分區域有細胞外基質形成.結論:轉染堿性成纖維生長因子基因的骨髓間充質榦細胞在珊瑚支架材料上生長狀況較未轉染組好,珊瑚人工骨不影響骨髓間充質榦細胞的增殖,可以作為骨髓間充質榦細胞支架材料構建組織工程骨.
배경:합면골결손시림상상상우도적문제,심조이상적충자세포동지가재료복합구건조직공정화인공골성위해류질병치료적발전추세.목적:장전염감성성섬유생장인자기인적골수간충질간세포여산호골적복합배양,관찰전염감성성섬유생장인자기흉적골수간충질간세포재산호지가재료상생장상황.설계、시간급단위:골조직공정실험,우2006-0312008-06 재중산대학구강의학연구소완성.재료:선용해남성천해탄산석두상빈산호위원료,장기제성8mm×8mm×2mm적산호인공골소괴.방법:채용밀도제도리심법분리신서란대백토골수간충질간세포,채용첩벽사선법대분리출적골수간충질하세포진행순화,이용지질체전염bFGF-pcDNA3도골수간충질간세포.취생장량호적전염감성성섬유생장인자기인골수간충질간세포화미전염적골수간충질간세포,분별접충우불동산호표면.주요관찰지표:MTT법관찰세포-지가연합배양골수간충질간세포적증식정황화이용소묘전경관찰산호지가상세포적생장상황.결과:MTT법검측현시세포-지가연합배양전염조세포여연합배양미전염조세포증식상비차이유현저성의의(P＜0.05),연합배양전염조세포생장증식강우미전염조세포.이연합배양적전염조세포동단순배양전염조세포증식상비차이무현저성의의(P＞0.05).소묘전경관찰현시복합골수간충질간세포세포첩부재산호상,병재재료상완전포전,형태다양,세포향공내장입혹과월미공표면,부분구역유세포외기질형성.결론:전염감성성섬유생장인자기인적골수간충질간세포재산호지가재료상생장상황교미전염조호,산호인공골불영향골수간충질간세포적증식,가이작위골수간충질간세포지가재료구건조직공정골.
BACKGROUND: Jaw defects are common clinically. It is desirable to find ideal seed cells combined with scaffolds to construct tissue engineered jaws for curing these diseases.
OBJECTIVE: To investigate the growth characteristics of bone marrow mesenchymal stem cells (BMSCs) transfected with basic fibroblast growth factor (bFGF) gene after seeded on coral scaffold in vitro.
DESIGN, TIME AND SETTING: An experimental study of bone tissue engineering was performed in the Research Institute of Stomatology, Sun Yat-sen University between March 2006 and June 2008.
MATERIALS: Natural coral from China Hainan bench was made into pieces of 8 mm×8 mm×2 mm.
METHODS: BMSCs were isolated from New England rabbits by density gradient centrifugation and then purified by adherent separation. bFGF-pcDNA3 gene was transfected into BMSCs using Lipofectamine TM 2000. bFGF gene-transfected (transfected group) or untransfected (untransfected group)BMSCs were seeded on different coral scaffolds. In addition, bFGF gene-transfected BMSCs were simply cultured but not on the coral scaffold for control (simple culture group).
MAIN OUTCOME MEASURES: BMSC proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay and BMSC growth on coral scaffold was observed under the scanning electron microscope.
RESULTS: MTT assay showed that the BMSC proliferation rate was significantly higher in the transfected group than in the untransfected group (P < 0.05) and that there was no significant difference in BMSC proliferation between the transfected and simple culture groups (P > 0.05). Scanning electron microscope results displayed that BMSCs adhered to and spread over the coral scaffold, exhibiting various appearances, with some cells had grown into scaffold micropores or spanned micropore surface, and some extracellular matrix secreted by BMSCs were found.
CONCLUSION: The transfected group exhibited better growth of BMSCs transfected by bFGF gene than the untransfected group. These findings indicate that coral skeleton does not influence BMSC proliferation and can be used as a scaffold of BMSCs to construct tissue-engineered bone.
