中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2014年
2期
87-93
,共7页
王林%马真胜%雷伟%胡蕴玉%王臻%冯亚非%张扬%裴国献
王林%馬真勝%雷偉%鬍蘊玉%王臻%馮亞非%張颺%裴國獻
왕림%마진성%뢰위%호온옥%왕진%풍아비%장양%배국헌
骨代用品%生物反应器%细胞骨架
骨代用品%生物反應器%細胞骨架
골대용품%생물반응기%세포골가
Bone substitutes%Bioreactors%Cytoskeleton
目的 探讨人胚成骨细胞在多孔β-磷酸三钙(β-TCP)支架内灌注性接种及培养的影响因素及其作用机制,对一体化灌注法体外构建活化人工骨进行优化研究.方法 应用自行设计的灌注式生物反应器进行多孔β-TCP支架内人胚成骨细胞的一体化灌注性接种和培养,以静态接种为对照.通过细胞活力(MTT法)测定、活细胞接种率、组织形态学观察和计量学分析等分别检测细胞接种时间、接种密度、灌注速率等因素对支架内细胞黏附和生长的作用.结果 细胞灌注接种效果优于静态接种;灌注接种-灌注培养法细胞生长及分布优于静态接种-灌注培养法.接种时间、接种密度、灌注速率等因素均显著影响细胞接种及培养效果.灌注法构建活化人工骨的最优条件:接种时间12h～24h,接种密度2×105/ml～5×105/ml,灌注接种速率1 ml/min,灌注培养速率0.5 ml/min～2ml/min(灌注初期24 h)及2 ml/min(灌注24 h后).结论 一体化灌注法利于多孔β-TCP支架内人胚成骨细胞的黏附和生长.接种时间、接种密度、灌注速率等因素均影响活化人工骨体外构建效果,对其进行优化有助于人工骨移植物的临床转化和推广应用.
目的 探討人胚成骨細胞在多孔β-燐痠三鈣(β-TCP)支架內灌註性接種及培養的影響因素及其作用機製,對一體化灌註法體外構建活化人工骨進行優化研究.方法 應用自行設計的灌註式生物反應器進行多孔β-TCP支架內人胚成骨細胞的一體化灌註性接種和培養,以靜態接種為對照.通過細胞活力(MTT法)測定、活細胞接種率、組織形態學觀察和計量學分析等分彆檢測細胞接種時間、接種密度、灌註速率等因素對支架內細胞黏附和生長的作用.結果 細胞灌註接種效果優于靜態接種;灌註接種-灌註培養法細胞生長及分佈優于靜態接種-灌註培養法.接種時間、接種密度、灌註速率等因素均顯著影響細胞接種及培養效果.灌註法構建活化人工骨的最優條件:接種時間12h～24h,接種密度2×105/ml～5×105/ml,灌註接種速率1 ml/min,灌註培養速率0.5 ml/min～2ml/min(灌註初期24 h)及2 ml/min(灌註24 h後).結論 一體化灌註法利于多孔β-TCP支架內人胚成骨細胞的黏附和生長.接種時間、接種密度、灌註速率等因素均影響活化人工骨體外構建效果,對其進行優化有助于人工骨移植物的臨床轉化和推廣應用.
목적 탐토인배성골세포재다공β-린산삼개(β-TCP)지가내관주성접충급배양적영향인소급기작용궤제,대일체화관주법체외구건활화인공골진행우화연구.방법 응용자행설계적관주식생물반응기진행다공β-TCP지가내인배성골세포적일체화관주성접충화배양,이정태접충위대조.통과세포활력(MTT법)측정、활세포접충솔、조직형태학관찰화계량학분석등분별검측세포접충시간、접충밀도、관주속솔등인소대지가내세포점부화생장적작용.결과 세포관주접충효과우우정태접충;관주접충-관주배양법세포생장급분포우우정태접충-관주배양법.접충시간、접충밀도、관주속솔등인소균현저영향세포접충급배양효과.관주법구건활화인공골적최우조건:접충시간12h～24h,접충밀도2×105/ml～5×105/ml,관주접충속솔1 ml/min,관주배양속솔0.5 ml/min～2ml/min(관주초기24 h)급2 ml/min(관주24 h후).결론 일체화관주법리우다공β-TCP지가내인배성골세포적점부화생장.접충시간、접충밀도、관주속솔등인소균영향활화인공골체외구건효과,대기진행우화유조우인공골이식물적림상전화화추엄응용.
Objective To investigate the influencing factors and mechanisms of human fetal osteoblasts seeding and culture in porous β-tricalcium phosphate (β-TCP) particles,and to optimize the bioartifical bone graft construction in vitro.Methods A newly designed perfusion bioreactor was employed for human fetal osteoblasts seeding and culture in porous β-TCP particles.Static seeding was served as the negative control.The effects of different conditions,including the time of seeding,density of cell suspension,flow rate of cell seeding and culture,on cell adhesion and growth were evaluated by the cell viability assay (MTT) and measurement of the cell seeding efficiency,histological and histomorphometric study.Results Perfusion seeding technique was superior to static seeding.Perfusion seeding and culture of fetal cells in β-TCP scaffolds resulted in significantly higher cell viability and growth rates than those by static seeding followed by perfusion culture.The time of seeding,density of cell suspension and flow rate of seeding and culture were determinants of the cell seeding and growth.The optimal conditions for constructing bioartificial bone included the time of seeding between 12 and 24 hours,the density of cell suspension between 2× 105/ml and 5× 105/ml,the flow rate of seeding 1 ml/min and the perfusion flow rate for incubation between 0.5 ml/min and 2 ml/min within the first 24 hours and 2 ml/min thereafter.Conclusions The in vitro perfusion technique is effective for seeding and culture of human fetal osteoblasts in porous β-TCP particles.The time of seeding,cell density and flow rate markedly impact on the outcomes of bioartificial bone construction in vitro.The optimization of conditions for bioartificial bone construction may facilitate clinical translation and promotion.
