CAJ | 학술논문

目的探讨应用自行设计的灌注接种环路对4种医用多孔材料进行体外人胚成骨细胞动态接种的可行性,筛选较优的支架材料并初步探讨其作用机制. 方法选取人同种多孔磷酸三钙(β-TCP)、异体松质骨粒、明胶海绵、胶原蛋白海绵4种医用材料作为支架载体,应用自行设计的灌注接种环路进行体外人胚成骨细胞动态接种(灌注接种组),以静态接种作为对照(静态接种组),通过检测支架上的细胞活力、活细胞接种率、细胞计数、细胞分布均匀度及组织学观察,比较不同接种方法和支架材料间细胞接种效果的差异.结果 β-TCP、明胶海绵:灌注接种组细胞活力、活细胞接种率和细胞计数均优于静态接种组,差异有统计学意义(P＜0.05);松质骨粒:两接种组间差异均无统计学意义(P＞0.05);胶原蛋白海绵:灌注接种组细胞活力、活细胞接种率和细胞计数均低于静态接种组(P＜0.05),细胞分布均匀度差异无统计学意义(P＞0.05).灌注接种:β-TCP、明胶海绵细胞接种率、细胞计数和细胞分布均匀度优于松质骨粒和胶原蛋白海绵,差异均有统计学意义(P＜0.05);静态接种:胶原蛋白海绵的细胞接种率和细胞计数最高,差异均有统计学意义(P＜0.05). 结论灌注接种法可明显提高β-TCP和明胶海绵的细胞接种效率和细胞分布均匀性,可用于快速复合成骨细胞;支架材料显著影响细胞的灌注接种效果.
목적 탐토응용자행설계적관주접충배로대4충의용다공재료진행체외인배성골세포동태접충적가행성,사선교우적지가재료병초보탐토기작용궤제. 방법 선취인동충다공린산삼개(β-TCP)、이체송질골립、명효해면、효원단백해면4충의용재료작위지가재체,응용자행설계적관주접충배로진행체외인배성골세포동태접충(관주접충조),이정태접충작위대조(정태접충조),통과검측지가상적세포활력、활세포접충솔、세포계수、세포분포균균도급조직학관찰,비교불동접충방법화지가재료간세포접충효과적차이.결과 β-TCP、명효해면:관주접충조세포활력、활세포접충솔화세포계수균우우정태접충조,차이유통계학의의(P＜0.05);송질골립:량접충조간차이균무통계학의의(P＞0.05);효원단백해면:관주접충조세포활력、활세포접충솔화세포계수균저우정태접충조(P＜0.05),세포분포균균도차이무통계학의의(P＞0.05).관주접충:β-TCP、명효해면세포접충솔、세포계수화세포분포균균도우우송질골립화효원단백해면,차이균유통계학의의(P＜0.05);정태접충:효원단백해면적세포접충솔화세포계수최고,차이균유통계학의의(P＜0.05). 결론 관주접충법가명현제고β-TCP화명효해면적세포접충효솔화세포분포균균성,가용우쾌속복합성골세포;지가재료현저영향세포적관주접충효과.
Objective To determine optimal scaffold materials by evaluating the dynamic seeding of human fetal bone cells into 4 kinds of medical material in vitro using a self-designed perfusion loop.Methods A novel flow perfusion system with multi-loops was designed for dynamic cell seeding in vitro into three-dimensional (3D) porous scaffolds made of 4 different kinds of medical materials:β-tricalcium phosphate (β-TCP),human allogenic cancellous bone particles,gelatin sponge and collagen sponge.Static seeding scaffolds were used as controls.The seeding efficiency and spatial distribution of human fetal bone cells were compared between different seeding methods and scaffold materials using quantitative biochemical and image analyses.Results Perfusion seeding into β-TCP and gelatin sponge yielded significantly better seeding efficiency and more homogeneous distribution than static seeding (P ＜ 0.05).There were no significant differences regarding cell load and cell distribution in allogenic cancellous bone particles between the perfusion and static groups.Cell viability,cell seeding efficiency and cell counting in collagen sponge were significantly higher in the static group than in the perfusion group (P ＜ 0.05),but the uniformity of seeding was similar in both groups.In addition,perfusion seeding into β-TCP and gelatin sponge was much more effective than into allogenic cancellous particles and collagen sponge; static seeding into collagen sponge yields the highest cell seeding efficiency and cell counting.Conclusions Since perfusion seeding technique may yield higher seeding efficiency and more homogeneous distribution in β-TCP and gelatin sponge,it can be used to facilitate rapid cell seeding.Scaffold materials may have an obvious impact on the seeding efficiency and spatial distribution of cells.