在多孔胶原支架上构建三维工程的心肌组织

背景目前已有转化的SV40细胞系、C2C12成肌细胞、心肌细胞、骨骼肌细胞以及胚胎干细胞用于修复哺乳动物心脏的细胞移植治疗,然而细胞移植治疗对严重缺失或损伤心肌结构的治疗的作用却不大.组织工程技术为此类心肌疾病的研究及治疗提供了新的可能性,目前已有可替代和改善骨、软骨、肾、肝、神经及皮肤等组织和器官的生物合成结构物.目的探索多孔胶原作为构建三维工程心肌组织支架材料的可行性.设计非随机、对照的实验研究.地点和对象实验地点军事医学科学院生物工程研究所.动物清洁级1日龄Wistar大鼠400～500只,雌雄不限,体质量8～10g;成年Wistar大鼠五六只,雌雄不限,体质量250 g,由军事医学科学院实验动物中心提供,饲养温度22～26℃,湿度40%～60%.干预在无菌条件下,将经胰蛋白酶消化获得的新生鼠心肌细胞分别接种于三维多孔胶原支架和培养板上,比较心肌细胞在两种培养模式的代谢差异.主要观察指标检测葡萄糖比消耗率、乳酸比产率、乳酸转化率、肌酸激酶及乳酸脱氢酶活力变化.结果培养于多孔胶原支架上的心肌细胞的代谢特性近似于心肌细胞在培养板上培养的二维生长模式.结论心肌细胞与多孔胶原支架形成了具有自律性同步收缩的三维工程心肌组织,多孔胶原作为心肌组织工程的支架材料有良好的应用前景.

Reestablishment ofthree-dimensional cardiac tissue architecture in porous collagen scaffolds

BACKGROUND: There have been a number of cell transplantation studies in which cells of various types, including SV40-transformed cell lines, C2C12 myoblasts, isolated cardiomyocytes, skeletal myoblasts, and embryonic stem cells, have been implanted into mammalian hearts. However, the effect of cell transplantation may be of little benefit in cases where the local cardiac structure is missing or seriously damaged. Tissue engineering offers the possibility of creating functional tissue equivalent for scientific studies and tissue repair. Biosynthetic constructs have been described that would replace or augment bone, cartilage, kidney, liver, neuronal tissue and skin.OBJECTIVE: To explore feasibility of porous collagen as three-dimensional cardiac scaffolds for tissue architecture.DESIGN: A non-randomized controlled experimental study.SETTING and PARTICIPANTS: The study was completed in the Institute of Biotechnology, Academy of Military Medical Sciences. Subjects were neonatal Wistar rats(1 day old, 8 - 10 g, clean grade) and adult Wistar (250 g) from the Laboratory Animal Center, Academy of Military Medical Sciences. Temperature and humidity of breeding were 22 - 26 ℃ and 40%- 60% respectively.INTERVENTIONS: Cardiac cells enzymatically digested from neonatal rats were seeded to three-dimensional porous collagen scaffolds and plastic plates. Metabolic characteristics of cardiac cells cultured between two culture modes were compared.MAIN OUTCOME MEASURES: The specific consumption rate of glucose(qg1c), specific production rate of lactate(q1ac), lactate transform rate(Y1aac/g1c), creatine kinase(CK) and lactate dehydrogenase(LDH) activities of cardiac cells.RESULTS: The metabolic characteristic of cardiac cells cultured in collagen scaffolds was similar to that cultured in polystyrene plates in two-dimensional mode.CONCLUSION: Synchronous and rhythmical contractile tissue constructs in vitro can be formed by cultivation of cardiac cells on porous collagen scaffolds. Porous collagen can be used as scaffolds in cardiac tissue engineering.