Characteristics of cartilage engineered from human pediatric auricular cartilage
Characteristics of cartilage engineered from human pediatric auricular cartilage
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DOI：
10.1097/00006534-199904040-00001
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发表时间：
1999-04-01
影响因子：
3.6
通讯作者：
Vacanti, CA
中科院分区：
文献类型：
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作者：
Rodriguez, A;Cao, YL;Vacanti, CA
在软骨缺损的修复中，自体组织具有持久生物相容性的优势。已经证明，从透明软骨中分离出的牛软骨细胞能够生成预定形状（例如人耳）的组织工程软骨；然而，从人类弹性软骨中分离出的软骨细胞的潜力仍然未知。在这项研究中，作者检查了人耳廓软骨细胞的增殖特征，以及这些细胞生成新弹性软骨的能力与它们在体外维持时间长度的函数关系。从 5 至 17 岁患者身上采集人耳软骨，用胶原酶消化，分离软骨细胞并在体外培养长达 12 周。每两周对细胞进行胰蛋白酶消化、计数和传代。每次传代时均创建软骨细胞聚合物（聚乙醇酸）构建体，然后植入无胸腺小鼠体内 8 周。研究了细胞在体外繁殖的能力以及它们产生新软骨的能力作为它们在体外维持时间的函数。总共植入了来自 12 名患者的 31 个实验结构，并与不含软骨细胞的对照组进行了比较。同时，对代表性细胞样本进行评估以确定胶原蛋白的存在。在研究群体中，人耳软骨细胞的体外倍增率保持恒定。 31 个实验植入物中有 22 个产生了新组织。该组织在肉眼检查中表现出耳软骨的物理特征。在组织学上，标本表现出密集的细胞结构和嵌入嗜碱性基质中的含腔细胞。这些标本类似于未成熟的软骨，并且部分不含构成该结构的合成材料。胶原蛋白、蛋白多糖和弹性蛋白的分析与弹性软骨一致。在对照植入物中未检测到软骨。人耳廓软骨细胞在体外繁殖良好，并且当接种到三维支架上时具有形成新软骨的能力。这些生长特征有一天可能使从小耳廓活检中分离出的软骨细胞在体外扩增，产生大型的定制形状的自体移植物，用于软骨缺损的临床重建，例如先天性小耳畸形。
In the repair of cartilage defects, autologous tissue offers the advantage of lasting biocompatibility. The ability of bovine chondrocytes isolated from hyaline cartilage to generate tissue-engineered cartilage in a predetermined shape, such as a human ear, has been demonstrated; however, the potential of chondrocytes isolated from human elastic cartilage remains unknown. In this study, the authors examined the multiplication characteristics of human auricular chondrocytes and the ability of these cells to generate new elastic cartilage as a function of the length of time they are maintained in vitro.Human auricular cartilage, harvested from patients 5 to 17 years of age, was digested in collagenase, and the chondrocytes were isolated and cultured in vitro for up to 12 weeks. Cells were trypsinized, counted, and passaged every 2 weeks. Chondrocyte-polymer (polyglycolic acid) constructs were created at each passage and then implanted into athymic mice for 8 weeks. The ability of the cells to multiply in vitro and their ability to generate new cartilage as a function of the time they had been maintained in vitro were studied. A total of 31 experimental constructs from 12 patients were implanted and compared with a control group of constructs without chondrocytes. In parallel, a representative sample of cells was evaluated to determine the presence of collagen.The doubling rate of human auricular chondrocytes in vitro remained constant within the population studied. New tissue developed in 22 of 31 experimental implants. This tissue demonstrated the physical characteristics of auricular cartilage on gross inspection. Histogically, specimens exhibited dense cellularity and lacunae-containing cells embedded in a basophilic matrix. The specimens resembled immature cartilage and were partially devoid of the synthetic material of which the construct had been composed. Analyses for collagen, proteoglycans, and elastin were consistent with elastic cartilage. No cartilage was detected in the control implants.Human auricular chondrocytes multiply well in vitro and possess the ability to form new cartilage when seeded onto a three-dimensional scaffold. These growth characteristics might some day enable chondrocytes isolated from a small auricular biopsy to be expanded in vitro to generate a large, custom-shaped, autologous graft for clinical reconstruction of a cartilage defect, such as for congenital microtia.