Yang W D, Chen S J, Mao T Q, Chen F L, Lei D L, Tao K, Tang L H, Xiao M G
Lab for Tissue Engineering, Department of Oral and Maxillofacial Surgery, Qin Du School of Stomatology, Fourth Military Medical University (FMMU), Kang Fu Street No. 7, Xi'an, Shanxi 710032, P.R. China. yangwdwy#pub.xaonline.com
Chin J Dent Res. 2000 Dec;3(4):10-5.
To test the effectiveness of the new techniques of tissue-engineered cartilage.
Chondrocytes were harvested through type II collagenase digestion from the auricle of New Zealand rabbits. The cells were mixed with alginate to generate chondrocytes/alginate composites with final cellular density of 50 x 10(6) per mL. Calcium chloride was used as the cross-linking agent to gel the aqueous alginate solution. The chondrocytes/alginate composites were injected into the dorsal subcutaneous tissue of New Zealand rabbits through autologous cells grafts. The specimens were observed during cartilage formation at 4, 8, and 12 weeks after injection.
Prior to harvesting, chondrocytes/alginate composites were easily visualized under the dorsal skin of animals. The appearance of experimental specimens was similar to that of native cartilage in gross morphology. Using a standard hematoxylin and eosin stain, the histologic features of all experimental specimens demonstrated new cartilage formation. With a Masson's trichrome and safranin O stain, the presence of collagen and glycosaminoglycan (GAG) was observed at 8 and 12 weeks.
This study demonstrated that polymerization of alginate hydrogel can be controlled to allow injection of chondrocytes that produce new autologous cartilage at subcutaneous dorsal site of rabbits. Injectable tissue-engineered autologous cartilage is promising for potential use in oral and maxillofacial surgery.
测试组织工程软骨新技术的有效性。
通过II型胶原酶消化从新西兰兔耳廓获取软骨细胞。将细胞与藻酸盐混合，制成细胞密度为每毫升50×10⁶的软骨细胞/藻酸盐复合物。使用氯化钙作为交联剂使藻酸盐水溶液凝胶化。通过自体细胞移植将软骨细胞/藻酸盐复合物注入新西兰兔背部皮下组织。在注射后4周、8周和12周观察软骨形成过程中的标本。
在收获前，软骨细胞/藻酸盐复合物在动物背部皮肤下易于观察到。实验标本的外观在大体形态上与天然软骨相似。使用标准苏木精和伊红染色，所有实验标本的组织学特征显示有新的软骨形成。用Masson三色染色和番红O染色，在8周和12周时观察到胶原和糖胺聚糖（GAG）的存在。
本研究表明，可以控制藻酸盐水凝胶的聚合，以便在兔背部皮下部位注射能产生新的自体软骨的软骨细胞。可注射的组织工程自体软骨在口腔颌面外科有潜在应用前景。
