深低温冻存组织工程化软骨在喉狭窄功能重建中的应用

目的：探讨低温保存组织工程化软骨在喉狭窄功能重建中的应用价值。方法：取3周龄新西兰兔关节软骨细胞，体外培养，取第2代对数生长期培养细胞，制成细胞悬液，调整软骨细胞悬液浓度约为5×10^7个／ml左右，接种于PGA三维支架材料上，复合物体外培养2周后冻存，冻存6个月后解冻复苏，再行体外培养观察，2周后接种于已建立的喉甲状软骨缺损模型的软骨缺损处，并设对照组。术后12周取材，行大体及组织学观察。结果：经低温冻存的组织工程化软骨生长良好，组织学观察有软骨形成，与周围软骨组织结合紧密，与非冻存组相比差异无统计学意义。结论：深低温冻存对组织工程化软骨的生物活性无明显的影响，低温冻存的组织工程化软骨可用于喉软骨缺损的修复，重建喉功能。

Laryngeal Function Reconstruction Using Cryogenically Frozen Tissue Engineering Cartilage

Objective： To study the biological function of cryogenically preserved tissue engineering cartilage and the value of Reconstruction of Laryngeal function. Methods： In this experiment, the chondrocytes were isolated and collected from articular cartilages of New Zealand white rabbits. Cartilage defects were created in both side of thyroid cartilage with 30 rabbits. Complexes ofchodrocytes and PGA was cryogenically preserved for 6 months, After 6 months,cryogenically frozen tissue engineering cartilages were grafted into the defect of left thyroid cartilage at once as experimental group, and no cryogenically frozen tissue engineering cartilages were served as control.General and histological examination were respeetively performed in both group at 12 weeks after surgery. Results： Histologic sections stained with hematoxylin-eosin and Alcian blue （for acidic proteoglycans） and masson （for collagen ）confirmed the presence of hyaline cartilage in the cartilage constructs. Ultrastructural examination using transmission electron microscopy demonstrated matrix for- mation and chondrocyte viability.There was no difference in cryopreserved and noncryopreserved tissue engineering cartilage. Conclu- sions： This study proves that tissue engineering cartilage that are cryogenically stored for extended periods can be used to grow cartilage. Cryogenically preserved tissue engineering cartilage retains their ability to grow and can effectively improve reconstruction of Laryngeal function.