Division of Orthopedic Surgery, Department of Surgery, University of Alberta, Edmonton, Alberta, Canada.
Department of Orthopedic Surgery, First Affiliated Hospital, Shantou University Medical College, Shantou, Guangdong, China.
Cartilage. 2021 Dec;13(2_suppl):1688S-1699S. doi: 10.1177/1947603520967077. Epub 2020 Oct 26.
Successful preservation of articular cartilage will increase the availability of osteochondral allografts to treat articular cartilage defects. We compared the effects of 2 methods for storing cartilage tissues using 10-mm diameter osteochondral dowels or femoral condyles at -196°C: (a) storage with a surrounding vitrification solution versus (b) storage without a surrounding vitrification solution. We investigated the effects of 2 additives (chondroitin sulfate and ascorbic acid) for vitrification of articular cartilage.
Healthy porcine stifle joints ( = 11) from sexually mature pigs were collected from a slaughterhouse within 6 hours after slaughtering. Dimethyl sulfoxide, ethylene glycol, and propylene glycol were permeated into porcine articular cartilage using an optimized 7-hour 3-step cryoprotectant permeation protocol. Chondrocyte viability was assessed by a cell membrane integrity stain and chondrocyte metabolic function was assessed by alamarBlue assay. Femoral condyles after vitrification were assessed by gross morphology for cartilage fractures.
There were no differences in the chondrocyte viability (70%) of 10-mm osteochondral dowels after vitrification with or without the surrounding vitrification solution. Chondrocyte viability in porcine femoral condyles was significantly higher after vitrification without the surrounding vitrification solution (70%) compared to those with the surrounding vitrification solution (8% to 36%). Moreover, articular cartilage fractures were not seen in femoral condyles vitrified without surrounding vitrification solution compared to fractures seen in condyles with surrounding vitrification solution.
Vitrification of femoral condyle allografts can be achieved by our optimized approach. Removing the surrounding vitrification solution is advantageous for vitrification outcomes of large size osteochondral allografts.
成功保存关节软骨将增加使用 10mm 直径骨软骨移植物治疗关节软骨缺损的同种异体骨软骨的可用性。我们比较了在-196°C 下使用 10mm 直径骨软骨栓或股骨髁对软骨组织进行 2 种储存方法的效果：（a）储存时周围有玻璃化溶液与（b）储存时无周围玻璃化溶液。我们研究了 2 种添加剂（硫酸软骨素和抗坏血酸）对关节软骨玻璃化的影响。
从屠宰场在屠宰后 6 小时内收集来自性成熟猪的健康猪膝关节（=11）。二甲亚砜、乙二醇和丙二醇使用优化的 7 小时 3 步渗透冷冻保护剂渗透方案渗透到猪关节软骨中。通过细胞膜完整性染色评估软骨细胞活力，并通过 alamarBlue 测定评估软骨细胞代谢功能。通过大体形态学评估玻璃化后的股骨髁有无软骨骨折。
在有或没有周围玻璃化溶液的情况下，10mm 骨软骨栓的玻璃化后软骨细胞活力（70%）没有差异。与周围玻璃化溶液相比，没有周围玻璃化溶液的猪股骨髁的软骨细胞活力明显更高（70%）。此外，与周围玻璃化溶液相比，没有周围玻璃化溶液的股骨髁未见关节软骨骨折，而有周围玻璃化溶液的髁可见骨折。
我们优化的方法可实现股骨髁同种异体移植物的玻璃化。去除周围玻璃化溶液有利于大尺寸骨软骨同种异体移植物的玻璃化结果。
