Malinin Theodore, Temple H Thomas, Buck Bill E
Department of Orthopaedics and Rehabilitation, University of Miami School of Medicine. P.O. Box 016960, Miami, FL 33101, USA.
J Bone Joint Surg Am. 2006 Apr;88(4):762-70. doi: 10.2106/JBJS.D.02991.
Transplantation of fresh osteochondral allografts stored at hypothermia into knee cartilage defects is a common procedure; however, the length of time that allografts can be stored prior to transplantation is controversial and has been determined, in part, by the results of vital stain uptake by chondrocytes. This study was performed to further define the limits of allograft storage.
Articular cartilage from six cadavers was stored for up to fifty-one days in tissue-culture media, and histologic sections were evaluated histomorphometrically to quantify the loss of chondrocytes. Samples of the cartilage were also placed into tissue culture to assess cell growth. Animal studies were performed in parallel on sixteen adult baboons with osteochondral allografts transplanted into the medial femoral condyle. Prior to transplantation, all allografts were stored in RPMI-1640 with 10% fetal calf serum at 4 degrees to 6 degrees C for up to eighty-five days. The transplants were graded on their gross and histological appearance, as well as their histochemical properties.
Many of the human samples stored at hypothermia in culture media for up to forty days retained some recognizable chondrocytes, but morphometry showed a gradual, significant decrease in the number of chondrocytes after nine days (p = 0.001). In addition, the cell outgrowth occurred from all specimens stored for up to fifteen days but not in samples stored for longer than thirty-four days. In animal studies, transplanted allograft cartilage that had been stored for less than eighteen days looked smooth and glistening, but grafts stored for over twenty-one days were pale, pitted, fragmented, or yellow, and chondrocytes were absent.
Time-dependent loss of chondrocytes in articular cartilage stored at hypothermia, especially in specimens stored for longer than fifteen to twenty days, was observed in this study. Cartilage allografts transplanted into nonhuman primates after twenty-one days of storage underwent more severe degenerative changes than allografts that had been stored for less than twenty-one days. These findings suggest caution when transplanting cartilage stored at hypothermia for over twenty days.
将低温保存的新鲜异体骨软骨移植到膝关节软骨缺损处是一种常见的手术;然而,异体移植物在移植前可保存的时间长度存在争议,部分是由软骨细胞对活体染色摄取的结果所决定的。进行本研究是为了进一步明确异体移植物储存的限度。
取自六具尸体的关节软骨在组织培养基中储存长达51天,对组织学切片进行组织形态计量学评估以量化软骨细胞的损失。软骨样本也被放入组织培养中以评估细胞生长。对16只成年狒狒进行了平行的动物研究,将异体骨软骨移植到股骨内侧髁。在移植前,所有异体移植物在含有10%胎牛血清的RPMI - 1640培养基中于4℃至6℃下储存长达85天。根据移植物的大体和组织学外观以及组织化学特性对其进行分级。
许多在培养基中低温保存长达40天的人体样本仍保留一些可识别的软骨细胞,但形态计量学显示9天后软骨细胞数量逐渐显著减少(p = 0.001)。此外,所有储存长达15天的标本都有细胞生长,但储存超过34天的样本则没有。在动物研究中,储存少于18天的移植异体软骨看起来光滑有光泽,但储存超过21天的移植物则苍白、有凹坑、破碎或发黄,且没有软骨细胞。
本研究观察到低温储存的关节软骨中软骨细胞随时间的损失,特别是在储存超过15至20天的标本中。储存21天后移植到非人灵长类动物体内的软骨异体移植物比储存少于21天的移植物发生更严重的退行性变化。这些发现提示在移植低温储存超过20天的软骨时要谨慎。