Department of Orthopedic Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Am J Sports Med. 2024 Jul;52(9):2384-2390. doi: 10.1177/03635465241254401.
Reduced viability in the deepest zones of osteochondral allografts (OCAs) can weaken the subchondral interface, potentially increasing the risk of failure. This reduction may result from nutritional imbalances due to uneven media distribution or interference from bone marrow elements.
To investigate whether culturing OCAs using a rotary shaker or removing the bone marrow elements would increase graft cellular viability.
Controlled laboratory study.
Bovine osteochondral explants were stored for 28 days at 4°C under 3 different conditions (n = 6 explants per group): static (control group), rotary shaker at 150 rpm (shaker group), and static after removal of bone marrow elements using a Waterpik device (Waterpik group). Chondrocyte viability was assessed using live/dead staining across the entire tissue and in each zone (superficial, middle, deep). Subchondral bone viability was assessed using TUNEL (terminal deoxynucleotidal transferase-mediated biotin-deoxyuridine triphosphate nick-end labeling) staining to detect apoptotic cells.
Both shaker (64.2%; = .010) and Waterpik (65.6%; = .005) conditions showed significantly higher chondrocyte viability compared with control (49.8%). When samples were analyzed by zone, the shaker and Waterpik groups displayed higher cellular viability at the middle zone (shaker = 60.6%, < .001; Waterpik = 56.1%, < .001) and deep zone (shaker = 63.1%, = .018; Waterpik = 61.5%, = .025) than the control group (25.6% at middle zone; 32.8% at deep zone). Additionally, shaker (56.7%; = .018) and Waterpik (51.4%; = .007) groups demonstrated a lower percentage of apoptotic cells in subchondral bone compared with control (88.0%). No significant differences were observed between the shaker and Waterpik groups in any of the analyses.
Both rotary shaking and removal of bone marrow elements during storage of osteochondral explants led to higher chondrocyte viability at the middle and deep zones of the graft compared with the static storage condition. Enhancing nutrition delivery to the graft could improve its quality, potentially improving outcomes of OCA transplantation.
The use of a rotary shaker or the removal of bone marrow elements may significantly improve the culture conditions, increasing graft viability and integrity after OCA storage.
骨软骨同种异体移植物（OCA）深层区域活力降低会削弱软骨下界面，从而增加失败风险。这种减少可能是由于营养物质分布不均导致的营养失衡，或者是骨髓成分的干扰造成的。
研究使用旋转摇床或去除骨髓成分培养 OCA 是否会增加移植物细胞活力。
对照实验室研究。
将牛骨软骨标本在 4°C 下储存 28 天，在 3 种不同条件下（每组 6 个标本）：静态（对照组）、以 150rpm 旋转摇床（摇床组）、使用 Waterpik 设备去除骨髓成分后的静态（Waterpik 组）。使用活/死染色法评估整个组织和每个区域（浅层、中层、深层）的软骨细胞活力。使用末端脱氧核苷酸转移酶介导的生物素-dUTP 缺口末端标记法（TUNEL）染色评估软骨下骨的活力以检测凋亡细胞。
摇床（64.2%， =.010）和 Waterpik（65.6%， =.005）条件下的软骨细胞活力均显著高于对照组（49.8%）。当按区域进行分析时，摇床组和 Waterpik 组在中层（摇床组=60.6%， <.001；Waterpik 组=56.1%， <.001）和深层（摇床组=63.1%， =.018；Waterpik 组=61.5%， =.025）区域的细胞活力更高，而对照组在中层（25.6%）和深层（32.8%）区域的细胞活力更低。此外，与对照组（88.0%）相比，摇床（56.7%， =.018）和 Waterpik（51.4%， =.007）组的软骨下骨中凋亡细胞的比例更低。在任何分析中，摇床组和 Waterpik 组之间均无显著差异。
与静态储存相比，在储存骨软骨标本时使用旋转摇床或去除骨髓成分可使移植物的中层和深层区域的软骨细胞活力更高。改善移植物的营养供应可能会提高其质量，从而改善 OCA 移植的效果。
使用旋转摇床或去除骨髓成分可显著改善培养条件，增加 OCA 储存后的移植物活力和完整性。
