Comet assay for quantification of the increased DNA damage burden in primary human chondrocytes with aging and osteoarthritis.
Comet assay for quantification of the increased DNA damage burden in primary human chondrocytes with aging and osteoarthritis.
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作者：
已知来自骨关节炎（OA）关节的软骨细胞表现出高水平的DNA损伤，但软骨细胞在“正常老化”期间积累DNA损伤的程度尚未确定。本研究的目的是量化从广泛年龄范围的尸体供体中获得的软骨细胞中存在的DNA损伤，并比较这种损伤与OA软骨细胞的程度。碱性彗星试验用于测量来自尸体供体的踝关节（距骨）和膝关节（股骨）的正常软骨以及在全膝关节置换术时获得的OA软骨细胞中的DNA损伤。根据彗星“尾巴”中DNA的百分比评估，来自年轻供体（<45岁）的软骨细胞比老年供体（>70岁）的DNA损伤更少。与尸体相比，在50至60岁之间的捐赠者中，OA软骨的软骨细胞DNA损伤增加。23例年龄在34 ~ 78岁之间的供体的距骨软骨细胞显示DNA损伤随年龄线性增加（R2 = 0.865，p < 0.0001）。使用“双尾”彗星试验来证明大部分累积损伤是以链断裂的形式，而不是碱不稳定的碱基损伤。来自年轻供体的软骨细胞需要10戈伊照射以重现来自老年供体的软骨细胞中存在的DNA损伤。鉴于DNA损伤可能导致软骨细胞功能障碍和衰老，本研究支持对低复制细胞类型积累高水平损伤的机制进行研究。软骨细胞在衰老过程中积累了大量的DNA损伤负担。如碱性彗星试验所测量的，从第4个到第8个十年的寿命呈线性增加。70岁以上捐赠者的组织损伤与骨关节炎关节置换时取出的软骨相似。
It is known that chondrocytes from joints with osteoarthritis (OA) exhibit high levels of DNA damage, but the degree to which chondrocytes accumulate DNA damage during “normal aging” has not been established. The goal of this study was to quantify the DNA damage present in chondrocytes obtained from cadaveric donors of a wide age range, and to compare the extent of this damage to OA chondrocytes. The alkaline comet assay was used to measure the DNA damage in normal cartilage from the ankle (talus) and the knee (femur) of cadaveric donors, as well as in OA chondrocytes obtained at the time of total knee replacement. Chondrocytes from younger donors (<45 years) had less DNA damage than older donors (>70 years) as assessed by the percentage of DNA in the comet “tail”. In donors between 50 and 60 years old, there was increased DNA damage in chondrocytes from OA cartilage as compared to cadaveric. Talar chondrocytes from 23 donors between the ages of 34 and 78 revealed a linear increase in DNA damage with age (R 2 = 0.865, p < 0.0001). A “two‐tailed” comet assay was used to demonstrate that most of the accumulated damage is in the form of strand breaks as opposed to alkali‐labile base damage. Chondrocytes from young donors required 10 Gy irradiation to recapitulate the DNA damage present in chondrocytes from older donors. Given the potential for DNA damage to contribute to chondrocyte dysfunction and senescence, this study supports the investigation of mechanisms by which hypo‐replicative cell types accumulate high levels of damage. Chondrocytes accumulate a substantial burden of DNA damage during aging. As measured by the alkaline comet assay, there is a linear increase from 4th to 8th decade of life. The damage in tissue from donors >70 years old is similar to cartilage taken at the time of joint replacement for osteoarthritis.
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影响因子：
4
作者：
Enciso, Maria;Sarasa, Jonas;Gosalvez, Jaime
通讯作者：
Gosalvez, Jaime
影响因子：
14.8
作者：
Olive, Peggy L.;Banath, Judit P.
通讯作者：
Banath, Judit P.
DOI：
10.1126/science.abb9032
发表时间：
2021-04-02
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
通讯作者：
--
影响因子：
--
作者：
Forsyth, CB;Pulai, J;Loeser, RF
通讯作者：
Loeser, RF
影响因子：
3.4
作者：
Anderson, CW;Dunn, JJ;Allalunis-Turner, MJ
通讯作者：
Allalunis-Turner, MJ