Functional Tissue Analysis Reveals Successful Cryopreservation of Human Osteoarthritic Synovium.
Functional Tissue Analysis Reveals Successful Cryopreservation of Human Osteoarthritic Synovium.
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DOI：
10.1371/journal.pone.0167076
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
van de Loo FA
中科院分区：
文献类型：
--
作者：
Broeren MG;de Vries M;Bennink MB;van Lent PL;van der Kraan PM;Koenders MI;Thurlings RM;van de Loo FA
骨关节炎是一种影响软骨的退行性关节疾病，是全世界最常见的关节炎形式。三分之一的骨性关节炎患者有严重的滑膜炎，只有不到10%的患者没有滑膜炎的证据。此外，滑膜炎预示着更严重的疾病进展。这为治疗提供了一个靶点，但需要对这些患者滑膜组织的病理生理过程进行更多的研究。当这种通过关节置换手术获得的材料可以储存起来供以后使用时，对滑膜组织进行的功能研究将更加容易。我们开始研究人类骨性关节炎滑膜组织缓慢冷冻的后果。因此，我们验证了一种可以应用于每个常规实验室的方法，并对五种冷冻保护剂(CPA)溶液进行了比较研究。研究了滑膜组织结构、代谢活性、RNA质量、冻存相关应激基因的表达以及骨性关节炎特有疾病基因的表达，以确定冷冻-解冻对滑膜活性可能产生的有害影响。此外，通过检测TLR配体脂多糖和Pam3Cys诱导的细胞因子分泌来检测冻存组织的生物学活性。与未冷冻的滑膜相比，CS10、标准和CryoSFM CPA液冷冻保存的滑膜在细胞和组织形态上没有明显差异。然而，我们观察到Biofreeze和CS2培养液对组织形态的保存率显著降低。其他活力分析显示出相同的趋势，但不够敏感，无法检测出不同条件之间的显着差异。在所测试的所有检测中，在没有CPA溶液的情况下，滑膜冷冻的情况下检测到的存活率明显较低。这一详细的分析表明，骨性关节炎滑膜组织外植体可以被冷冻保存，同时保持解冻后的形态、活性和表型反应，为人类体外研究提供了更多的机会。
Osteoarthritis (OA) is a degenerative joint disease affecting cartilage and is the most common form of arthritis worldwide. One third of OA patients have severe synovitis and less than 10% have no evidence of synovitis. Moreover, synovitis is predictive for more severe disease progression. This offers a target for therapy but more research on the pathophysiological processes in the synovial tissue of these patients is needed. Functional studies performed with synovial tissue will be more approachable when this material, that becomes available by joint replacement surgery, can be stored for later use. We set out to determine the consequences of slow-freezing of human OA synovial tissue. Therefore, we validated a method that can be applied in every routine laboratory and performed a comparative study of five cryoprotective agent (CPA) solutions. To determine possible deleterious cryopreservation-thaw effects on viability, the synovial tissue architecture, metabolic activity, RNA quality, expression of cryopreservation associated stress genes, and expression of OA characteristic disease genes was studied. Furthermore, the biological activity of the cryopreserved tissue was determined by measuring cytokine secretion induced by the TLR ligands lipopolysaccharides and Pam3Cys. Compared to non frozen synovium, no difference in cell and tissue morphology could be identified in the conditions using the CS10, standard and CryoSFM CPA solution for cryopreservation. However, we observed significantly lower preservation of tissue morphology with the Biofreeze and CS2 media. The other viability assays showed trends in the same direction but were not sensitive enough to detect significant differences between conditions. In all assays tested a clearly lower viability was detected in the condition in which synovium was frozen without CPA solution. This detailed analysis showed that OA synovial tissue explants can be cryopreserved while maintaining the morphology, viability and phenotypical response after thawing, offering enhanced opportunities for human in vitro studies.
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DOI：
10.1073/pnas.011404098
发表时间：
2001-01-02
影响因子：
11.1
作者：
Li, C;Wong, WH
通讯作者：
Wong, WH
DOI：
10.1016/s1387-2656(05)11004-7
发表时间：
2005-01-01
期刊：
BIOTECHNOLOGY ANNUAL REVIEW, VOL 11
影响因子：
--
作者：
Berridge, MV;Herst, PM;Tan, AS
通讯作者：
Tan, AS
影响因子：
64.8
作者：
Relling MV;Evans WE
通讯作者：
Evans WE
影响因子：
3.1
作者：
Gholami, Mohammadreza;Hemadi, Masoud;Mohammadi-asl, Javad
通讯作者：
Mohammadi-asl, Javad
影响因子：
3.2
作者：
Guan, Na;Blomsma, Sylvia A.;de Graaf, Inge A. M.
通讯作者：
de Graaf, Inge A. M.