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短期酶处理对细胞迁移和软骨再生的影响:牛关节软骨的体外器官培养

Effect of short-term enzymatic treatment on cell migration and cartilage regeneration: in vitro organ culture of bovine articular cartilage.

作者信息

Seol Dongrim, Yu Yin, Choe Hyeonghun, Jang Keewoong, Brouillette Marc J, Zheng Hongjun, Lim Tae-Hong, Buckwalter Joseph A, Martin James A

机构信息

1 Department of Orthopaedics and Rehabilitation, The University of Iowa , Iowa City, Iowa.

出版信息

Tissue Eng Part A. 2014 Jul;20(13-14):1807-14. doi: 10.1089/ten.TEA.2013.0444. Epub 2014 May 20.

Abstract

Depending on the damage extent and adjacent tissue condition in traumatic cartilage injury, it is possible to heal the tissue by resident cells. Unlike autologous chondrocyte implantation, short-term enzymatic treatment is an effective single-step procedure without extra cell expansion. Moreover, this method has been shown to significantly increase cellularity in lesion edges, resulting in enhanced integration and interfacial strength. We hypothesize that the locally digested extracellular matrix by treatment allows effortless cell migration from the adjacent tissue. Full-thickness cartilage discs and osteochondral explants were prepared from mature bovine stifle joints. These specimens were treated with collagenase in a culture medium. Two concentrations, 0.25 and 0.5 mg/mL, were used with various treating time of 10, 30, and 180 min. The cartilages were subsequently washed and cultured with fibrin hydrogel. The effect of enzymatic treatment on cell migration was apparent in both experiments of the cartilage disc and full-thickness cartilage defect model. In the disc culture, the treatment resulted in an approximately three to four times higher number of migrated cells than nontreated control. In short-term collagenase-treated groups, the proteoglycan (PG) loss was localized in the edge of tissue with minimal cell death. The treatment also accelerated cell migration in the full-thickness cartilage defects and some cells differentiated into chondrocytes with the deposit of PG. Gene expression results could support the characteristics of migrated cells, which had migratory ability and chondrogenic differentiation potential with overexpression of collagen type I and II, respectively. Based on these results, short-term enzymatic treatment, which can accelerate cell migration into traumatically injured cartilage, has great potential for clinical application.

摘要

根据创伤性软骨损伤的损伤程度和相邻组织状况,受损组织有可能通过自身细胞实现愈合。与自体软骨细胞移植不同,短期酶处理是一种有效的单步操作,无需额外的细胞扩增。此外,该方法已被证明能显著增加损伤边缘的细胞数量,从而增强整合度和界面强度。我们推测,通过处理局部消化的细胞外基质可使细胞轻松地从相邻组织迁移过来。从成熟的牛膝关节制备全层软骨盘和骨软骨外植体。这些标本在培养基中用胶原酶处理。使用了0.25和0.5毫克/毫升两种浓度,处理时间分别为10、30和180分钟。随后对软骨进行冲洗,并用纤维蛋白水凝胶培养。在软骨盘和全层软骨缺损模型的实验中,酶处理对细胞迁移的影响都很明显。在盘培养中,处理后的迁移细胞数量比未处理的对照组高出约三到四倍。在短期胶原酶处理组中,蛋白聚糖(PG)的损失局限于组织边缘,细胞死亡极少。该处理还加速了全层软骨缺损处的细胞迁移,一些细胞分化为软骨细胞并伴有PG沉积。基因表达结果能够支持迁移细胞的特性,即分别通过I型和II型胶原的过表达而具有迁移能力和成软骨分化潜能。基于这些结果,能够加速细胞迁移至创伤性损伤软骨的短期酶处理在临床应用中具有巨大潜力。

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