Variation of Mesenchymal Cells in Polylactic Acid Scaffold in an Osteochondral Repair Model
Variation of Mesenchymal Cells in Polylactic Acid Scaffold in an Osteochondral Repair Model
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DOI：
10.1089/ten.tec.2008.0487
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发表时间：
2009-12-01
影响因子：
3
通讯作者：
Amiel, David
中科院分区：
文献类型：
--
作者：
Oshima, Yasushi;Harwood, Frederick L.;Amiel, David
目的：利用软骨系细胞和适当的支架移植实现骨软骨再生，对移植细胞在修复过程中的行为进行表征至关重要。本研究的目的是阐明间充质细胞（MCs）的存活。在聚乳酸（PLA）支架中，评估MC/PLA结构用于骨软骨修复的可能性。设计：将成熟雄性兔骨髓培养2周，获得含有间充质干细胞（MSCs）的成纤维细胞样MCs。使用旋转装置制备了100万个mc和可生物降解的PLA核心的细胞/支架结构。培养1周后，将该结构体移植到母兔股骨内侧髁骨软骨缺损处，组织学观察其愈合过程。为了检测移植MCs的存活能力，采用聚合酶链反应（PCR）技术评估了男性来源的性别决定区Y （SRY）基因作为MCs缺陷的标志物。结果：未作任何处理的缺损组和不含细胞的聚乳酸移植组缺损均未得到透明软骨修复。红花素O染色可识别软骨基质，免疫组化染色可识别II型胶原，但移植后24周缺损内仍存在聚乳酸基质。随着时间的推移，移植的MCs数量从1周的7.8 × 105个减少到4周的3.5 × 105个，12周的3.8 × 104个。24周时未检测到移植的MCs。结论：表达软骨基质的MCs参与骨软骨修复，但随着时间的延长（即24周），MCs的数量逐渐减少。这些结果可能是通过生长因子和/或基因治疗的细胞移植策略实现软骨再生所必需的。
Objective: To achieve osteochondral regeneration utilizing transplantation of cartilage-lineage cells and adequate scaffolds, it is essential to characterize the behavior of transplanted cells in the repair process. The objectives of this study were to elucidate the survival of mesenchymal cells (MCs). In a polylactic acid (PLA) scaffold and assess the possibility of MC/PLA constructs for osteochondral repair.Design: Bone marrow from mature male rabbits was cultured for 2 weeks, and fibroblast-like MCs, which contain mesenchymal stem cells (MSCs), were obtained. A cell/scaffold construct was prepared with one million MCs and a biodegradable PLA core using a rotator device. One week after culturing, the construct was transplanted into an osteochondral defect in the medial femoral condyle of female rabbits and the healing process examined histologically. To examine the survivability of transplanted MCs, the male-derived sex-determining region Y (SRY) gene was assessed as a marker of MCs in the defect by polymerase chain reaction (PCR).Results: In the groups of defects without any treatment, and the transplantation of PLA without cells, the defects were not repaired with hyaline cartilage. The cartilaginous matrix by safranin O staining and type II collagen by immunohistochemical staining were recognized, however the PLA matrix was still present in the defects at 24 weeks after transplantation of the construct. During the time passage, transplanted MCs numbers decreased from 7.8 x 105 at 1 week, to 3.5 x 105 at 4 weeks, and to 3.8 x 104 at 12 weeks. Transplanted MCs were not detectable at 24 weeks.Conclusions: MCs contribute to the osteochondral repair expressing the cartilaginous matrix, however the number of MCs were decreasing with time (i.e. 24 weeks). These results could be essential for achieving cartilage regeneration by cell transplantation strategies with growth factors and/or gene therapy.