Pomahac Bohdan, Zuhaili Baraa, Kudsi Yusef, Aflaki Pejman, Eriksson Elof
Division of Plastic Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Eplasty. 2009 Aug 28;9:e35.
Autologous chondrocyte transplantation (ACT) is the most commonly used cell-based surgical procedure for repair of articular cartilage defects. The challenges of this technique include dedifferentiation of chondrocytes following several in vitro passages, invasive means of transplantation, and inadequate cell retention leading to washout of transplanted cells. To overcome these obstacles, we developed a novel technique of transplanting high-density chondrocyte pellets seeded on a prefabricated, resorbable, rigid, 2-dimensional template amenable to minimally invasive implantation.
Chondrocytes were obtained from the costal cartilage of New Zealand white rabbits and expanded in vitro in monolayer culture. After 2 passages, chondrocyte suspension was centrifuged and a total of 1 x 10(6) cells condensed on the surface of a prefabricated, resorbable template of LactoSorb plate (0.5-mm thick, 4-mm diameter). The construct was incubated for 24 hours in a culture medium before transplantation into circular 4-mm diameter, 0.5-mm deep defects in a non-weight-bearing part of the femoral condyle. Control defects were left empty or implanted with LactoSorb alone. Macroscopic and histological evaluation was performed 4 weeks posttransplantation.
Macroscopically, boundaries of all defects were demarcated and distinguishable from adjacent intact cartilage. Regenerative tissue in experimental group appeared white, smooth, and uniform showing more resemblance to hyaline cartilage. Control groups revealed absent cartilaginous tissue and defects were filled with soft, fibrous tissue with an irregular surface. Histologically, the repair tissue in the control groups was fibroinflammatory with irregular surface and no evidence of continuous chondrocytic regeneration. Cartilage regeneration in the experimental defects revealed a continuous, high-density layer of chondrocytes surrounding the LactoSorb plates. Consistently with chondrocyte pellets grown for 4 weeks only, the amount of extracellular matrix deposition in the transplanted group was less than the normal cartilage.
We have developed a novel approach for ACT, utilizing high-density chondrocyte pellets seeded on a prefabricated, rigid, 2-dimensional resorbable carrier. Our study can serve as a model for further minimally invasive development of this technique and evaluating its potential role as an alternative in ACT.
自体软骨细胞移植(ACT)是修复关节软骨缺损最常用的基于细胞的外科手术。该技术面临的挑战包括软骨细胞在多次体外传代后去分化、侵入性移植方式以及细胞保留不足导致移植细胞被冲走。为克服这些障碍,我们开发了一种新技术,即移植接种在预制的、可吸收的、刚性二维模板上的高密度软骨细胞团,该模板适合微创植入。
从新西兰白兔的肋软骨获取软骨细胞,并在单层培养中进行体外扩增。传代2次后,将软骨细胞悬液离心,使总共1×10⁶个细胞聚集在预制的LactoSorb板可吸收模板(0.5毫米厚,4毫米直径)表面。构建物在培养基中孵育24小时后,移植到股骨髁非负重部位直径4毫米、深0.5毫米的圆形缺损处。对照缺损处不植入或仅植入LactoSorb。移植后4周进行宏观和组织学评估。
宏观上,所有缺损边界清晰,与相邻完整软骨可区分。实验组的再生组织呈白色、光滑且均匀,更类似于透明软骨。对照组未见软骨组织,缺损处充满表面不规则的柔软纤维组织。组织学上,对照组的修复组织为纤维炎性,表面不规则,无连续软骨细胞再生迹象。实验缺损处的软骨再生显示LactoSorb板周围有连续的高密度软骨细胞层。与仅培养4周的软骨细胞团一致,移植组细胞外基质沉积量少于正常软骨。
我们开发了一种新的ACT方法,利用接种在预制刚性二维可吸收载体上的高密度软骨细胞团。我们的研究可作为该技术进一步微创开发以及评估其作为ACT替代方法潜在作用的模型。