Mouthuy P-A, El-Sherbini Y, Cui Z, Ye H
Institute of Biomedical Engineering, Department of Engineering Science, University of Oxford, UK.
J Tissue Eng Regen Med. 2016 Apr;10(4):E263-74. doi: 10.1002/term.1765. Epub 2013 Jun 11.
It is now widely acknowledged that implants that have been designed with an effort towards reconstructing the transition between tissues might improve their functionality and integration in vivo. This paper contributes to the development of improved treatment for articular cartilage repair by exploring the potential of the combination of electrospinning technology and cell sheet engineering to create cartilage tissue. Poly(lactic-co-glycolic acid) (PLGA) was used to create the electrospun membranes. The focus being on the cartilage-bone transition, collagen type I and hydroxyapatite (HA) were also added to the scaffolds to increase the histological biocompatibility. Human mesenchymal stem cells (hMSCs) were cultured in thermoresponsive dishes to allow non-enzymatic removal of an intact cell layer after reaching confluence. The tissue constructs were created by layering electrospun membranes with sheets of hMSCs and were cultured under chondrogenic conditions for up to 21 days. High viability was found to be maintained in the multilayered construct. Under chondrogenic conditions, reverse-transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry have shown high expression levels of collagen type X, a form of collagen typically found in the calcified zone of articular cartilage, suggesting an induction of chondrocyte hypertrophy in the PLGA-based scaffolds. To conclude, this paper suggests that layering electrospun scaffolds and cell sheets is an efficient approach for the engineering of tissue transitions, and in particular the cartilage-bone transition. The use of PLGA-based scaffold might be particularly useful for the bone-cartilage reconstruction, since the differentiated tissue constructs seem to show characteristics of calcified cartilage.
现在人们普遍认识到，经过精心设计以重建组织间过渡的植入物可能会改善其在体内的功能和整合。本文通过探索静电纺丝技术与细胞片工程相结合创造软骨组织的潜力，为改进关节软骨修复治疗方法做出了贡献。聚乳酸-羟基乙酸共聚物（PLGA）被用于制造静电纺丝膜。由于重点是软骨-骨过渡，还向支架中添加了I型胶原蛋白和羟基磷灰石（HA）以提高组织学相容性。人间充质干细胞（hMSCs）在热响应培养皿中培养，汇合后可通过非酶法去除完整的细胞层。通过将静电纺丝膜与hMSCs片层叠来构建组织，并在软骨形成条件下培养长达21天。发现多层构建体中保持了高活力。在软骨形成条件下，逆转录-聚合酶链反应（RT-PCR）和免疫组织化学显示X型胶原蛋白表达水平很高，X型胶原蛋白是一种通常在关节软骨钙化区发现的胶原蛋白形式，这表明基于PLGA的支架中诱导了软骨细胞肥大。总之，本文表明将静电纺丝支架和细胞片层叠是组织过渡工程，特别是软骨-骨过渡工程的有效方法。基于PLGA的支架的使用可能对骨-软骨重建特别有用，因为分化的组织构建体似乎显示出钙化软骨的特征。
