Pettersson Sofia, Wetterö Jonas, Tengvall Pentti, Kratz Gunnar
Laboratory for Reconstructive Plastic Surgery, Department of Clinical and Experimental Medicine, Linköping University, Linköping SE-581 85, Sweden.
J Tissue Eng Regen Med. 2009 Aug;3(6):450-60. doi: 10.1002/term.179.
Biodegradable macroporous gelatin microcarriers fixed with blood-derived biodegradable glue are proposed as a delivery system for human autologous chondrocytes. Cell-seeded microcarriers were embedded in four biological glues-recalcified citrated whole blood, recalcified citrated plasma with or without platelets, and a commercially available fibrin glue-and cultured in an in vitro model under static conditions for 16 weeks. No differences could be verified between the commercial fibrin glue and the blood-derived alternatives. Five further experiments were conducted with recalcified citrated platelet-rich plasma alone as microcarrier sealant, using two different in vitro culture models and chondrocytes from three additional donors. The microcarriers supported chondrocyte adhesion and expansion as well as extracellular matrix (ECM) synthesis. Matrix formation occurred predominantly at sample surfaces under the static conditions. The presence of microcarriers proved essential for the glues to support the structural takeover of ECM proteins produced by the embedded chondrocytes, as exclusion of the microcarriers resulted in unstable structures that dissolved before matrix formation could occur. Immunohistochemical analysis revealed the presence of SOX-9- and S-100-positive chondrocytes as well as the production of aggrecan and collagen type I, but not of the cartilage-specific collagen type II. These results imply that blood-derived glues are indeed potentially applicable for encapsulation of chondrocyte-seeded microcarriers. However, the static in vitro models used in this study proved incapable of supporting cartilage formation throughout the engineered constructs.
固定有血液衍生的可生物降解胶水的可生物降解大孔明胶微载体被提议作为人自体软骨细胞的递送系统。接种细胞的微载体被嵌入四种生物胶中——重新钙化的枸橼酸盐全血、含或不含血小板的重新钙化的枸橼酸盐血浆，以及一种市售纤维蛋白胶——并在体外模型中在静态条件下培养16周。在市售纤维蛋白胶和血液衍生的替代物之间未发现差异。另外进行了五项实验，单独使用重新钙化的富含血小板的枸橼酸盐血浆作为微载体密封剂，使用两种不同的体外培养模型和来自另外三名供体的软骨细胞。微载体支持软骨细胞的黏附、扩增以及细胞外基质（ECM）的合成。在静态条件下，基质形成主要发生在样品表面。微载体的存在被证明对于胶水支持由嵌入的软骨细胞产生的ECM蛋白的结构接管至关重要，因为排除微载体导致结构不稳定，在基质形成之前就溶解了。免疫组织化学分析显示存在SOX-9和S-100阳性软骨细胞以及聚集蛋白聚糖和I型胶原蛋白的产生，但未发现软骨特异性II型胶原蛋白的产生。这些结果表明血液衍生的胶水确实有可能适用于封装接种软骨细胞的微载体。然而，本研究中使用的静态体外模型被证明无法支持整个工程构建体中的软骨形成。
