Möller Thomas, Amoroso Matteo, Hägg Daniel, Brantsing Camilla, Rotter Nicole, Apelgren Peter, Lindahl Anders, Kölby Lars, Gatenholm Paul
3D Bioprinting Centre, Department of Chemistry and Chemical Engineering, Chalmers University of Technology, Göteborg, Sweden; Gothenburg University, Sahlgrenska Academy, Institute of Clinical Sciences, Department of Plastic Surgery, Sahlgrenska University Hospital, Göteborg, Sweden; Department of Clinical Chemistry and Transfusion Medicine, Institute of Biomedicine, Sahlgrenska University Hospital, Göteborg, Sweden; and University Medical Center Ulm, Department of Otorhinolaryngology, Frauensteige 12, 89075 Ulm, Germany.
Plast Reconstr Surg Glob Open. 2017 Feb 15;5(2):e1227. doi: 10.1097/GOX.0000000000001227. eCollection 2017 Feb.
The three-dimensional (3D) bioprinting technology allows creation of 3D constructs in a layer-by-layer fashion utilizing biologically relevant materials such as biopolymers and cells. The aim of this study is to investigate the use of 3D bioprinting in a clinically relevant setting to evaluate the potential of this technique for in vivo chondrogenesis.
Thirty-six nude mice (Balb-C, female) received a 5- × 5- × 1-mm piece of bioprinted cell-laden nanofibrillated cellulose/alginate construct in a subcutaneous pocket. Four groups of printed constructs were used: (1) human (male) nasal chondrocytes (hNCs), (2) human (female) bone marrow-derived mesenchymal stem cells (hBMSCs), (3) coculture of hNCs and hBMSCs in a 20/80 ratio, and (4) Cell-free scaffolds (blank). After 14, 30, and 60 days, the scaffolds were harvested for histological, immunohistochemical, and mechanical analysis.
The constructs had good mechanical properties and keep their structural integrity after 60 days of implantation. For both the hNC constructs and the cocultured constructs, a gradual increase of glycosaminoglycan production and hNC proliferation was observed. However, the cocultured group showed a more pronounced cell proliferation and enhanced deposition of human collagen II demonstrated by immunohistochemical analysis.
In vivo chondrogenesis in a 3D bioprinted human cell-laden hydrogel construct has been demonstrated. The trophic role of the hBMSCs in stimulating hNC proliferation and matrix deposition in the coculture group suggests the potential of 3D bioprinting of human cartilage for future application in reconstructive surgery.
三维（3D）生物打印技术能够利用生物聚合物和细胞等生物相关材料，以逐层方式创建3D构建体。本研究的目的是在临床相关环境中研究3D生物打印的应用，以评估该技术在体内软骨形成方面的潜力。
36只裸鼠（雌性，Balb-C）在皮下囊袋中植入一块5×5×1毫米的生物打印载细胞纳米纤维纤维素/藻酸盐构建体。使用了四组打印构建体：（1）人（男性）鼻软骨细胞（hNCs），（2）人（女性）骨髓间充质干细胞（hBMSCs），（3）hNCs和hBMSCs以20/80比例共培养，以及（4）无细胞支架（空白）。在14天、30天和60天后，收获支架进行组织学、免疫组织化学和力学分析。
构建体具有良好的力学性能，植入60天后仍保持其结构完整性。对于hNC构建体和共培养构建体，均观察到糖胺聚糖产量和hNC增殖逐渐增加。然而，免疫组织化学分析表明，共培养组显示出更明显的细胞增殖和人胶原蛋白II沉积增强。
已证明在3D生物打印的载人类细胞水凝胶构建体中可实现体内软骨形成。hBMSCs在共培养组中刺激hNC增殖和基质沉积的营养作用表明，人类软骨的3D生物打印在未来重建手术中的应用潜力巨大。
