Bioengineering vascularized tissue constructs using an injectable cell-laden enzymatically crosslinked collagen hydrogel derived from dermal extracellular matrix.
Bioengineering vascularized tissue constructs using an injectable cell-laden enzymatically crosslinked collagen hydrogel derived from dermal extracellular matrix.
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DOI：
10.1016/j.actbio.2015.09.002
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发表时间：
2015-11
影响因子：
9.7
通讯作者：
Chen YC
中科院分区：
文献类型：
--
作者：
Kuo KC;Lin RZ;Tien HW;Wu PY;Li YC;Melero-Martin JM;Chen YC
组织工程有望通过创造功能性和可移植的人工组织来恢复或替换患病或受损的组织。具有超过扩散限制的大尺寸的人造组织的开发将需要营养物和氧气经由灌注而不是在短时间段内单独扩散来递送。灌注的一种方法是使工程化组织血管化，在组织构建体内创建从头三维（3D）微血管网络。这显著缩短了体内吻合、灌注和移植物与宿主整合的时间。在这项研究中，我们的目的是开发可注射同种异体胶原蛋白-酚羟基（胶原蛋白-Ph）水凝胶，能够控制广泛的物理化学性质，包括刚度，吸水性和降解性。我们测试了胶原蛋白-Ph水凝胶是否可以支持人血来源的内皮细胞集落形成细胞（ECFC）和骨髓来源的间充质干细胞（MSC）在体内形成血管化的工程组织移植物。首先，我们研究了贴壁ECFC和MSC在水凝胶上或水凝胶中的生长。为了检查体内功能性血管网络的潜在形成，将含有人ECFC和MSC、辣根过氧化物酶和过氧化氢的胶原蛋白-Ph的液体预聚物溶液注射到免疫缺陷小鼠的皮下空间或腹部肌肉缺损中，然后凝胶化，以形成3D载有细胞的聚合构建体。这些结果表明，可以在7天内产生广泛的人ECFC内衬的血管网络，胶原蛋白-Ph水凝胶构建体内的工程化血管密度可以通过可精制的机械性能和蛋白水解降解性来操纵，并且这些网络可以与现有的血管系统形成功能性的血管网，以进一步支持宿主肌肉组织的存活。最后，优化条件的细胞负载的胶原蛋白-Ph水凝胶导致不仅改善移植的MSC向矿化成骨细胞的长期分化，而且胶原蛋白-Ph水凝胶还改善了在植入1个月后小鼠中血管化生物工程组织内脂肪细胞的增加。
Tissue engineering promises to restore or replace diseased or damaged tissue by creating functional and transplantable artificial tissues. The development of artificial tissues with large dimensions that exceed the diffusion limitation will require nutrients and oxygen to be delivered via perfusion instead of diffusion alone over a short time period. One approach to perfusion is to vascularize engineered tissues, creating a de novo three-dimensional (3D) microvascular network within the tissue construct. This significantly shortens the time of in vivo anastomosis, perfusion and graft integration with the host. In this study, we aimed to develop injectable allogeneic collagen-phenolic hydroxyl (collagen-Ph) hydrogels that are capable of controlling a wide range of physicochemical properties, including stiffness, water absorption and degradability. We tested whether collagen-Ph hydrogels could support the formation of vascularized engineered tissue graft by human blood-derived endothelial colony-forming cells (ECFCs) and bone marrow-derived mesenchymal stem cells (MSC) in vivo. First, we studied the growth of adherent ECFCs and MSCs on or in the hydrogels. To examine the potential formation of functional vascular networks in vivo, a liquid pre-polymer solution of collagen-Ph containing human ECFCs and MSCs, horseradish peroxidase and hydrogen peroxide was injected into the subcutaneous space or abdominal muscle defect of an immunodeficient mouse before gelation, to form a 3D cell-laden polymerized construct. These results showed that extensive human ECFC-lined vascular networks can be generated within 7 days, the engineered vascular density inside collagen-Ph hydrogel constructs can be manipulated through refinable mechanical properties and proteolytic degradability, and these networks can form functional anastomoses with the existing vasculature to further support the survival of host muscle tissues. Finally, optimized conditions of the cell-laden collagen-Ph hydrogel resulted in not only improving the long-term differentiation of transplanted MSCs into mineralized osteoblasts, but the collagen-Ph hydrogel also improved an increased of adipocytes within the vascularized bioengineered tissue in a mouse after 1 month of implantation.