Vascular smooth muscle cell optimization of vasculogenesis within naturally derived, biodegradable, hybrid hydrogel scaffolds.
Vascular smooth muscle cell optimization of vasculogenesis within naturally derived, biodegradable, hybrid hydrogel scaffolds.
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DOI：
10.1097/prs.0b013e3182a805df
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发表时间：
2013-12
影响因子：
3.6
通讯作者：
Spector JA
中科院分区：
文献类型：
--
作者：
Golas AR;Perez JL;Fullerton N;Lekic N;Hooper RC;Spector JA
由于血管化是永久结合基于水凝胶的组织再生模板的限速步骤，我们试图确定能够优化内皮细胞粘附和侵入定制水凝胶结构的材料化学。我们进一步研究了生长因子补充和旁分泌刺激对内皮小管形成的诱导作用。将海藻酸盐、I型胶原和壳聚糖组成的水凝胶支架植入人脐静脉内皮细胞（HUVEC），在标准条件下维持14d。然后评估细胞密度和侵袭情况。加入碱性成纤维细胞生长因子（bFGF）或与人主动脉平滑肌细胞（HASMC）共培养后，评估小管形成情况。海藻酸盐+胶原10:1 w/w支架的HUVECs细胞表面密度和浸润体积最大（p<0.05）。补充bFGF增加了表面密度，但既没有侵入也没有形成小管。随着HASMC:HUVEC比例的增加，微管含量/组织显著增加。与所研究的其他组合相比，海藻酸盐+胶原10:1支架允许最大的细胞化。补充生长因子不影响HUVEC侵袭或形态。通过与HASMC共培养的旁分泌信号刺激内皮小管形成和血管原网络组织。这些发现有助于指导我们未来的努力，以制造预血管化组织结构。
As vascularization represents the rate-limiting step in permanent incorporation of hydrogel-based tissue-regeneration templates, we sought to identify the material chemistry that would optimize endothelial cell adhesion and invasion into custom hydrogel constructs. We further investigated induction of endothelial tubule formation by growth factor supplementation and paracrine stimulation. Hydrogel scaffolds consisting of combinations of alginate, collagen type I, and chitosan were seeded with human umbilical vein endothelial cells (HUVEC) and maintained under standard conditions for 14d. Cell density and invasion were then evaluated. Tubule formation was evaluated following basic fibroblast growth factor (bFGF) addition or co-culture with human aortic smooth muscle cells (HASMC). HUVECs demonstrated greatest cell-surface density and invasion volumes with alginate+collagen 10:1 w/w scaffolds (p<0.05). Supplementation with bFGF increased surface-density but neither invasion nor tubule formation. A significant increase in tubule content/organization was observed with increasing HASMC:HUVEC ratio co-culture. Alginate+collagen 10:1 scaffolds allow for maximal cellularization compared with other combinations studied. Growth factor supplementation did not affect HUVEC invasion or morphology. Paracrine signaling via co-culture with HASMC stimulated endothelial tubule formation and vascular proto-network organization. These findings serve to guide our future endeavors towards fabrication of pre-vascularized tissue constructs.