Tissue-Engineered Endothelial Cells Induce Sustained Vascular Healing Through Early Induction of Vascular Repair

Regenerative Engineering and Translational Medicine - 2023
Sahil A. Parikh1, Elazer R. Edelman2
1Columbia University Vagelos College of Physicians and Surgeons, 161 Fort Washington Avenue, 6th Floor, New York, NY, 10032, USA
2Harvard-MIT Division of Health Sciences and Technology, MIT E25-438, 77 Massachusetts Avenue, Cambridge, MA, 02139, USA

Tóm tắt

Abstract Background Perivascular implantation of tissue-engineered endothelial cells (TEEC) after vascular injury profoundly inhibits neointimal hyperplasia. However, the time course and mechanism by which this effect occurs remain unknown. By developing genetically modified TEEC that express a “suicide gene,” we can control the time during which the TEEC could exert their effect and determine the length of time TEEC need to be present following vascular injury to exert their inhibitory effect on long-term neointimal hyperplasia. Methods Bovine aortic endothelial cells (BAE) were transfected with the human herpes simplex virus thymidine kinase (tk) gene to render them sensitive to ganciclovir (GCV). These BAE+tk were grown to confluence on Gelfoam and shown to have the same growth kinetics and biologic potency as control cells but were sensitive to GCV at low concentrations. The BAE+tk were grown on Gelfoam and placed in the perivascular space around balloon-injured rat carotid arteries. Rats randomly received BAE-tk, BAE+tk, or nothing (control) after balloon injury. GCV was administered early (days 1–7), late (days 5–11), or not at all. Results Two weeks after injury, extensive neointimal hyperplasia was observed in control animals with an intima:media (I:M) area ratio of 0.80 ± 0.19. Early administration of GCV killed the BAE in constructs with TK sensitivity and eliminated the impact of TEEC regulation of intimal hyperplasia (0.45 ± 0.06). Intimal hyperplasia was still effectively reduced in animals with implants containing BAE-tk or BAE+tk which received GCV late (0.11 ± 0.04 and 0.19 ± 0.05). Immunohistochemistry demonstrated the lethal effect of GCV on TK-sensitive cells. Conclusions The application of perivascular TEEC for only the first few days after injury had a significant inhibitory effect on intimal hyperplasia. This is in contrast to the results obtained in this same animal model with the infusion of isolated anti-smooth muscle cell proliferative agents. This suggests that the mechanism of action of TEEC may be upstream from smooth muscle cell proliferation. Moreover, the use of this technique to further elucidate biologic mechanisms will prove invaluable in the tissue engineering field. Lay Summary We report a novel, genetically altered tissue-engineered endothelial cell (TEEC) implant that inhibits neointimal hyperplasia after experimental vascular injury. The viability of these implants can be carefully controlled and suggest a putative mechanism by which TEEC recapitulate control over the vascular response to injury.

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Tài liệu tham khảo

Langer R, Vacanti JP. Tissue engineering. Science. 1993;260(5110):920–6.

Nugent HM, Edelman ER. Tissue engineering therapy for cardiovascular disease. Circ Res. 2003;92(10):1068–78.

Nathan A, Nugent MA, Edelman ER. Tissue engineered perivascular endothelial cell implants regulate vascular injury. Proc Natl Acad Sci U S A. 1995;92(18):8130–4.

Han RO, et al. Heparin/heparan sulfate chelation inhibits control of vascular repair by tissue-engineered endothelial cells. Am J Physiol. 1997;273(6 Pt 2):H2586–95.

Nugent HM, Rogers C, Edelman ER. Endothelial implants inhibit intimal hyperplasia after porcine angioplasty. Circ Res. 1999;84(4):384–91.

Nugent MA, et al. Perlecan is required to inhibit thrombosis after deep vascular injury and contributes to endothelial cell-mediated inhibition of intimal hyperplasia. Proc Natl Acad Sci U S A. 2000;97(12):6722–7.

Nugent HM, Edelman ER. Endothelial implants provide long-term control of vascular repair in a porcine model of arterial injury. J Surg Res. 2001;99(2):228–34.

Nugent HM, et al. Perivascular endothelial implants inhibit intimal hyperplasia in a model of arteriovenous fistulae: a safety and efficacy study in the pig. J Vasc Res. 2002;39(6):524–33.

Garasic J, Rogers C, Edelman ER. In: Beyar R, et al., editors. Stent design and the biologic response, in Frontiers in Interventional Cardiology. London: Marin Dunitz Ltd.; 1997. p. 95–100.

Ozaki K, et al. Use of von Willebrand factor promoter to transduce suicidal gene to human endothelial cells, HUVEC. Hum Gene Ther. 1996;7(13):1483–90.

Yang Z, et al. Gene transfer approaches to the regulation of vascular cell proliferation. Semin Interv Cardiol. 1996;1(3):181–4.

Chang MW, et al. Adenovirus-mediated transfer of the herpes simplex virus thymidine kinase gene inhibits vascular smooth muscle cell proliferation and neointima formation following balloon angioplasty of the rat carotid artery. Mol Med. 1995;1(2):172–81.

Ohno T, et al. Gene therapy for vascular smooth muscle cell proliferation after arterial injury. Science. 1994;265(5173):781–4.

Clowes AW, Reidy MA, Clowes MM. Kinetics of cellular proliferation after arterial injury. I. Smooth muscle growth in the absence of endothelium. Lab Invest. 1983;49(3):327–33.

Clowes AW, Clowes MM. Kinetics of cellular proliferation after arterial injury. II. Inhibition of smooth muscle growth by heparin. Lab Invest. 1985;52(6):611–6.

Clowes AW, Schwartz SM. Significance of quiescent smooth muscle migration in the injured rat carotid artery. Circ Res. 1985;56(1):139–45.

Clowes AW, Clowes MM. Kinetics of cellular proliferation after arterial injury. IV. Heparin inhibits rat smooth muscle mitogenesis and migration. Circ Res. 1986;58(6):839–45.

Clowes AW, Clowes MM, Reidy MA. Kinetics of cellular proliferation after arterial injury. III. Endothelial and smooth muscle growth in chronically denuded vessels. Lab Invest. 1986;54(3):295–303.

Clowes AW, et al. Kinetics of cellular proliferation after arterial injury. V. Role of acute distension in the induction of smooth muscle proliferation. Lab Invest. 1989;60(3):360–4.

Costa MA, Simon DI. Molecular basis of restenosis and drug-eluting stents. Circulation. 2005;111(17):2257–73.

Methe H, et al. Matrix embedding alters the immune response against endothelial cells in vitro and in vivo. Circulation. 2005;112(9 Suppl):I89–95.

Methe H, Edelman ER. Tissue engineering of endothelial cells and the immune response. Transplant Proc. 2006;38(10):3293–9.

Methe H, Edelman ER. Cell-matrix contact prevents recognition and damage of endothelial cells in states of heightened immunity. Circulation. 2006;114(1 Suppl):I233–8.

Methe H, Hess S, Edelman ER. The effect of three-dimensional matrix-embedding of endothelial cells on the humoral and cellular immune response. Semin Immunol. 2008;20(2):117–22.

Zani BG, et al. Tissue-engineered endothelial and epithelial implants differentially and synergistically regulate airway repair. Proc Natl Acad Sci U S A. 2008;105(19):7046–51.

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Regenerative Engineering and Translational Medicine

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