We developed a reliable and quantitative method for measuring the dynamic process of unidirectional two-dimensional (2-D) tissue formation of endothelial cells (ECs) in vitro. The culturing of bovine ECs in an assembled culture chamber provided a square monolayered cell sheet with a linear margin when disassembled at the confluency. The cell sheet maintained in culture showed a unidirectional endothelialization in vitro. The cell population-distance histogram, which was determined from the daily observation of tissue, allowed us to determine quantitatively the dynamic process of unidirectional endothelialization in vitro. The endothelialized distance and the endothelializing zone on a glass slide were found to be nearly 500 μm/day and 750 μm in width, respectively. Thus, the method developed here provided information of the 2-D tissue formation process. This model would be useful as an in vitro model which simulates the anastomotic endothelialization of an artificial vascular graft.
ClowesA.W.GownA.M.HansonS.R.ReidyM.A.Mechanisms of arterial graft failure: I.Role of cellular proliferation in early healing of PTFE prostheses. Am. J. Pathol.118: 43–54; 1985.
2.
ClowesA.W.KirkmanT.R.ReidyM.A.Mechanisms of arterial graft healing.Rapid transmural capillary ingrowth provides a source of intimal endothelium and smooth muscle in porous PTFE prostheses. Am. J. Pathol.123: 220–230; 1986.
3.
DekkerA.ReitsmaK.BeugelingT.BantjesA.FeijenJ.van AkenW.G.Adhesion of endothelial cells and adsorption of serum proteins on gas plasma-treated polytetrafluoroethylene.Biomaterials12: 130–138; 1990.
4.
FishmanJ.A.RyanG.B.KarnovskyM.J.Endothelial regeneration in the rat carotid artery and the significance of endothelial denudation in the pathogenesis of myointimal thickening.Lab. Invest.32: 339–351; 1975.
5.
FloreyH.W.GreerS.J.PooleJ.C.F.WerthessenN.T.The development of the pseudointima lining fabric grafts of the aorta.Br. J. Exp. Pathol.43: 655–660; 1962.
6.
GotliebA.I.SpectorW.Migration into an in vitro wound.A comparison of porcine aortic endothelial and smooth muscle cells and the effect of culture irradiation. Am. J. Pathol.103: 271–282; 1981.
7.
GrahamL.M.BurkelW.E.FordJ.W.VinterD.W.KahnR.H.StanleyJ.C.Immediate seeding of enzymatically derived endothelium in Dacron vascular grafts.Early experimental studies with autogenous canine cells. Arch. Surg.115: 1289–1294; 1980.
8.
HerringM.GardnerA.GloverJ.A single-staged technique for seeding vascular grafts with autogenous endothelium.Surgery84: 498–504; 1978.
9.
HodgkinJ.H.HeqthG.R.NorrisW.D.DonaldG.S.JohnsonG.Endothelial cell response to polyvinyl chloride-packaged GORETEX: Effect of surface contamination.Biomaterials11: 9–12; 1990.
10.
KaehlerJ.ZillaP.FasolR.DeutschM.KadletzM.Precoating substrate and surface configuration determine adherence and spreading of seeded endothelial cells on polytetrafluoroethylene grafts.J. Vase. Surg.9: 535–541; 1989.
11.
KeslerK.A.HerringM.B.ArnoldM.P.GloverJ.L.ParkH.HelmusM.N.BendickP.J.Enhanced strength of endothelial attachment on polyester elastomer and polytetrafluoroethylene graft surfaces with fibronectin substrate.J. Vase. Surg.3: 58–64: 1986.
12.
MadriJ.A.PrattB.M.Yannariello-BrownJ.Matrix-driven cell size change modulates aortic endothelial cell proliferation and sheet migration.Am. J. Pathol.132: 18–27; 1985.
13.
MadriJ.A.StennK.S.Aortic endothelial cell migration. I. Matrix requirements and composition.Am. J. Pathol.106: 180–186; 1982.
14.
NiuS.MatsudaT.OkaT.Endothelialization on various segmented polyurethanes: Cellular behavior and its substrate dependency.Trans. Am. Soc. Artif. Intern. Organs36: M164–M168; 1990.
15.
NiuS.MatsudaT.OkaT.In vitro Model of endothelialization at anastomotic sites.Trans. Am. Soc. Artif. Intern. Organs36: M757–M760; 1990.
16.
PedersonD.C.BowyerD.E.Endothelial injury and healing in vitro.Studies using an organ culture system. Am. J. Pathol.119: 264–272; 1985.
17.
RamalanjaonaG.KempczinskiR.F.RosenmanJ.E.DouvilleE.C.SilbersteinE.B.The effect of fibronectin coating on endothelial cell kinetics in polytetrafluoroethylene grafts.J. Vase. Surg.3: 264–272; 1986.
18.
SauvageL.R.Biologic behavior of grafts in arterial system. In: HaimoviciH.CallowA.D.DePalmaR.G.ErnstC.B.HollierL.H., eds. Haimovici's vascular surgery. Principles and technique, third ed.Norwalk, CT: Appleton & Lange;1989: 136–160.
19.
SholleyM.M.GimbroneM.A.CotranR.S.Cellular migration and replication in endothelial regeneration.A study using irradiated endothelial cultures. Lab. Invest.36: 18–25; 1977.
20.
van WachemP.B.BeugelingT.FeijenJ.BantjesA.DetmersJ.P.van AkenW.G.Interaction of the cultured human endothelial cells with polymeric surfaces of different wettabilities.Biomaterials6: 403–408; 1985.
21.
van WachemP.B.SchakenraadJ.M.FeijenJ.BeugelingT.van AkenW.G.BlaauwE.H.NieuwenhuisP.MolenaarI.Adhesion and spreading of cultured endothelial cells on modified and unmodified poly(ethylene terephthalate): A morphological study.Biomaterials10: 532–539; 1989.
