Morphological Reconstruction of Main Arteries by Perivascular Implantation of Sulfated Chitosan in Experimental Atherosclerosis

The aim of the study is to demonstrate morphological reconstruction of the main arteries of rabbit hind limbs by perivascular implantation of sulfated chitosan in an atherosclerosis model.

Materials and Methods. The study was performed on 24 rabbits divided into four groups of 6 animals. The rabbits in the experimental group kept on intensive cholesterol diet for 110 days were implanted 1% water-soluble sulfated chitosan gel in the perivascular fascial compartment of the saphenous artery and the femoral artery of the left hind limb. Some animals that were also kept on cholesterol diet did not undergo implantation. The group of rabbits fed with normal vivarium diet were subjected to perivascular implantation of the biopolymer, while a group of 6 rabbits also kept on normal diet served as the intact control. We estimated wall thickness of the femoral artery and the saphenous artery, their average lumen diameter, the mean lumen area of the newly formed vessels in the adventitia.

Results. Daily intensive cholesterol diet for 3.5 months in rabbits leads to vivid signs of atherogenic inflammation forming in the intimal and medial layers of the main arteries of the hind limbs. Introduction of 1% water-soluble sulfated chitosan gel into the paravasal compartment of the saphenous and femoral arteries promotes formation of a large number of microvessels at the polymer resorption site increasing the specific area of new vessels. Morphological reconstruction of the main arteries is achieved through reducing the vascular wall thickness, increasing the vessel lumen and the number of para-adventitial microvessels.

Conclusion. Implantation of sulfated chitosan into the perivascular compartment allows achieving the effect of therapeutic paravasal angiogenesis and inhibiting the early signs of atherogenic inflammation.

1. Aronov D.M. Social aspects of atherosclerosis and methods of its treatment. Russkiy meditsinskiy zhurnal 2000; 7: 276.
2. Andozhskaya Yu.S., Girina M.B, Vasina E.Yu. Present-day methods of microcirculation estimate in efferent therapy of patients with atherosclerosis. Regionarnoe krovoobrashchenie i mikrotsirkulyatsiya 2002; 1(1): 52–59.
3. Karpov Yu.A., Sorokin E.V. Intensive medical treatment of patients with atherosclerosis. Kardiologiya 2005; 45(8): 4–7.
4. Klimenko E.D., Kobozeva L.P., Michunskaia A.B., Khabarina I.Iu. Function of the microcirculatory system during the long-term regression of the early stages of atherogenesis. Biull Eksp Biol Med 1988; 105(3): 365–368.
5. Dua A., Lee J.C. Epidemiology of peripheral arterial disease and critical limb ischemia. Tech Vasc Interv Radiol 2016; 19(2): 91–95, https://doi.org/10.1053/j.tvir.2016.04.001 .
6. Auger F.A., D’Orléans-Juste P., Germain L. Adventitial contribution to vascular contraction: hints provided by tissue-engineered substitutes. Cardiovasc Res 2007; 75(4): 669–678, https://doi.org/10.1016/j.cardiores.2007.06.001.
7. Laflamme K., Roberge C.J., Grenier G., Rémy-Zolghadri M., Pouliot S., Baker K., Labbé R., D’Orléans-Juste P., Auger F.A., Germain L. Adventitia contribution in vascular tone: insights from adventitia-derived cells in a tissue-engineered human blood vessel. FASEB J 2006; 20(8): 1245–1247, https://doi.org/10.1096/fj.05-4702fje.
8. Ni W., Kitamoto S., Ishibashi M., Usui M., Inoue S., Hiasa K., Zhao Q., Nishida K., Takeshita A., Egashira K. Monocyte chemoattractant protein-1 is an essential inflammatory mediator in angiotensin II-induced progression of established atherosclerosis in hypercholesterolemic mice. Arterioscler Thromb Vasc Biol 2004; 24(3): 534–539, https://doi.org/10.1161/01.atv.0000118275.60121.2b.
9. Nugent H.M., Sjin R.T., White D., Milton L.G., Manson R.J., Lawson J.H., Edelman E.R. Adventitial endothelial implants reduce matrix metalloproteinase-2 expression and increase luminal diameter in porcine arteriovenous grafts. J Vasc Surg 2007; 46(3): 548–556, https://doi.org/10.1016/j.jvs.2007.04.074.
10. Pagano P.J., Gutterman D.D. The adventitia: the outs and ins of vascular disease. Cardiovasc Res 2007; 75(4): 636–639, https://doi.org/10.1016/j.cardiores.2007.07.006.
11. Rey F.E., Pagano P.J. The reactive adventitia: fibroblast oxidase in vascular function. Arterioscler Thromb Vasc Biol 2002; 22(12): 1962–1971, https://doi.org/10.1161/01.atv.0000043452.30772.18.
12. Bolshakov I.N., Shestakova L.A., Kotikov A.R., Kaptyuk G.I. Experimental atherosclerosis in rats. morphological reconstruction of the main artery wall with the polyssacharide biopolymers. Fundamental’nye issledovaniya 2013; 10–3: 557–563.
13. Kim S., Kawai T., Wang D., Yang Y. Engineering a dual-layer chitosan-lactide hydrogel to create endothelial cell aggregate-induced microvascular networks in vitro and increase blood perfusion in vivo. ACS Appl Mater Interfaces 2016; 8(30): 19245–19255, https://doi.org/10.1021/acsami.6b04431.
14. Lee S., Valmikinathan C.M., Byun J., Kim S., Lee G., Mokarram N., Pai S.B., Um E., Bellamkonda R.V., Yoon Y.S. Enhanced therapeutic neovascularization by CD31-expressing cells and embryonic stem cell-derived endothelial cells engineered with chitosan hydrogel containing VEGF-releasing microtubes. Biomaterials 2015; 63: 158–167, https://doi.org/10.1016/j.biomaterials.2015.06.009.
15. Bolshakov I.N., Dolgikh O.A., Kirichenko A.K., Kotikov A.R., Gorbunova V.O. Lipid spectrum and microcirculation when using biopolymers in an atherogenesis model. Fundamental’nye issledovaniya 2009; S7: 41–42.
16. Klimov A.N., Parfenova N.S., Golikov Y.P. One century of the cholesterol model of atherosclerosis. Biomeditsinskaya khimiya 2012; 58(1): 5–11.
17. Bolshakov I.N., Shestakova L.A., Kotikov A.R., Kaptyuk G.I. The experimental atherosclerotic inflammation of the main arteries in rabbits. Low traumatic technology of morphological reconstruction of the vascular wall at the early atherosclerotic stages. Fundamental’nye issledovaniya 2013; 8–2: 343–350.
18. Dzyak G.V., Koval’ E.L. Atherosclerosis and inflammation. Problema stareniya i dolgoletiya 1999; 3: 316–326.
19. Liu M.H., Tang Z.H., Li G.H., Qu S.L., Zhang Y., Ren Z., Liu L.S., Jiang Z.S. Janus-like role of fibroblast growth factor 2 in arteriosclerotic coronary artery disease: аtherogenesis and angiogenesis. Atherosclerosis 2013; 229(1): 10–17, https://doi.org/10.1016/j.atherosclerosis.2013.03.013.

Kirichenko A.K., Shulmin A.V., Sharkova A.F., Patlataya N.N., Bolshakov I.N. Morphological Reconstruction of Main Arteries by Perivascular Implantation of Sulfated Chitosan in Experimental Atherosclerosis. Sovremennye tehnologii v medicine 2017; 9(4): 115, https://doi.org/10.17691/stm2017.9.4.14