Regeneration of Cartilaginous Tissue After Transplantation of Allofibroblasts

Aim of investigation. To prove the usefulness of the use of embryonic fibroblasts for the regeneration of cartilaginous tissue.

Material and methods. New experimental model of symphysis pubis rupture followed by its recovery was employed in 24 male Shinshilla rabbits (2.3—3.5 kg body mass) in 3 series of experiments. In the first series, reconstruction was not employed. In the 2nd series, the reconstruction of symphisis was performed with the aid of tantal wired cerclage (d=1.2 mm). In the 3d series, the reconstitution was the same as in the previous series of experiments with addition of grown embrional allofibroblast tissue pieces. Duration of experiments was 7, 14, 21and 30 days. After each experiment, pieces of cartilaginous tissue were fixed with 10% neutral formalin and embedded in paraffin. Histological sections were stained with hematoxylin, eosin and VanGieson.

Results. After the rupture of symphysis pubis of the pelvic bone, the cartilaginous tissue undergone alterations followed by formation of chondromatous nodules of different sizes and sclerosis of surrounding muscular and osseous tissues. In the second series of experiment after pelvis symphysis pubis rupture and symphysis recovery with the help of cerclage, the morphologic investigation revealed the formation of mixed initial callus consisting of connective and chondromatous tissue elements. In the third series of experiment it has been shown that within 4 weeks of experiment, the connective tissue with intensive basophil stain is formed in the area of symphysis pubisrupture. Connective tissue formation in surrounding soft tissue structures was associated with the formation of cartilaginous cells.

Conclusion. Process of formation of cartilaginous tissue in the area of symphysis pubis rupture as evident by the experimental observation demonstrates the stimulating influence of allogeneic fibroblasts on process of cartilaginous tissue regeneration that ensures premises for clinical development of suggested method of regeneration.

1. Avrushchenko M.S., Ostrova I.V., Volkov A.V. Postreanimatsionnye izmeneniya ekspressii glialnogo neirotroficheskogo faktora (GDNF): vzaimosvyaz s povrezhdeniem kletok Purkinye mozzhechka (eksperimentalnoe issledovanie). Obshchaya Reanimatologiya. [Postresuscitation changes in the expression of Glial Derived Neurotrophic Factor (GDNF): association with cerebellar Purkinje cell damage (an experimental study). General Reanimatology]. 2014; 10 (5): 59—68. http://dx.doi.org/10.15360/18139779201455968. [In Russ.]

2. Savilov P.N., Molchanov D.V. Vliyanie giperbaricheskoi oksigenatsii na ammiakekskretiruyushchuyu funktsiyu pochek pri rezektsii pecheni v eksperimente. Obshchaya Reanimatologiya. [Impact of hyperbaric oxygenation on renal ammonia excretion during experimental liver resection. General Reanimatology]. 2015; 11 (2): 56—63. http://dx.doi.org/10.15360/18139779201525663. [In Russ.]

3. Mahmoudi Rad M., Talebpour Amiri F., Mirhoseini M., Ghasemi M., Mirzaei M., Mosaffa N. Application of allogeneic fibroblast cultured on acellular amniotic membrane for fullthickness wound healing in rats. Wounds. 2016; 28 (1): 14—19. PMID: 26779806

4. Larsen M., Willems W.F., Pelzer M., Friedrich P.F., Dadsetan M., Bishop A.T. Fibroblast growth factor2 and vascular endothelial growth factor mediated augmentation of angiogenesis and bone formation in vascularized bone allotransplants. Microsurgery. 2014; 34 (4): 301—307. http://dx.doi.org/10.1002/micr.22221. PMID: 24395434

5. Sindberg G.M., Lindborg B.A., Wang Q., Clarkson C., Graham M., Donahue R., Hering B.J., Verfaillie C.M., BansalPakala P., O’Brien T.D. Comparisons of phenotype and immunomodulatory capacity among rhesus bonemarrowderived mesenchymal stem/stromal cells, multipotent adult progenitor cells, and dermal fibroblasts. J. Med. Primatol. 2014; 43 (4): 231—241. http://dx.doi.org/10.1111/jmp.12122. PMID: 24825538

6. Larijani B., Ghahari A., Warnock G.L., Aghayan H.R., Goodarzi P., Falahzadeh K., Arjmand B. Human fetal skin fibroblasts: Extremely potent and allogenic candidates for treatment of diabetic wounds. Med. Hypotheses. 2015; 84 (6): 577—579. http://dx.doi.org/10.1016/j.mehy.2015.03.004. PMID: 25784640

7. Teklemariam T., Zhao L., Hantash B.M. Heterologous expression of mutated HLAG1 reduces alloreactivity of human dermal fibroblasts. Regen. Med. 2014; 9 (6): 775—784. http://dx.doi.org/10.2217/rme.14.58. PMID: 25431913

8. Arsenyev I.G., Berchenko D.S., Urazgildeyev G.N. Aktualnye voprosy klinicheskoi i eksperimentalnoi meditsiny. [Actual problems of clinical and experimental medicine]. SanktPeterburg; 2007: 379—381. [In Russ.]

9. Bobro L.I. Fibroblasty i ikh znachenie v tkanevykh reaktsiyakh. [Fibroblasts and their significance in tissue reactions]. Arkhiv Patologii. 1990; 52 (12): 65—68. PMID: 2091587. [In Russ.]

10. Gololobov V.G. Regeneratsiya kostnoi tkani pri zazhivlenii ognestrelnykh perelomov. [Regeneration of bone tissue in the healing of gunshot fractures]. SanktPeterburg: Peterburg — XXI vek; 1997: 160. [In Russ.]

11. Eremeyev A.V., Svetlakov A.V., Bolshakov I.N., Vlasov A.A., Arapova V.A. Funktsii kultiviruemykh embrionalnykh kletok na kollagenkhi tozanovoi matritse. [Functions of cultured embryonic stem cells on collagenchitosan matrix]. Kletochnaya Transplantologiya i Tkanevaya Inzheneriya. 2009; 4 (2): 55—62. [In Russ.]

12. Ozerskaya O.S., Shchegolev V.V. Eksperimentalnye podkhody k obosnovaniyu primeneniya kletochnykh kompozitsii na osnove fibrob lastov v dermatokosmetologii. [Experimental approaches to the justification of the use of cellbased compositions of fibroblasts in dermatology]. Kletochnaya Transplantologiya i Tkanevaya Inzheneriya. 2008; 3 (2): 66—67. [In Russ.]

13. Shved Yu.L., Kukhareva L.V., Zorin I.M., Solovyev A.Yu., Blinova M.I., Bilibin A.Yu., Pinaev G.P. Razrabotka polimernoi podlozhki dlya kultivirovaniya fibroblastov kozhi cheloveka. [Elaboration of biodegrad able polymer substrate for cultivation of human dermal fibroblasts]. Tsitologiya. 2006; 2: 161—168. PMID: 16737184. [In Russ.]

14. Kassin V.Yu., Aboyants R.K., Mironova L.L., Konyushko O.I., Nikolaev M.P., Vasilyev A.V., Zhitkov I.L. Eksperimentalnoe izuchenie reaktsii soedinitelnoi tkani na implantatsiyu gidrogelei s inkorporirovannymi fibroblastami. [Experimental study on reaction of connective tissue to implanted fibroblast containing hydrogels]. Biotechnology in Russia. 2004; 6: 99—106. [In Russ.]

15. Rasulov M.F. Ispolzovanie mezenkhimalnykh stvolovykh kletok kostnogo mozga i embrionalnykh fibroplastov v lechenii ozhogovykh ran. [Application of bone marrow mesenchimal stem cells and embryonal fibroblasts during burn wounds treatment]. Tikhookeansky Meditsinsky Zhurnal. 2004; 1: 7—9. [In Russ.]

16. Sonobe M., Hattori K., Tomita N., Yoshikawa Т., Aoki H., Takakura Y., Suguro T. Stimulatory effects of statins on bone marrowderived mesenchymal stem cells. Study of a new therapeutic agent for fracture. Biomed. Mater. Eng. 2005; 15 (4): 261—267. PMID: 16010034

17. Lin C., Sun J.S., Hou S.M. External fixation with or without supple mentary intramedullary Kirschner wires in the treatment of distalradial fractures. Can. J. Surg. 2004; 47 (6): 431—437. PMID: 15646442

18. Mangano C., Bartolucci E.G., Mazzocco C. A new porous hydroxyapatite for promotion of bone regeneration in maxillary sinus augmentation: clinical and histologic study in humans. Int. J. Oral Maxillofac. Implants. 2003; 18 (1): 23—30. PMID: 12608665

19. Qu S.X., Guo X., Weng J., Cheng J.C.Y., Feng В., Yeung H.Y., Zhang X.D. Evaluation of the expression of collagen type I in porous calcium phosphate ceramics implanted in an extraosseous site. Biomaterials. 2004; 25 (4): 659—667. PMID: 14607504

20. Urazmetova M.D., Shek A.A. Sposob lecheniya ozhogov i ran. Patent Respubliki Uzbekistan №4995. [A method of treating burns and wounds. Patent of the Republic of Uzbekistan №4995]. 1998. [In Russ.]