Summary. The aim of our study was to determine the effectiveness of cultured articular chondrocytes in agarose hydrogel (early follow-up, 3 months) in the restoration of simulated cartilaginous defects of the knee of critical size in large animals.
Materials and Methods. The experimental series consisted of 4 dogs (right hind limb) with created defects of articular cartilage deeper than the subchondral bone in the area of the intercondylar notch of the femur with a diameter of 3 mm and 5 mm deep. At the second stage of the experiment, agarose hydrogel with cultured allogeneic chondrocytes was implanted (sealed) into the injured joint intraarticularly. The control series consisted of 4 animals (left hind limb) without agarose hydrogel with cultured allogeneic chondrocytes implantation to the intercondylar notch of the femur. Agarose hydrogel with cultured allogeneic chondrocytes was implanted by application into the defects of the articular surfaces after curettage of the bottom and walls of bone and cartilage defects of the articular surfaces. Cell preparations and cultures were microscopically magnified at 40, 100, 200, and 320 times. Histological sections 10-15 μm thick, made using a cryotome, were stained with hematoxylin-eosin and hematoxylin-picrofuxin according to van Gizon.
Results. The technique of enzymatic isolation of chondrocytes articular cartilage of a dog was optimized. The satisfactory efficacy of cultured allogeneic articular chondrocytes in agarose hydrogel (early observation) in the treatment of simulated bone and cartilage defects of critical size in large animals within the planned observation period is shown.
Conclusions. The experimental study explored the effectiveness of the use of cultured allogenic articular chondrocytes embedded in agarose hydrogel (early follow-up period) in the restoration of modeled knee cartilage defects of critical size in dogs within the planned follow-up period. For the selected follow-up period (3 months), the signs of regenerate formation, which was characterized by its immaturity and the presence of a mixture of fibrous connective tissue and fibrous cartilage with the chondroid structure inclusions, were revealed in experimental group.
Brittberg M, Lindahl A, Nilsson A, Ohlsson C, Isaksson O, Peterson L. Treatment of deep cartilage defects in the knee with autologous chondrocyte transplantation N. Engl. J. Med. 1994;331(14):889-895. DOI: 10.1056/NEJM199410063311401.
Brittberg M. Gersoff W. Cartilage surgery: an operative manual. Elsevier Saunders; 2011. 320 p.
Deev RV. Market analysis of cell preparations for the correction of skeletal tissue pathology. Klet. transpl. i tkan. inzhener. 2006;2(4):78-83. [in Russian].
Wakitani S, Goto T, Pineda SJ, Young RG, Mansour JM, Caplan AI, et al. Mesenchymal cell-based repair of large, full-thickness defects of articular artilage J. Bone Joint Surg. Am. 1994;76(4):579-592. DOI: 10.2106/00004623-199404000-00013.
Wakitani S, Imoto K, Yamamoto T, Saito M, Murata N, Yoneda M, et al. Human autologous culture expanded bone marrow mesenchymal cell transplantation for repair of cartilage defects in osteoarthritic knees. Osteoarthritis Cartilage. 2002;10(3):199-206. DOI: 10.1053/joca.2001.0504.
Quantavalla J, Uzeil-Fusi S, Yin J. Boehnlein E, Pastor G, Blancuzzi V, et al. Fluorescently labeled mesenchymal stem cells (MSCs) maintain multilineage potential and can be detected following implantation into articular cartilage defects Biomaterials. 2002;23(1):109-19. DOI: 10.1016/S0142-9612(01)00086-2.
Prockop DJ, Phinney DG, Bunnell BA. Mesenchymal stem cells: methods and protocols. Totowa, NJ: Humana Press; 2008. 192 p.
Freshni Ya. Animal cell culture: a practical guide. Per. 5-go angl. izd. M.: BINOM. Laboratoriya znaniy; 2010. 691 s. [in Russian].
Hurtig MB, Buschmann MD, Fortier LA, Hoemann CD, Hunziker EB, Jurvelin JS, et al. Preclinical studies for cartilage repair: Recommendations from the International Cartilage Repair Society (ICRS Recommendation Papers). Cartilage. 2011;2(2):137-152. DOI: 10.1177%2F1947603511401905.
This work is licensed under a Creative Commons Attribution 4.0 International License.