Reparation of chondral defects in rabbits by autologous and allogenous chondrocytes seeded on collagen/hyaluronan scaffold or suspended in fibrin glue

Abstract

The topic of the study was to verify in vivo survival of in vitro cultured autologous and allogenous chondrocytes suspended in a fi brin glue Beriplast® or seeded on Collagen type I-Hyaluronan (Col type I-HYA) scaffolds for the regeneration of articular cartilage defects in rabbits. The study was carried out on 15 domestic rabbits randomly assigned to fi ve groups (n = 3 in each) with different treatments of artifi cially created chondral defects (ChD ́s). These defects were made in a non-load-bearing area of medial condyle of the distal femur, and were treated as follows: 1st and 3rd group: the ChD ́s were fi lled with autologous or allogenous chondrocytes seeded on Col type I-HYA scaffolds, respectively. The scaffolds were fi xed to the ChD ́s by fi brin glue Beriplast®; 2nd and 4th group: the ChD ́s were fi lled with a suspension of autologous or allogenous chondrocytes in fi brin glue Beriplast®, respectively, and they were immediately covered by unseeded Col type I-HYA scaffolds; Control group: the ChD ́s were left to heal spontaneously without any treatment. Macroscopical, histological and immunohistochemical analyses of the ChD ́s were performed 12 months after the treatment. In all treated groups, the chondrocytes were capable to proliferate and produce the cartilage extracellular matrix, including proteoglycans and type II collagen, as compared to the control “untreated” group. On the other hand, the production of hyaline-like cartilage tissue confi rmed that both therapeutic methods using autologous chondrocytes can be applied successfully for the treatment of chondral defects in rabbits.

Cilj studije je bio da se verifi kuje in vivo preživljavanje in vitro kultivisanih autolognih i alogenih hondrocita suspendovanih u fi brinski Beriplast® lepak ili na kolagen Hyaluronan tip I (Col type I-HYA) osnovu radi regeneracije oštećenja zglobne hrskavice kod kunića. Ispitivanje je obavljeno na 15 domaćih kunića, odabranih slučajnim izborom koji su podeljeni u pet grupa (3 po grupi) kod kojih su veštački izazvana oštećenja hrskavice (ChD), tretirana na različite načine. Oštećenja su napravljena na površinama koje nisu noseće na medijalnom kondilusu distalnog femura, a tretman je podrazumevao: prva i treća grupa: ChD su punjene sa autolognim ili alogenim hondrocitima zasejanim na Col tip I-HYA osnovu. Osnove su fi ksirane na ChD pomoću fi brinskog lepka Beriplast®; druga i četvrta grupa: oštećenja su punjena suspenzijom autolognih ili alogenih hondrocita u fi brinskom Beriplast® lepku, a neposredno posle toga, prekriveni su sa nezasejanom Col tip I-HYA osnovom; kontrolna grupa: oštećenja su ostavljena da spontano zarastu, bez tretmana. Dvanaest meseci posle tretmana obavljene su makroskopske, histološke i imunohistološke analize promena. Kod svih životinja tretiranih grupa, hondrociti su bili sposobni da proliferišu i proizvode ekstracelularni matriks hrskavice, uključujući proteoglikane i II tip kolagena u poređenju sa kontrolnom “netretriranom” grupom. Sa druge strane, proizvodnja hrskavičavog tkiva nalik na hijalin potvrđuje da se oba terapijska metoda koji koriste autologne hondrocite mogu da uspešno primene u tretiranju defekata hrskavice kod kunića.