Matrix-assisted Autologous Chondrocyte Transplantation for Remodeling and Repair of Chondral Defects in a Rabbit Model

Articular cartilage defects are considered a major health problem because articular cartilage has a limited capacity for self-regeneration 1. Untreated ...

Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

The treatment of osteochondral articular defects has been challenging physicians for many years. The better understanding of interactions of articular ...

Klinikum rechts der Isar der Technischen Universität München

Matrix-assisted chondrocyte transplantation (m-ACI) still lacks any standardization in its execution in terms of cell passage (P), cell yield (C) and in vitro membrane-holding time (T). It was the goal of this study to analyze the effect of shifting cell culture parameters (P, C, T) on the in vitro as well as in vivo effort of a regulated animal m-ACI. Autologous rabbit knee articular chondrocytes were seeded within bilayer collagen I/III 3-D matrices in variation of P, C and T. Each time, 2 PCT-identical by 2 PCT-identical cell-matrix-constructs (CMC)/animal were created. Simultaneously 2 (PCT-distinct) were re-implanted (CMC-e) autologous into artificial trochlear pristine chondral defects in vivo to remain for 12 weeks while the remaining 2 were harvested (CMC-i) for immediate in vitro analysis at the time of transplantation of their identical twins. mRNA of both, CMC-e regenerates and CMC-i membranes, was analyzed for Collagen-1,-2,-10, COMP, Aggrecan, Sox9 expression by use of a mixed linear model, multiple regression analysis. Generally, CMC-i values were higher than CMC-e values for differentiation targets; the opposite was true for dedifferentiation targets. Regarding individual gene expression, in vivo regenerate cell-matrix properties were significantly dependent on initial cell-matrix in vitro values as a sign of linearity. The parameter membrane-holding time (T) had strongest effects on the resulting mRNA expression with slightly less impact of the parameter passage (P), whereas cell yield (C) had clearly less effects. Noting differences between in vitro and in vivo data, in general, optimal expression patterns concerning chondrogenic differentiation were achieved by few passages, medium cellular yield, short membrane-holding time. Clinical m-ACI may benefit from optimal orchestration of the cell culture parameters passage, yield and time.

The dependence of autologous chondrocyte transplantation on varying cellular passage, yield and culture duration.

Greffe de chondrocytes assist&eacute;e par matrice (m-ACI) manque encore toute normalisation dans son ex&eacute;cution en termes de passage (P) de la cellule, rendement cellulaire (C) et temps de membrane-retenue in vitro (T). C&#39;&eacute;tait le but de cette &eacute;tude d&#39;analyser l&#39;effet de d&eacute;placer les param&egrave;tres de culture cellulaire (P, C, T) sur l&#39;effort in vitro comme in vivo d&#39;une animale r&eacute;glement&eacute;e m-ACI. Lapin autologue genou chondrocytes articulaires ont &eacute;t&eacute; ensemenc&eacute;es dans le collag&egrave;ne de la bicouche je / matrices 3D III en variation de P, C et T. Chaque fois, 2 PCT identiques par 2 PCT-identique cellule-matrice-constructions (CMC) / animal ont &eacute;t&eacute; cr&eacute;&eacute;s. En m&ecirc;me temps 2 (PCT-distinctes) ont &eacute;t&eacute; r&eacute;implant&eacute;s (MCC-e) autologues d&eacute;fauts artificiels chondrales vierges trochl&eacute;en in vivo de rester pour 12 semaines alors que les 2 autres ont &eacute;t&eacute; r&eacute;colt&eacute;s (MCC-i) pour l&#39;analyse in vitro imm&eacute;diate au moment de la transplantation de leurs jumeaux identiques. ARNm des deux, r&eacute;g&eacute;n&egrave;re MCC-e et les membranes de CMC-i, a &eacute;t&eacute; analys&eacute; pour collag&egrave;ne-1, -2, -10, COMP, AGGRECANE, Sox9 expression par l&#39;utilisation d&#39;un mod&egrave;le lin&eacute;aire mixte, analyse de r&eacute;gression multiple. G&eacute;n&eacute;ralement, valeurs de CMC-i &eacute;taient plus &eacute;lev&eacute;s que les valeurs de MCC-e pour les cibles de diff&eacute;renciation ; le contraire est vrai pour les cibles de d&eacute;diff&eacute;renciation. Au sujet de l&#39;expression des g&egrave;nes individuels, propri&eacute;t&eacute;s de la matrice cellulaire in vivo r&eacute;g&eacute;n&eacute;rer d&eacute;pendaient significativement cellule-matrice initiale en vitro valeurs comme un signe de la lin&eacute;arit&eacute;. Le temps de membrane-retenue de param&egrave;tre (T) ont des effets plus forts sur l&#39;expression des ARNm obtenue avec l&eacute;g&egrave;rement moins de cons&eacute;quences du passage du param&egrave;tre (P), alors que le rendement de la cellule (C) avait nettement moins d&#39;effets. Tenant compte des diff&eacute;rences entre les donn&eacute;es in vitro et in vivo, en g&eacute;n&eacute;ral, les profils d&#39;expression optimale concernant chondrog&eacute;niques distinction ont &eacute;t&eacute; r&eacute;alis&eacute;es par quelques passages, moyen rendement cellulaire, membrane-tenue peu de temps. Clinique m-ACI peut-&ecirc;tre b&eacute;n&eacute;ficier d&#39;une orchestration optimale du passage de param&egrave;tres cell culture, rendement et le temps.

La d&eacute;pendance de la greffe de chondrocytes autologues adapter la dur&eacute;e de passage, de rendement et de la culture cellulaire.

Trapianto di condrociti Matrix-assisted (m-ACI) manca ancora qualsiasi standardizzazione nella sua esecuzione in termini di passaggio cella (P), rendimento di cella (C) e in vitro membrana-azienda tempo (T). E &#39; stato l&#39;obiettivo di questo studio per analizzare l&#39;effetto di spostare i parametri di coltura delle cellule (P, C, T) sullo sforzo oltre in vitro che in vivo di un animale regolamentato m-ACI. Coniglio autologo ginocchio condrociti articolari sono stati seminati all&#39;interno di collagene doppio strato I / matrici tridimensionali III variazione di P, C e T. Ogni volta, 2 PCT-identici da 2 PCT-identica cellula-matrice-costrutti (CMC) / animale sono stati creati. Contemporaneamente 2 (PCT-distinti) erano re-implanted (CMC-e) autologo in artificiale trocleare incontaminate condrali in vivo di rimanere per 12 settimane, mentre i restanti 2 sono stati raccolti (CMC-i) per l&#39;analisi in vitro immediata al momento del trapianto dei loro gemelli identici. mRNA di entrambi, rigenera e CMC e membrane CMC-i, &egrave; stato analizzato per collagene-1, -2, -10, COMP, Aggrecan, Sox9 espressione dall&#39;uso di un modello lineare misto, analisi di regressione multipla. In genere, CMC-i valori erano superiori valori di CMC-e per gli obiettivi di differenziazione; l&#39;opposto &egrave; stato vero per le destinazioni di indifferenziazione. Per quanto riguarda l&#39;espressione del gene individuali, rigenera in vivo delle cellule-matrice propriet&agrave; erano significativamente dipendente iniziale cellula-matrice in vitro valori come un segno della linearit&agrave;. Il tempo di tenuta di membrana di parametro (T) aveva pi&ugrave; forti effetti sull&#39;espressione del mRNA risultante con leggermente meno impatto del passaggio di parametri (P), considerando che il rendimento della cella (C) aveva chiaramente meno effetti. Notando le differenze tra i dati in vitro e in vivo, in generale, i modelli di espressione ottimale relativa differenziazione chondrogenic sono stati raggiunti da alcuni passaggi, resa media cellulare, breve tempo di membrana-azienda. Clinica m-ACI possono beneficiare di orchestrazione ottimale del passaggio di parametri cultura cellulare, resa e tempo.

Markus T. Berninger

Department of Orthopaedic Sports Medicine,
Department of Radiology

Klinikum rechts der Isar der Technischen Universität München

Matrix-assisted Autologous Chondrocyte Transplantation for Remodeling and Repair of Chondral Defects in a Rabbit Model

Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

Markus T. Berninger

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Orthopaedic Sports Medicine,Department of Radiology

Klinikum rechts der Isar der Technischen Universität München

Matrix-assisted Autologous Chondrocyte Transplantation for Remodeling and Repair of Chondral Defects in a Rabbit Model

Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

Department of Orthopaedic Sports Medicine,
Department of Radiology