Nehrer S, Breinan H A, Ramappa A, Hsu H P, Minas T, Shortkroff S, Sledge C B, Yannas I V, Spector M
Department of Orthopedic Surgery, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Biomaterials. 1998 Dec;19(24):2313-28. doi: 10.1016/s0142-9612(98)00143-4.
The objective of our study was to evaluate reparative tissues formed in chondral defects in an adult canine model implanted with cultured autologous articular chondrocytes seeded in type I and II collagen GAG matrices. Two defects were produced in the trochlea grooves of the knees of 21 dogs, with cartilage removed down to the tidemark. This study includes the evaluation of 36 defects distributed among five treatment groups: Group A, type II collagen matrix seeded with autologous chondrocytes under a sutured type II collagen flap; Group B, type I collagen matrices seeded with chondrocytes under a sutured fascia flap; Group C, unseeded type I collagen matrix implanted under a sutured fascia flap; Group D, fascia lata flap alone; and Group E, untreated defects. All animals were killed 15 weeks after implantation. Six other defects were created at the time of death and evaluated immediately after production as 'acute defect controls'. In three additional defects, unseeded matrices were sutured to the defect and the knee closed and reopened after 30 min to determine if early displacement of the graft was occurring; these defects served as 'acute implant controls'. The areal percentages of four tissue types in the chondral zone of the original defect were determined histomorphometrically: fibrous tissue (FT); hyaline cartilage (HC); transitional tissue (TT, including fibrocartilage); and articular cartilage (AC). New tissue formed in the remodeling subchondral bone underlying certain defects was also assessed. Bonding of the repair tissue to the subchondral plate and adjacent cartilage, and degradation of the adjacent tissues were evaluated. There were no significant differences in the tissues filling the original defect area of the sites treated with chondrocyte-seeded type I and type II matrices. Most of the tissue in the area of the original defect in all of the groups was FT and TT. The areal percentage of HC plus AC was highest in group E, with little such tissue in the cell-seeded groups, and none in groups C and D. The greatest total amount of reparative tissue, however, was found in the cell-seeded type II matrix group. Moreover, examination of the reparative tissue formed in the subchondral region of defects treated with the chondrocyte-seeded collagen matrices (Groups A and B) demonstrated that the majority of the tissue was positive for type II collagen and stained with safranin O. These results indicate an influence of the exogenous chondrocytes on the process of chondrogenesis in this site. In all groups with implants (A-D), 30(50% of the FT and TT was bonded to the adjacent cartilage. Little of this tissue (6-22%) was attached to the subchondral plate, which was only about 50% intact. Remarkable suture damage was found in sections from each group in which sutures were used. Harvest sites showed no regeneration of normal articular cartilage, 18 weeks after the biopsy procedure. Future studies need to investigate other matrix characteristics, and the effects of cell density and incubation of the seeded sponges prior to implantation on the regenerative response.
我们研究的目的是评估在植入接种于I型和II型胶原糖胺聚糖基质中的培养自体关节软骨细胞的成年犬模型中,软骨缺损处形成的修复组织。在21只犬的膝关节滑车沟处制造两个缺损,将软骨去除至潮标处。本研究包括对分布在五个治疗组中的36个缺损的评估:A组,在缝合的II型胶原皮瓣下接种自体软骨细胞的II型胶原基质;B组,在缝合的筋膜皮瓣下接种软骨细胞的I型胶原基质;C组,在缝合的筋膜皮瓣下植入未接种的I型胶原基质;D组,单独的阔筋膜张肌皮瓣;E组,未治疗的缺损。所有动物在植入后15周处死。在处死时制造另外六个缺损,并在制造后立即作为“急性缺损对照”进行评估。在另外三个缺损中,将未接种的基质缝合到缺损处,膝关节闭合,30分钟后重新打开以确定移植物是否发生早期移位;这些缺损用作“急性植入对照”。通过组织形态计量学确定原始缺损软骨区域中四种组织类型的面积百分比:纤维组织(FT);透明软骨(HC);过渡组织(TT,包括纤维软骨);和关节软骨(AC)。还评估了某些缺损下方重塑软骨下骨中形成的新组织。评估修复组织与软骨下板和相邻软骨的结合以及相邻组织的降解。接种软骨细胞的I型和II型基质治疗部位的原始缺损区域填充的组织没有显著差异。所有组中原始缺损区域的大部分组织是FT和TT。E组中HC加AC的面积百分比最高,接种细胞的组中这种组织很少,C组和D组中没有。然而,在接种细胞的II型基质组中发现的修复组织总量最大。此外,对用接种软骨细胞的胶原基质(A组和B组)治疗的缺损软骨下区域形成的修复组织的检查表明,大部分组织II型胶原呈阳性并被番红O染色。这些结果表明外源性软骨细胞对该部位软骨形成过程有影响。在所有植入组(A-D)中,30%(FT和TT的50%)与相邻软骨结合。这种组织很少(6-22%)附着在软骨下板上,软骨下板仅约50%完整。在使用缝线的每组切片中都发现了明显的缝线损伤。活检程序18周后,取材部位未显示正常关节软骨再生。未来的研究需要研究其他基质特性,以及植入前接种海绵的细胞密度和孵育对再生反应的影响。