Martins Edivaldo A N, Michelacci Yara M, Baccarin Raquel Y A, Cogliati Bruno, Silva Luis C L C
Departamento de Bioquímica, Escola Paulista de Medicina, UNIFESP, Rua Três de Maio, 100, São Paulo, 04044-020, SP, Brazil.
BMC Vet Res. 2014 Aug 27;10:197. doi: 10.1186/s12917-014-0197-4.
Articular cartilage, because of its avascular nature, has little capacity for spontaneous healing, and tissue engineering approaches, employing different biomaterials and cells, are under development. Among the investigated biomaterials are the chitosan-based hydrogels. Although thoroughly studied in other mammalian species, studies are scarce in equines. So, the aim of the present study was to investigate the biocompatibility of chitosan-GP in horse joints submitted to high mechanical loads.
An osteochondral defect was created by arthroscopy in the medial surface of lateral trochlea of talus of left or right leg, randomly selected, from six healthy geldings. The defect was filled up with chitosan-GP. The contralateral joint received an identical defect with no implant. The chondral fragment removed to produce the defect was collected, processed and used as the "Initial" sample (normal cartilage) for histology, immunohistochemistry, and metabolic labelling of PGs. After 180 days, the repair tissues were collected, and also analyzed. At the end of the experiment (180 days after lesion), the total number of cells per field in repair tissues was equal to control, and macrophages and polymorphonuclear cells were not detected, suggesting that no significant inflammation was present. These cells were able to synthesize type II collagen and proteoglycans (PGs). Nevertheless, the cell population in these tissues, both in presence of chitosan-GP and in untreated controls, were heterogeneous, with a lower proportion of type II collagen-positives cells and some with a fibroblastic aspect. Moreover, the PGs synthesized in repair tissues formed in presence or absence of chitosan-GP were similar to those of normal cartilage. However, the chitosan-GP treated tissue had an disorganized appearance, and blood vessels were present.
Implanted chitosan-GP did not evoke an important inflammatory reaction, and permitted cell growth. These cells were able to synthesize type II collagen and PGs similar to those synthesized in normal cartilage and in healing tissue without implant, indicating its chondrocyte nature.
关节软骨由于其无血管的特性,自发愈合能力有限,目前正在研发采用不同生物材料和细胞的组织工程方法。在已研究的生物材料中,基于壳聚糖的水凝胶备受关注。尽管在其他哺乳动物物种中已进行了深入研究,但在马类中的研究却很少。因此,本研究的目的是探讨壳聚糖 - GP在承受高机械负荷的马关节中的生物相容性。
从六匹健康的去势公马中随机选择左或右下肢距骨外侧滑车内侧表面,通过关节镜制造骨软骨缺损。缺损处填充壳聚糖 - GP。对侧关节制造相同缺损但不植入任何物质。移除制造缺损时取下的软骨碎片,进行处理后用作组织学、免疫组织化学和蛋白聚糖代谢标记的“初始”样本(正常软骨)。180天后,收集修复组织并进行分析。在实验结束时(损伤后180天),修复组织中每视野的细胞总数与对照组相等,未检测到巨噬细胞和多形核细胞,表明不存在明显炎症。这些细胞能够合成II型胶原蛋白和蛋白聚糖(PGs)。然而,这些组织中的细胞群体,无论是存在壳聚糖 - GP还是未处理的对照组,都是异质性的,II型胶原蛋白阳性细胞比例较低,有些细胞呈现成纤维细胞形态。此外,在有无壳聚糖 - GP的情况下形成的修复组织中合成的PGs与正常软骨中的相似。然而,壳聚糖 - GP处理的组织外观紊乱,且有血管存在。
植入的壳聚糖 - GP未引发重要的炎症反应,并允许细胞生长。这些细胞能够合成与正常软骨和无植入物的愈合组织中合成的类似II型胶原蛋白和PGs,表明其具有软骨细胞性质。