Baker B M, Nathan A S, Huffman G Russell, Mauck R L
McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, University of Pennsylvania, Philadelphia, PA 19104, United States.
Osteoarthritis Cartilage. 2009 Mar;17(3):336-45. doi: 10.1016/j.joca.2008.08.001. Epub 2008 Oct 10.
Injuries to the avascular regions of the meniscus fail to heal and so are treated by resection of the damaged tissue. This alleviates symptoms but fails to restore normal load transmission in the knee. Tissue engineering functional meniscus constructs for re-implantation may improve tissue repair. While numerous studies have developed scaffolds for meniscus repair, the most appropriate autologous cell source remains to be determined. In this study, we hypothesized that the debris generated from common meniscectomy procedures would possess cells with potential for forming replacement tissue. We also hypothesized that donor age and the disease status would influence the ability of derived cells to generate functional, fibrocartilaginous matrix.
Meniscus derived cells (MDCs) were isolated from waste tissue of 10 human donors (seven partial meniscectomies and three total knee arthroplasties) ranging in age from 18 to 84 years. MDCs were expanded in monolayer culture through passage 2 and seeded onto fiber-aligned biodegradable nanofibrous scaffolds and cultured in a chemically defined media. Mechanical properties, biochemical content, and histological features were evaluated over 10 weeks of culture.
Results demonstrated that cells from every donor contributed to increasing biochemical content and mechanical properties of engineered constructs. Significant variability was observed in outcome parameters (cell infiltration, proteoglycan and collagen content, and mechanical properties) amongst donors, but these variations did not correlate with patient age or disease condition. Strong correlations were observed between the amount of collagen deposition within the construct and the tensile properties achieved. In scaffolds seeded with particularly robust cells, construct tensile moduli approached maxima of approximately 40 MPa over the 10-week culture period.
This study demonstrates that cells derived from surgical debris are a potent cell source for engineered meniscus constructs. Results further show that robust growth is possible in MDCs from middle-aged and elderly patients, highlighting the potential for therapeutic intervention using autologous cells.
半月板无血管区域的损伤无法自愈,因此通过切除受损组织进行治疗。这可缓解症状,但无法恢复膝关节的正常负荷传递。用于重新植入的组织工程功能性半月板构建体可能会改善组织修复。虽然众多研究已开发出用于半月板修复的支架,但最合适的自体细胞来源仍有待确定。在本研究中,我们假设常见半月板切除术产生的碎片会含有具有形成替代组织潜力的细胞。我们还假设供体年龄和疾病状态会影响所衍生细胞生成功能性纤维软骨基质的能力。
从10名年龄在18至84岁的人类供体(7例部分半月板切除术和3例全膝关节置换术)的废弃组织中分离出半月板衍生细胞(MDCs)。MDCs在单层培养中传代至第2代并接种到纤维排列的可生物降解纳米纤维支架上,然后在化学成分明确的培养基中培养。在10周的培养过程中评估其力学性能、生化成分和组织学特征。
结果表明,每个供体的细胞都有助于增加工程构建体的生化成分和力学性能。在供体之间观察到结果参数(细胞浸润、蛋白聚糖和胶原蛋白含量以及力学性能)存在显著差异,但这些差异与患者年龄或疾病状况无关。在构建体内观察到胶原蛋白沉积量与所达到的拉伸性能之间存在强相关性。在接种了特别强壮细胞的支架中,构建体的拉伸模量在10周培养期内接近约40 MPa的最大值。
本研究表明,手术碎片衍生的细胞是工程化半月板构建体的有效细胞来源。结果进一步表明,中年和老年患者的MDCs有可能实现强劲生长,突出了使用自体细胞进行治疗干预的潜力。