Ashinsky Beth G, Gullbrand Sarah E, Bonnevie Edward D, Wang Chao, Kim Dong Hwa, Han Lin, Mauck Robert L, Smith Harvey E
Translational Musculoskeletal Research Center, Corporal Michael J. Crescenz VA Medical Center, Philadelphia, PA; McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, University of Pennsylvania, Philadelphia, PA; School of Biomedical Engineering and Sciences, Drexel University, Philadelphia, PA.
Translational Musculoskeletal Research Center, Corporal Michael J. Crescenz VA Medical Center, Philadelphia, PA; McKay Orthopaedic Research Laboratory, Department of Orthopaedic Surgery, University of Pennsylvania, Philadelphia, PA.
Acta Biomater. 2020 Jul 15;111:232-241. doi: 10.1016/j.actbio.2020.05.019. Epub 2020 May 22.
Tissue-engineered replacement discs are an area of intense investigation for the treatment of end-stage intervertebral disc (IVD) degeneration. These living implants can integrate into the IVD space and recapitulate native motion segment function. We recently developed a multiphasic tissue-engineered disc-like angle-ply structure (DAPS) that models the micro-architectural and functional features of native tissue. While these implants resulted in functional restoration of the motion segment in rat and caprine models, we also noted deficiencies in cell infiltration and homogeneity of matrix deposition in the electrospun poly(ε-caprolactone) outer region (annulus fibrosus, AF) of the DAPS. To address this limitation, here, we incorporated a sacrificial water-soluble polymer, polyethylene oxide (PEO), as a second fiber fraction within the AF region to increase porosity of the implant. Maturation of these PEO-modified DAPS were evaluated after 5 and 10 weeks of in vitro culture in terms of AF biochemical content, MRI T2 values, overall construct mechanical properties, AF micromechanical properties and cell and matrix distribution. To assess the performance of the PEO-modified DAPS in vivo, precultured constructs were implanted into the rat caudal IVD space for 10 weeks. Results showed that matrix distribution was more homogenous in PCL/PEO DAPS, as evidenced by more robust histological staining, organized collagen deposition and micromechanical properties, compared to standard PCL-only DAPS in vitro. Cell and matrix infiltration were also improved in vivo, but no differences in macromechanical properties and a trend towards improved micromechanical properties were observed. These findings demonstrate that the inclusion of a sacrificial PEO fiber fraction in the DAPS AF region improves cellular colonization, matrix elaboration, and in vitro and in vivo function of an engineered IVD implant. STATEMENT OF SIGNIFICANCE: This work establishes a method for improving cell infiltration and matrix distribution within tissue-engineered dense fibrous scaffolds for intervertebral disc replacement. Tissue-engineered whole disc replacements are an attractive alternative to the current gold standard (mechanical disc arthroplasty or vertebral fusion) for the clinical treatment of patients with advanced disc degeneration.
组织工程化替代椎间盘是治疗终末期椎间盘(IVD)退变的一个深入研究领域。这些活植入物能够整合到IVD空间并重现天然运动节段的功能。我们最近开发了一种多相组织工程化盘状角向铺层结构(DAPS),它模拟了天然组织的微观结构和功能特征。虽然这些植入物在大鼠和山羊模型中实现了运动节段的功能恢复,但我们也注意到在DAPS的电纺聚(ε-己内酯)外层区域(纤维环,AF)中细胞浸润和基质沉积的均匀性存在不足。为了解决这一局限性,在此我们在AF区域引入了一种牺牲性水溶性聚合物聚环氧乙烷(PEO)作为第二种纤维成分,以增加植入物的孔隙率。在体外培养5周和10周后,从AF生化成分、MRI T2值、整体构建体力学性能、AF微观力学性能以及细胞和基质分布方面评估这些PEO修饰的DAPS的成熟情况。为了评估PEO修饰的DAPS在体内的性能,将预培养的构建体植入大鼠尾侧IVD空间10周。结果表明,与仅含标准PCL的DAPS相比,在体外PCL/PEO DAPS中的基质分布更均匀,更强烈的组织学染色、有序的胶原沉积和微观力学性能证明了这一点。在体内细胞和基质浸润也得到了改善,但未观察到宏观力学性能的差异,且微观力学性能有改善的趋势。这些发现表明,在DAPS的AF区域引入牺牲性PEO纤维成分可改善工程化IVD植入物的细胞定植、基质形成以及体外和体内功能。重要性声明:这项工作建立了一种改善组织工程化致密纤维支架内细胞浸润和基质分布以用于椎间盘置换的方法。组织工程化全椎间盘置换是晚期椎间盘退变患者临床治疗中当前金标准(机械性椎间盘置换术或椎体融合术)的一种有吸引力的替代方案。