Regenerative Potential of Decellularized Porcine Nucleus Pulposus Hydrogel Scaffolds: Stem Cell Differentiation, Matrix Remodeling, and Biocompatibility Studies
Regenerative Potential of Decellularized Porcine Nucleus Pulposus Hydrogel Scaffolds: Stem Cell Differentiation, Matrix Remodeling, and Biocompatibility Studies
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DOI：
10.1089/ten.tea.2012.0088
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发表时间：
2013-04-01
影响因子：
4.1
通讯作者：
Simionescu, Dan T.
中科院分区：
文献类型：
--
作者：
Mercuri, Jeremy J.;Patnaik, Sourav;Simionescu, Dan T.
髓核（NP）组织再生已被提议作为对抗椎间盘退变的早期介入疗法。我们以前报道了一种新型的仿生脱细胞猪NP（APNP）水凝胶的开发和表征。在此，我们的目的是评估这种材料作为一个合适的支架NP组织再生。将人脂肪来源的干细胞（hADSC）在化学成分确定的分化培养基中在APNP水凝胶上培养14天，并分析NP细胞样mRNA表达谱、水凝胶重塑的证据（包括水凝胶收缩测量）、细胞外基质产生和压缩动态力学性质。还通过在缺乏可溶性分化因子的培养基中培养来检查水凝胶本身诱导干细胞分化的固有能力。此外，在大鼠皮下模型中评价了非交联和乙基二甲基氨基丙基碳二亚胺/N-羟基琥珀酰亚胺和五镓酰葡萄糖交联水凝胶的体内生物相容性。结果表明，当在APNP水凝胶上培养时，hADSC表达推定的NP细胞阳性基因转录标志物。此外，接种hADSC的水凝胶的糖胺聚糖和胶原蛋白含量显著大于未接种的对照，接种细胞的水凝胶表现出收缩和金属蛋白酶-1产生的组织抑制剂的证据。与非细胞接种的对照相比，hADSC接种的水凝胶的动态机械性能随着培养时间的推移而增加，并且接近天然NP组织报告的值。移植物的免疫组织化学分析表明，植入4周后，植入物间隙内存在单核细胞（包括巨噬细胞和成纤维细胞）以及血管浸润和胶原沉积。总之，这些结果表明，APNP水凝胶，与自体ADSC，可以作为一个合适的支架NP组织再生。
Nucleus pulposus (NP) tissue regeneration has been proposed as an early stage interventional therapy to combat intervertebral disc degeneration. We have previously reported on the development and characterization of a novel biomimetic acellular porcine NP (APNP) hydrogel. Herein, we aimed to evaluate this material for use as a suitable scaffold for NP tissue regeneration. Human-adipose-derived stem cells (hADSCs) were cultured for 14 days on APNP hydrogels in chemically defined differentiation media and were analyzed for an NP-cell-like mRNA expression profile, evidence of hydrogel remodeling including hydrogel contraction measurements, extracellular matrix production, and compressive dynamic mechanical properties. The innate capacity of the hydrogel itself to induce stem cell differentiation was also examined via culture in media lacking soluble differentiation factors. Additionally, the in vivo biocompatibility of non-crosslinked and ethyldimethylaminopropyl carbodiimide/N-hydroxysuccinimide and pentagalloyl glucose crosslinked hydrogels was evaluated in a rat subdermal model. Results indicated that hADSCs expressed putative NP-cell-positive gene transcript markers when cultured on APNP hydrogels. Additionally, glycosaminoglycan and collagen content of hADSC-seeded hydrogels was significantly greater than nonseeded controls and cell-seeded hydrogels exhibited evidence of contraction and tissue inhibitors of metalloproteinase-1 production. The dynamic mechanical properties of the hADSC-seeded hydrogels increased with time in culture in comparison to noncell-seeded controls and approached values reported for native NP tissue. Immunohistochemical analysis of explants illustrated the presence of mononuclear cells, including macrophages and fibroblasts, as well as blood vessel infiltration and collagen deposition within the implant interstices after 4 weeks of implantation. Taken together, these results suggest that APNP hydrogels, in concert with autologous ADSCs, may serve as a suitable scaffold for NP tissue regeneration.