The Tissue-Engineered Tendon-Bone Interface: In Vitro and In Vivo Synergistic Effects of Adipose-Derived Stem Cells, Platelet-Rich Plasma, and Extracellular Matrix Hydrogel
The Tissue-Engineered Tendon-Bone Interface: In Vitro and In Vivo Synergistic Effects of Adipose-Derived Stem Cells, Platelet-Rich Plasma, and Extracellular Matrix Hydrogel
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DOI：
10.1097/prs.0000000000003840
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发表时间：
2017-12-01
影响因子：
3.6
通讯作者：
Chang, James
中科院分区：
文献类型：
--
作者：
McGoldrick, Rory;Chattopadhyay, Arhana;Chang, James
背景：肌腱-骨界面次优愈合仍然是一个未解决的问题。作者假设(1)富含血小板的血浆和长时间的体外孵育将产生更多可再生脂肪干细胞的界面支架；(2)当植入细胞外基质水凝胶时，构建的支架将显示出良好的体内强度修复和生物相容性。方法：取30只Wistar大鼠的跟腱-跟腱-骨复合界面支架。经过物理化学脱细胞和冷冻干燥后，支架在大鼠血浆或100%激活的大鼠富血小板血浆中活化，并重新接种有活力的脂肪干细胞。在研究的第二部分，90只SD大鼠接受了五种脱细胞、冷冻干燥的支架振兴/再种植条件之一的重建：(1)磷酸盐缓冲液；(2)冷冻干燥的100%激活的富血小板血浆；(3)富血小板血浆和细胞外基质水凝胶；(4)富血小板血浆和ASC-Luc2-EGFP细胞再种植14天；结果：在第一部分中，富含血小板的血浆活化的移植物在第3、7和14天显示了更多的存活脂肪干细胞负载，在第7和14天显示出总的脂肪来源干细胞负载，并明显增加了活体表面细胞密度、分层、迁移和穿透。在第二部分中，生物发光成像证实细胞存活到植入后第22天。生物力学强度测试显示，在2周时，再植组的极限破坏载荷显著高于其他所有组，而在4周和8周时，只有水凝胶再植组的肌腱细胞侵袭和纤维软骨再生明显增加。在8周时，Masson三色染色显示支架结构持续存在，盲法ImageJ分析显示，补种/水凝胶组在2、4和8周时III型胶原明显增多。结论：脱细胞冻干同种异体肌腱-骨界面支架可以通过在富血小板血浆中活化，重新种植有活力的脂肪干细胞，并在植入时补充细胞外基质肌腱水凝胶来优化。当这样做时，它们表现出更大的修复强度和生物相容性。
Background: Suboptimal healing of the tendon-bone interface remains an unsolved problem. The authors hypothesized that (1) platelet-rich plasma and prolonged in vitro incubation will produce interface scaffolds with greater reseeding of viable adipose-derived stem cells; and (2) when implanted with extracellular matrix hydrogel, constructs will display superior in vivo strength repair and biocompatibility.Methods: Achilles-calcaneal composite tendon-bone interface scaffold grafts were harvested from 30 Wistar rats. After physicochemical decellularization and lyophilization, scaffolds were revitalized in rat plasma or 100% activated rat platelet-rich plasma and reseeded with viable adipose-derived stem cells. For part 2 of the study, 90 Sprague-Dawley rats underwent reconstruction with one of five decellularized, lyophilized scaffold revitalization/reseeding conditions: (1) phosphate-buffered saline; (2) lyophilized, 100% activated platelet-rich plasma; (3) platelet-rich plasma and extracellular matrix hydrogel; (4) platelet-rich plasma and 14-day reseeding with ASC-luc2-eGFP cells; and (5) plasma, reseeding, and hydrogel.Results: In part 1, platelet-rich plasma-revitalized grafts demonstrated greater live viable adipose-derived stem cell loads at 3, 7, and 14 days and total adipose-derived stem cell loads at 7 and 14 days with visibly greater live surface cellularity, layering, migration, and penetration. In part 2, bioluminescence imaging confirmed cell viability to day 22 after implantation. Biomechanical strength testing demonstrated a significant increase in ultimate failure load for reseeded groups compared with all other groups at week 2, whereas only reseeded grafts with hydrogel remained significantly stronger at weeks 4 and 8. Histologic examination demonstrated most increased tendinous cellular invasion and fibrocartilage repopulation at 8 weeks in the reseeded group with hydrogel. Masson trichrome staining demonstrated persistence of the scaffold structure at week 8 and blinded ImageJ analysis demonstrated significantly more type III collagen in the reseeded/hydrogel group at 2, 4, and 8 weeks.Conclusions: Decellularized lyophilized allogeneic tendon-bone interface scaffolds can be optimized by revitalization in platelet-rich plasma, reseeding with viable adipose-derived stem cells, and supplemented by an extracellular matrix tendon hydrogel at the time of implantation. When this is done, they display greater repair strength and biocompatibility.