3D bioprinting of photo-crosslinkable silk methacrylate (SilMA)-polyethylene glycol diacrylate (PEGDA) bioink for cartilage tissue engineering
3D bioprinting of photo-crosslinkable silk methacrylate (SilMA)-polyethylene glycol diacrylate (PEGDA) bioink for cartilage tissue engineering
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DOI：
10.1002/jbm.a.37336
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发表时间：
2021-12-16
影响因子：
4.9
通讯作者：
Bhardwaj, Nandana
中科院分区：
文献类型：
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作者：
Bandyopadhyay, Ashutosh;Mandal, Biman B.;Bhardwaj, Nandana
由于关节软骨固有的再生能力极弱，关节软骨损伤给全球医疗保健部门带来了巨大的负担。使用复合生物墨水开发软骨模拟构建体的三维（3D）生物打印作为一个新兴的视角。然而，开发合适的生物墨水的困难和化学交联相关的固有毒性阻碍了该技术的广泛采用。为了避免这一点，有助于重现复杂软骨微环境的基于光可聚合水凝胶的生物墨水是相关的。在本文中，将含有不同浓度的丝甲基丙烯酸酯（SilMA）和聚乙二醇二丙烯酸酯（PEGDA）的可光交联的生物墨水与软骨细胞混合，用于3D生物打印软骨构建体的生物制造。沿着软骨组织形成，检查生物墨水的流变学性质、可印刷性和印刷的水凝胶构建体的物理化学表征。印刷的SilMA-PEGDA水凝胶构建体具有适当的内部多孔结构，并表现出最可靠的流变性质、印刷适性沿着良好的机械性质和适于软骨再生的降解性质。活/死染色显示3D生物打印的SilMA-PEGDA构建体的细胞相容性。此外，如细胞活力和DNA含量定量所示，在软骨组织内观察到细胞数量和DNA含量显著增加。生物化学评估证实了SilMA-PEGDA生物打印的构建体内的新软骨形成，如软骨特异性细胞外基质硫酸化GAG（sGAG）和II型胶原蛋白随时间的沉积增强（>2倍增加，p < 0.001）所揭示的。最后，免疫组化分析表明，II型胶原蛋白和聚集蛋白聚糖的表达与软骨组织形成相证实。综上所述，我们得出结论，SilMA-PEGDA生物墨水可能是生物打印软骨细胞以支持软骨组织修复和再生的合适候选者。
Articular cartilage damage poses huge burden on healthcare sector globally due to its extremely weak inherent regenerative ability. Three-dimensional (3D) bioprinting for development of cartilage mimic constructs using composite bioinks serves as an emerging perspective. However, difficulty in development of suitable bioink and chemical crosslinking associated inherent toxicity hamper widespread adoption of this technique. To circumvent this, a photo-polymerizable hydrogel-based bioink which helps in recapitulation of the complex cartilage microenvironment is pertinent. Herein, a photo-crosslinkable bioink containing different concentrations of silk methacrylate (SilMA) and polyethylene glycol diacrylate (PEGDA) was mixed with chondrocytes for biofabrication of 3D bioprinted cartilage constructs. The rheological properties, printability of bioink and physico-chemical characterization of printed hydrogel constructs were examined along with cartilaginous tissue formation. The printed SilMA-PEGDA hydrogel constructs possessed proper internal porous structure and demonstrated most reliable rheological properties, printability along with good mechanical, and degradation properties suitable for cartilage regeneration. Live/dead staining showed cytocompatibility of the 3D-bioprinted SilMA-PEGDA constructs. Moreover, a marked increase in cell number and DNA content was observed within the cartilaginous tissue as indicated by cell viability and DNA content quantitation. Biochemical evaluation confirmed the neocartilage formation within SilMA-PEGDA bioprinted constructs as revealed by enhanced deposition of cartilage specific extracellular matrix-sulphated GAG (sGAG) and collagen type II (>2-fold increase, p < 0.001) with time. Finally, immunohistochemical analysis indicated expression of collagen type II and aggrecan which corroborated with cartilaginous tissue formation. Taken together, we conclude that SilMA-PEGDA bioink could be suitable candidate for bioprinting chondrocytes to support cartilage tissue repair and regeneration.