Biomaterials and Tissue Engineering Laboratory, Department of Biosciences and Bioengineering, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India.
Centre for Nanotechnology, Indian Institute of Technology Guwahati, Guwahati, Assam, 781039, India.
Adv Healthc Mater. 2024 Jun;13(15):e2303312. doi: 10.1002/adhm.202303312. Epub 2024 Mar 26.
Physiologically-relevant in vitro skin models hold the utmost importance for efficacy assessments of pharmaceutical and cosmeceutical formulations, offering valuable alternatives to animal testing. Here, an advanced immunocompetent 3D bioprinted human skin model is presented to assess skin sensitization. Initially, a photopolymerizable bioink is formulated using silk fibroin methacrylate, gelatin methacrylate, and photoactivated human platelet releasate. The developed bioink shows desirable physicochemical and rheological attributes for microextrusion bioprinting. The tunable physical and mechanical properties of bioink are modulated through variable photocuring time for optimization. Thereafter, the bioink is utilized to 3D bioprint "sandwich type" skin construct where an artificial basement membrane supports a biomimetic epidermal layer on one side and a printed pre-vascularized dermal layer on the other side within a transwell system. The printed construct is further cultured in the air-liquid interface for maturation. Immunofluorescence staining demonstrated a differentiated keratinocyte layer and dermal extracellular matrix (ECM)-remodeling by fibroblasts and endothelial cells. The biochemical estimations and gene-expression analysis validate the maturation of the printed model. The incorporation of macrophages further enhances the physiological relevance of the model. This model effectively classifies skin irritative and non-irritative substances, thus establishing itself as a suitable pre-clinical screening platform for sensitization tests.
生理相关的体外皮肤模型对于药物和化妆品制剂的功效评估至关重要,为动物试验提供了有价值的替代方法。在这里,我们提出了一种先进的免疫活性 3D 生物打印人类皮肤模型,用于评估皮肤致敏性。首先,我们使用丝素蛋白甲基丙烯酯、明胶甲基丙烯酯和光激活的人血小板释放物来配制可光聚合的生物墨水。所开发的生物墨水具有用于微挤出生物打印的理想的物理化学和流变学特性。通过改变光固化时间来调节生物墨水的可调物理和机械性能以进行优化。然后,我们使用该生物墨水来 3D 生物打印“三明治型”皮肤结构,其中人工基底膜在 Transwell 系统的一侧支撑仿生表皮层,在另一侧支撑打印的预血管化真皮层。打印的结构进一步在气液界面中进行培养以实现成熟。免疫荧光染色显示出分化的角质形成细胞层和由成纤维细胞和内皮细胞重塑的真皮细胞外基质 (ECM)。生化估算和基因表达分析验证了打印模型的成熟。巨噬细胞的掺入进一步提高了模型的生理相关性。该模型能够有效地区分皮肤刺激性和非刺激性物质,因此成为一种合适的用于致敏测试的临床前筛选平台。
Nat Protoc. 2021-12
Adv Healthc Mater. 2019-2-6
Micromachines (Basel). 2025-4-30
Theranostics. 2025-1-1
Adv Healthc Mater. 2024-10