Minne Dekker, Luke Hipwood, Akhilandeshwari Ravichandran, Dietmar W Hutmacher, Christoph Meinert, Jacqui McGovern
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引用次数: 0
摘要
脱细胞细胞外基质(dECM)为基础的生物材料模拟天然ECM和支持三维细胞培养。一种可光交联的猪骨源性dECM水凝胶(dECM- ma)具有可调的机械性能,可用于组织特异性体外模型。用10% EDTA对小梁骨进行脱矿处理,并使用3.4 m NaCl通过渗透休克进行脱细胞处理,在保留关键ECM蛋白的同时降低了94%的DNA含量。蛋白质组学分析鉴定出81种基质蛋白,其中76种在原生和脱细胞组织中共享。dECM通过胃蛋白酶消化溶解,并与甲基丙烯酰基进行功能化,实现87-98%的功能化。光交联的dECM-MA水凝胶的杨氏模量(0.5-120 kPa)取决于聚合物浓度(0.25-2% w/v)和交联时间(8-120 s)。包裹在dECM-MA(5、10和20 kPa)中的原代人成骨细胞(hOBs)仍然具有活力,并表现出成骨形态。在10 kPa水凝胶中,hOBs显示出代谢活性增加,碱性磷酸酶升高,矿物质沉积(µCT,茜素红)。DMP-1和骨钙素的表达表明细胞成熟和ECM重塑。这项研究证明了为3D培养创建可调的骨特异性dECM水凝胶的可行性。dECM-MA提供了一个可控的基质环境,代表了一个在组织特异性微环境中进行疾病建模和药物筛选的通用平台。
Bone-Derived dECM Hydrogels Support Tunable Microenvironments for In Vitro Osteogenic Differentiation.
Decellularized extracellular matrix (dECM)-based biomaterials mimic native ECM and support 3D cell culture. A photocrosslinkable porcine bone-derived dECM hydrogel (dECM-MA) is developed with tunable mechanical properties for tissue-specific in vitro models. Trabecular bone is demineralized with 10% EDTA and decellularized via osmotic shock using 3.4 m NaCl, reducing DNA content by 94% while preserving key ECM proteins. Proteomic analysis identifies 81 matrisome proteins, with 76 shared between native and decellularized tissue. The dECM is solubilized by pepsin digestion and functionalized with methacryloyl groups, achieving 87-98% functionalization. Photocrosslinked dECM-MA hydrogels shows tunable Young's moduli (0.5-120 kPa) depending on polymer concentration (0.25-2% w/v) and crosslinking duration (8-120 s). Primary human osteoblasts (hOBs) encapsulated in dECM-MA (5, 10, and 20 kPa) remains viable and exhibits osteogenic morphology. In 10 kPa hydrogels, hOBs shows increased metabolic activity, elevated alkaline phosphatase, and mineral deposition (µCT, Alizarin Red). Expression of DMP-1 and osteocalcin indicates cell maturation and ECM remodeling. This study demonstrates the feasibility of creating tunable, bone-specific dECM hydrogels for 3D culture. dECM-MA provides a controllable matrix environment and represents a versatile platform for disease modeling and drug screening in tissue-specific microenvironments.
期刊介绍:
Advanced Healthcare Materials, a distinguished member of the esteemed Advanced portfolio, has been dedicated to disseminating cutting-edge research on materials, devices, and technologies for enhancing human well-being for over ten years. As a comprehensive journal, it encompasses a wide range of disciplines such as biomaterials, biointerfaces, nanomedicine and nanotechnology, tissue engineering, and regenerative medicine.