Wisdom E Cate, Lamont Andrew, Martinez Hannah, Rockovich Michael, Lee Woojin, Gilchrist Kristin H, Ho Vincent B, Klarmann George J
USU Center for Biotechnology (4DBio3), Department of Radiology, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road, Bethesda, MD 20814, USA.
The Geneva Foundation, 917 Pacific Ave, Tacoma, WA 98402, USA.
Bioengineering (Basel). 2024 Aug 8;11(8):804. doi: 10.3390/bioengineering11080804.
Skin wounds often form scar tissue during healing. Early intervention with tissue-engineered materials and cell therapies may promote scar-free healing. Exosomes and extracellular vesicles (EV) secreted by mesenchymal stromal cells (MSC) are believed to have high regenerative capacity. EV bioactivity is preserved after lyophilization and storage to enable use in remote and typically resource-constrained environments. We developed a bioprinted bandage containing reconstituted EVs that can be fabricated at the point-of-need. An alginate/carboxymethyl cellulose (CMC) biomaterial ink was prepared, and printability and mechanical properties were assessed with rheology and compression testing. Three-dimensional printed constructs were evaluated for Young's modulus relative to infill density and crosslinking to yield material with stiffness suitable for use as a wound dressing. We purified EVs from human MSC-conditioned media and characterized them with nanoparticle tracking analysis and mass spectroscopy, which gave a peak size of 118 nm and identification of known EV proteins. Fluorescently labeled EVs were mixed to form bio-ink and bioprinted to characterize EV release. EV bandages were bioprinted on both a commercial laboratory bioprinter and a custom ruggedized 3D printer with bioprinting capabilities, and lyophilized EVs, biomaterial ink, and thermoplastic filament were deployed to an austere Arctic environment and bioprinted. This work demonstrates that EVs can be bioprinted with an alginate/CMC hydrogel and released over time when in contact with a skin-like substitute. The technology is suitable for operational medical applications, notably in resource-limited locations, including large-scale natural disasters, humanitarian crises, and combat zones.
皮肤伤口在愈合过程中常形成瘢痕组织。早期采用组织工程材料和细胞疗法进行干预可能促进无瘢痕愈合。间充质基质细胞(MSC)分泌的外泌体和细胞外囊泡(EV)被认为具有很高的再生能力。EV的生物活性在冻干和储存后得以保留,以便在偏远且通常资源有限的环境中使用。我们开发了一种含有重构EV的生物打印绷带,可在需要时现场制作。制备了藻酸盐/羧甲基纤维素(CMC)生物材料墨水,并通过流变学和压缩测试评估其可打印性和机械性能。对三维打印结构进行了杨氏模量相对于填充密度和交联的评估,以获得具有适合用作伤口敷料刚度的材料。我们从人MSC条件培养基中纯化了EV,并通过纳米颗粒跟踪分析和质谱对其进行表征,结果显示峰值尺寸为118nm,并鉴定出已知的EV蛋白。将荧光标记的EV混合形成生物墨水并进行生物打印以表征EV释放。EV绷带在商业实验室生物打印机和具有生物打印功能的定制坚固3D打印机上进行生物打印,冻干的EV、生物材料墨水和热塑性长丝被部署到北极严酷环境中并进行生物打印。这项工作表明,EV可以与藻酸盐/CMC水凝胶进行生物打印,并在与皮肤样替代品接触时随时间释放。该技术适用于实际医疗应用,特别是在资源有限的地区,包括大规模自然灾害、人道主义危机和战区。