Hacettepe University, Graduate School of Science and Engineering, Bioengineering Department, Beytepe, Ankara, Turkey.
Hacettepe University, Chemical Engineering Department, Beytepe, Ankara, Turkey.
J Photochem Photobiol B. 2022 Jul;232:112476. doi: 10.1016/j.jphotobiol.2022.112476. Epub 2022 May 20.
Conventional wound healing treatments are insufficient for chronic wounds caused by factors such as senescence of fibroblasts, reduced growth factor synthesis, and poor angiogenesis. Recently, tissue engineering approaches have been investigated to develop effective therapies. In this study, a biochemical/biophysical stimulant-based 3D system was developed for the healing of chronic wounds. In this direction, genipin crosslinked chitosan (CHT)/gelatin (GEL) scaffolds were fabricated by freeze-drying and loaded with platelet-rich plasma (PRP). The scaffolds were seeded with human dermal fibroblasts and then, polychromatic light in near infrared region (NIR) was applied to the scaffolds for activating the platelets and stimulating the fibroblasts (photoactivation, PAC). Thus, fibroblasts were stimulated both chemically and physically by PRP and light, respectively. Cell migration, proliferation, morphology, gene expressions and reactive oxygen species (ROS) activity were evaluated in-vitro. Laminin and collagen 4 expressions that are important for extracellular matrix (ECM) formation, and PDGF (Platelet-derived growth factor) and VEGF (Vascular endothelial growth factor) expressions that are important for vascularization significantly increased in the presence of both PRP and light. Besides, PRP and light improved cell migration in 3D core-and shell model synergistically. Hydrogen peroxide content decreased in both PRP and light, indicating inhibition of ROS production. It was concluded that the stimulation of platelets with light in the NIR has a great potential to use for both platelets activation and stimulation of fibroblasts. As a result, an effective therapy can be developed for chronic wounds by using scaffold-based 3D systems together with PRP and photostimulation.
传统的伤口愈合治疗方法对于由成纤维细胞衰老、生长因子合成减少和血管生成不良等因素引起的慢性伤口效果有限。最近，人们研究了组织工程方法来开发有效的治疗方法。在本研究中，开发了一种基于生化/生物物理刺激的 3D 系统来治疗慢性伤口。在这个方向上，通过冷冻干燥制备了京尼平交联壳聚糖（CHT）/明胶（GEL）支架，并负载富血小板血浆（PRP）。将人真皮成纤维细胞接种到支架上，然后用近红外区域（NIR）的多色光激活血小板并刺激成纤维细胞（光激活，PAC）。因此，成纤维细胞分别受到 PRP 和光的化学和物理刺激。在体外评估了细胞迁移、增殖、形态、基因表达和活性氧（ROS）活性。重要的细胞外基质（ECM）形成相关基因表达，如层粘连蛋白和胶原 4，以及重要的血管生成相关基因表达，如血小板衍生生长因子（PDGF）和血管内皮生长因子（VEGF），在 PRP 和光的存在下显著增加。此外，PRP 和光协同促进了 3D 核壳模型中的细胞迁移。PRP 和光都降低了过氧化氢含量，表明 ROS 产生受到抑制。结果表明，NIR 中的光对血小板的刺激具有很大的潜力，可用于血小板激活和刺激成纤维细胞。因此，通过使用基于支架的 3D 系统和 PRP 与光刺激，可以为慢性伤口开发有效的治疗方法。
