Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Trauma Research Center, Vienna, Austria; Austrian Cluster for Tissue Regeneration, Vienna, Austria.
Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Trauma Research Center, Vienna, Austria; Austrian Cluster for Tissue Regeneration, Vienna, Austria.
J Photochem Photobiol B. 2021 Aug;221:112243. doi: 10.1016/j.jphotobiol.2021.112243. Epub 2021 Jun 18.
Due to their capacity to differentiate into the chondrogenic lineage, adipose-derived stromal/stem cells (ASC) are a promising source of therapeutically relevant cells for cartilage tissue regeneration. Their differentiation potential, however, varies between patients. In our study, we aim to stimulate ASC towards a more reliable chondrogenic phenotype using photobiomodulation (PBM). LED devices of either blue (475 nm), green (516 nm) or red (635 nm) light were used to treat human ASC from donors of varying chondrogenic potential. The treatment was applied either once during the 2D expansion phase or repeatedly during the 3D differentiation phase. Chondrogenic differentiation was assessed via pellet size, GAG/DNA content, histology and gene expression analysis. Reactions to PBM were found to be wavelength-dependent and more pronounced when the treatment was applied during expansion. Donors were assigned to responder categories according to their response to the treatment during expansion, whereby good responders were mainly donors with low intrinsic chondrogenic potential. Exposed to light, they revealed a particularly high relative increase in pellet size (more than twice the size of untreated controls after red light PBM), intense collagen type II immunostaining (low/absent in untreated controls) and activation of otherwise absent COL2A1 expression. Conversely, on a donor with high intrinsic chondrogenic potential, light had adverse effects. When applied with shorter wavelengths (blue, green), it led to reduced pellet size, GAG/DNA content and collagen type II immunostaining. However, when PBM was applied in 3D, the same donor was the only one to react with increased differentiation to all three wavelengths. We were able to demonstrate that PBM can be used to enhance or hamper chondrogenesis of ASC, and that success depends on treatment parameters and intrinsic cellular potential. The improvement of chondrogenesis in donors with low intrinsic potential highlights PBM as potent tool for cell-based cartilage regeneration. Its cost-effectiveness and ease of use make for an attractive treatment option to enhance the performance of ASC in cartilage tissue engineering.
由于其向软骨谱系分化的能力，脂肪来源的基质/干细胞（ASC）是一种很有前途的治疗相关细胞来源，可用于软骨组织再生。然而，它们的分化潜能在不同患者之间存在差异。在我们的研究中，我们旨在通过光生物调节（PBM）来刺激 ASC 向更可靠的软骨表型分化。使用来自不同成软骨潜能供体的人 ASC 进行实验，实验中使用了蓝色（475nm）、绿色（516nm）或红色（635nm）的 LED 设备进行治疗。该治疗要么在 2D 扩增阶段进行一次，要么在 3D 分化阶段重复进行。通过球状体大小、GAG/DNA 含量、组织学和基因表达分析来评估软骨分化。发现 PBM 的反应与波长有关，在扩增阶段进行治疗时更为明显。根据其在扩增阶段对治疗的反应，将供体分为反应者类别，其中良好反应者主要是成软骨潜能低的供体。暴露于光线下，它们的球状体大小相对增加特别明显（红光 PBM 后比未经处理的对照组大两倍多），强烈的 II 型胶原免疫染色（未经处理的对照组中低/无），以及 COL2A1 表达的激活（否则无表达）。相反，在成软骨潜能高的供体中，光有不良影响。当使用较短波长（蓝色、绿色）时，会导致球状体大小、GAG/DNA 含量和 II 型胶原免疫染色减少。然而，当在 3D 中进行 PBM 时，只有同一位供体对所有三种波长都表现出增加的分化反应。我们能够证明 PBM 可用于增强或抑制 ASC 的软骨生成，其成功取决于治疗参数和内在细胞潜能。在内在潜能低的供体中改善软骨生成突出了 PBM 作为基于细胞的软骨再生的有力工具。其成本效益和易用性使其成为增强 ASC 在软骨组织工程中性能的有吸引力的治疗选择。
