Center for Regenerative Medicine and Orthopaedics, Danube University Krems, Dr. Karl-Dorrek-Str. 30, 3500, Krems, Austria.
Division of Orthopaedics and Traumatology, University Hospital Krems, Karl Landsteiner University of Health Sciences, Dr. Karl-Dorrek-Straße 30, 3500, Krems, Austria.
Int Orthop. 2024 Jan;48(1):279-290. doi: 10.1007/s00264-023-05930-7. Epub 2023 Aug 30.
Mesenchymal stem cells/medicinal signaling cells (MSCs) possess therapeutic potential and are used in regenerative orthopaedics. The infra-patellar fat pad (IFP) is partially resected during knee arthroscopy (KASC) and contains MSCs. Heat, irrigation, and mechanical stress during KASC may decrease MSC's therapeutic potential. This study assessed MSCs' regenerative potential after arthroscopic IFP harvest and potential effects of two blood products (BP) (platelet-rich plasma (PRP), hyperacute serum (HAS)) on MSCs' viability and chondrogenic differentiation capacity.
IFP was arthroscopically harvested, isolated, and counted (n = 5). Flow cytometry was used to assess cell viability via staining with annexin V/7-AAD and stemness markers via staining for CD90, CD73, and CD105. MSCs were incubated with blood products, and metabolic activity was determined via an XTT assay. Deposition of cartilage extracellular matrix was determined in histologic sections of chondrogenically differentiated 3D pellet cultures via staining with Alcian Blue. Expression of cartilage-specific genes (SOX9, MMP3/13, ACAN, COL1/2) was analyzed via quantitative PCR.
MSC isolation from IFP yielded 2.6610 ± 1.4910 viable cells from 2.7 (0.748) g of tissue. MSC markers (CD 90/105/73) were successfully detected and annexin V staining showed 81.5% viable cells. XTT showed increased metabolic activity. Within the BP groups, this increase was significant (days 0-14, p < 0.05). PCR showed expression of cartilage-specific genes in each group. COL2 (p < 0.01) as well as ACAN (p < 0.001) expression levels were significantly higher in the HAS group. Histology showed successful differentiation.
Arthroscopic harvest of IFP-MSCs yields sufficient cells with maintained regenerative potential and viability. Blood products further enhance MSCs' viability.
间充质干细胞/药用信号细胞（MSCs）具有治疗潜力，用于再生骨科。在膝关节镜检查（KASC）期间，部分切除髌下脂肪垫（IFP），其中包含 MSCs。KASC 过程中的热量、冲洗和机械应力可能会降低 MSC 的治疗潜力。本研究评估了关节镜 IFP 采集后 MSC 的再生潜力，以及两种血液制品（PRP、HAS）对 MSC 活力和软骨分化能力的潜在影响。
关节镜下采集、分离和计数 IFP（n=5）。通过用膜联蛋白 V/7-AAD 染色评估细胞活力，并用 CD90、CD73 和 CD105 染色评估干细胞标志物来评估间充质干细胞的活力。将 MSCs 与血液制品孵育，并通过 XTT 测定代谢活性。通过对软骨分化 3D 球培养物的组织学切片进行阿尔辛蓝染色，确定软骨细胞外基质的沉积。通过定量 PCR 分析软骨特异性基因（SOX9、MMP3/13、ACAN、COL1/2）的表达。
从 2.7（0.748）g 组织中分离出 IFP 获得 2.6610±1.4910 个有活力的细胞。成功检测到 MSC 标志物（CD90/105/73），膜联蛋白 V 染色显示 81.5%的活细胞。XTT 显示代谢活性增加。在血液制品组中，这种增加在第 0-14 天是显著的（p<0.05）。PCR 显示各组均表达软骨特异性基因。COL2（p<0.01）和 ACAN（p<0.001）表达水平在 HAS 组显著更高。组织学显示成功分化。
关节镜下采集 IFP-MSCs 可获得具有维持再生潜力和活力的足够细胞。血液制品进一步增强了 MSC 的活力。
