Infra-patellar fat pad-derived mesenchymal stem cells maintain their chondrogenic differentiation potential after arthroscopic harvest with blood-product supplementation

Markus Neubauer, Alexander Otahal, Olga Kuten, Seth Lawrence Sherman, Lukas Moser, Karina Kramer, Andrea DeLuna, Johannes Neugebauer, Dietmar Dammerer, Thomas Muellner, Stefan Nehrer

Research output: Journal article (peer-reviewed)Journal article

Abstract

PURPOSE: 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.

METHODS: 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.

RESULTS: MSC isolation from IFP yielded 2.66*106 ± 1.49*106 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.

CONCLUSION: Arthroscopic harvest of IFP-MSCs yields sufficient cells with maintained regenerative potential and viability. Blood products further enhance MSCs' viability.

Original languageEnglish
Pages (from-to)279-290
Number of pages12
JournalInternational Orthopaedics
Volume48
Issue number1
Early online date30 Aug 2023
DOIs
Publication statusPublished - Jan 2024

Keywords

  • Blood products
  • Cartilage regeneration
  • Infrapatellar fat pad
  • Mesenchymal stem cells
  • Humans
  • Cells, Cultured
  • Annexin A5/metabolism
  • Cell Differentiation
  • Adipose Tissue
  • Chondrogenesis
  • Dietary Supplements
  • Mesenchymal Stem Cells

ASJC Scopus subject areas

  • Surgery
  • Orthopedics and Sports Medicine

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