Fibrin Scaffolds Perfused with Transforming Growth Factor-β1 as an In Vitro Model to Study Healthy and Tendinopathic Human Tendon Stem/Progenitor Cells

• Published in: International journal of molecular sciences 2024-09, Vol.25 (17), p.9563
• Main Authors: Ciardulli, Maria Camilla, Lovecchio, Joseph, Parolini, Ornella, Giordano, Emanuele, Maffulli, Nicola, Della Porta, Giovanna
• Format: Article
• Language: English
• Subjects: 3D advanced in vitro model; Basic Helix-Loop-Helix Transcription Factors - genetics; Basic Helix-Loop-Helix Transcription Factors - metabolism; Biomechanics; Biopsy; Cell culture; Cell Survival - drug effects; Cells, Cultured; Collagen; Collagen Type I - genetics; Collagen Type I - metabolism; Collagen Type I, alpha 1 Chain - metabolism; Collagen Type III - genetics; Collagen Type III - metabolism; Cytokines; dynamic culture; Fibrin - metabolism; Gene expression; Growth factors; human tendon/stem progenitor cells; Humans; Hydrogels; Injuries; Investigations; Male; Membrane Proteins; Middle Aged; Morphology; Polyethylene glycol; Polyethylene terephthalate; Repair & maintenance; Stem cells; Stem Cells - cytology; Stem Cells - metabolism; Tendinopathy - metabolism; Tendinopathy - pathology; Tendons; Tendons - cytology; Tendons - metabolism; tenogenesis; Tissue engineering; Tissue Engineering - methods; Tissue Scaffolds - chemistry; Transforming Growth Factor beta1 - metabolism
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms25179563

A limited understanding of tendon cell biology in healthy and pathological conditions has impeded the development of effective treatments, necessitating in vitro biomimetic models for studying tendon events. We established a dynamic culture using fibrin scaffolds, bioengineered with tendon stem/progenitor cells ( TSPCs) from healthy or diseased human biopsies and perfused with 20 ng/mL of human transforming growth factor-β1 for 21 days. Both cell types showed long-term viability and upregulated Scleraxis (SCX-A) and Tenomodulin (TNMD) gene expressions, indicating tenogenic activity. However, diseased TSPCs underexpressed collagen type I and III (COL1A1 and COL3A1) genes and exhibited lower SCX-A and TNMD protein levels, but increased type I collagen production, with a type I/type III collagen ratio > 1.5 by day 14, matching healthy cells. Diseased TSPCs also showed constant high levels of pro-inflammatory cytokines, such as IL-8 and IL-6. This biomimetic environment is a valuable tool for studying tenogenic and inflammatory events in healthy and diseased tendon cells and identifying new therapeutic targets.