Development of three-layer collagen scaffolds to spatially direct tissue-specific cell differentiation for enthesis repair. (April 2023)

• Record Type: Journal Article
• Authors: Pugliese, Eugenia; Sallent, Ignacio; Ribeiro, Sofia; Trotier, Alexandre; Korntner, Stefanie H.; Bayon, Yves; Zeugolis, Dimitrios I.
• Is Part Of: Materials today bio. Volume 19(2023)
• Publication Date: 2023-04
• Subjects: Stem cell differentiation -- Functionalisation -- Bioactive molecules -- Growth factors -- Tendon-to-bone interface; 3-L Three-layer -- α-MEM Alpha-minimal essential medium -- ALP Alkaline phosphatase -- B-L Bone-like layer -- BGLAP Bone Gamma-Carboxyglutamate Protein -- BMP-2 bone morphogenetic protein 2 -- COL I Collagen type I (protein) -- COL1A1 Collagen type I (gene) -- COL II Collagen type II (protein) -- COL2A1 Collagen type II (gene) -- COL III Collagen type III (protein) -- COL3A1 Collagen type III (gene) -- COL X Collagen type X (protein) -- COL10A1 Collagen type X (gene) -- CS Chondroitin sulphate -- DMEM Dulbecco's modified Eagle medium -- EDTA Ethylenediaminetetraacetic acid -- ECM Extracellular matrix -- FC-L Fibrocartilage-like layer -- FGF-2 Fibroblast growth factor 2 -- FBS Foetal bovine serum -- FOV Field of view -- GAG Glycosaminoglycan -- GDF-5 Growth differentiation factor 5 -- GFD-7 Growth differentiation factor 7 -- HAp Hydroxyapatite -- hBMSCs Human bone marrow mesenchymal stromal cells -- IGF-1 Insulin-like growth factor 1 -- ITS+1 Insulin -- transferrin sodium selenite -- linoleic-bovine serum albumin MKX -- Mohawk PDGF-bb -- Platelet-derived growth factor bb PEG-SG -- Polyethylene glycol succinimidyl glutarate PS -- Penicillin/streptomycin PBS -- Phosphate-buffered saline RUNX2 -- Runt-related transcription factor 2 SCXA -- Scleraxis SOX9 -- Sex-determining region Y-box 9 SPP1 -- Secreted phosphoprotein 1 T-L -- Tendon-like layer TGF-β3 -- Transforming growth factor β3 TNC -- Tenascin C TNMD, Tenomodulin; Materials science -- Periodicals; Biomedical engineering -- Periodicals; Biomedical materials -- Periodicals; 620.1
• DOI: 10.1016/j.mtbio.2023.100584
• Languages: English
• ISSNs: 2590-0064

Abstract

Abstract: Enthesis repair remains a challenging clinical indication. Herein, a three-layer scaffold composed of a tendon-like layer of collagen type I, a fibrocartilage-like layer of collagen type II and a bone-like layer of collagen type I and hydroxyapatite, was designed to recapitulate the matrix composition of the enthesis. To aid tenogenic and fibrochondrogenic differentiation, bioactive molecules were loaded in the tendon-like layer or the fibrocartilage-like layer and their effect was assessed in in vitro setting using human bone marrow derived mesenchymal stromal cells and in an ex vivo model. Seeded human bone marrow mesenchymal stromal cells infiltrated and homogeneously spread throughout the scaffold. As a response to the composition of the scaffold, cells differentiated in a localised manner towards the osteogenic lineage and, in combination with differentiation medium, towards the fibrocartilage lineage. Whilst functionalisation of the tendon-like layer did not improve tenogenic cell commitment within the time frame of this work, relevant fibrochondrogenic markers were detected in the fibrocartilage-like layer when scaffolds were functionalised with bone morphogenetic protein 2 or non-functionalised at all, in vitro and ex vivo, respectively. Altogether, our data advocate the use of compartmentalised scaffolds for the repair and regeneration of interfacial tissues, such as enthesis. Graphical abstract: 3-L collagen scaffolds that recapitulate the matrix composition of the enthesis are able to promote zonal-specific differentiation of seeded human bone marrow derived mesenchymal stromal cells towards an osteogenic lineage and, with the aid of bioactive molecules, towards a fibrocartilage lineage, in an in vitro and ex vivo scenario (graphical abstract created with BioRender.com ). Image 1 Highlights: A three-layer enthesis-composition inspired collagen scaffold was developed. The porous network of the tendon-like layer supported initial cellular alignment. The bone-like layer promoted local cell commitment towards osteogenic lineage. Growth factor functionalisation enhanced spatial fibrochondrogenic cell commitment. The formation of an osteo/fibrochondrogenic interface was observed ex vivo.