Osmolarity modulates the de-differentiation of horse articular chondrocytes during cell expansion in vitro: implications for tissue engineering in cartilage repair.
Abstract: Due to the importance of joint disease and ostearthritis (OA) in equine athletes, new regenerative treatments to improve articular cartilage repair after damage are gaining relevance. Chondrocyte de-differentiation, an important pathogenetic mechanism in OA, is a limiting factor when differentiated articular chondrocytes are used for cell-based therapies. Current research focuses on the prevention of this de-differentiation and/or on the re-differentiation of chondrocytes by employing different strategies in vitro and in vivo. Articular chondrocytes normally live in a condition of higher osmolarity (350-450 mOsm/L) compared to normal physiological fluids (~ 300 mOsm/L) and some studies have demonstrated that osmolarity has a chondroprotective effect in vitro and in vivo. Therefore, the response of horse articular chondrocytes to osmolarity changes (280, 380, and 480 mOsm/L) was studied both in proliferating, de-differentiated chondrocytes grown in adhesion, and in differentiated chondrocytes grown in a 3D culture system. To this aim, cell proliferation (cell counting), morphology (optical microscopy), and differentiation (gene expression of specific markers) were monitored along with the expression of osmolyte transporters involved in volume regulation [betaine-GABA transporter (BGT-1), taurine transporter (SLC6A6), and neutral amino acid transporter (SNAT)] real-time qPCR. Proliferating chondrocytes cultured under hyperosmolar conditions showed low proliferation, spheroidal morphology, a significant reduction of de-differentiation markers [collagen type I (Col1) and RUNX2] and an increase of differentiation markers [collagen type II (Col2) and aggrecan]. Notably, a persistently high level of BGT-1 gene expression was maintained in chondrocyte cultures at 380 mOsm/L, and particularly at 480 mOsm/L both in proliferating and differentiated chondrocytes. These preliminary data encourage the study of osmolarity as a microenvironmental co-factor to promote/maintain chondrocyte differentiation in both 2D and 3D in vitro culture systems.
© 2023. The Author(s), under exclusive licence to Springer Nature B.V.
Publication Date: 2023-05-19 PubMed ID: 37202645DOI: 10.1007/s11259-023-10140-yGoogle Scholar: Lookup
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Summary
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The research article explores how osmolarity impacts the de-differentiation of horse articular chondrocytes during cell expansion in vitro, providing key insights for cartilage repair in tissue engineering.
Research Objective
- The study aims to investigate how changes in osmolarity, a condition related to the concentration of a solution, influence the de-differentiation of horse articular chondrocytes – the cells that make up cartilage. This is critical for understanding joint diseases and osteoarthritis in equine athletes and for developing new regenerative treatments.
Research Methodology
- The response of horse articular chondrocytes to varying osmolarity levels (280, 380, and 480 mOsm/L) was studied.
- Both proliferating, de-differentiated chondrocytes grown in an adherent state and differentiated chondrocytes grown in a 3D culture system were studied.
- Cell proliferation, morphology, and differentiation were monitored, including the gene expression of specific markers. The expression of osmolyte transporters involved in volume regulation was also studied.
Key Findings
- Chondrocytes cultured under hyperosmolar conditions showed reduced proliferation, spheroidal morphology, a significant decrease of de-differentiation markers, and an increase of differentiation markers.
- Specifically, hyperosmolar conditions led to a decline in collagen type I (Col1) and RUNX2 (markers of de-differentiation), and an increase in collagen type II (Col2) and aggrecan (markers of differentiation).
- A persistently high level of betaine-GABA transporter (BGT-1) gene expression, involved in volume regulation, was maintained in chondrocyte cultures at higher osmolarity levels (380 and 480 mOsm/L).
Implications of the Research
- The study provides encouraging preliminary data for further research into the role of osmolarity as a key factor influencing chondrocyte differentiation in both 2D and 3D in vitro culture systems.
- Understanding how osmolarity impacts chondrocyte de-differentiation could improve regenerative therapies aimed at promoting articular cartilage repair after damage.
Cite This Article
APA
De Angelis E, Barilli A, Saleri R, Rotoli BM, Ravanetti F, Ferrari F, Ferrari L, Martelli P, Dall'Asta V, Borghetti P.
(2023).
Osmolarity modulates the de-differentiation of horse articular chondrocytes during cell expansion in vitro: implications for tissue engineering in cartilage repair.
Vet Res Commun.
https://doi.org/10.1007/s11259-023-10140-y Publication
Researcher Affiliations
- Department of Veterinary Science, University of Parma, Strada del Taglio 10, Parma, 43126, Italy.
- Laboratory of General Pathology, Department of Medicine and Surgery, University of Parma, Via Volturno 39, Parma, 43125, Italy.
- Department of Veterinary Science, University of Parma, Strada del Taglio 10, Parma, 43126, Italy.
- Laboratory of General Pathology, Department of Medicine and Surgery, University of Parma, Via Volturno 39, Parma, 43125, Italy.
- Department of Veterinary Science, University of Parma, Strada del Taglio 10, Parma, 43126, Italy. francesca.ravanetti@unipr.it.
- Laboratory of General Pathology, Department of Medicine and Surgery, University of Parma, Via Volturno 39, Parma, 43125, Italy.
- Department of Veterinary Science, University of Parma, Strada del Taglio 10, Parma, 43126, Italy.
- Department of Veterinary Science, University of Parma, Strada del Taglio 10, Parma, 43126, Italy.
- Laboratory of General Pathology, Department of Medicine and Surgery, University of Parma, Via Volturno 39, Parma, 43125, Italy.
- Department of Veterinary Science, University of Parma, Strada del Taglio 10, Parma, 43126, Italy.
References
This article includes 28 references
- Arabiyat AS, Chen H, Erndt-Marino J. Hyperosmolar Ionic Solutions modulate inflammatory phenotype and sGAG loss in a cartilage explant model. Cartilage 13.
- Barilli A, Visigalli R, Ferrari F. Organic cation transporters (OCTs/OCTNs) in human primary alveolar epithelial cells. Biochem Biophys Res Commun 576.
- Bush PG, Hall AC. The osmotic sensitivity of isolated and in situ bovine articular chondrocytes. J Orthop Res 19.
- Capito NM, Cook JL, Yahuaca B. Safety and efficacy of hyperosmolar irrigation solution in shoulder arthroscopy. J Shoulder Elb Surg 26.
- Charlier E, Céline D, Ciregia F. Chondrocyte dedifferentiation and osteoarthritis. OA) Biochemical Pharmacology .
- De Angelis E, Giorgio Petronini P, Borghetti P. Induction of betaine-γ-aminobutyric acid transport activity in porcine chondrocytes exposed to hypertonicity. J Physiol 518.
- De Angelis E, Grolli S, Saleri R, Conti V, Andrani M, Berardi M, Cavalli V, Passeri B, Ravanetti F, Borghetti P. Platelet lysate reduces the chondrocyte dedifferentiation during in vitro expansion: Implications for cartilage tissue engineering.. Res Vet Sci 2020 Dec;133:98-105.
- Franchi-Gazzola R, Dall’Asta V, Sala R. The role of the neutral amino acid transporter SNAT2 in cell volume regulation. Acta Physiol (Oxf) May-Jun 187(1–2):273–283.
- Gibson JS, Milner PI, White R. Oxygen and reactive oxygen species in articular cartilage: modulators of ionic homeostasis. Pflugers Arch. Eur. J. Physiol. 455.
- Guo J, Jourdian GW, Maccallum DK. Culture and growth characteristics of chondrocytes encapsulated in alginate beads. Connect Tissue Res 19.
- Hall AC. Volume-sensitive taurine transport in bovine articular chondrocytes. J Physiol 484.
- Hing WA, Sherwin AF, Poole CA. The influence of the pericellular microenvironment on the chondrocyte response to osmotic challenge. Osteoarthr Cartil Apr 10(4):297–307.
- Karim A, Hall AC. Hyperosmolarity normalises serum-induced changes to chondrocyte properties in a model of cartilage injury. Eur Cells Mater 31.
- Koo J, Kim K, Il, Min BH, Lee GM. Controlling medium osmolality improves the expansion of human articular chondrocytes in serum-free media. Tissue Eng - Part C Methods 16.
- Kyriacos AA, Darling EM, Du Raine GD. Articular cartilage. Hydrated materials. Jenny Stanford Publishing pp 43–50.
- Medvedeva EV, Grebenik EA, Gornostaeva SN. Repair of damaged articular cartilage: current approaches and future directions. Int. J. Mol. Sci. 19.
- n Der Windt AE, Haak E, Das RHJ. Physiological tonicity improves human chondrogenic marker expression through nuclear factor of activated T-cells 5 in vitro. Arthritis Res Ther 12.
- Negoro K, Kobayashi S, Takeno K. Effect of osmolarity on glycosaminoglycan production and cell metabolism of articular chondrocyte under three-dimensional culture system. Clin Exp Rheumatol 26.
- Ortved KF, Nixon AJ. Cell-based cartilage repair strategies in the horse. Vet. J. 208.
- Oswald ES, Ahmed HS, Kramer SP. Effects of hypertonic (NaCl) two-dimensional and three-dimensional culture conditions on the properties of cartilage tissue engineered from an expanded mature bovine chondrocyte source. Tissue Eng - Part C Methods 17.
- Parameswaran R, Kachroo U, Amirtham SM. An in vitro analysis of the effect of hyperosmolarity on the chondrogenic potential of human articular cartilage derived chondroprogenitors. Tissue Cell 72.
- Rakic R, Bourdon B, Hervieu M. RNA interference and BMP-2 stimulation allows equine chondrocytes redifferentiation in 3D-Hypoxia cell culture model: application for Matrix-Induced autologous chondrocyte implantation. Int J Mol Sci 18.
- Ravanetti F, Saleri R, Martelli P, Andrani M, Ferrari L, Cavalli V, Conti V, Rossetti AP, De Angelis E, Borghetti P. Hypoxia and platelet lysate sustain differentiation of primary horse articular chondrocytes in xeno-free supplementation culture.. Res Vet Sci 2022 Dec 20;152:687-697.
- Schmittgen TD, Livak KJ. Analyzing real-time PCR data by the comparative C(T) method.. Nat Protoc 2008;3(6):1101-8.
- Sieber S, Michaelis M, Gühring H. Importance of osmolarity and oxygen tension for cartilage tissue engineering. Biores Open Access 9.
- Urban JPG, Hall AC, Gehl KA. Regulation of matrix synthesis rates by the ionic and osmotic environment of articular chondrocytes. J Cell Physiol 154.
- van der Caron MMJ, Emans PJ. Osmolarity determines the in vitro chondrogenic differentiation capacity of progenitor cells via nuclear factor of activated T-cells 5. Bone 53.
- Zhang Z. Chondrons and the pericellular matrix of chondrocytes. Tissue Eng Part B Rev Jun 21(3):267–277.
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