Cellular and molecular events during chondrogenesis of human mesenchymal stromal cells grown in a three-dimensional hyaluronan based scaffold

• Published in: Biomaterials Vol. 26; no. 28; pp. 5677 - 5686
• Main Authors: Lisignoli, Gina, Cristino, Sandra, Piacentini, Anna, Toneguzzi, Stefania, Grassi, Francesco, Cavallo, Carola, Zini, Nicoletta, Solimando, Liliana, Mario Maraldi, Nadir, Facchini, Andrea
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 01.10.2005
• Subjects: Biocompatible Materials - chemistry; Biomaterial; Cell Differentiation - physiology; Cell Proliferation; Cell Survival - physiology; Cells, Cultured; Chondrocytes - cytology; Chondrocytes - physiology; Chondrogenesis; Chondrogenesis - physiology; Humans; Hyaluronic Acid - chemistry; Materials Testing; Mesenchymal stromal cells; Mesenchymal Stromal Cells - cytology; Mesenchymal Stromal Cells - physiology; Stromal Cells - cytology; Stromal Cells - physiology; Tissue engineering; Tissue Engineering - methods; Mesenchymal stromal cells; Biomaterial; Chondrogenesis; Tissue engineering
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2005.02.031

Mesenchymal stromal cells (MSCs) seem to be a good alternative to chondrocytes for cartilage regeneration. To obtain new information on the sequence of cellular and molecular events during in vitro chondrogenic differentiation we analysed MSCs on a widely used hyaluronic acid biomaterial (Hyaff ®-11). Cellular differentiation was induced using two different concentrations of TGF β 1 (10 and 20 ng/ml) and the process was analysed at different time points (24 h, and 7, 14, 21 and 28 days) using techniques of light and electron microscopy, real-time PCR and immunohistochemistry. We found that without TGF β MSCs did not survive while in the presence of TGF β the cells significantly proliferated from day 7 until day 28. Light and electron microscopy showed that TGF β at 20 ng/ml better induced the formation of cartilage-like tissue. Real-time PCR showed an increased expression of collagen type II, IX and aggrecan associated to a down-regulation of collagen type I. Immunohistochemical analysis confirmed that collagen type I was down-modulated while collagen type II increased from day 14 to day 28. These data clearly showed that higher concentrations of TGF β (20 ng/ml) induce chondrogenesis of MSCs on Hyaff ®-11 scaffold better than 10 ng/ml of TGF β . This process is characterized by a sequence of cellular and molecular events that deal with the in vitro formation of a cartilage-like tissue.