Human chondrocyte migration behaviour to guide the development of engineered cartilage

Abstract

Tissue-engineering techniques have been successful in developing cartilage-like tissues in vitro using cells from animal sources. The successful translation of these strategies to the clinic will likely require cell expansion to achieve sufficient cell numbers. Using a two-dimensional (2D) cell migration assay to first identify the passage at which chondrocytes exhibited their greatest chondrogenic potential, the objective of this study was to determine a more optimal culture medium for developing three-dimensional (3D) cartilage-like tissues using human cells. We evaluated combinations of commonly used growth factors that have been shown to promote chondrogenic growth and development. Human articular chondrocytes (AC) from osteoarthritic (OA) joints were cultured in 3D environments, either in pellets or encapsulated in agarose. The effect of growth factor supplementation was dependent on the environment, such that matrix deposition differed between the two culture systems. ACs in pellet culture were more responsive to bone morphogenetic protein (BMP2) alone or combinations containing BMP2 (i.e. BMP2 with PDGF or FGF). However, engineered cartilage development within agarose was better for constructs cultured with TGFβ3. These results with agarose and pellet culture studies set the stage for the development of conditions appropriate for culturing 3D functional engineered cartilage for eventual use in human therapies.

Dexamethasone Release from Within Engineered Cartilage as a Chondroprotective Strategy Against Interleukin-1α

“…Recent work demonstrates the successful translation of this dex microsphere strategy to engineered constructs seeded with adult human chondrocytes 95 from three donors (Fig. 8), supporting the need for continued research efforts pursuing dex-loaded MS.…”

High seeding density of human chondrocytes in agarose produces tissue-engineered cartilage approaching native mechanical and biochemical properties

“…Relatively few studies have utilized hACs in CTE (Adkisson et al, 2010; Dehne et al, 2009; Dell’Accio et al, 2001; Fernandes et al, 2013; Kafienah et al, 2002; Lehmann et al, 2013; Saha et al, 2013; Sittinger et al, 1994; Wang et al, 2006; Wenger et al, 2006), due perhaps to logistical challenges of obtaining donor cells in sufficient numbers and encouraging them to produce sufficient matrix. Even fewer studies have characterized functional properties of human cartilage tissue constructs such as mechanical integrity (Murphy et al, 2015; O’Connell et al, 2015; Zhao et al, 2012). …”

“…Growth factor-aided monolayer expansion of hACs is generally necessary to obtain sufficient cell number for CTE studies (Barbero et al, 2003; Francioli et al, 2007; Jakob et al, 2001). We previously obtained E Y of 100 kPa and 2.2 %/ww GAG in agarose constructs cast with passaged chondrocytes from two osteoarthritic donors at 30 × 10 6 cells/mL for 56 days (O’Connell et al, 2015). While these results are promising for viable long-term 3D culture of hACs, functional properties were below native levels as well as levels in bovine constructs.…”

Nutrient Channels Aid the Growth of Articular Surface-Sized Engineered Cartilage Constructs

“…Cells cultured in monolayer were plated at 2 · 10 4 cells/cm 2 in expansion medium (high-glucose Dulbecco's modified Eagle's medium with 10% fetal bovine serum, 1% antibiotic-antimycotic, 10 ng/mL platelet-derived growth factor-bb, 5 ng/mL fibroblast growth factor-2, and 1 ng/mL TGF-b3). 45,48,51 After reaching confluence at second passage, P2 cells were trypsinized.…”

“…Thus, other studies have used passaged bovine, canine, or human chondrocytes. 34,[41][42][43][44][45][46][47][48][49] A direct comparison of passaged versus primary bovine chondrocytes in agarose could elucidate whether results derived with primary bovine cells are applicable for passaged cells.…”