Informing future cartilage repair strategies: a comparative study of three different human cell types for cartilage tissue engineering

Abstract

A major clinical need exists for cartilage repair and regeneration. Despite many different strategies having been pursued, the identification of an optimised cell type and of pre-treatment conditions remains a challenge. This study compares the cartilage-like tissue generated by human bone marrow stromal cells (HBMSCs) and human neonatal and adult chondrocytes cultured on three-dimensional (3D) scaffolds under various conditions in vitro and in vivo with the aim of informing future cartilage repair strategies based upon tissue-engineering approaches. After 3 weeks in vitro culture, all three cell types showed cartilage-like tissue formation on 3D poly (lactide-co-glycolide) acid scaffolds only when cultured in chondrogenic medium. After 6 weeks of chondro-induction, neonatal chondrocyte constructs revealed the most cartilage-like tissue formation with a prominent superficial zone-like layer, a middle zone-like structure and the thinnest fibrous capsule. HBMSC constructs had the thickest fibrous capsule formation. Under basal culture conditions, neonatal articular chondrocytes failed to form any tissue, whereas HBMSCs and adult chondrocytes showed thick fibrous capsule formation at 6 weeks. After in vivo implantation, all groups generated more compact tissues compared with in vitro constructs. Pre-culturing in chondrogenic media for 1 week before implantation reduced fibrous tissue formation in all cell constructs at week 3. After 6 weeks, only the adult chondrocyte group pre-cultured in chondrogenic media was able to maintain a more chondrogenic/less fibrocartilaginous phenotype. Thus, pre-culture under chondrogenic conditions is required to maintain a long-term chondrogenic phenotype, with adult chondrocytes being a more promising cell source than HBMSCs for articular cartilage tissue engineering.

Bone tissue engineering by using a combination of polymer/Bioglass composites with human adipose-derived stem cells

“…5 m sections were prepared and stained using Weigert's haematoxylin solutions before staining with 0.5% Alcian blue. After treatment with 1% molybdophosphoric acid, sections were stained using 0.1% Sirius red (Saha et al 2013;Yang et al 2006). …”

“…ADSCs were trypsinised and seeded, in MEM, onto sterile scaffolds (n=4 for each scaffold: 0%, 5% and 40% Bioglass ® in PDLLA) for 24 h. Cell-scaffold constructs were then placed into diffusion chambers (Diffusion Chamber Kit, Millipore). Intraperitoneal transplantation of the diffusion chambers (Saha et al 2013) was carried out under anaesthesia using a mixture (1:1, vol/vol) of Hypnorm (1:4 in sterile water) and Hypnorval (1:1 in sterile water) administered as intraperitoneal injections at 7-10 mL/kg body weight. After 8 weeks, the animals were sacrificed and the diffusion chambers were retrieved and fixed in 10% NBF for histological analysis.…”

Comparative Potential of Juvenile and Adult Human Articular Chondrocytes for Cartilage Tissue Formation in Three-Dimensional Biomimetic Hydrogels

“…Previous studies have compared adult and neonatal human chondrocytes, or adult and osteoarthritic (OA) human chondrocytes using two-dimensional (2D) or 3D culture platforms. [11][12][13] However, how chondrocytes from different stages (juvenile, adult, and diseased) compare in their potential for forming cartilage remains poorly understood. The aim of this study was to compare the potential of human adult and juvenile articular chondrocytes for cartilage formation and contrast it with OA chondrocytes.…”

Application of cell and biomaterial-based tissue engineering methods in the treatment of cartilage, menisci and ligament injuries

“…At the 10th week, meniscal regeneration was observed and collagen and PG content in the newly-formed tissue was similar to that of native tissue. Saha et al [106] studied three different cell types cultured under various conditions in vitro and in vivo on the PLGA-based scaffolds. Human bone marrow MSCs, human neonatal chondrocytes and adult chondrocytes were cultured in the chondrogenic medium on the cartilage phase of a commercial osteochondral scaffold.…”