Human chondroprogenitors in alginate-collagen hybrid scaffolds produce stable cartilagein vivo

Abstract

The goal of this study was to evaluate human epiphyseal chondroprogenitor cells (ECPs) as a potential new cell source for cartilage regeneration. ECPs were compared to human bone marrow stromal cells (MSCs) and human adult articular chondrocytes (ACs) for their chondrogenic potential and phenotypic stability in vitro and in vivo. The cells were seeded in Optimaix-3D scaffolds at 5 × 10 cells/mm and gene expression, matrix production and mechanical properties were analysed up to 6 weeks. In vitro, ECPs synthesized consistently high collagen 2 and low collagen 10. AC-seeded constructs exhibited high donor variability in GAG/DNA values as well as in collagen 2 staining, but showed low collagen 10 production. MSCs, on the other hand, expressed high levels of collagen 2 but also of collagens 1 and 10, and were therefore not considered further. In vivo, there was considerable loss of matrix proteins in ECPs compared to in vitro cultured samples. To overcome this, a second implantation study investigated the effect of mixing cells with alginate prior to seeding in the scaffold. ECPs in alginate maintained their cartilage matrix and resisted mineralization and vessel infiltration better 6 weeks after subcutaneous implantation, whereas ACs lost their chondrogenic matrix completely. This study shows the great potential of ECPs as an off-the-shelf, highly chondrogenic cell type that produces stable cartilage in vivo. Copyright © 2016 John Wiley & Sons, Ltd.

A comparative study of cartilage engineered constructs in immunocompromised, humanized and immunocompetent mice

“…We investigated how differences in the innate and adaptive immune systems of the mouse strain affect the quality and amount of extracellular matrix (ECM) produced and maintained by the cells. hCCs are a newly proposed cell source [24], which have previously shown to produce phenotypically stable cartilage in combination with HA-TG in vitro [22] and in different collagen scaffolds in an in vivo subcutaneous mouse model [25]. Allogeneic cells derived from fetal or juvenile tissue have the additional benefit of not only having a more stable phenotype and higher chondrogenic potential than adult articular chondrocytes [26], but also lower immunogenicity [27].…”

Holistic Approach of Swiss Fetal Progenitor Cell Banking: Optimizing Safe and Sustainable Substrates for Regenerative Medicine and Biotechnology

“…developmental efforts (Vrahas et al, 2004;Flanigan et al, 2010;Makris et al, 2015;Carluccio et al, 2020). The avascular and alymphatic nature of cartilage tissues confers relative immune privileges (i.e., isolation from antigen-presenting cells, migratory macrophages, and dendritic cells) and renders allogenic cell therapy approaches possible for tissue regeneration chaperoning (Quintin et al, 2010;Studer et al, 2017). Autologous cartilage cell therapy implementation remains hindered or delayed due to the induction of hypertrophic tissue phenotypes, fibrocartilage formation, high-cost cell expansions, in vitro de-differentiation, two-step surgery, donor-site morbidity, and high variability in functional outcomes (Brittberg et al, 1994;Horas et al, 2003;Lu et al, 2006;Katopodi et al, 2009;Vinardell et al, 2012).…”

“…High phenotypic stability and chondrogenic potential (i.e., elevated sulfated GAG content, Sox9:Scleraxis ratios, IHH and PTH1R gene expression, TGF-β3-induced production of aggrecan, types I+II collagen) of cartilage FPCs are differential advantages supporting their application in tissue engineering (Broguiere et al, 2016;Studer et al, 2017). Despite expressing stem cell surface markers, cartilage FPCs present relatively lower adipogenic and osteogenic differentiation capacities on a site-specific basis (Stokes et al, 2002;Quintin et al, 2010).…”

“…Despite expressing stem cell surface markers, cartilage FPCs present relatively lower adipogenic and osteogenic differentiation capacities on a site-specific basis (Stokes et al, 2002;Quintin et al, 2010). Conjugation with alginate optimally stimulates and maintains ECM production, while resisting mineralization and circulatory vessel infiltration in vivo, thereby drastically improving stability and therapeutic potential of cartilage FPCs, along with optimal structural parameters (Häuselmann et al, 1994;Mellor et al, 2014;Mhanna et al, 2014;Studer et al, 2017). Polyethylene glycol, chitosan, albumin, or hyaluronan scaffolds have been investigated as functional cell vectors for injectable applications, yielding adhesive, chondrogenic, and mitogenic properties (Madeira et al, 2015;Mardones et al, 2015).…”

“…Various dynamic cell seeding protocols for the induction of active infiltration (e.g., perfusion, centrifugation, orbital shaking, spinner flasks) allow cell distribution uniformity and optimal preservation of cellular integrity and function (Burg et al, 2000;Alvarez-Barreto et al, 2007;Roh et al, 2007;Thevenot et al, 2008). An equilibrium must be reached between cell proliferation and adequate chondrogenesis (i.e., responsiveness versus stability) following homogeneous scaffold seeding, directly defining adequate seeding density, methods for construct obtention, and preculture conditions (Roche et al, 2001;Moretti et al, 2005;Hasegawa et al, 2010;Erickson et al, 2012;Nasrollahzadeh et al, 2017;Studer et al, 2017). This ultimately results in the integration of structural and mass transport properties with the functional chondrogenesis components FIGURE 10 | Clinical case-reports illustrating the use and efficacy of Progenitor Biological Bandages for the management of human pediatric burns and donor-site wounds.…”