In Vivo Cartilage Tissue Engineering Using a Cell‐Derived Extracellular Matrix Scaffold

Abstract

Abstract:We have observed in our previous study that a cell-derived extracellular matrix (ECM) scaffold could assure the growth of a cartilage tissue construct in vitro.The purpose of the present study was to evaluate the feasibility of a chondrocyte-seeded cell-derived ECM scaffold by implanting it in vivo in nude mouse. A porous cell-derived ECM scaffold was prepared with a freeze-drying protocol using porcine chondrocytes. Rabbit articular chondrocytes were seeded onto the scaffold and cultured for 2 days in vitro, and then implanted into the nude mouse subcutaneously. They were retrieved at 1, 2, and 3 weeks postimplantation. Under macroscopic analysis, the cartilage-like tissue formation matured with time and developed a smooth, white surface.Contrary to the control (in which no cells were seeded), the size of the neocartilage tissue increased slightly by the third week and remained more stable. Total glycosaminoglycan (GAG) content and the GAG/DNA ratio increased significantly with time in the chemical analysis. The histology exhibited a sustained accumulation of newly synthesized sulfated proteoglycans. Immunohistochemistry, Western blot, and reverse transcriptase-polymerase chain reaction (RT-PCR) clearly identified type II collagen at all time points. Compressive strength of in vivo neocartilage increased from 0.45 Ϯ 0.06 MPa at 1 week to 1.18 Ϯ 0.17 MPa at 3 weeks. In conclusion, this study demonstrated that the cell-derived ECM scaffold could provide chondrocytes with favorable in vivo environment to produce a hyaline-like cartilage tissue.

The chondrogenic differentiation of mesenchymal stem cells on an extracellular matrix scaffold derived from porcine chondrocytes

“…In particular, Hyaff, a hyaluronanbased scaffold, was also shown to support cartilage formation of chondrocytes better in a fiber type than in a sponge type [29]. In our previous studies, however, a sponge type ECM scaffold derived from porcine chondrocytes showed significantly better cartilage tissue formation of chondrocytes in vitro and in vivo than the fiber type PGA scaffold, which is superior in porosity, pore size and flexibility [13,14]. This result suggested that the biological composition of scaffolds could also be a critical factor in cartilage tissue formation.…”

“…The addition of chondroitin sulfate (CS) in the polyethylene glycol (PEG)-based hydrogel also improved chondrogenic differentiation of goat MSCs and, notably, reduced the expression of collagen type X and Cbfa-1/Runx-2, which are involved in hypertrophic change of cartilage [33]. The ECM scaffold consists of a mixture of various cartilage ECM molecules, predominantly of type II collagen and sulfated GAGs, and thereby provides an environmental context very similar to that of native cartilage tissue [13,14]. Therefore, the dynamic interplay between cells and the cartilage matrix environment may be a main contributing factor to the beneficial effect of the ECM scaffold used in this study.…”

“…This ECM scaffold was biodegradable, biocompatible and highly porous in nature. When compared with polyglycolic acid (PGA) scaffolds, the ECM scaffold was more efficient at producing high quality hyaline cartilage-like tissues using rabbit chondrocytes both in vitro and in vivo [13,14]. In contrast, the influence of the ECM scaffold on chondrogenic differentiation and in turn cartilage tissue formation of MSCs has not been addressed.…”

“…The ECM scaffold was prepared from porcine chondrocytes by our proprietary technique as previously described [13]. In brief, primary chondrocytes from porcine knee joints were first expanded in monolayer culture for 3 weeks and further cultured in a 3 dimensional (3-D) pellet for another 3 weeks.…”

Cartilage fragments from osteoarthritic knee promote chondrogenesis of mesenchymal stem cells without exogenous growth factor induction

“…6,7 Recently, more and more ECM-derived scaffolds have been examined and fabricated to offer a specific environment for cartilage repair or regeneration. [8][9][10][11][12] A previous study has revealed that cartilage extract matrix can promote the chondrogenesis of embryonic stem cells. 7 This suggests that articular cartilage ECM may play an important role in guiding the chondrogenic differentiation of MSCs.…”