Elsevier

Biomaterials

Volume 24, Issue 19, August 2003, Pages 3255-3263
Biomaterials

Cross-linked type I and type II collagenous matrices for the repair of full-thickness articular cartilage defects—A study in rabbits

https://doi.org/10.1016/S0142-9612(03)00143-1Get rights and content

Abstract

The physico-chemical properties of collagenous matrices may determine the tissue response after insertion into full-thickness articular cartilage defects. In this study, cross-linked type I and type II collagen matrices, with and without attached chondroitin sulfate, were implanted into full-thickness defects in the femoral trochlea of adolescent rabbits. The tissue response was evaluated 4 and 12 weeks after implantation by general histology and two semi-quantitative histological grading systems.

Four weeks after implantation, type I collagenous matrices were completely filled with cartilage-like tissue. By contrast, type II collagenous matrices revealed predominantly cartilaginous tissue only at the superficial zone and at the interface of the matrix with the subchondral bone, leaving large areas of the matrix devoid of tissue. Attachment of chondroitin sulfate appeared to promote cellular ingrowth and cartilaginous tissue formation in both types of collagen matrices.

Twelve weeks after implantation, the differences between the matrices were less pronounced. The deep parts of the subchondral defects were largely replaced by new bone with a concomitant degradation of the matrices. The original cartilage contours in defects with type I collagen-based matrices were repaired with fibro-cartilaginous tissue. Defects containing type II matrices showed an increase in the amount of superficial cartilage-like tissue. The original contour, however, was not completely restored in all animals, occasionally leaving a central depression or fissure.

It is concluded that different types of collagen matrices induce different tissue responses in full-thickness articular cartilage defects. Type I collagen-based matrices are superior to guide progenitor cells from a subchondral origin into the defect. In type II collagen-based matrices cell migration is less, but invading cells are directed into a chondrocyte phenotype. Based on these observations it is suggested that a composite matrix consisting of a deep layer of type I collagen and a more superficial layer of type II collagen may be the matrix of choice for cartilage regeneration.

Introduction

The highly limited potential of articular cartilage to regenerate has led to various procedures to restore cartilage defects. Classical techniques, such as Pridy drilling [1] and microfracture [2], use the potential of non-differentiated subchondral pluripotent cells to migrate into the defect and to differentiate locally into cartilage cells [3], [4]. Recently, methods have been developed for the repair of articular cartilage on the basis of autologous cartilage–bone transplants [5], [6], [7] or transplantation of cultured autologous chondrocytes [4], [8], [9], [10]. A major concern of all cartilage repair techniques, however, is that the produced cartilage repair tissue tends to be of the mechanical and biological inferior fibro-cartilaginous type [11], [12], [13], [14], [15], [16]. So far it is not known if the results of the more recent techniques are superior to the classical ones, with respect to clinical outcome and the nature of the repair tissue.

An extra dimension to cartilage repair techniques is the use of porous matrices. These matrices may function as vehicles for the transplantation of chondrocytes and offer temporary support to the cells. Matrices can also be used to guide the infiltration of progenitor cells from the bone marrow and to stimulate these cells to adopt a cartilage phenotype [10], [17], [18]. Cell behavior like migration, proliferation and differentiation may be mediated by the physico-chemical properties of the matrices [10], [11], [12], [13], [14], [15], [16], [17], [18], [19].

Various matrices have been used in orthopedic applications. These include matrices based on polylactic acid [9], [20], [21], polyglycolic acid [22], fibrin glue [23], [24], alginate [25], [26], [27], collagen [28], [29], [30], [31], [32] and hyaluronic acid [25], [28], [29]. Particularly, collagen-based matrices may be promising in this respect due to their biocompatibility, biodegradability and mechanical integrity.

In this study, cross-linked type I and type II collagen matrices, with and without attached chondroitin sulfate (CS), were implanted into full-thickness articular cartilage defects in the trochlea of rabbits. Cartilage and subchondral bone remodeling was evaluated 4 and 12 weeks after implantation using histology, and two semi-quantitative histological grading systems.

Section snippets

Preparation, cross-linking and characterization of collagen matrices

Insoluble type I collagen was isolated and purified from bovine Achilles tendon using neutral salt and dilute acid extractions [33]. Reconstituted type II collagen was isolated and purified from bovine tracheal cartilage using pepsin digestions, specific salt precipitation and dialysis against phosphate buffer [34]. CS was isolated and purified from bovine tracheal cartilage using extensive papain digestion, mild alkaline borohydride treatment and DEAE ion exchange chromatography [35]. Porous

Physicochemical characteristics of matrices

The physico-chemical characteristics of the different collagenous matrices are presented in Table 1. EDC treatment of matrices increases the denaturation temperature, and decreases the amine group content, indicating that cross-linking occurred. The amount of CS attached to type I and type II collagen matrices is 13% (w/w) and 18% (w/w), respectively.

Clinical evaluation and gross morphology

No difference in wound repair was observed between rabbits with and without scaffolds. After wound repair there were no clear changes in walking

Discussion

Partial-thickness defects in which damage is limited to the cartilage, and which do not extend into the subchondral bone, never repair spontaneously. This is due to the absence of an appropriate source of cells for repair, and indicates the value of the subchondral bone marrow as a pool for non-differentiated mesenchymal cells. The implantation of collagenous matrices in full-thickness defects may guide the infiltration, proliferation, and differentiation of these progenitor cells, and improve

Acknowledgments

The study was made possible by financial contributions of the foundations “Stichting De Drie Lichten” and the “Stichting Anna Fonds”. The authors wish to thank Mrs. D. Versleyen for skilful technical support with the histology.

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