Changes in the avascular area of the meniscus using mesenchymal stem cells and growth plate chondrocytes in a pig model

Abstract

Menisci are wedge-shaped cartilage discs that are divided into two parts: the avascular and vascular regions. They are formed by fibrocartilage tissue, which contains round cartilage-like cells and extracellular matrix. Meniscus injury in animals is a common orthopedic problem, but data on the natural healing process mainly deals with the vascular zone. The healing processes in the avascular zone of the meniscus are significantly limited. Thus, this study aimed to evaluate autologous growth plate chondrocytes' impact on the healing process of a damaged meniscus in the avascular zone based on a growing animal model. The study group consisted of 10 pigs at about three months of age. From each animal, chondrocytes from the iliac growth plate and from concentrated bone marrow were taken. Knee joints were divided into right (R) and left (L). The medial meniscus of the R knee joint was treated with a hyaluronic acid based scaffold incubated with bone marrow cells from marrow aspirates (nCHON). The medial meniscus of the L knee joint was treated with a hyaluronic acid based scaffold incubated with bone marrow cells from marrow aspirates supplemented with immature chondrocytes isolated from growth plates (wCHON). The meniscus was damaged in the avascular zone in both knee joints. Followingly, the damaged part of the meniscus was filled with a scaffold with cells from the concentrated bone marrow and from growth plate chondrocytes. In the control group, a scaffold with concentrated bone marrow cells was used. After three months the animals were euthanized and preparations (microscopic slides) were made from the meniscus' damaged part. A qualitative and quantitative analysis have been prepared. The wCHON group in comparison with the nCHON group showed a statistically significantly higher number of fusiform cells on the surface of the graft as well as better healing of the graft. In addition, the degree of vascularization was higher in specimens from the wCHON group than in the nCHON group. The results of our research on immature pig knees revealed that mesenchymal stem cell and growth plate chondrocytes could be treated as the cell source for meniscus reconstruction, and growth plate chondrocytes enhance healing processes in the avascular zone of the injured meniscus.

Keywords: growth plate chondrocytes; meniscus tear; mesenchymal stem cells; pigs; scaffold.

FIGURE 1

nCHON group—experiment scheme

FIGURE 2

wCHON group—experiment scheme

FIGURE 3

Histological assessment of meniscus using HE staining, Masson staining, and Safranin O staining. (a–c) meniscus nCHON—severe with no graft healing, (d–f) meniscus wCHON – normal healing with normal fusiform cells, but moderate residual scaffold, (g–i) meniscus wCHON with normal graft healing, normal fusiform cells, severe fibroblasts in graft, severe Masson staining, severe Safranin O staining, (j–l) meniscus nCHON – severe graft healing, major fibroblast graft, major residual scaffold, minor Masson staining, moderate Safranin O staining. (m) meniscus, (sc) scaffold. Light microscope, 200×

FIGURE 4

Boxplots for selected quantifiers (according to Wilcoxon test) influenced by the applied treatment (no cells; nCHON and cells; wCHON; p < .05), corresponding to Table 2