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. 2017 Aug 9;10(8):924.
doi: 10.3390/ma10080924.

Histological and Immunohistochemical Analyses of Repair of the Disc in the Rabbit Temporomandibular Joint Using a Collagen Template

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Free PMC article

Histological and Immunohistochemical Analyses of Repair of the Disc in the Rabbit Temporomandibular Joint Using a Collagen Template

Kuo-Hwa Wang et al. Materials (Basel). .
Free PMC article

Abstract

A previous study demonstrated that the reconstituted type I collagen matrix extracted from rabbit tendons enabled the TMJ disc to regenerate in the rabbit. The aim of this study was to investigate changes in the extracellular matrix (ECM) and mechanisms of regeneration in the TMJ disc. In 36 New Zealand rabbits that underwent a partial discectomy, discs were replaced with reconstituted collagen templates for 3 months. A histological analysis showed that moderate to severe degeneration appeared in partially discectomized joints without implantation. In contrast, discs experienced regeneration of reconstituted collagen template implantation and the joint returned to normal function. Cells in the regenerative tissue expressed ECM, and fibers became regular and compact due to tissue remodeling over time. Reparative cells differentiated into chondroblasts, and showed highly dense pericellular fibers. The morphology and collagen composition of the disc and condyle in the 3-month experimental group were similar to those of normal tissues. In conclusion, the reconstituted collagen template facilitated the regeneration of surgically discectomized discs. Type I and type II collagens play a crucial role in the regeneration of articular discs.

Keywords: reconstituted collagen template; temporomandibular joint disc; tissue regeneration.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Analysis of the intact control. (A) Immunohistochemistry of type I collagen on normal temporomandibular joint (TMJ) cartilage, 800×. Type I collagen was expressed on the cartilage surface, bone, and proliferation zone (black arrow head); (B) Immunohistochemistry of type II collagen on normal TMJ cartilage, 800×. Type II collagen was expressed at the hyaline zone (white arrowhead) and proliferation zone (black arrow head); (C) hematoxylin and eosin (HE) analysis of a normal TMJ, 160×. Normal architecture of the disc, condyle, and cartilage; (D) HE analysis of the normal TMJ disc, 1600×. Chondrocytes, surrounded by wavy fibers, are scattered in the disc; (E) Immunohistochemistry of type I collagen in the normal TMJ disc, 1600×. Type I collagen was noted in wavy fibers; (F) Immunohistochemistry of type II collagen on a normal TMJ disc, 1600×. Type II collagen was expressed around chondrocyte-like cells; (G) Immunohistochemistry of type I collagen in the normal TMJ disc, 3200×. Magnification of (E); (H) Immunohistochemistry of type II collagen in the normal TMJ disc, 3200×. Magnification of (F).
Figure 2
Figure 2
Histological analysis of a 1-month discectomied TMJ without implantation. (A) One-month group, 320×. Mild chondroblast proliferation in the cartilage was evident. The condyle surface appears erosive and moderately deformed (black arrowhead). The remainder of the disc shows fibrous degeneration; (B) Immunohistochemistry of type I collagen on 1-month untreated TMJ cartilage, 800×. Type I collagen expression increased and surrounded proliferating cells on the cartilage surface; (C) Immunohistochemistry of type II collagen on 1-month untreated TMJ cartilage, 800×. Type II collagen, like type I, surrounded proliferating cells; (D) Immunohistochemistry of type I collagen on a 1-month untreated TMJ disc, 1600×. Type I collagen showed nearly normal expression; (E) Immunohistochemistry of type II collagen on a 1-month untreated TMJ disc, 1600×. Minimal expression was observed.
Figure 3
Figure 3
Histological analysis of the discectomied TMJ without implantation. (A) Two-month group, 320×. The remainder of the disc is characterized by fibrillary degeneration, and it was covered by a synovial lining. Marked cartilage enlargement and fragmentation, and fibrous degeneration of both the condyle (black arrowhead) and tympanic fossa was found; (B) Three-month group, 320×. The condyle shows complete erosion of cartilage from the subchondral bone; (C) Immunohistochemistry of type I collagen in 2-month untreated TMJ cartilage, 800×. Type I collagen was greatly expressed in enlarged cartilage; (D) Immunohistochemistry of type II collagen in 2-month untreated TMJ cartilage, 800×. The expression of type II collagen had significantly decreased in enlarged cartilage; (E) Immunohistochemistry of type I collagen in a 2-month untreated TMJ disc, 1600×. Like the normal disc, type I collagen expression was found in fibers; (F) Immunohistochemistry of type II collagen in the 2-month untreated TMJ disc, 1600×. Minimal type II collagen expression surrounds cells.
Figure 4
Figure 4
Discectomied TMJ with implantation in 1 month. (A) HE analysis of the temporomandibular joint (TMJ) with a 1-month reconstituted collagen template, 320×. Cartilage exhibited mild chondroblast proliferation with minimal erosion (black arrow). The disc shows fibrous degeneration; (B) Immunohistochemistry of type I collagen in a 1-month treated TMJ, 320×; (C) Immunohistochemistry of type II collagen in a 1-month treated condyle; 320× (D) Immunohistochemistry of type I collagen in 1-month treated TMJ cartilage, 800×. Type I collagen was increasingly expressed on the cartilage surface; (E) Immunohistochemistry of type II collagen in 1-month treated TMJ cartilage, 800×. Mild type II collagen expression surrounds proliferating cells on the surface; (F) Immunohistochemistry of type I collagen in a 1-month treated TMJ disc, 1600×. Type I collagen showed increasing expression; (G) Immunohistochemistry of type II collagen in a 1-month treated TMJ disc, 1600×. Decreased expression was observed.
Figure 5
Figure 5
Discectomied TMJ with implantation after 2 months. (A) HE stain, 320×. The disc appears to be somewhat thicker than that of the 1-month group. No erosion is present on either the tympanic fossa or condyle (black arrowhead); (B) Immunohistochemistry of type I collagen on 2-month treated TMJ cartilage, 800×. Type I collagen showed a slightly increasing expression; (C) Immunohistochemistry of type II collagen in 2-month treated TMJ cartilage, 800×. Expression of type II collagen was decreased compared to the intact control; (D) Immunohistochemistry of type I collagen in a 2-month treated TMJ disc, 1600×. Type I collagen was expressed on wavy fibers; (E) Immunohistochemistry of type II collagen in a 2-month treated TMJ disc, 1600×. Type II collagen was expressed surrounding chondrocyte-like cells; (F) Immunohistochemistry of type I collagen in a 2-month treated TMJ disc tip, 1600×. Type I collagen was expressed in chondrocyte-like cells; (G) Immunohistochemistry of type II collagen in a 2-month treated TMJ disc tip, 1600×. Less type II collagen was observed.
Figure 6
Figure 6
Discectomied TMJ with implantation in 3 months. (A) HE analysis of the temporomandibular joint (TMJ) with a 3-month reconstituted collagen template, 320×. New collagen bundles appeared in the disc and had adhered to the tympanic fossa; the condyle appears normal with a smooth condylar surface (black arrowhead) (B) HE analysis of the TMJ disc with a 3-month reconstituted collagen template, 1600×. The direction and arrangement of new fibers differed from those of old fibers; (C) Immunohistochemistry of type I collagen in 3-month treated TMJ cartilage, 1600×. Type I collagen had recovered to a normal state; (D) Immunohistochemistry of type II collagen in 3-month treated TMJ cartilage, 1600×. Expression of type II collagen was normal; (E) Immunohistochemistry of type I collagen in a 3-month treated TMJ disc, 3200×. Type I collagen was expressed in fibers; (F) Immunohistochemistry of type II collagen in a 3-month treated TMJ disc, 3200×. Type II collagen was expressed randomly in fibers adjacent to cells.

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