Preparation and Characterization of a Novel Decellularized Fibrocartilage “Book” Scaffold for Use in Tissue Engineering

Abstract

At the tendon-to-bone insertion, there is a unique transitional structure: tendon, non-calcified fibrocartilage, calcified fibrocartilage, and bone. The reconstruction of this special graded structure after defects or damage is an important but challenging task in orthopedics. In particular, reconstruction of the fibrocartilage zone has yet to be successfully achieved. In this study, the development of a novel book-shape scaffold derived from the extracellular matrix of fibrocartilage was reported. Specifically, fibrocartilage from the pubic symphysis was obtained from rabbits and sliced into the shape of a book (dimensions: 10 mm × 3 mm × 1 mm) with 10 layers, each layer (akin to a page of a book) with a thickness of 100-μm. These fibrocartilage “book” scaffolds were decellularized using sequentially 3 freeze-thaw cycles, 0.1% Triton X-100 with 1.5 M KCl, 0.25% trypsin, and a nuclease. Histology and DNA quantification analysis confirmed substantial removal of cells from the fibrocartilage scaffolds. Furthermore, the quantities of DNA, collagen, and glycosaminoglycan in the fibrocartilage were markedly reduced following decellularization. Scanning electron microscopy confirmed that the intrinsic ultrastructure of the fibrocartilage tissue was well preserved. Therefore, the results of this study suggest that the novel “book” fibrocartilage scaffold could have potential applications in tissue engineering.

Preparation of Decellularized Triphasic Hierarchical Bone‐Fibrocartilage‐Tendon Composite Extracellular Matrix for Enthesis Regeneration

“…Another limitation to preparing D‐BFT composites is that fibrocartilage presents low vascularity and lacks direct cell–cell connections, allowing for solution infiltration of few chemicals. Triton‐X100 and SDS are typically used in existing protocols for preparing tendon‐to‐bone composites or for fibrocartilage decellularization . However, the decellularized tendon‐to‐bone composites did not assess their bio‐components, structures, biocompatibility, and ability to repair bone defects.…”

“…However, the boundary between the tendon and bone was still observed after 16 weeks . Moreover, to date, few methods are available for thoroughly removing cells from the enthesis scaffold or decellularization of the whole composite . Recently, Xu et al developed several protocols for preparing porcine Achilles tendon to bone composite.…”

Characterization of the Content and Distribution of Collagen and Proteoglycan in the Decellularized Book-Shaped Enthesis Scaffolds by SR-FTIR

“…The quanti cations of DNA in the decellularized book-shaped enthesis scaffolds were performed using DNeasy Blood & Tissue protocol according to the manufacturer's instructions [12]. Speci cally, the decellularized book-shaped enthesis scaffolds (N=3) were weighed and minced after freeze-dried for 24 h using a lyophilizer (SIM International Group, USA).…”

“…With the development of tissue engineering technology, decellularized bioscaffolds have received great attention due to their ability to enhance tissue regeneration and repair damaged tissues [10][11][12]. Previous studies indicated that organ-speci c extracellular matrix scaffolds derived from site-speci c homologous tissues may be better suited for constructive tissue remodeling than no site-speci c tissue sources.…”

Site-dependent acellularisation effects explain altered tissue mechanics: ultrastructural insights

“…Being freed of cells and cellular components, these scaffolds produce minute antigenicity [2,6,9] and provide a biologic environment for cellular ingrowth and subsequent remodelling. However, there is a lack of morphologic and functional data on how exactly the commonly used methods of acellularisation impact extracellular matrix integrity, including commonly used chemical and physical protocols [3,10].…”