Synthesis of Human Cartilage Using Organotypic Cell Culture

Abstract

The limited supply of fresh autologous cartilage tissue for use in reconstructive surgery necessitates the use of vital banked allografts. A feasible in vitro production of cartilage tissue composed of living cells requires the use of modern tissue culture techniques retaining the phenotypic characteristics of chondrocytes. With this purpose in mind, human chondrocytes were isolated and cultured using different culture procedures: monolayer, suspension and agar gel. The differentiation state of chondrocytes as well as proteoglycan and collagen syntheses were assessed by histochemical and immunohistochemical methods. Whereas chondrocytes in monolayer displayed an unstable phenotype and tended to dedifferentiate, in three-dimensional culture the chondrocytes remained morphologically, phenotypically and functionally differentiated. Furthermore, an accumulation of matrix products pericellularly was observed in the agar gel. The results suggest that three-dimensional cultures in agar gel may allow the in vitro production of bioartificial cartilage for transplantation.

Poroviscoelasto-plasticity of agarose-based hydrogels

“…1–3 For example, chitin is important for medical devices and wound-healing dressings, 4 carrageenan films play an important role in extending the shelf life of foods, 5 alginate is used to prevent dehydration of meats, 6 and agar is common for culturing cells. 7–9 Biopolymers are selected for their ease of manufacture from natural precursor materials as well as for their ability to bear loads over long time scales. However, biopolymers also exhibit complex behaviors that may influence their durability and function.…”

Perichondrium-Wrapped Collagenous Matrices to Induce Chondroneogenesis

“…Bean and coworkers 6,7 have shown that demineralized bovine bone matrix can be transformed into a suitable cartilage substitute by exposing this material to a vascularized perichondrial flap. 6,7 In vitro studies by Bujía et al 8,9 showed that it is possible to obtain a 3-dimensional cartilaginous structure from polylactic acid fleeces and human chondrocytes in agarose gel. Kim et al 10 engineered nasoseptal cartilage replacements through a process of neomorphogenesis of cartilage by culturing bovine chondrocytes in close relation to biodegradable polymers composed of polyglycolic acid and poly-L-lactic acid.…”

“…The goal of the present study was to find a model for cartilage regeneration in vitro. 8,9,11 For this purpose, different collagenous matrices were wrapped in a perichondrium layer. 7,12,13 The matrices serve as a carrier for migration of cells from the perichondrium into the matrix.…”