The culture of articular chondrocytes in hydrogel constructs within a bioreactor enhances cell proliferation and matrix synthesis

Abstract

Bovine and human articular chondrocytes were seeded in 2% alginate constructs and cultured for up to 19 days in a rotating-wall-vessel (RWV) and under static conditions. Culture within the RWV enhanced DNA levels for bovine chondrocyte-seeded constructs when compared with static conditions but did not produce enhancement for human cells. There was a significant enhancement of glycosaminoglycans and hydroxyproline synthesis for both bovine and human chondrocytes. In all cases, histological analysis revealed enhanced Safranin-O staining in the peripheral regions of the constructs compared with the central region. There was an overall increase in staining intensity after culture within the RWV compared with static conditions. Type-II collagen was produced by both bovine and human chondrocytes in the peripheral and central regions of the constructs and the staining intensity was enhanced by culture within the RWV. A capsule of flattened cells containing type-I collagen developed around the constructs maintained under static conditions when seeded with either bovine or human chondrocytes, but not when cultured within the RWV bioreactor.

Engineering of Large Cartilaginous Tissues Through the Use of Microchanneled Hydrogels and Rotational Culture

“…These applied dynamic culturing conditions (perfusion/fluid flow, hydrostatic pressure or dynamic compressive loading) enhance developing neo-tissues F o r P e e r R e v i e w 4 through enhanced nutrient delivery and metabolic waste removal (15)(16)(17) and/or by providing a mechanical stimulus to the cells (6,(18)(19)(20)(21)(22)(23)(24)(25)(26)(27)(28)(29). For example, Akmal et.…”

“…al. (17), showed that bovine encapsulated alginate hydrogels cultured in a rotating-wall vessel (RWV) enhanced DNA, glycosaminoglycan and collagen levels. However, histological analysis revealed enhanced Safranin-O staining in the peripheral regions of the constructs compared with the central region.…”

In vitro resistance of articular chondrocytes to 5‐Aminolevulinic acid based photodynamic therapy

“…The alginate cell culture system was chosen since only a three-dimensional construct is able to maintain the phenotype of the chondrocytes: maintenance of the production of cartilage specific matrix proteins such as collagen type II and aggrecan and a low proliferation rate [26][27][28]. The diameter of the alginate beads measures 2-3 mm imposing the question whether sufficient oxygen is delivered to the cells for phototoxic reactions to occur [29].…”

Application of cell and biomaterial-based tissue engineering methods in the treatment of cartilage, menisci and ligament injuries

“…To date, no consensus has been reached concerning optimal cell densities in cell-based therapeutic methods. In different reports cell numbers ranged from 2.5 × 10 5 [53] to 1 × 10 7 per cm 2 [54]. Another study reported that MSCs require an even higher cell density during the chondrogenesis process [55].…”