Gelatin-based haemostyptic Spongostan as a possible three-dimensional scaffold for a chondrocyte matrix?

Abstract

The gelatin-based haemostyptic compound Spongostan was tested as a three-dimensional (3D) chondrocyte matrix in an in vitro model for autologous chondrocyte transplantation using cells harvested from bovine knees. In a control experiment of monolayer cultures, the proliferation or de-differentiation of bovine chondrocytes was either not or only marginally influenced by the presence of Spongostan (0.3 mg/ml). In monolayers and 3-D Minusheet culture chambers, the cartilage-specific differentiation markers aggrecan and type-II collagen were ubiquitously present in a cell-associated fashion and in the pericellular matrix. The Minusheet cultures usually showed a markedly higher mRNA expression than monolayer cultures irrespective of whether Spongostan had been present or not during culture. Although the de-differentiation marker type-I collagen was also present, the ratio of type-I to type-II collagen or aggrecan to type-I collagen remained higher in Minusheet 3-D cultures than in monolayer cultures irrespective of whether Spongostan had been included in or excluded from the monolayer cultures. The concentration of GAG in Minusheet cultures reached its maximum after 14 days with a mean of 0.83 +/- 0.8 microg/10(6) cells; mean +/-, SEM, but remained considerably lower than in monolayer cultures with/without Spongostan. Our results suggest that Spongostan is in principle suitable as a 3-D chondrocyte matrix, as demonstrated in Minusheet chambers, in particular for a culture period of 14 days. Clinically, differentiating effects on chondrocytes, simple handling and optimal formability may render Spongostan an attractive 3-D scaffold for autologous chondrocyte transplantation.

Cell-Type Specific Four-Component Hydrogel

“…Such an adhesion-function segregation is also evident in our endothelial and neuronal cultures in which, despite cell adhesion on quattroGels, angiogenesis and neurite formation were essentially abrogated. Notably, concerning cell proliferation, both gel types were essentially indistinguishable from each other, which is consistent with published data on mitotic indices on pure gelatin and fibrin hydrogels [26], [28]. Thus, a defined combination of two permissive gels resulted in a cell-type specificity with regard to cell differentiation but leaving proliferation essentially unaffected.…”

“…Despite the widespread application of fibrin- and gelatin matrices together with the crosslinking enzymes thrombin and transglutaminase [9], [21], [22], [23], [24], [25], [26], [27], no reports can be found describing the combination of these components to create a four-component blend with unique biological features. Based on a conceptual error, that the integration of fibrin-based proteolytic sites into a gelatin matrix would further accelerate neuronal infiltration, we observed just the opposite and identified a cell-type specificity that could be of interest for engineering indications that aim at fostering sessile rather than motile cells.…”

A novel in vitro bovine cartilage punch model for assessing the regeneration of focal cartilage defects with biocompatible bacterial nanocellulose

“…These included an increase of the mRNA for aggrecan/collagen type II over time and substantially decreased levels of collagen type I mRNA compared to those in condrocytes on the cartilage surface [76]. This suggests that BNC, as already observed for other biomaterials [75,77-79], is capable of stabilizing the chondrocytic phenotype. This was further supported by a substantial initial deposition of proteoglycan and collagen type II by the cells on the BNC surface in long-term high-density pellet cultures.…”

“…Chondrocytes emigrated onto the BNC surface showed certain signs of dedifferentiation, such as a fibroblastic phenotype, as well as higher expression of collagen type I mRNA and lower mRNA expression for aggrecan/collagen type II mRNA than in fresh cartilage [59,60,72-75]. It has to be taken into account, however, that a transient dedifferentiation may be beneficial for the recruitment of the cells from the cartilage matrix [64].…”

Is Macroporosity Absolutely Required for Preliminary in Vitro Bone Biomaterial Study? A Comparison Between Porous Materials and Flat Materials

“…Besides, increasing the HAC seeding density in the scaffolds (from 25 × 10 3 to 66 × 10 3 cells/mm 3 ) significantly improved chondrogenesis (up to 3.3-fold higher GAG accumulation and up to 9.3-fold higher type II collagen mRNA).[87]ChondrocyteAutologous chondrocytes and mesenchymal stem cells (MSCs)Monolayer3D pellet or fibrin-sealant constructThere was a proliferative effect for MSCs exposed to PRP in monolayer culture and an increase in the expression of chondrogenic markers when cells are exposed to a 3D environment.[88]ChondrocyteBovine chondrocytesMonolayer in flasks3D MinusheetThe Minusheet cultures usually showed a markedly higher mRNA expression than monolayer cultures. Besides, the ratio of type-I to type-II collagen or aggrecan to type-I collagen remained higher in Minusheet 3D cultures than in monolayer cultures.[89]Endothelial cellPrimary human artery-derived fibroblasts and human umbilical vein endothelial cellsMonolayer cell sheetsSelf-assembled cell-based microtissuesThe microtissue has significant enhancement of ECM expression and maturation.[90]FibroblastFibroblast cell line GD25b1Monolayer3D matrices derived mouse embryo sections or naturally deposited ECMThe relative content of unsaturated fatty acids, which serve as targets of oxidative attack, was observed to be higher in major phospholipids in plasma membranes of 3D cells.[91]FibroblastHuman foreskin fibroblastsGlass cover slips coated with different proteins or 3D matrix compressed to form localised 2D matrix3D matrices derived either from detergent-extracted mouse embryo sections or naturally deposited 3D ECM3D-matrix adhesions differ from focal and fibrillar adhesions characterized on 2D substrates in their content of α 5 β 1 and α v β 3 integrins, paxillin, other cytoskeletal components, and tyrosine phosphorylation of focal adhesion kinase (FAK). Relative to 2D substrates, 3D-matrix interactions also display enhanced cell biological activities and narrowed integrin...…”