Stiffness Variation of 3D Collagen Networks by Surface Functionalization of Network Fibrils with Sulfonated Polymers

Fibrillar collagen is the most prominent protein in the mammalian extracellular matrix. Therefore, it is also widely used for cell culture research and clinical therapy as a biomimetic 3D scaffold. Charged biopolymers, such as sulfated glycosaminoglycans, occur in vivo in close contact with collagen fibrils, affecting many functional properties such as mechanics and binding of growth factors. For in vitro application, the functions of sulfated biopolymer decorations of fibrillar collagen materials are hardly understood. Herein, we report new results on the stiffness dependence of 3D collagen I networks by surface functionalization of the network fibrils with synthetic sulfonated polymers, namely, poly(styrene sulfonate) (PSS) and poly(vinyl sulfonate) (PVS). A non-monotonic stiffness dependence on the amount of adsorbed polymer was found for both polymers. The stiffness dependence correlated to a transition from mono- to multilayer adsorption of sulfonated polymers on the fibrils, which was most prominent for PVS. PVS mono- and multilayers caused a network stiffness change by a factor of 0.3 and 2, respectively. A charge-dependent weakening of intrafibrillar salt bridges by the adsorbed sulfonated polymers leading to fibrillar softening is discussed as the mechanism for the stiffness decrease in the monolayer regime. In contrast, multilayer adsorption can be assumed to induce interfibrillar bridging and an increase in network stiffness. Our in vitro results have a strong implication on in vivo characteristics of fibrillar collagen I, as sulfated glycosaminoglycans frequently attach to collagen fibrils in various tissues, calling for an up to now overlooked impact on matrix and tendon mechanics.

Collagen Scaffolds Treated by Hydrogen Peroxide for Cell Cultivation

Collagen in the body is exposed to a range of influences, including free radicals, which can lead to a significant change in its structure. Modeling such an effect on collagen fibrils will allow one to get a native structure in vitro, which is important for modern tissue engineering. The aim of this work is to study the effect of free radicals on a solution of hydrogen peroxide with a concentration of 0.006–0.15% on the structure of collagen fibrils in vitro, and the response of cells to such treatment. SEM measurements show a decrease in the diameter of the collagen fibrils with an increase in the concentration of hydrogen peroxide. Such treatment also leads to an increase in the wetting angle of the collagen surface. Fourier transform infrared spectroscopy demonstrates a decrease in the signal with wave number 1084 cm−1 due to the detachment of glucose and galactose linked to hydroxylysine, connected to the collagen molecule through the -C-O-C- group. During the first day of cultivating ASCs, MG-63, and A-431 cells, an increase in cell adhesion on collagen fibrils treated with H2O2 (0.015, 0.03%) was observed. Thus the effect of H2O2 on biologically relevant extracellular matrices for the formation of collagen scaffolds was shown.

An Overview of in vivo Functions of Chondroitin Sulfate and Dermatan Sulfate Revealed by Their Deficient Mice

Chondroitin sulfate (CS), dermatan sulfate (DS) and heparan sulfate (HS) are covalently attached to specific core proteins to form proteoglycans in their biosynthetic pathways. They are constructed through the stepwise addition of respective monosaccharides by various glycosyltransferases and maturated by epimerases as well as sulfotransferases. Structural diversities of CS/DS and HS are essential for their various biological activities including cell signaling, cell proliferation, tissue morphogenesis, and interactions with a variety of growth factors as well as cytokines. Studies using mice deficient in enzymes responsible for the biosynthesis of the CS/DS and HS chains of proteoglycans have demonstrated their essential functions. Chondroitin synthase 1-deficient mice are viable, but exhibit chondrodysplasia, progression of the bifurcation of digits, delayed endochondral ossification, and reduced bone density. DS-epimerase 1-deficient mice show thicker collagen fibrils in the dermis and hypodermis, and spina bifida. These observations suggest that CS/DS are essential for skeletal development as well as the assembly of collagen fibrils in the skin, and that their respective knockout mice can be utilized as models for human genetic disorders with mutations in chondroitin synthase 1 and DS-epimerase 1. This review provides a comprehensive overview of mice deficient in CS/DS biosyntheses.

Time-varying regularity of changes in biomechanical properties of the corneas after removal of anterior corneal tissue

Background The corneal biomechanical properties with the prolongation of time after corneal refractive surgery are important for providing a mechanical basis for the occurrence of clinical phenomena such as iatrogenic keratectasia and refractive regression. The aim of this study was to explore the changes of corneal elastic modulus, and stress relaxation properties from the 6-month follow-up observations of rabbits after a removal of anterior corneal tissue in simulation to corneal refractive surgery. Methods The anterior corneal tissue, 6 mm in diameter and 30–50% of the original corneal thickness, the left eye of the rabbit was removed, and the right eye was kept as the control. The rabbits were normally raised and nursed for 6 months, during which corneal morphology data, and both of corneal hysteresis (CH) and corneal resistance factor (CRF) were gathered. Uniaxial tensile tests of corneal strips were performed at months 1, 3, and 6 from 7 animals, and corneal collagen fibrils were observed at months 1, 3, and 6 from 1 rabbit, respectively. Results Compared with the control group, there were statistical differences in the curvature radius at week 2 and month 3, and both CH and CRF at months 1, 2, and 6 in experiment group; there were statistical differences in elastic modulus at 1, 3, and month 6, and stress relaxation degree at month 3 in experiment group. The differences in corneal elastic modulus, stress relaxation degree and the total number of collagen fibrils between experiment and control groups varied gradually with time, and showed significant changes at the 3rd month after the treatment. Conclusions Corneas after a removal of anterior corneal tissue undergo dynamic changes in corneal morphology and biomechanical properties. The first 3 months after treatment could be a critical period. The variation of corneal biomechanical properties is worth considering in predicting corneal deformation after a removal of anterior corneal tissue.