Serum albumin is the most prominent protein in blood, and it aids in bone fracture healing, though the manner through which enhanced healing occurs is not well understood. This study investigates the influence of calcium on the bioactivity of albumin due to the prevalence of calcium at bone injury sites. Bovine serum albumin (BSA) was exposed to varying concentrations of calcium, adsorbed to tissue culture polystyrene, and the subsequent BSA-coated surfaces were evaluated with calcium titration, and cell adhesion, viability, and binding inhibition studies. Calcium-modified BSA improved overall MC3T3-E1 osteoblast-like cell adhesion, although high calcium concentrations induced cell death. Inhibiting specific integrins revealed that without calcium exposure, cell binding to BSA was primarily mediated by integrins that typically bind to the GFOGER sequence of collagen. As calcium exposure increases, the primary binding interaction transitioned to integrins known to bind RGD. However, cell binding to calciummodified BSA was not completely eliminated during the inhibition studies indicating additional unidentified binding interactions occur. Overall, these results suggest that the exposure to calcium induces conformational changes that affect the cell-binding bioactivity of BSA, which may explain the beneficial impact of albumin in bone tissue.
Polyampholytes are a class of polymers made up of positively and negatively charged monomer subunits. Polyampholytes offer a unique tunable set of properties driven by the interactions between the charged monomer subunits. Some tunable properties of polyampholytes include mechanical properties, nonfouling characteristics, swelling due to changes in pH or salt concentration, and drug delivery capability. These characteristics lend themselves to multiple biomedical applications, and this review paper will summarize applications of polyampholyte polymers demonstrated over the last five years in tissue engineering, cryopreservation and drug delivery.
Tissue-engineered scaffolds encounter many challenges including poor integration with native tissue. Nonspecific protein adsorption can trigger the foreign body response leading to encapsulation and isolation from the native injured tissue. This concern is mitigated with nonfouling polymer scaffolds. This study investigates the long-term biocompatibility of a nonfouling polyampholyte system composed of positively charged [2-(acryloyloxy)ethyl]trimethylammonium chloride monomers and negatively charged 2-carboxyethyl acrylate monomers, cross-linked with triethylene glycol dimethacrylate. This system has previously shown resistance to nonspecific protein adsorption and short-term cell attachment via conjugated proteins. However, longer-term cell survival has not been evaluated with this system. First, the environmental pH was monitored with varying amounts of counter ions present in the hydrogel synthesis buffer. The lowest level (3 M NaOH) and the level that resulted in pH values closest to physiological conditions (6.7 M NaOH) were chosen for further investigation. These two formulations were then compared in terms of their contact angle, qualitative protein adsorption and conjugation capacity, and quantitative cell adhesion, proliferation, and viability. The 3 M NaOH formulation showed higher initial protein conjugation and cell adhesion compared to the 6.7 M NaOH formulation. However, the 3 M NaOH hydrogels had low cell viability after 24 h due to the acidic component release into the culture environment. The 6.7 M NaOH formulation showed a lower initial conjugation and cell adhesion but overcame this limitation by providing a stable environment that maintained cell viability for over 5 days. The 6.7 M NaOH polyampholyte hydrogel formulation shows increased biocompatibility, while maintaining resistance to nonspecific protein adsorption, as demonstrated by the targeted cell adhesion and proliferation. Therefore, this polyampholyte formulation demonstrates strong potential as a tissue-engineered scaffold.
Polymeric tissue engineering scaffolds have shown promise to aid in regeneration and repair of damaged tissue. In particular, nonfouling polymers have been proposed for eliminating biomaterial-induced concerns such as infection, scarring, and rejection by the immune system. Polyampholyte polymers are one class of nonfouling polymers that are composed of an equimolar mixture of positively and negatively charged monomer subunits. They possess nonfouling properties, bioactive molecule conjugation capabilities, and tunable mechanical properties. In this study, the influence of the cross-linker species on the degradation behavior, mechanical strength, and nonfouling properties of polyampholytes composed of a 1:1 molar ratio of [2-(acryloyloxy)ethyl] trimethylammonium chloride (positively charged) and 2-carboxyethyl acrylate (negatively charged) monomers was investigated. Specifically, the impact of ethylene glycol repeat units on the overall material performance was evaluated by synthesizing and characterizing hydrogels containing di-, tri-, and tetra-ethylene glycol dimethacrylate cross-linker species. The degradation studies were conducted for over 100 days in Sorenson's buffer with pH values of 4.5, 7.4, and 9.0 by tracking the swelling behavior and weight change over time. The mechanical properties were assessed using compression and tensile testing to failure. The retention of the nonfouling and protein conjugation capabilities was demonstrated using fluorescently labeled bovine serum albumin. The results demonstrate the tunability of both degradation behavior and mechanical properties through the cross-linker selection, without impacting the underlying nonfouling and biomolecule delivery capabilities. Therefore, it is concluded that polyampholyte hydrogels represent a promising platform for tissue engineering.
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