Effects of Hydroxyapatite Additions on Alginate Gelation Kinetics During Cross-Linking.

Abstract

Alginate hydrogels have gathered significant attention in biomedical engineering due to their remarkable biocompatibility, biodegradability, and ability to encapsulate cells and bioactive molecules, but much less has been reported on the kinetics of gelation. Scarce experimental data are available on cross-linked alginates (AL) with bioactive components. The present study addressed a novel method for defining the crosslinking mechanism using rheological measurements for aqueous mixtures of AL and calcium chloride (CaCl₂) with the presence of hydroxyapatite (HAp) as filler particles. The time-dependent crosslinking behaviour of these mixtures was exploited using a plate-plate rheometer, when crosslinking occurs due to calcium ions (Ca²⁺) binding to the guluronic acid blocks within the AL polymer, forming a stable "egg-box" structure. To reveal the influence of HAp particles as filler on crosslinked sample morphology, after rheological measurement and crosslinking, crosslinked samples were freeze-dried and their morphology was assessed using an optical microscope and SEM. It was found that the addition of HAp particles, which are known to enhance the mechanical properties and biocompatibility of crosslinked AL gels, significantly decreased (usually rapidly) the interaction between the Ca²⁺ and AL chains. In this research, the physical "shielding" effect of HAp particles on the crosslinking of AL with Ca²⁺ ions has been observed for the first time, and its crosslinking behaviour was defined using rheological methods. After crosslinking and rheometer measurements, the samples were further evaluated for morphological properties and the observations were correlated with their dewatering properties. While the presence of HAp particles led to a slower crosslinking process and a more uniform development of the rheological parameters, it also led to a more uniform porosity and improved dewatering properties. The observed effects allow for a better understanding of the crosslinking process kinetics, which directly affects the physical and chemical properties of the AL gels. The shielding behaviour (retardation) of filler particles occurs when they physically or chemically block certain components in a mixture, delaying their interaction with other reactants. In hydrogel formulations, filler particles like hydroxyapatite (HAp) can act as barriers, adsorbing onto reactive components or creating physical separation, which slows the reaction rate and allows for controlled gelation or delayed crosslinking. This delayed reactivity is beneficial for precise control over the reaction timing, enabling the better manipulation of material properties such as crosslinking distribution, pore structure, and mechanical stability. In this research, the physical shielding effect of HAp particles was observed through changes in rheological properties during crosslinking and was dependent on the HAp concentration. The addition of HAp also enabled more uniform porosity and improved dewatering properties. The observed effects allow for a better understanding of the crosslinking process kinetics, which directly affects the physical and chemical properties of the AL gels.