Elaboration and Biocompatibility of an Eggshell-Derived Hydroxyapatite Material Modified with Si/PLGA for Bone Regeneration in Dentistry

Abstract

Hydroxyapatite (HAp) is the most commonly used biomaterial in modern bone regeneration studies because of its chemical similarity to bone, biocompatibility with different polymers, osteoconductivity, low cost, and lack of immune response. However, to overcome the disadvantages of HAp, which include fragility and low mechanical strength, current studies typically focus on property modification through the addition of other materials. Objective. To develop and evaluate the biocompatibility of a HAp material extracted from eggshells and modified with silicon (Si) and poly(lactic-co-glycolic) acid (PLGA). Materials and Methods. An in vitro experimental study in which a HAp material prepared from eggshells was synthesized by wet chemical and conventional chemical precipitation. Subsequently, this material was reinforced with Si/PLGA using the freezing/lyophilization method, and then osteoblast cells were seeded on the experimental material (HAp/Si/PLGA). To analyse the biocompatibility of this composite material, scanning electron microscopy (SEM) and fluorescence confocal microscopy (FCM) techniques were used. PLGA, bovine bone/PLGA (BB/PLGA), and HAp/PLGA were used as controls. Results. A cellular viability of 96% was observed for the experimental HAp/Si/PLGA material as well as for the PLGA. The viability for the BB/PLGA material was 90%, and the viability for the HAp/PLGA was 86%. Cell adhesion was observed on the exterior surface of all materials. However, a continuous monolayer and the presence of filopodia were observed over both external and internal surface of the experimental materials. Conclusions. The HAp/Si/PLGA material is highly biocompatible with osteoblastic cells and can be considered promising for the construction of three-dimensional scaffolds for bone regeneration in dentistry.

In Vitro Degradation and Cytotoxicity of Eggshell-Based Hydroxyapatite: A Systematic Review and Meta-Analysis

“…Out of 71 relevant article titles collected from SCOPUS, PubMed, and Google Scholar, only 36 were included after screening the title, abstract, and content for in vitro degradation and cytotoxicity testing for the biocompatibility of eggshell-based hydroxyapatite. The articles came from several countries, such as India [ 11 , 14 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 ], Bangladesh [ 41 , 42 ], China [ 43 ], Korea [ 25 , 44 , 45 ], Turkey [ 46 , 47 , 48 ], Thailand [ 49 , 50 ], Brazil [ 26 ], Italy [ 51 ], USA [ 52 ], Romania [ 2 ], Vietnam [ 53 , 54 ], Colombia [ 55 ], and Egypt [ 56 ]. The study selection of the articles included in the systematic review is shown in Figure 1 .…”

“…The statistical significance of the difference between the control and treatment groups was evaluated using a one-way analysis of variance (ANOVA) [ 45 ]. The quantification of viable cells was evaluated with the help of fluorescence microscopy [ 2 , 25 , 26 ], field emission scanning electron microscopy (FESEM) [ 52 ], and scanning electron microscopy [ 55 ]. The optical density test results from articles [ 2 , 20 , 49 , 50 ] are displayed in Figure 4 with a comparison of the control samples and treatment material variations, which can be used to determine the value of cell viability.…”

“…The addition of Si and Mg to the hydroxyapatite composite can increase the viability and proliferation of cells in osteoblasts [ 55 ]. Yilmaz et al [ 47 ] also tested HA/CS/GO composites, in which the addition of graphite was shown to increase the osteoconductivity of the materials.…”

Biocompatibility of Polylactic Acid as a Bone Substitute: An In Vitro Study

“…Each test is carried out thrice and the IC50 of the test samples as the percentage survival of the cells are calculated. (Gutiérrez-Prieto et al, 2019) Invitro Bromodeoxyuridine (BrDu) cell proliferation assay: The 10 mM stock solution of BrdU assay is prepared by dissolving 3 mg of BrdU in 1 mL of water. Then, the 10 mM BrdU stock solution is diluted into a cell culture medium which forms a 10 µM BrdU of labeling solution.…”

Preparation of Nano-hydroxyapatite Obtained from Lates Calcarifer Fish Bone by Alkaline Hydrolysis Method

“…HAp can be obtained by two main ways: (1) synthesis from compounds containing calcium and phosphorus [6]; (2) extraction from natural sources including corals [11][12][13], cuttlefish bones [14][15][16][17][18], seashells from clams, oysters, snails [19][20][21][22][23], eggshell [24,25], mammalian bone (e.g. bovine [26,27], camel and horse [28]), fish scales [29][30][31][32], and fish bones [33][34][35][36].…”