Simvastatin-Enriched Macro-Porous Chitosan-Calcium-Aluminate Scaffold for Mineralized Tissue Regeneration

Cassiano, Fernanda Balestrero; Soares, Diana Gabriela; Bordini, Ester Alves Ferreira; Anovazzi, Giovana; Hebling, Josimeri; Costa, Carlos Alberto de Souza

doi:10.1590/0103-6440202003252

Cited by 8 publications

(7 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The released components of this composite biomaterial demonstrated improved cell proliferation and ALP activity, upregulating the mineralization by approximately four-fold compared to that displayed by the released components from single-component formulations. In our prior research on human pulp cells, the synergistic effect of bioactive mineral phases with low-dosage SV has been observed for chitosan-calcium hydroxide 4,5 and chitosancalcium aluminate formulations, 36 wherein an increase in odontoblastic markers was observed in the presence of adsorbed 1 µM SV, indicating a beneficial action of this association for the cell differentiation and deposition of the mineralized matrix. The incorporation of SV in CH-Ca scaffolds also has a chemotactic effect on human pulp cells.…”

Section: Discussionmentioning

confidence: 86%

Assessment of the regenerative potential of macro-porous chitosan-calcium simvastatin scaffolds on bone cells

et al. 2023

Self Cite

View full text Add to dashboard Cite

This study evaluated the bioactive potential of a macro-porous chitosan scaffold incorporated with calcium hydroxide (CH-Ca) and functionalized with bioactive doses of simvastatin (SV) for bone tissue regeneration. Initially, the bioactive dose of SV in osteoblastic cells (SAOS-2) was determined. For the direct contact experiment, SAOS-2 cells were plated on scaffolds to assess cell viability and osteogenic differentiation. The second assay was performed at a distance using extracts from scaffolds incubated in culture medium to assess the effect of conditioned medium on viability and osteogenic differentiation. The initial screening showed that 1 μM SV presented the best biostimulating effects, and this dose was selected for incorporation into the CH-Ca and pure chitosan (CH) scaffolds. The cells remained viable throughout the direct contact experiment, with the greatest cell density in the CH-Ca and CH-Ca-SV scaffolds because of their higher porosity. The CH-Ca-SV scaffold showed the most intense bio-stimulating effect in assays in the presence and absence of osteogenic medium, leading to an increased deposition of mineralized matrix. There was an increase in the viability of cells exposed to the extracts for CH-Ca, CH-SV, and CH-Ca-SV during the one-day period. There was an increase in ALP activity in the CH-Ca and CH-Ca-SV; however, the CH-Ca-SV scaffold resulted in an intense increase in the deposition of mineralized nodules, approximately 56.4% at 7 days and 117% at 14 days, compared with CH (control). In conclusion, functionalization of the CH-Ca scaffold with SV promoted an increase in bioactivity, presenting a promising option for bone tissue regeneration.

show abstract

Section: Discussionmentioning

confidence: 86%

Assessment of the regenerative potential of macro-porous chitosan-calcium simvastatin scaffolds on bone cells

et al. 2023

Self Cite

View full text Add to dashboard Cite

show abstract

“…Chitosan (CH) and chitosan-calcium-hydroxide (CH-Ca) scaffolds were prepared by phase separation, following the technique previously established by our group (Soares et al 2020). Simvastatin (SV; ≥97% high-performance liquid chromatography, solid; Sigma-Aldrich) at 1 µM was incorporated by scaffold immersion and drying at 37°C, as proposed by Soares, Anovazzi, et al (2018) and Cassiano et al (2020). Refer to Appendix Figure 1 for details.…”

Section: Methodsmentioning

confidence: 99%

“…In our previous experiments, incorporation of 1 μM SV on chitosan and chitosan-calcium-aluminate scaffolds had better results than lower dosages, increasing alkaline phosphatase (ALP) activity and mineralized matrix deposition. This dosage also had a homing effect, by attracting human dental pulp cells from a 3-dimensional (3D) culture to scaffold surface and improving the gene expression of relevant odontoblastic markers (Soares, Anovazzi, et al 2018; Cassiano et al 2020).…”

Section: Introductionmentioning

confidence: 99%

Chitosan-Calcium-Simvastatin Scaffold as an Inductive Cell-Free Platform

Soares¹,

Bordini

Bronze‐Uhle³

et al. 2021

J Dent Res

Self Cite

View full text Add to dashboard Cite

The development of biomaterials based on the combination of biopolymers with bioactive compounds to develop delivery systems capable of modulating dentin regeneration mediated by resident cells is the goal of current biology-based strategies for regenerative dentistry. In this article, the bioactive potential of a simvastatin (SV)–releasing chitosan-calcium-hydroxide (CH-Ca) scaffold was assessed. After the incorporation of SV into CH-Ca, characterization of the scaffold was performed. Dental pulp cells (DPCs) were seeded onto scaffolds for the assessment of cytocompatibility, and odontoblastic differentiation was evaluated in a microenvironment surrounded by dentin. Thereafter, the cell-free scaffold was adapted to dentin discs positioned in artificial pulp chambers in direct contact with a 3-dimensional (3D) culture of DPCs, and the system was sealed to simulate internal pressure at 20 cm/H2O. In vivo experiments with cell-free scaffolds were performed in rats’ calvaria defects. Fourier-transform infrared spectroscopy spectra proved incorporation of Ca and SV into the scaffold structure. Ca and SV were released upon immersion in a neutral environment. Viable DPCs were able to spread and proliferate on the scaffold over 14 d. Odontoblastic differentiation occurred in the DPC/scaffold constructs in contact with dentin, in which SV supplementation promoted odontoblastic marker overexpression and enhanced mineralized matrix deposition. The chemoattractant potential of the CH-Ca scaffold was improved by SV, with numerous viable and dentin sialoprotein–positive cells from the 3D culture being observed on its surface. Cells at 3D culture featured increased gene expression of odontoblastic markers in contact with the SV-enriched CH-Ca scaffold. CH-Ca-SV led to intense mineralization in vivo, presenting mineralization foci inside its structure. In conclusion, the CH-Ca-SV scaffold induces differentiation of DPCs into a highly mineralizing phenotype in the presence of dentin, creating a microenvironment capable of attracting pulp cells to its surface and inducing the overexpression of odontoblastic markers in a cell-homing strategy.

show abstract

“…Drug repurposing, another clinically useful approach to identify compounds that may be effective at healing the dentine–pulp complex, was also explored. Examples of drugs investigated in the repurposing studies included simvastatin (Cassiano et al, 2020 ; Karanxha et al, 2013 ) and fluocinolone acetonide (Liu et al, 2013 ). Furthermore, a relatively large number of studies explored the use of scaffolds such as hydrogels of chitosan and collagen, and compared different cell types, such as DPSCs, adipose tissue and bone marrow‐derived stem cells, for their use in dentine–pulp complex tissue engineering (Jin et al, 2020 ; Kim et al, 2009 ; Vagropoulou et al, 2021 ; Wang et al, 2016 ).…”

Section: Purpose Of Pulp Cell In Vitro Mineralizat...mentioning

confidence: 99%

A critical review ofin vitroresearch methodologies used to study mineralization in human dental pulp cell cultures

et al. 2022

View full text Add to dashboard Cite

Background:The pulp contains a resident population of stem cells which can be stimulated to differentiate in order to repair the tooth by generating a mineralized extracellular matrix. Over recent decades there has been considerable interest in utilizing in vitro cell culture models to study dentinogenesis, with the aim of developing regenerative endodontic procedures, particularly where some vital pulp tissue remains. Objectives:The purpose of this review is to provide a structured oversight of in vitro research methodologies which have been used to study human pulp mineralization processes. Method:The literature was screened in the PubMed database up to March 2021 to identify manuscripts reporting the use of human dental pulp cells to study mineralization. The dataset identified 343 publications initially which were further screened and consequently 166 studies were identified and it was methodologically mined for information on: i) study purpose, ii) source and characterization of cells, iii) mineralizing supplements and concentrations, and iv) assays and markers used to characterize mineralization and differentiation, and the data was used to write this narrative review.Results: Most published studies aimed at characterizing new biological stimulants for mineralization as well as determining the effect of scaffolds and dental (bio) materials. In general, pulp cells were isolated by enzymatic digestion, although the pulp explant technique was also common. For enzymatic digestion, a range of enzymes and concentrations were utilized, although collagenase type I and dispase were the most frequent. Isolated cells were not routinely characterized using either fluorescence-activated cell sorting (FACS) and magnetic-activated cell sorting (MACS) approaches and there was little consistency in terming cultures as dental pulp cells or dental pulp stem cells. A combination of media supplements, at a range of concentrations, of dexamethasone, ascorbic acid and beta-glycerophosphate, were frequently applied as the basis for the experimental conditions. Alizarin Red S

show abstract

Simvastatin-Enriched Macro-Porous Chitosan-Calcium-Aluminate Scaffold for Mineralized Tissue Regeneration

Cited by 8 publications

References 21 publications

Assessment of the regenerative potential of macro-porous chitosan-calcium simvastatin scaffolds on bone cells

Assessment of the regenerative potential of macro-porous chitosan-calcium simvastatin scaffolds on bone cells

Chitosan-Calcium-Simvastatin Scaffold as an Inductive Cell-Free Platform

A critical review ofin vitroresearch methodologies used to study mineralization in human dental pulp cell cultures

Contact Info

Product

Resources

About