Evaluation of the In Vitro Cytotoxicity of Cross-Linked Biomaterials

Abstract

This study evaluated the in vitro cytotoxicity of poly(propylene fumarate) (PPF). PPF is an aliphatic biodegradable polymer that has been well characterized for use in bone tissue engineering scaffolds. Four different cell types, human mesenchymal stem cells (hMSC), fibroblasts (L929), pre-osteoblasts (MC3T3), and canine mesenchymal stem cells (cMSC), were used to evaluate the cytotoxicity of PPF. These cell types represent the tissues that PPF would interact with in vivo as a bone tissue scaffold. The sol fraction of the PPF films was measured and then utilized to estimate crosslinking density. Cytotoxicity was evaluated using XTT assay and fluorescence imaging. Results showed that PPF supported similar cell metabolic activities of hMSC, L929, MC3T3 and cMSC compared to the non-cytotoxic control, high density polyethylene (HDPE) and were statistically different than those cultured with the cytotoxic control, a polyurethane film containing 0.1% zinc diethyldithiocarbamate (ZCF). Results showed differing cellular responses to ZCF, the cytotoxic control. The L929 cells had the lowest cell metabolic activity levels after exposure to ZCF compared to the cell metabolic activity levels of the MC3T3, hMSC or cMSC cells. Qualitative verification of the results using fluorescence imaging demonstrated no change in cell morphology, vacuolization, or detachment when cultured with PPF compared to HDPE or blank media cultures. Overall the cytotoxicity response of the cells to PPF was demonstrated to be similar to the cytotoxic response of cells to known non-cytotoxic materials (HDPE).

Evaluating Changes in Structure and Cytotoxicity DuringIn VitroDegradation of Three-Dimensional Printed Scaffolds

“…These results are in line with our previous work that showed good cellular responses to highly crosslinked PPF thin films. 1 The novelty of this work is that we evaluated the potential cytotoxicity elicited during the absorption of PPF scaffolds. PPF is absorbed slower than other absorbable polyesters, such as the saturated aliphatic polyesters poly(glycolic) acid (PGA) or poly(l-lactic) acid (PLA).…”

“…PPF has already been shown to be noncytotoxic; however, the cytotoxicity of PPF during degradation has not been evaluated. 1 Therefore, we investigated the potential for a cytotoxic response to the byproducts of degradation.…”

“…The individual resin components have also been shown to be noncytotoxic; however, the cytotoxicity of the combined materials as well as during polymer absorption has not been evaluated. 1,7,9,10 In addition, we hypothesize that due to the slow degradation of PPF, the reduction in pH from the formation of the degradation byproduct fumaric acid would have a little to no detrimental impact as the cells would metabolize the fumaric acid. To test this hypothesis, we modified testing recommendations in ISO Standard 10993-5, ''Tests for In vitro Cytotoxicity '' 11 to evaluate polymer degradation cytotoxicity.…”

Reinforced Pericardium as a Hybrid Material for Cardiovascular Applications

“…For this purpose, the synthetic polymer poly(propylene fumarate) (PPF) was selected for its demonstrated biocompatibility, biodegradability, and strength in other biomedical applications. 18,19 PPF is an unsaturated linear polyester that is crosslinkable through UV radiation with itself or with other compatible crosslinkers through the double bonds in fumarate. PPF is biodegradable by hydrolysis of ester bonds and forms the naturally occurring byproducts fumaric acid and propylene glycol upon degradation.…”

Toxicity Analysis of RGD-Chitosan From Shrimp Shell Scaffold Membranes Toward Human Dental Pulp Cells

“…Currently, the Indonesia National Atomic Energy Agency (BATAN) has been developing chitosan scaffolds from shrimp shells in the form of a membrane combined with RGD. However, based on the ISO standard 10993, before the scaffold can be transplanted or used in tissue engineering, that scaffold should undergo a biocompatibility test [15]. The test should be performed to avoid any negative effect that may arise after the scaffold comes in contact with body tissue.…”