Full factorial design of experiment-based and response surface methodology approach for evaluating variation in uniaxial compressive mechanical properties, and biocompatibility of photocurable PEGDMA-based scaffolds

The goal of this study is to fabricate biocompatible and minimally invasive bone tissue engineering scaffolds that allowin situphotocuring and further investigate the effect on the mechanical properties of the scaffold due to the prevailing conditions around defect sites, such as the shift in pH from the physiological environment and swelling due to accumulation of fluids during inflammation. A novel approach of incorporating a general full factorial design of experiment (DOE) model to study the effect of the local environment of the tissue defect on the mechanical properties of these injectable and photocurable scaffolds has been formulated. Moreover, the cross-interaction between factors, such as pH and immersion time, was studied as an effect on the response variable. This study encompasses the fabrication and uniaxial mechanical testing of polyethylene glycol dimethacrylate (PEGDMA) scaffolds for injectable tissue engineering applications, along with the loss in weight of the scaffolds over 72 h in a varying pH environment that mimicsin vivoconditions around a defect. The DOE model was constructed with three factors: the combination of PEGDMA and nano-hydroxyapatite referred to as biopolymer blend, the pH of the buffer solution used for immersing the scaffolds, and the immersion time of the scaffolds in the buffer solution. The response variables recorded were compressive modulus, compressive strength, and the weight loss of the scaffolds over 72 h of immersion in phosphate-buffered saline at respective pH. The statistical model analysis provided adequate information in explaining a strong interaction of the factors on the response variables. Further, it revealed a significant cross-interaction between the factors. The factors such as the biopolymer blend and pH of the buffer solution significantly affected the response variables, compressive modulus and strength. At the same time, the immersion time had a strong effect on the loss in weight from the scaffolds over 72 h of soaking in the buffer solution. The biocompatibility study done using a set of fluorescent dyes for these tissue scaffolds highlighted an enhancement in the pre-osteoblasts (OB-6) cell attachment over time up to day 14. The representative fluorescent images revealed an increase in cell attachment activity over time. This study has opened a new horizon in optimizing the factors represented in the DOE model for tunable PEGDMA-based injectable scaffold systems with enhanced bioactivity.