Development of 3D-printed PLGA/TiO2 nanocomposite scaffolds for bone tissue engineering applications.

Porous scaffolds were 3D-printed using poly lactic-co-glycolic acid (PLGA)/TiO2 composite (10:1 weight ratio) for bone tissue engineering applications. Addition of TiO2 nanoparticles improved the compressive modulus of scaffolds. Differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) revealed an increase in both glass transition temperature and thermal decomposition onset of the composite compared to pure PLGA. Furthermore, addition of TiO2 was found to enhance the wettability of the surface evidenced by reducing the contact angle from 90.5 ±â€¯3.2 to 79.8 ±â€¯2.4 which is in favor of cellular attachment and activity. The obtained results revealed that PLGA/TiO2 scaffolds significantly improved osteoblast proliferation compared to pure PLGA (p < 0.05). Furthermore, osteoblasts cultured on PLGA/TiO2 nanocomposite showed significantly higher ALP activity and improved calcium secretion compared to pure PLGA scaffolds (p < 0.05).

The efficacy of commercial tooth storage media for maintaining the viability of human periodontal ligament fibroblasts.

AIM: To evaluate Save-A-Tooth (SAT), EMT Toothsaver (EMT) and Hank's Balanced Salt Solution (HBSS) for their influence on the viability and proliferative capacity of human periodontal ligament fibroblasts (HPDLFs). METHODOLOGY: Primary HPDLFs were seeded into 96-well cell culture plates and exposed to SAT, EMT, HBSS and water (negative control) for 0.5, 1, 3, 6, 12 and 24 h at room temperature (22 °C). After each exposure time, cell viability was measured through quantifying adenosine triphosphate (ATP) using a luminescent dye. The proliferative capacity was also quantified using the PrestoBlue assay after 12 or 24 h storage in each medium. The data were analysed statistically by two-way anova and post hoc Least Significant Difference (LSD) test (P < 0.05). The morphology of the cells after 12 h storage was also investigated through live/dead viability/cytotoxicity kit together with fluorescence microscopy. RESULTS: There was no significant difference in cell viability amongst HBSS, SAT and EMT groups up to 6 h. SAT was effective in maintaining cell viability only up to 12 h and then became detrimental to HPDLF; after 24 h, the effectiveness of SAT in maintaining cell viability was similar to that of water (P > 0.05). Amongst all the media, only EMT could maintain the proliferative capacity of HPDLFs significantly higher than the negative control, that is water (P < 0.05) after 24 h storage. CONCLUSION: EMT maintained the proliferative capacity of HPDLFs after 24 h storage.

3D printed TCP-based scaffold incorporating VEGF-loaded PLGA microspheres for craniofacial tissue engineering.

OBJECTIVE: Vascularization is a critical process during bone regeneration/repair and the lack of tissue vascularization is recognized as a major challenge in applying bone tissue engineering methods for cranial and maxillofacial surgeries. The aim of our study is to fabricate a vascular endothelial growth factor (VEGF)-loaded gelatin/alginate/ß-TCP composite scaffold by 3D printing method using a computer-assisted design (CAD) model. METHODS: The paste, composed of (VEGF-loaded PLGA)-containing gelatin/alginate/ß-TCP in water, was loaded into standard Nordson cartridges and promptly employed for printing the scaffolds. Rheological characterization of various gelatin/alginate/ß-TCP formulations led to an optimized paste as a printable bioink at room temperature. RESULTS: The in vitro release kinetics of the loaded VEGF revealed that the designed scaffolds fulfill the bioavailability of VEGF required for vascularization in the early stages of tissue regeneration. The results were confirmed by two times increment of proliferation of human umbilical vein endothelial cells (HUVECs) seeded on the scaffolds after 10 days. The compressive modulus of the scaffolds, 98±11MPa, was found to be in the range of cancellous bone suggesting their potential application for craniofacial tissue engineering. Osteoblast culture on the scaffolds showed that the construct supports cell viability, adhesion and proliferation. It was found that the ALP activity increased over 50% using VEGF-loaded scaffolds after 2 weeks of culture. SIGNIFICANCE: The 3D printed gelatin/alginate/ß-TCP scaffold with slow releasing of VEGF can be considered as a potential candidate for regeneration of craniofacial defects.