Effect of extracellular matrix and dental pulp stem cells on bone regeneration with 3D printed PLA/HA composite scaffolds (original) (raw)
Related papers
Journal of The Mechanical Behavior of Biomedical Materials, 2020
3D printing of polylactic acid (PLA) and hydroxyapatite (HA) or bioglass (BG) bioceramics composites is the most promising technique for artificial bone construction. However, HA and BG have different chemical composition as well as different bone regeneration inducing mechanisms. Thus, it is important to compare differentiation processes induced by 3D printed PLA þ HA and PLA þ BG scaffolds in order to evaluate the strongest osteoconductive and osteoinductive properties possessing bioceramics. In this study, we analysed porous PLA þ HA (10%) and PLA þ BG (10%) composites' effect on rat's dental pulp stem cells fate in vitro. Obtained results indicated, that PLA þ BG scaffolds lead to weaker cell adhesion and proliferation than PLA þ HA. Nevertheless, osteoinductive and other biofriendly properties were more pronounced by PLA þ BG composites. Overall, the results showed a strong advantage of bioceramic BG against HA, thus, 3D printed PLA þ BG composite scaffolds could be a perspective component for patient-specific, cheaper and faster artificial bone tissue production.
Journal of Stem Cell Research & Therapy, 2016
Objective: The aim of this study was to test specific stem cells that could enhance bone formation in combination with specific scaffolds. Methods: Dental Pulp Stem Cells (DPSCs) were seeded with Granular Deproteinized Bovine Bone (GDPB) or Beta-Tricalcium Phosphate (ß-TCP) in a rat model of calvarial "critical size" defect. DPSCs were isolated from permanent human teeth, obtained and characterized using specific stem cells markers (Nanog and Oct-4) by real time-PCR and immunofluorescence. Cells were differentiated for 10-15 days towards the osteoblastic phenotype with 100μM L-ascorbic acid, added every day in culture medium and 20 vol. percentage of FBS in α-MEM medium. Osteogenic commitment was evaluated with real time-PCR by measuring the expression of specific markers (osteonectin and runx2). When a sufficient cell number was obtained, DPSCs were trypsinized, washed in culture medium and seeded onto the GDPB and ß-TCP scaffold sat a density of 0.5-1×10 6 cells/scaffold. Two bilateral critical-size circular defects (5 mm diameter; 1 mm thickness) were created from the parietal bone of the 8 athymic T-cell deficient nude rats. One cranial defect for each rat was filled with the scaffold alone and the other defect with the scaffold seeded with stem cells. After 12 weeks post-surgery animals were euthanized and histomorphometric analysis was performed. Differences between groups were analyzed by one-way analysis of variance (ANOVA) followed by Fisher's Protected Least Significant Difference (PLSD) post-hoc test. A p-value <0.05 was considered statistically significant. Results: GDPB group presented higher percentage of lamellar bone than that of GDPB/DPSC, ß-TCP alone had lower levels as compared to ß-TCP/DPSC. The addition of stem cells significantly increased woven bone formation in both scaffold-based implants, although still higher in GDPB based implants. Conclusion: Our findings indicate that GDPB and ß-TCP used as scaffold to induce bone regeneration may benefit from adding DPSC to tissue-engineered constructs.
Journal of Biomedical Materials Research Part B: Applied Biomaterials, 2013
Bone regeneration strategies in dentistry utilize biodegradable scaffolds seeded with stem cells able to induce bone formation. However, data on regeneration capacity of these tissue engineering constructs are still deficient. In this study micro-Computed tomography (micro-CT) and positron emission tomography (PET) analyses were used to investigate bone regeneration induced by two scaffolds [Granular deproteinized bovine bone (GDPB) and Beta-tricalcium phosphate (b-TCP)] used alone or in combination with dental pulp stem cells (DPSC) in a tissue engineered construct implanted in a rat critical calvarial defect. Bone mineral density (BMD) and standard uptake value (SUV) of tracer incorporation were measured after 2, 4, 8, and 12 weeks post-implant. The results showed that: (1) GDPB implants were mostly well positioned, as compared to ß-TCP; (2) GDPB induced higher BMD and SUV values within the cranial defect as compared to ß-TCP, either alone or in combination with stem cells; (3) addition of DPSC to the grafts did not significantly induce an increase in BMD and SUV values as compared to the scaffolds grafted alone, although a small tendency to increase was observed. Thus our study demonstrates that GDPB, when used to fill critical calvarial defects, induces a greater percentage of bone formation as compared to ß-TCP. Moreover, this study shows that addition of DPSC to pre-wetted scaffolds has the potential to ameliorate bone regeneration process, although the set of optimal conditions requires further investigation. V
Polymers, 2021
The search of suitable combinations of stem cells, biomaterials and scaffolds manufacturing methods have become a major focus of research for bone engineering. The aim of this study was to test the potential of dental pulp stem cells to attach, proliferate, mineralize and differentiate on 3D printed polycaprolactone (PCL) scaffolds. A 100% pure Mw: 84,500 ± 1000 PCL was selected. 5 × 10 × 5 mm3 parallelepiped scaffolds were designed as a wood-pilled structure composed of 20 layers of 250 μm in height, in a non-alternate order ([0,0,0,90,90,90°]). 3D printing was made at 170 °C. Swine dental pulp stem cells (DPSCs) were extracted from lower lateral incisors of swine and cultivated until the cells reached 80% confluence. The third passage was used for seeding on the scaffolds. Phenotype of cells was determined by flow Cytometry. Live and dead, Alamar blue™, von Kossa and alizarin red staining assays were performed. Scaffolds with 290 + 30 μm strand diameter, 938 ± 80 μm pores in the a...
International Journal of Molecular Sciences
Background: Recently a greater interest in tissue engineering for the treatment of large bone defect has been reported. The aim of the present systematic review and meta-analysis was to investigate the effectiveness of dental pulp stem cells and synthetic block complexes for bone defect treatment in preclinical in vivo articles. Methods: The electronic database and manual search was conducted on Pubmed, Scopus, and EMBASE. The papers identified were submitted for risk-of-bias assessment and classified according to new bone formation, bone graft characteristics, dental pulp stem cells (DPSCs) culture passages and amount of experimental data. The meta-analysis assessment was conducted to assess new bone formation in test sites with DPSCs/synthetic blocks vs. synthetic block alone. Results: The database search identified a total of 348 papers. After the initial screening, 30 studies were included, according to the different animal models: 19 papers on rats, 3 articles on rabbits, 2 man...
Iranian Red Crescent Medical Journal, 2017
Background: Tissue engineering mainly focuses on creating appropriate conditions for the regeneration of tissues. Scaffolds, signal molecules, and stem cells interact with each other and compose the essential components of this field. Objectives: This study aimed at investigating the osteogenic induction ability of PLA Poly Lactic Acid (PLA) scaffolds and comparing the osteogenic differentiation behavior of Stem Cells from Human Exfoliated Deciduous Teeth (hSHEDs) in standard culture medium and on PLA scaffolds. Methods: The current clinical experimental study was conducted between April 2016 and October 2016 at the Near East University cell culture laboratory located in North Cyprus. The pulp tissues of deciduous teeth (non-decayed and in the absence of abscess, fistula or periapical lesion) were sampled from 10 healthy children aged between 6 and 11 years. The isolated hSHEDs were divided to 4 groups. The control group/Group1 consisted of cells, which were cultivated in standard culture medium, and Group2 cells were differentiated into an osteogenic lineage using osteogenic differentiation medium. Group 3 represented the non-differentiated group, which was transferred onto three dimensional (3D) printed PLA scaffolds and Group 4 cells were differentiated to the osteogenic lineage and transferred onto 3D printed PLA scaffolds. All groups were analyzed immunohistochemically and by immune-labeling, and were evaluated semi-quantitatively using the HSCORE. Results: Cultivation of hSHEDS on PLA scaffolds was assessed for 14 and 21 days; osteogenic differentiation was detected both histochemically and immunohistochemically. Generally, Osteocalcin (OCN) immunoreactivities were higher than Osteonectin (ON) immunoreactions in all groups. Despite higher OCN immunoreactivities, the intensities of OCN between 14 days and 21 days in group 4 (497.3 ± 0.57% and 486.7 ± 5.77%, respectively) were similar (P > 0.05). While the intensity of ON was 280.0 ± 10% in group 4, in group 2 the intensity of ON was 206.7 ± 5.77%, and on the 14th day the results were statistically significant (P < 0.0001). Conclusions: Poly lactic acid is a suitable scaffold material for osteogenic induction of the hSHEDs. The expression patterns of both markers showed that a 14-day cultivation period is adequate for hSHEDs with/without PLA scaffolds to differentiate into osteoblasts.
BMC Oral Health, 2021
Background: Mesenchymal stem cells isolated from the dental pulp of primary and permanent teeth can be differentiated into different cell types including osteoblasts. This study was conducted to compare the morphology and osteogenic potential of stem cells from exfoliated deciduous teeth (SHED) and dental pulp stem cells (DPSC) in granular hydroxyapatite scaffold (gHA). Preosteoblast cells (MC3T3-E1) were used as a control group. Methodology: The expression of stemness markers for DPSC and SHED was evaluated using reverse transcriptase-polymerase chain reaction (RT-PCR). Alkaline phosphatase assay was used to compare the osteoblastic differentiation of these cells (2D culture). Then, cells were seeded on the scaffold and incubated for 21 days. Morphology assessment using field emission scanning electron microscopy (FESEM) was done while osteogenic differentiation was detected using ALP assay (3D culture). Results: The morphology of cells was mononucleated, fibroblast-like shaped cells with extended cytoplasmic projection. In RT-PCR study, DPSC and SHED expressed GAPDH, CD73, CD105, and CD146 while negatively expressed CD11b, CD34 and CD45. FESEM results showed that by day 21, dental stem cells have a round like morphology which is the morphology of osteoblast as compared to day 7. The osteogenic potential using ALP assay was significantly increased (p < 0.01) in SHED as compared to DPSC and MC3T3-E1 in 2D and 3D cultures. Conclusion: gHA scaffold is an optimal scaffold as it induced osteogenesis in vitro. Besides, SHED had the highest osteogenic potential making them a preferred candidate for tissue engineering in comparison with DPSC. © 2021, The Author(s).
Journal of Biomedical Materials Research Part A, 2019
Stereolithography (SLA) is an interesting manufacturing technology to overcome limitations of commercially available particulated biomaterials dedicated to intra-oral bone regeneration applications. The purpose of this study was to evaluate the in vitro and in vivo biocompatibility and osteoinductive properties of two CaP-based scaffolds manufactured by SLA 3D printing. Pellets and macro-porous scaffolds were manufactured in pure hydroxyapatite (HA) and in biphasic CaP (HA:60-TCP:40). Physicochemical characterization was performed using Micro X-ray Fluorescence (µXRF), Scanning Electron Microscopy (SEM), optical interferometry and microtomography (µCT) analyses. Osteoblast-like MG-63 cells were used to evaluate the biocompatibility of the pellets in vitro with MTS assay and the cell morphology and growth characterized by SEM and DAPI-actin staining showed similar early behavior. For in vivo biocompatibility, newly formed bone and biodegradability of the experimental scaffolds were evaluated in a subperiosteal cranial rat model using µCT and descriptive histology. The histological analysis has not indicated evidences of inflammation but highlighted close contacts between newly formed bone and the experimental biomaterials revealing an excellent scaffold osseointegration. This study emphasizes the relevance of SLA 3D printing of CaP-based biomaterials for intra-oral bone regeneration even if manufacturing accuracy has to be improved and further experiments using biomimetic scaffolds should be conducted.
Response of Dental Pulp Stem Cells to Synthetic, Allograft, and Xenograft Bone Scaffolds
The International journal of periodontics & restorative dentistry, 2016
Different degrees of clinical success have been reported for synthetic, allograft, and xenograft bone substitutes in human trials. Although these substitutes have been clinically investigated, their in vitro effects on cell differentiation remain unclear. Proliferation, differentiation, and attachment of dental pulp stem cells (DPSCs) to β-tricalcium phosphate (β-TCP), freeze-dried bone allograft (FDBA), and deproteinized bovine bone mineral (DBBM) were compared in this study. MTT assay, measurement of total DNA, and reverse transcriptase polymerase chain reaction were performed. β-TCP had the highest potential for DPSC attachment and proliferation, while FDBA induced osteoblastic differentiation of DPSCs. Further in vivo investigations are necessary to select a clinically appropriate scaffold.
Comparison of the Efficacy of Three Different Bone Regeneration Materials: An Animal Study
European Journal of Dentistry, 2019
Objective The proposed study aimed to evaluate and compare the bone regeneration between commercially available hydroxyapatite–β-tricalcium phosphate (Ossifi; Equinox, the Netherlands), powdered polylactic acid (powdered PLA; Sigma-Aldrich, United States), and three-dimensionally printed PLA (3D-printed PLA; Cubex, SC, United States) using 3D printer (Cube X trio) in an animal model. Materials and Methods Eighteen New Zealand rabbits were divided into three groups with six animals each. Platelet-rich fibrin (PRF) was collected from the venous blood and preserved. Bone defect (4 mm × 2 mm) without disturbing the bone marrow was created and filled with bone graft material (group 1–Ossifi, group 2–powdered PLA, and group 3–3D-printed PLA), over which PRF membranes were placed. The graft material and the barrier were stabilized using resorbable sutures, and all the animals were maintained for 4, 8, and 12 weeks, after which they were euthanized, and bone samples were retrieved. Retrieve...