Dental-derived mesenchymal stem cells (MSCs) provide an advantageous therapeutic option for tissue engineering due to their high accessibility and bioavailability. However, delivering MSCs to defect sites while maintaining a high MSC survival rate is still a critical challenge in MSC-mediated tissue regeneration. Here, we tested the osteogenic and adipogenic differentiation capacity of dental pulp stem cells (DPSCs) in a thermoreversible Pluronic F127 hydrogel scaffold encapsulation system in vitro. DPSCs were encapsulated in Pluronic (®) F-127 hydrogel and stem cell viability, proliferation and differentiation into adipogenic and osteogenic tissues were evaluated. The degradation profile and swelling kinetics of the hydrogel were also analyzed. Our results confirmed that Pluronic F-127 is a promising and non-toxic scaffold for encapsulation of DPSCs as well as control human bone marrow MSCs (hBMMSCs), yielding high stem cell viability and proliferation. Moreover, after 2 weeks of differentiation in vitro, DPSCs as well as hBMMSCs exhibited high levels of mRNA expression for osteogenic and adipogenic gene markers via PCR analysis. Our histochemical staining further confirmed the ability of Pluronic F-127 to direct the differentiation of these stem cells into osteogenic and adipogenic tissues. Furthermore, our results revealed that Pluronic F-127 has a dense tubular and reticular network morphology, which contributes to its high permeability and solubility, consistent with its high degradability in the tested conditions. Altogether, our findings demonstrate that Pluronic F-127 is a promising scaffold for encapsulation of DPSCs and can be considered for cell delivery purposes in tissue engineering.

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Pluronic F-127 hydrogel as a promising scaffold for encapsulation of dental-derived mesenchymal stem cells

https://www.materialstoday.com/download/51100/

Hydrogels in calcium phosphate moldable and injectable bone substitutes: Sticky excipients or advanced 3-D carriers?

Background: Despite significant surgical advances over the last decades, segmental mandibular bone repair remains a challenge. In light of this, tissue engineering might offer a next step in the evolution of mandibular reconstruction.

Purpose: The purpose of the present report was to (1) systematically review preclinical in vivo as well as clinical literature regarding bone tissue engineering for mandibular continuity defects, and (2) to analyze their effectiveness.

Materials and Methods: An electronic search in the databases of the National Library of Medicine and ISI Web of Knowledge was carried out. Only publications in English were considered, and the search was broadened to animals and humans. Furthermore, the reference lists of related review articles and publications selected for inclusion in this review were systematically screened. Results of histology data and amount of bone bridging were chosen as primary outcome variables. However, for human reports, clinical radiographic evidence was accepted for defined primary outcome variable. The biomechanical properties, scaffold degradation, and clinical wound healing were selected as co-outcome variables.

Results: The electronic search in the databases of the National Library of Medicine and ISI Web of Knowledge resulted in the identification of 6727 and 5017 titles, respectively. Thereafter, title assessment and hand search resulted in 128 abstracts, 101 full-text articles, and 29 scientific papers reporting on animal experiments as well as 11 papers presenting human data on the subject of tissue-engineered reconstruction of mandibular continuity defects that could be included in the present review.

Conclusions: It was concluded that (1) published preclinical in vivo as well as clinical data are limited, and (2) tissue-engineered approaches demonstrate some clinical potential as an alternative to autogenous bone grafting.

Tissue-Engineered Mandibular Bone Reconstruction for Continuity Defects: A Systematic Approach to the Literature

Pluronic® F127 is a biocompatible, injectable, and thermoresponsive polymer with promising biomedical applications. In this study, a chemically modified form, i.e., Pluronic ALA-L with tailored degradation rate, was tested as an encapsulation vehicle for osteoblastic cells. UV cross-linking of the modified polymer results in a stable hydrogel with a slower degradation rate. Toxicological screening showed no adverse effects of the modified Pluronic ALA-L on the cell viability. Moreover, high viability of embedded cells in the cross-linked Pluronic ALA-L was observed with life/death fluorescent staining during a 7-day-culture period. Cells were also cultured on macroporous, cross-linked gelatin microbeads, called CultiSpher-S® carriers, and encapsulated into the modified cross-linked hydrogel. Also, in this situation, good cell proliferation and migration could be observed in vitro. Preliminary in vivo tests have shown the formation of new bone starting from the injected pre-loaded CultiSpher-S® carriers.

Cell survival and proliferation after encapsulation in a chemically modified Pluronic® F127 hydrogel

In an ever-aging society the demand for bone-defect filling grafts continues to gain in importance. While autologous grafting still prevails as the gold standard, allografts and xenografts present viable alternatives with promising results. Physiochemical properties of a graft strongly depend on the processing method such as the decellularization protocol. In addition, the physiochemical characteristics are critical factors for a successful integration of the graft after the implantation and might influence mesenchymal stem cell function in therapeutic approaches combining grafts and autologous mesenchymal stem cells (MSCs). Several decellularization methods have been proposed, however it still remains unclear which method results in favorable physiochemical properties or might be preferred in stem cell applications. In the first part of this study we compared two decellularization approaches resulting in chemically processed allografts (CPAs) or sonication-based processed allografts (SPAs). Each decellularization approach was compared for its decellularization efficacy and its influence on the grafts’ surface texture and composition. In the second part of this study biocompatibility of grafts was assessed by testing the effect of extraction medium on MSC viability and comparing them to commercially available allografts and xenografts. Additionally, grafts’ performance in terms of MSC functionality was assessed by reseeding with MSCs pre-differentiated in osteogenic medium and determining cell adhesion, proliferation, as well as alkaline phosphatase (ALP) activity and the degree of mineralization. In summary, results indicate a more effective decellularization for the SPA approach in comparison to the CPA approach. Even though SPA extracts induced a decrease in MSC viability, MSC performance after reseeding was comparable to commercially available grafts based on DNA quantification, alkaline phosphatase activity and quantification of mineralization. Commercial Tutoplast allografts showed overall the best effects on MSC functionality as indicated by extraction biocompatibility testing as well as by comparing proliferation and osteogenic differentiation.

Evaluation of bone allograft processing methods: Impact on decellularization efficacy, biocompatibility and mesenchymal stem cell functionality.

Purpose: The purpose of this project was to compare alveolar bone repair by allogeneic mesenchymal stem cells using bioglass or synthetic hydroxyapatite (HA)/tricalcium phosphate (TCP) granular scaffolds delivered in a thermoplastic polymeric carrier.



Materials and Methods:  Canine mesenchymal stem cells were obtained from iliac crest bone marrow of beagle dogs and expanded without differentiation. Cells were resuspended at a final concentration of 5 × 106 cells/ml in a thermoplastic polymeric carrier (30% w/v Pluronic F-127) and mixed with an equal volume of synthetic HA/TCP or bioglass scaffold and placed into surgically created 5 mm cylindrical defects in the edentulous premolar region of beagle dogs. After 4 weeks or 7 weeks, tissue healing was evaluated by standard histomorphometric methods (Bioquant Nova, Bioquant Image Analysis Corporation, Nashville, TN) by measurement of bone formation within five random sites from each biopsy.



Results:  After 4 weeks, sites treated with or without mesenchymal stem cells contained 58.25 ±18.43% or 43.35 ± 17.68% bone area (p= 0.049), respectively. After 7 weeks, sites treated with or without mesenchymal stem cells contained 62.73 ± 19.10% or 60.39 ± 21.32% bone area. Bone formation occurred without inflammation in defects treated using Pluronic F-127 carrier with and without mesenchymal stem cells. There was no difference in percent bone area when bioglass or HA/TCP scaffolds were compared at either time point.



Conclusions: The thermoplastic polymeric carrier did not limit alveolar bone repair in the canine mandible. The combination of a thermoplastic, viscous carrier with a granular scaffold allowed for the delivery of allogeneic mesenchymal stem cells in a clinically manageable form that enhanced bone formation at early stages of alveolar repair.

Investigation of a thermoplastic polymeric carrier for bone tissue engineering using allogeneic mesenchymal stem cells in granular scaffolds.

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Investigation of a thermoplastic polymeric carrier for bone tissue engineering using allogeneic mesenchymal stem cells in granular scaffolds.