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Influence of highly porous electrospun PLGA/PCL/nHA fibrous scaffolds on the differentiation of tooth bud cells in vitro.

ABSTRACT: In this study, we investigated the use of three-dimensional electrospun poly(lactic-co-glycolic acid)/poly(?-caprolactone) (PLGA/PCL) scaffolds seeded and cultured with postnatal dental cells, for improved dental tissue regeneration. Wet-electrospinning combined with ultrasonic treatment was studied as a method to enhance scaffold porosity and to promote cell-cell interactions. We also investigated whether nano-hydroxyapatite (nHA) incorporation could enhance dental cell differentiation. All scaffolds were seeded with human tooth pulp-derived dental mesenchymal (hDM) cells, or a combination of hDM and pig dental epithelial (pDE) cells, cultured for up to 28 days. Developmentally staged samples were assessed using scanning electron microscopy, histological, immunohistochemical, DNA and alkaline phosphatase activity assays, and quantitative-PCR for ameloblastic, odontoblastic, and osteogenic related gene expression. Results showed that electrospun scaffolds exhibited sufficient porosity to support robust cell ingrowth. Additional ultrasonic treatment led to a less homogeneous scaffold porosity, resulting in evident cell clustering and enhanced hDM-pDE cell-cell interactions. Finally, nHA incorporation was found to enhance dental cell differentiation. However, it also resulted in smaller fiber diameter and reduced scaffold porosity, and inhibited cell ingrowth and proliferation. In conclusion, ultrasonically treated wet-electrospun PLGA/PCL scaffolds are a suitable material for dental tissue engineering, and support future in vivo evaluations of this model. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2597-2607, 2017.

SUBMITTER: Cai X

PROVIDER: S-EPMC5695215 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature

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Influence of highly porous electrospun PLGA/PCL/nHA fibrous scaffolds on the differentiation of tooth bud cells in vitro.

Cai Xinjie X Ten Hoopen Sofie S Zhang Weibo W Yi Charles C Yang Wanxun W Yang Fang F Jansen John A JA Walboomers X Frank XF Yelick Pamela C PC

Journal of biomedical materials research. Part A 20170615 9

In this study, we investigated the use of three-dimensional electrospun poly(lactic-co-glycolic acid)/poly(ε-caprolactone) (PLGA/PCL) scaffolds seeded and cultured with postnatal dental cells, for improved dental tissue regeneration. Wet-electrospinning combined with ultrasonic treatment was studied as a method to enhance scaffold porosity and to promote cell-cell interactions. We also investigated whether nano-hydroxyapatite (nHA) incorporation could enhance dental cell differentiation. All sca ...[more]

PMID: 28544201

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Project description:Whole tooth regeneration approaches currently are limited by our inability to bioengineer full-sized, living replacement teeth. Recently, decellularized organ scaffolds have shown promise for applications in regenerative medicine by providing a natural extracellular matrix environment that promotes cell attachment and tissue-specific differentiation leading to full-sized organ regeneration. We hypothesize that decellularized tooth buds (dTBs) created from unerupted porcine tooth buds (TBs) can be used to guide reseeded dental cell differentiation to form whole bioengineered teeth, thereby providing a potential off-the-shelf scaffold for whole tooth regeneration. Porcine TBs were harvested from discarded 6-mo-old pig jaws, and decellularized by successive sodium dodecyl sulfate/Triton-X cycles. Four types of replicate implants were used in this study: 1) acellular dTBs; 2) recellularized dTBs seeded with porcine dental epithelial cells, human dental pulp cells, and human umbilical vein endothelial cells (recell-dTBs); 3) dTBs seeded with bone morphogenetic protein (BMP)-2 (dTB-BMPs); and 4) freshly isolated nondecellularized natural TBs (nTBs). Replicate samples were implanted into the mandibles of host Yucatan mini-pigs and grown for 3 or 6 mo. Harvested mandibles with implanted TB constructs were fixed in formalin, decalcified, embedded in paraffin, sectioned, and analyzed via histological methods. Micro-computed tomography (CT) analysis was performed on harvested 6-mo samples prior to decalcification. All harvested constructs exhibited a high degree of cellularity. Significant production of organized dentin and enamel-like tissues was observed in dTB-recell and nTB implants, but not in dTB or dTB-BMP implants. Micro-CT analyses of 6-mo implants showed the formation of organized, bioengineered teeth of comparable size to natural teeth. To our knowledge, these results are the first to describe the potential use of dTBs for functional whole tooth regeneration.

Project description:BackgroundIt is known that a number of parameters can influence the post-printing properties of bone tissue scaffolds. Previous research has primarily focused on the effect of parameters associated with scaffold design (e.g., scaffold porosity) and specific scaffold printing processes (e.g., printing pressure). To our knowledge, no studies have investigated variations in post-printing properties attributed to the techniques used to synthesize the materials for printing (e.g., melt-blending, powder blending, liquid solvent, and solid solvent).MethodsFour material preparation techniques were investigated to determine their influence on scaffold properties. Polycaprolactone/nano-hydroxyapatite 30% (wt.) materials were synthesized through melt-blending, powder blending, liquid solvent, and solid solvent techniques. The material printability and the properties of printed scaffolds, in terms of swelling/degradation, mechanical strength, morphology, and thermal properties, were examined and compared to one another using Kruskal-Wallis nonparametric statistical analysis.ResultsMaterial prepared through the liquid solvent technique was found to have limited printability, while melt-blended material demonstrated the highest degree of uniformity and lowest extent of swelling and degradation. Scaffolds prepared with powder-blended material demonstrated the highest Young's modulus, yield strength, and modulus of resilience; however, they also demonstrated the highest degree of variability. The higher degree of inhomogeneity in the material was further supported by thermal gravimetric analysis. While scaffolds printed from melt-blended, powder-blended, and solid solvent materials demonstrated a high degree of micro-porosity, the liquid solvent material preparation technique resulted in minimal micro-porosity.ConclusionsStudy results indicate that specific techniques used to prepare materials influence the printing process and post-printing scaffold properties. Among the four techniques examined, melt-blended materials were found to be the most favorable, specifically when considering the combination of printability, consistent mechanical properties, and efficient preparation. Techniques determined to be favourable based on the properties investigated should undergo further studies related to biological properties and time-dependent properties beyond 21-days.

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Influence of highly porous electrospun PLGA/PCL/nHA fibrous scaffolds on the differentiation of tooth bud cells in vitro.

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Influence of highly porous electrospun PLGA/PCL/nHA fibrous scaffolds on the differentiation of tooth bud cells in vitro.

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