The objective of the present study was to investigate the effect of a fabricated combination of poly-ɛ-caprolactone (PCL)-biphasic calcium phosphate (BCP) with the modified melt stretching and multilayer deposition (mMSMD) technique on human dental pulp stem cell (hDPSC) differentiation to be osteogenic like cells for bone regeneration of calvarial defects in rabbit models. hDPSCs extracted from human third molars were seeded onto mMSMD PCL-BCP scaffolds and the osteogenic gene expression was tested prior to implantation in vivo. Two standardized 11 mm in diameter circular calvarial defects were created in 18 adult male New Zealand white rabbits. The rabbits were divided into 4 groups: (1) hDPSCs seeded in mMSMD PCL-BCP scaffolds; (2) mMSMD PCL-BCP scaffolds alone, (3) empty defects and (4) autogenous bone (n = 3 site/time point/groups). After two, four and eight weeks after the operation, the specimens were harvested for micro-CT including histological and histomorphometric analysis. The explicit results presented an interesting view of the bioengineered constructs of hDPSCs in PCL-BCP scaffolds that increased the newly formed bone compared to the empty defect and scaffold alone groups. The results demonstrated that hDPSCs combined with mMSMD PCL-BCP scaffolds may be an augmentation material for bony defect.
Fabrication of polycaprolactone (PCL)-chitosan (CS) three-dimensional (3D) scaffolds using the novel technique of melt stretching and multilayer deposition was introduced. In brief, firstly, the PCL-CS monofilaments containing 0% (pure PCL), 10%, 20% and 30% CS by weight were fabricated by melting and stretching processes. Secondly, the desired multilayer (3D) scaffolds were fabricated by arranging and depositing the filaments. Physical properties of the filaments and the scaffolds were evaluated. MC3T3-E1 cell lines were seeded on the scaffolds to assess their proliferation. A typical micro-groove pattern was found on the surfaces of pure PCL filaments due to stretching. The filaments of PCL-30%CS had the highest tendency of fracture during stretching and could not be used to form the scaffold. Increasing CS proportions tended to reduce the micro-groove pattern, surface roughness, tensile strength and elasticity of the filaments, whilst compressive strength of the PCL-CS scaffolds was not affected. The average pore size and porosity of the scaffolds were 536.90 ± 17.91 µm and 45.99 ± 2.8% respectively. Over 60 days, degradation of the scaffolds gradually increased (p > 0.05). The more CS containing scaffolds were found to increase in water uptake, but decrease in degradation rate. During the culture period, the growth of the cells in PCL-CS groups was significantly higher than in the pure PCL group (p < 0.05). On culture-day 21, the growth in the PCL-20%CS group was significantly higher than the other groups (p < 0.05). In conclusion, the PCL-20%CS scaffolds obtained the optimum results in terms of physical properties and cellular response.
The PCL-20% BCP MSCM scaffolds were biocompatible and biodegradable in vivo. Their properties were comparable to those of the commercial PCL-20% TCP scaffolds.
Physical properties and biocompatibility of polycaprolactone (PCL)-biphasic calcium phosphate (BCP) scaffolds fabricated by the modified melt stretching and multilayer deposition (mMSMD) technique were evaluated in vitro. The PCL-BCP scaffold specimens included group A; PCL: BCP (wt%) = 80:20 and group B; 70:30. Mechanical properties of the scaffolds were assessed using a universal testing machine. Degradation behaviors of the scaffolds were assessed over 60 days. The amount of calcium and phosphate ions released from the scaffolds was detected over 30 days. Attachment and growth of osteoblasts on the scaffolds and indirect cytocompatibility to those cells were evaluated. The results showed that the scaffolds of both groups could withstand compressive forces on their superior aspect very well; however, their lateral aspect could only withstand light forces. Degradation of the scaffolds over 2 months was low (group A = 1.92 ± 0.47% and group B = 2.9 ± 1.3%,p > 0.05). The concentrations of calcium and phosphate ions released from the scaffolds of both groups significantly increased on day 7 (p < 0.05). Growth of the cells seemed to relate to accumulative increase in those ions. All results between the two ratios of the scaffolds were not statistically different.
The MSCM scaffolds are suitable for supporting attachment and growth of the osteoblasts. Additional BCP into the PCL-based scaffolds accelerate early differentiation of the cells in the constructs even without osteogenic-inductive condition.
The monolayer scaffolds were suitable for reconstruction of the orbital floor and mandibular defects under light load-bearing conditions. The 3-D scaffolds could be used in the high load bearing-areas only if the forces were directed at their superior aspects.
Craniofacial bone defects such as alveolar cleft affect the esthetics and functions that need bone reconstruction. The advanced techniques of biomaterials combined with stem cells have been a challenging role for maxillofacial surgeons and scientists. PCL-coated biphasic calcium phosphate (PCL-BCP) scaffolds were created with the modified melt stretching and multilayer deposition (mMSMD) technique and merged with human dental pulp stem cells (hDPSCs) to fulfill the component of tissue engineering for bone substitution. In the present study, the objective was to test the biocompatibility and biofunctionalities that included cell proliferation, cell viability, alkaline phosphatase activity, osteocalcin, alizarin red staining for mineralization, and histological analysis. The results showed that mMSMD PCL-BCP scaffolds were suitable for hDPSCs viability since the cells attached and spread onto the scaffold. Furthermore, the constructs of induced hDPSCs and scaffolds performed ALP activity and produced osteocalcin and mineralized nodules. The results indicated that mMSMD PCL-BCP scaffolds with hDPSCs showed promise in bone regeneration for treatment of osseous defects.
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