Polycaprolactone-tricalcium Phosphate Scaffolds Research Articles

Alveolar Ridge Augmentation with a Novel Combination of 3D-Printed Scaffolds and Adipose-Derived Mesenchymal Stem Cells-A Pilot Study in Pigs.

Alveolar ridge augmentation is an important dental procedure to increase the volume of bone tissue in the alveolar ridge before the installation of a dental implant. To meet the high demand for bone grafts for alveolar ridge augmentation and to overcome the limitations of autogenous bone, allografts, and xenografts, researchers are developing bone grafts from synthetic materials using novel fabrication techniques such as 3D printing. To improve the clinical performance of synthetic bone grafts, stem cells with osteogenic differentiation capability can be loaded into the grafts. In this pilot study, we propose a novel bone graft which combines a 3D-printed polycaprolactone-tricalcium phosphate (PCL-TCP) scaffold with adipose-derived mesenchymal stem cells (AD-MSCs) that can be harvested, processed and implanted within the alveolar ridge augmentation surgery. We evaluated the novel bone graft in a porcine lateral alveolar defect model. Radiographic analysis revealed that the addition of AD-MSCs to the PCL-TCP scaffold improved the bone volume in the defect from 18.6% to 28.7% after 3 months of healing. Histological analysis showed the presence of AD-MSCs in the PCL-TCP scaffold led to better formation of new bone and less likelihood of fibrous encapsulation of the scaffold. Our pilot study demonstrated that the loading of AD-MSCs improved the bone regeneration capability of PCL-TCP scaffolds, and our novel bone graft is suitable for alveolar ridge augmentation.

Synergistic Effect of PVDF-Coated PCL-TCP Scaffolds and Pulsed Electromagnetic Field on Osteogenesis.

Bone exhibits piezoelectric properties. Thus, electrical stimulations such as pulsed electromagnetic fields (PEMFs) and stimuli-responsive piezoelectric properties of scaffolds have been investigated separately to evaluate their efficacy in supporting osteogenesis. However, current understanding of cells responding under the combined influence of PEMF and piezoelectric properties in scaffolds is still lacking. Therefore, in this study, we fabricated piezoelectric scaffolds by functionalization of polycaprolactone-tricalcium phosphate (PCL-TCP) films with a polyvinylidene fluoride (PVDF) coating that is self-polarized by a modified breath-figure technique. The osteoinductive properties of these PVDF-coated PCL-TCP films on MC3T3-E1 cells were studied under the stimulation of PEMF. Piezoelectric and ferroelectric characterization demonstrated that scaffolds with piezoelectric coefficient d33 = −1.2 pC/N were obtained at a powder dissolution temperature of 100 °C and coating relative humidity (RH) of 56%. DNA quantification showed that cell proliferation was significantly enhanced by PEMF as low as 0.6 mT and 50 Hz. Hydroxyapatite staining showed that cell mineralization was significantly enhanced by incorporation of PVDF coating. Gene expression study showed that the combination of PEMF and PVDF coating promoted late osteogenic gene expression marker most significantly. Collectively, our results suggest that the synergistic effects of PEMF and piezoelectric scaffolds on osteogenesis provide a promising alternative strategy for electrically augmented osteoinduction. The piezoelectric response of PVDF by PEMF, which could provide mechanical strain, is particularly interesting as it could deliver local mechanical stimulation to osteogenic cells using PEMF.

Rabbit palatum-derived mesenchymal progenitor cells tri-lineage differentiation on 2D substrates and 3D printed constructs.

Hard palate, developed by embryo neural crest stem cells, is a tissue with strong regenerative abilities. It is considered an abundant source of progenitor cells, forming various mesenchymal tissues. Rabbits are more suitable models than murine animals for regenerative preclinical study of the head and neck, owing to their larger size. However, there are no reports of the existence or characteristics of neural crest stem cells in the hard palate of rabbits. In this study, we demonstrate for the first time the presence of nestin-, Sox2-, and p75-positive neural crest stem cells obtained from the hard palate of rabbits and the properties of these cells. Flow cytometry analysis revealed that CD29, CD44, and CD81 were positive; and CD11b, CD34, and CD90 were negative on the ex vivo expanded palatal progenitor cells. Finally, we differentiated them into cells of mesenchymal lineages (bone, cartilage, and fat) in vitro, and in three-dimensional fabricated polycaprolactone and polycaprolactone-tricalcium phosphate scaffolds. Taken together, our data showed the existence of rabbit palatum-derived mesenchymal progenitor cells, and successful fabrication of progenitor cell-loaded biodegradable scaffold using three-dimensional printing. This study will open avenues for new tissue engineering strategies for cell therapy using three-dimensional printing with scaffolds for reconstruction of head and neck defects.

Scapula pre-augmentation in sheep with polycaprolactone tricalcium phosphate scaffolds

A scapula free flap is a commonly used method to reconstruct intraoral defects of the mandible and maxilla. Despite its clear advantages, it shows some deficiencies concerning the amount and shape of the available bone, especially with respect to later implant placement. To overcome these limitations, we pre-augmented the scapula prior to a potential flap-raising procedure with polycaprolactone (PCL) tricalcium phosphate (TCP) scaffolds in a sheep model. In our study, the scapula angle was augmented with a block of PCL-TCP in three adult sheep. After 6 months, the amount of newly formed bone and scaffold degradation were evaluated using cone-beam computed tomography scans and histomorphometric analysis. All animals survived the study and showed no problems in the augmented regions. The scaffolds were attached firmly to the scapula and showed a bonelike consistency. A fair amount of the scaffold material was degraded and replaced by vital bone. Our method seems to be a valid approach to pre-augment the scapula in sheep. In further experiments, it will be interesting to determine whether it is possible to transplant a modified scapula flap to an intraoral defect site.

A humanised tissue-engineered bone model allows species-specific breast cancer-related bone metastasis in vivo.

Bone metastases frequently occur in the advanced stages of breast cancer. At this stage, the disease is deemed incurable. To date, the mechanisms of breast cancer-related metastasis to bone are poorly understood. This may be attributed to the lack of appropriate animal models to investigate the complex cancer cell-bone interactions. In this study, two established tissue-engineered bone constructs (TEBCs) were applied to a breast cancer-related metastasis model. A cylindrical medical-grade polycaprolactone-tricalcium phosphate scaffold produced by fused deposition modelling (scaffold 1) was compared with a tubular calcium phosphate-coated polycaprolactone scaffold fabricated by solution electrospinning (scaffold 2) for their potential to generate ectopic humanised bone in NOD/SCID mice. While scaffold 1 was found not suitable to generate a sufficient amount of ectopic bone tissue due to poor ectopic integration, scaffold 2 showed excellent integration into the host tissue, leading to bone formation. To mimic breast cancer cell colonisation to the bone, MDA-MB-231, SUM1315, and MDA-MB-231BO breast cancer cells were cultured in polyethylene glycol-based hydrogels and implanted adjacent to the TEBCs. Histological analysis indicated that the breast cancer cells induced an osteoclastic reaction in the TEBCs, demonstrating analogies to breast cancer-related bone metastasis seen in patients.

Enhanced osteogenic gene expression in microcarrier-bound scalably expanded fetal bone marrow MSCS

Mesenchymal stem cell (MSC) therapy is a promising strategy to address the unmet clinical need for improved bone regeneration. However, challenges to the scalable stem cell production hinder the clinical translation of MSC therapy. Furthermore, the effects of scalable microcarrier-based culture and enzymatic cell harvest on the phenotype and molecular identity of MSCs upon directed osteogenic differentiation are not well understood. In order to address these issues, we expanded fetal bone marrow MSCs on either commercially-available microcarriers in spinner flask culture or on control tissue culture plastic (TCP) monolayer surfaces. At the end of this expansion, we characterized the phenotype of microcarrier- or TCP monolayer-expanded MSCs in terms of cytokine, gene and surface marker expression. We then differentiated microcarrier-bound, microcarrier-harvested or plastic-harvested MSCs which were seeded onto collagen-coated plates or polycaprolactone-tricalcium phosphate scaffolds. In both cases, microcarrier-bound MSCs displayed increased expression of osteogenic genes such as RUNX2 and ALPL compared to microcarrier-harvested and plastic-harvested cells upon osteogenic induction. These results indicate that microcarrier-bound MSCs undergo enhanced osteogenic differentiation. Our findings also suggest that compared to conventional methods of delivering MSC cell suspensions that were harvested either from microcarriers cultured in agitated bioreactors or from TCP surfaces, in vivo delivery of MSC-bound microcarriers may present a promising method for bone regeneration.

A polycaprolactone-tricalcium phosphate composite scaffold as an autograft-free spinal fusion cage in a sheep model

Titanium (Ti) based spinal fusion cages are frequently used in the clinics for the treatment of spinal degeneration and related diseases, however, their further clinical application is generally harassed by several drawbacks such as stress shielding, non-biodegradability and additional bone grafting procedure. Our earlier work has demonstrated the efficacy of a biodegradable macro-porous polycaprolactone-tricalcium phosphate (PCL-TCP) composite scaffold in promoting bony tissue ingrowth as well as its ability to sustain mechanical loads upon implantation into an orthotopic defect site. In this study, we investigated the use of PCL-TCP scaffold as an autograft-free spinal fusion cage in a preclinical sheep model over 12 months, and compared the fusion efficacy against Ti cages incorporated with autografts. Results showed that despite PCL-TCP scaffold as an autograft-free cage attaining a slower fusion rate at early stage (6 month), it achieved similar degree of spinal fusion efficacy as Ti cages aided with autograft at 12 month post-operation as evidenced by the radiographic and histological evaluation. PCL-TCP cages alone demonstrated better bone ingrowth with 2.6 fold higher bone/interspace ratio (B/I) and more homogeneous bone tissue distribution compared with that of the Ti cages (88.10 ± 3.63% vs. 33.74 ± 2.78%, p < 0.05) as seen from the histological and micro-CT analysis. Moreover, besides the bone tissue ingrowth, a quantitative approach was illustrated to accurately evaluate the osteointegration of fusion cage with surrounding bone tissue, and showed a 1.36 fold higher degree of osteointegration occurred in PCL-TCP cage group than Ti cage group (CS/PC: 79.31 ± 3.15% vs 58.44 ± 2.43%, p < 0.05). Furthermore, biomechanical analysis showed comparable mechanical strength of fused segments in both groups in terms of the range of motion and stiffness at 12 month (p > 0.05). The degradation profile of the PCL-TCP cages was noted to increase in tandem with new bone ingrowth into the pores, while maintaining good structural integrity necessary for supporting the spinal interbody segments. Therefore, with the better osteointegration, more bone tissue ingrowth as well as its favorable biodegradable and radiolucent properties, PCL-TCP cage has been demonstrated to be a promising candidate as an autograft-free fusion cage for clinical application.

The use of a polycaprolactone–tricalcium phosphate scaffold for bone regeneration of tooth socket facial wall defects and simultaneous immediate dental implant placement in Macaca fascicularis

Bone regeneration and aesthetic outcomes may be compromised when immediate implants are placed at extraction sites with dehiscence defects. The aim of this study was to compare, in a monkey model, peri-implant bone regeneration and implant stability after immediate implant placement into tooth sockets with facial wall defects in two treatment groups. In eight control monkeys, the bony defect was reconstructed with autogenous particulate bone, whereas in 10 test monkeys a polycaprolactone-tricalcium phosphate (PCL-TCP) scaffold was used. The monkeys were sacrificed after 6 months and the specimens were analyzed by histology and histomorphometry. Better maintenance of facial bone contour was noted in the test group; however, bone regeneration was seen only at areas adjacent to a bony wall of the defect. The mean bone-to-implant contact was 27.6 ± 19.1% (control group) versus 6.8 ± 7.9% (test group). The mean bone area percentage was 11.8 ± 10.1% (control group) versus 6.8 ± 6.9% (test group). Implant survival was 100% at 6 months for both the groups. It was concluded that although the use of a PCL-TCP scaffold showed better maintenance of the alveolar contour as compared to autogenous particulate bone at 6 months, there was minimal bone regeneration within the defect.

The effect of PCL‐TCP scaffold loaded with mesenchymal stem cells on vertical bone augmentation in dog mandible: A preliminary report

Polycaprolactone-tricalcium phosphate (PCL-TCP), a new composite scaffold, has been shown to facilitate early revascularization and speed up bone regeneration process. The objective of this study was to evaluate the effect of PCL-TCP seeded with mesenchymal stem cells (MSCs) on healing of the vertical bone critical sized defect in dog's mandible. Bone marrow aspirate from dog humerous was cultured and the stemness of the cells was examined by differentiation staining methods and flow cytometric analysis. Third passage subculture cells (5 × 10⁵ cells) were loaded on 20 × 10 × 10 mm³ and incubated for 48 h. The presence of MSCs in the pores was evaluated by scanning electron microscope. Bilateral mandibular premolar teeth were extracted in four dogs and the buccal and lingual bone plates were reduced to make a vertical defect. Cell-loaded scaffolds were fixed in right side and left side received pure PCL-TCP scaffolds as a control side defects. Histomorphometric analysis after 8 weeks of the scaffold implantation showed higher amount of lamellar bone in the test side (48.63%) than control side (17.27%) (p < 0.05).The results suggest that PCL-TCP may be an appropriate scaffold for loading MSCs in bone regeneration.

A Tissue Engineering Solution for Segmental Defect Regeneration in Load-Bearing Long Bones

The reconstruction of large defects (>10 mm) in humans usually relies on bone graft transplantation. Limiting factors include availability of graft material, comorbidity, and insufficient integration into the damaged bone. We compare the gold standard autograft with biodegradable composite scaffolds consisting of medical-grade polycaprolactone and tricalcium phosphate combined with autologous bone marrow-derived mesenchymal stem cells (MSCs) or recombinant human bone morphogenetic protein 7 (rhBMP-7). Critical-sized defects in sheep--a model closely resembling human bone formation and structure--were treated with autograft, rhBMP-7, or MSCs. Bridging was observed within 3 months for both the autograft and the rhBMP-7 treatment. After 12 months, biomechanical analysis and microcomputed tomography imaging showed significantly greater bone formation and superior strength for the biomaterial scaffolds loaded with rhBMP-7 compared to the autograft. Axial bone distribution was greater at the interfaces. With rhBMP-7, at 3 months, the radial bone distribution within the scaffolds was homogeneous. At 12 months, however, significantly more bone was found in the scaffold architecture, indicating bone remodeling. Scaffolds alone or with MSC inclusion did not induce levels of bone formation comparable to those of the autograft and rhBMP-7 groups. Applied clinically, this approach using rhBMP-7 could overcome autograft-associated limitations.

Lateral ridge augmentation using a PCL‐TCP scaffold in a clinically relevant but challenging micropig model

In implant dentistry, there is a need for synthetic bone substitute blocks to support ridge augmentation in situations where large bone volumes are missing. Polycaprolactone-based scaffolds demonstrated excellent results in bone tissue engineering applications. The use of customized polycaprolactone-tricalcium phosphate (PCL-TCP) displayed promising results from recent rat femur and rabbit calvaria studies. However, data from clinically representative models in larger animals do not exist. To evaluate new bone formation in association with a novel PCL-TCP scaffold in comparison with an autogenous bone block graft for the reconstruction of large dentoalveolar defects in a clinically relevant but challenging pig jaw model. Chronic, non-contained one-wall defects were created in the mandible of micropigs and randomly assigned to receive one of the following guided bone regeneration (GBR) procedures for a period of 6 months. (A) Collagen membrane + autogenous block graft or (B) Collagen membrane + PCL-TCP scaffold. Micro computed tomography (μ-CT), histology and histomorphometry were used to assess new bone formation. Although μ-CT and histomorphometric analysis demonstrated a slight discrepancy between the measurements, the group utilizing autogenous bone grafts consistently reported superior new bone formation as compared to PCL-TCP scaffolds. When measured using μ-CT, the ratio of bone volume fraction for PCL-TCP scaffolds with respect to autografts yielded a mean efficacy of approximately 51%. Histological examination revealed that under favorable conditions, the new bone matrix and new bone marrow were in direct contact with the PCL-TCP scaffold rods and invading the interstices, suggesting good biocompatibility and high osteoconductivity. Autograft block grafts demonstrated 48.5-57.4% of pronounced resorption after 6 months following ridge augmentation. PCL-TCP scaffolds have demonstrated the potential application for lateral ridge augmentation following a healing period of 6 months in a micropig model.

A collagen network phase improves cell seeding of open-pore structure scaffolds under perfusion

Scaffolds with open-pore morphologies offer several advantages in cell-based tissue engineering, but their use is limited by a low cell-seeding efficiency. We hypothesized that inclusion of a collagen network as filling material within the open-pore architecture of polycaprolactone-tricalcium phosphate (PCL-TCP) scaffolds increases human bone marrow stromal cells (hBMSCs) seeding efficiency under perfusion and in vivo osteogenic capacity of the resulting constructs. PCL-TCP scaffolds, rapid prototyped with a honeycomb-like architecture, were filled with a collagen gel and subsequently lyophilized, with or without final crosslinking. Collagen-free scaffolds were used as controls. The seeding efficiency was assessed after overnight perfusion of expanded hBMSCs directly through the scaffold pores using a bioreactor system. By seeding and culturing freshly harvested hBMSCs under perfusion for 3 weeks, the osteogenic capacity of generated constructs was tested by ectopic implantation in nude mice. The presence of the collagen network, independently of the crosslinking process, significantly increased the cell seeding efficiency (2.5-fold), and reduced the loss of clonogenic cells in the supernatant. Although no implant generated frank bone tissue, possibly due to the mineral distribution within the scaffold polymer phase, the presence of a non-crosslinked collagen phase led to in vivo formation of scattered structures of dense osteoids. Our findings verify that the inclusion of a collagen network within open morphology porous scaffolds improves cell retention under perfusion seeding. In the context of cell-based therapies, collagen-filled porous scaffolds are expected to yield superior cell utilization, and could be combined with perfusion-based bioreactor devices to streamline graft manufacture.

Surface modification of PCL‐TCP scaffolds in rabbit calvaria defects: Evaluation of scaffold degradation profile, biomechanical properties and bone healing patterns

Traditionally, polycaprolactone (PCL) based scaffolds tend to degrade at a slow rate. Pretreatment of polycaprolactone-20% tricalcium phosphate (PCL-TCP) scaffolds under alkaline conditions can be utilized to increase the degradation rate and improve mechanical properties. Three groups of PCL-TCP scaffolds with varying pretreatment exposures with sodium hydroxide (NaOH) were studied in a rabbit calvaria defect model and analyzed at 2, 4, 8, 12, and 24 weeks. (Group A: Untreated, Group B: 3 M NaOH/ 48 h and Group C: 3 M NaOH/96 h). Micro-CT analysis demonstrated that scaffolds with increased surface roughness (Groups B and C) showed a greater impact on the overall volume loss during the early healing period between 2 and 8 weeks as compared to the untreated group. In addition, greater bone formation was detected in NaOH treated scaffolds as compared to the untreated group throughout the experiment. Scaffolds with increased surface roughness generally reported higher push out test and compressive strength values from 4 to 8 weeks of early healing. Interestingly, the mechanical properties displayed a decline in values from 12 weeks onwards in the modified groups suggesting a favorable breakdown or weakening of PCL-TCP scaffolds tailored for replacement by new bone formation.

Surface modification of PCL‐TCP scaffolds improve interfacial mechanical interlock and enhance early bone formation: An in vitro and in vivo characterization

Pretreatment of polycaprolactone-20% tricalcium phosphate (PCL-TCP) scaffolds under alkaline conditions can be utilized to alter surface characteristics for enhanced early bone formation. PCL-TCP scaffolds were treated with sodium hydroxide (NaOH) at various time intervals (group A: untreated, group B: 3M NaOH for 48 h, and group C: 3M NaOH for 96 h). In vitro results showed a greater degree of physical changes in the NaOH-treated scaffolds (B and C) than the untreated group (A). Clearly, the NaOH-treated scaffolds showed an increased surface roughness than the untreated ones. A significantly large number of "channel-like" pits and greater average pit sizes were detected in groups B (14.51 +/- 10.9 microm) and C (20.27 +/- 14.3 microm); and absent in group A. In addition, treated scaffolds had a significant reduction of the water contact angle (40.9-58.2%). Favorably, the pore dimensions and scaffold rod thickness remained unchanged throughout the experiment. When implanted in the calvaria of rabbits, NaOH-treated scaffolds reported greater early matrix deposition and bone formation from scanning electron images and Micro-computed tomography analyses. In conclusion, pretreatment of PCL-TCP scaffolds with NaOH increases the wettability and surface area for initial matrix deposition and early bone ingrowth.

Effects of Polycaprolactone-Tricalcium Phosphate, Recombinant Human Bone Morphogenetic Protein-2 and Dog Mesenchymal Stem Cells on Bone Formation: Pilot Study in Dogs

PurposeThe aim of this study was to evaluate the survival, proliferation, and bone formation of dog mesenchymal stem cells (dMSCs) in the graft material by using Polycaprolactone-tricalcium phosphate (PCL-TCP), auto-fibrin glue (AFG), recombinant human bone morphogenetic protein-2 (rhBMP-2), and dMSCs after a transplantation to the scapula of adult beagle dogs.Materials and MethodsThe subjects were two beagle dogs. Total dose of rhBMP-2 on each block was 10 µg with 50 µg/mg concentration. The cortical bone of the scapula of the dog was removed which was the same size of PCL-TCP block (Osteopore International Pte, Singapore; 5.0×5.0×8.0 mm in size), and the following graft material then was fixed with orthodontic mini-implant, Dual-top® (Titanium alloy, Jeil Co. Seoul, Korea). Four experimental groups were prepared for this study, Group 1: PCL-TCP + aFG; Group 2: PCL-TCP + aFG + dMSCs; Group 3: PCL-TCP + aFG + dMSCs + rhBMP-2; Group 4: PCL-TCP + aFG + dMSCs + rhBMP-2 + PCL membrane. The survival or proliferation of dMSCs cells was identified with an extracted tissue through a fluorescence microscope, H-E staining and Von-Kossa staining in two weeks and four weeks after the transplantation.ResultsThe survival and proliferation of dMSCs were identified through a fluorescence microscope from both Group 1 and Group 2 in two weeks and four weeks after the transplantation. Histological observation also found that the injected cells were proliferating well in the G2, G3, and G4 scaffolds.ConclusionThis study concluded that bone ingrowth occurred in PCL-TCP scaffold which was transplanted with rhBMP-2, and MSCs did not affect bone growth. More sufficient healing time would be needed to recognize effects of dMSCs on bone formation.

Customizing the degradation and load‐bearing profile of 3D polycaprolactone‐tricalcium phosphate scaffolds under enzymatic and hydrolytic conditions

The degradation of polycaprolactone-20% tricalcium phosphate (PCL-TCP) scaffolds was customized for dentoalveolar augmentation applications, where 5-6 months period is optimal. The scaffolds were treated with either 3M sodium hydroxide (NaOH) or 0.1% lipase solution for a total of 108 h. A greater degree of degradation and reduction in the physical properties of the scaffolds was observed in the lipase treated when compared with NaOH-treated scaffolds. After 108 h, increases in weight loss and average porosity of the scaffolds in the lipase-treated group measured 90.6% and 22.9%, respectively, when compared with 52.8% and 11.8% in the NaOH-treated group. The mechanical testing results revealed a similar trend, with a complete loss of compressive strength and modulus measured as early as 60 h in the lipase-treated group. The honeycomblike architecture was well preserved throughout the experiment only for the NaOH-treated scaffolds in addition to a favorable surface roughness ideal for bone-regeneration applications. In conclusion, pretreatment with NaOH demonstrates a simple approach for tailoring the physical properties and degradation rate of PCL-TCP scaffolds for the potential use as biomaterials targeted for dentoalveolar bone-regeneration procedures.

Polycaprolactone-20% Tricalcium Phosphate Scaffolds in Combination With Platelet-Rich Plasma for the Treatment of Critical-Sized Defects of the Mandible: A Pilot Study

Our group has recently fabricated 3-dimensional scaffolds of unique architecture to mediate favorable cell binding, proliferation, and differentiation. In this study, the osteoconductive and bioactive polycaprolactone-20% tricalcium phosphate (PCL-TCP) scaffolds were assessed for the treatment of critical-sized defects of the mandible, with respect to new bone formation. Platelet-rich plasma (PRP) was combined with some scaffolds to test if bone regeneration could be enhanced. Autologous PRP was prepared from whole blood collected from 8 mongrel dogs. In each dog, 3 defects (18 x 10 x 7 mm) were created at either the left or right mandible and treated with 1 of 2 treatment modalities: 1) grafting with scaffolds alone; or 2) grafting with scaffolds loaded with PRP. The scaffolds were stabilized using 2 dental implants each to prevent rotation. Micro-CT and histologic analysis were carried out on samples after 6 and 9 months. Micro-CT measurements showed that PRP-treated defects had 98.3% and 58.3% more bone volume fraction than defects grafted with scaffolds alone at 6 and 9 months, respectively (P < .05). No significant difference was noted between caudal and frontal situated PRP-treated defects, but a significant difference was observed between male and female dogs. Histologic analyses verified the deposition of osteoid and new bone trabeculae throughout the section at 6 months. The defect margins were filled with mature bone trabeculae at 9 months but the middle section of the scaffolds manifested disturbed mineralization. The scaffolds experienced 33% degradation from 6 to 9 months. PRP treatment had negligible effect on the degradation of the scaffolds. The pilot study showed that the treatment of critical-sized defects of the mandible with PCL-TCP scaffolds may be augmented by the addition of PRP.

Combination of platelet‐rich plasma with polycaprolactone‐tricalcium phosphate scaffolds for segmental bone defect repair

Porous scaffold biomaterials may offer a clinical alternative to bone grafts; however, scaffolds alone are typically insufficient to heal large bone defects. Numerous studies have demonstrated that osteoinductive growth factor or gene delivery significantly improves bone repair. However, given the important role of vascularization during bone regeneration, it may also be beneficial to incorporate factors that promote vascular ingrowth into constructs. In this study, a strategy combining structural polycaprolactone-20% tricalcium phosphate (PCL-TCP) composite scaffolds with platelet-rich plasma (PRP) was tested. Following bilateral implantation of constructs into 8 mm rat nonunion femoral defects, 3D vascular and bone ingrowth were quantified at 3 and 12 weeks using contrast-enhanced microcomputed tomography (micro-CT) imaging. At week 3, PRP-treated femurs displayed 70.3% higher vascular volume fraction than control femurs. Interestingly, bone volume fraction (BVF) was significantly higher for the empty scaffold group at the early time point. At 12 weeks, BVF measurements between the two groups were statistically equivalent. However, a greater proportion of PRP-treated femurs (83%) achieved bone union as compared to empty scaffold controls (33%). Consistent with this observation, biomechanical evaluation of functional integration also revealed a significantly higher torsional stiffness observed for PRP-treated defects compared to empty scaffolds. Ultimate torque at failure was not improved, however, perhaps due to the slow resorption profile of the scaffold material. Histological evaluation illustrated infiltration of vascularized connective tissue and bone in both groups. Given that bone ingrowth into untreated defects in this model is minimal, PCL-TCP scaffolds were clearly able to promote bone ingrowth but failed to consistently bridge the defect. The addition of PRP to PCL-TCP scaffolds accelerated early vascular ingrowth and improved longer-term functional integration. Taken together, the results of this study suggest that the use of PRP, alone or in combination with other bioactive components, may be an effective approach to augment the ability of porous biomaterial scaffolds to repair orthotopic defects.

Investigating the effects of preinduction on human adipose-derived precursor cells in an athymic rat model

The osteogenic potential of human adipose-derived precursor cells seeded on medical-grade polycaprolactone-tricalcium phosphate scaffolds was investigated in this in vivo study. Three study groups were investigated: (1) induced—stimulated with osteogenic factors only after seeding into scaffold; (2) preinduced—induced for 2 weeks before seeding into scaffolds; and (3) uninduced—cells without any introduced induction. For all groups, scaffolds were implanted subcutaneously into the dorsum of athymic rats. The scaffold/cell constructs were harvested at the end of 6 or 12 weeks and analyzed for osteogenesis. Gross morphological examination using scanning electron microscopy indicated good integration of host tissue with scaffold/cell constructs and extensive tissue infiltration into the scaffold interior. Alizarin Red histology and immunostaining showed a heightened level of mineralization and an increase in osteonectin, osteopontin, and collagen type I protein expression in both the induced and preinduced groups compared with the uninduced groups. However, no significant differences were observed in these indicators when compared between the induced and preinduced groups.

In vitro degradation of novel bioactive polycaprolactone—20% tricalcium phosphate composite scaffolds for bone engineering

Our group recently fabricated novel 3D polycaprolactone—20% tricalcium phosphate (PCL-TCP) composite scaffolds for applications in bone engineering. The bioactivity of such synthetic biomaterials can be evaluated by examining its ability to initiate the formation of apatite on its surface when immersed in simulated body fluids (SBF). In this study, the in vitro degradation behaviors of these scaffolds were systematically monitored for varying time periods of 1, 7, 14, 21 and 28 days post-immersion in SBF at 37 °C. Weight loss and water absorption of the samples indicated that PCL-TCP scaffolds were only slowly degraded. Biochemical assays and pH measurements revealed that hydroxyapatite, the main inorganic constituent of bone, commenced to form on the surface of the scaffolds after 17 days of immersion in SBF. Von Kossa assays demonstrated that calcium deposits increased progressively on the surface of the scaffolds after soaking in SBF for 2 weeks. Scanning electron microscopy verified the surface crystallization of the apatite layer formed over the entire period of time. In conclusion, the synergy of PCL with TCP in a composite scaffold confers both bioresorbability as well as bioactivity that offer an exciting approach for bone regeneration purposes.