Polycaprolactone Scaffolds Research Articles

Evaluation of a Bioabsorbable Scaffold and Interlocked Nail System for Segmental Bone Defect

In the current study, we designed and manufactured a scaffold and fixation system for the reconstruction of long-bone segmental defects in a rabbit tibia model. We used biocompatible and biodegradable materials, polycaprolactone (PCL) and PCL soaked with sodium alginate (PCL-Alg) to manufacture the scaffold, interlocking nail and screws using a phase separation casing method. Degradation and mechanical tests on the PCL and PCL-Alg scaffolds indicated that both were suitable for faster degradation and early weight-bearing capacity. PCL scaffold surface porosity facilitated the infiltration of alginate hydrogel through the scaffold. Cell viability results showed that the number of cells increased on Day 7 and decreased marginally by Day 14. For accurate placement of the scaffold and fixation system, a surgical jig was designed and 3D-printed using biocompatible resin in a stereolithography (SLA) 3D printer, then cured with UV light for increased strength. Our cadaver tests using New Zealand White rabbit confirmed our novel jigs' potential for accurate placement of the bone scaffold, intramedullary nail and the alignment of the fixation screws in future reconstructive surgeries on rabbit long-bone segmental defects. Additionally, the cadaver tests confirmed that our designed nails and screws were strong enough to carry the surgical insertion force. Therefore, our designed prototype has the potential for further clinical translational study using the rabbit tibia model.

Characterization of the Protein Corona of Three Chairside Hemoderivatives on Melt Electrowritten Polycaprolactone Scaffolds.

In tissue engineering, the relationship between a biomaterial surface and the host's immune response during wound healing is crucial for tissue regeneration. Despite hemoderivative functionalization of biomaterials becoming a common tissue-engineering strategy for enhanced regeneration, the characteristics of the protein-biomaterial interface have not been fully elucidated. This study characterized the interface formed by the adsorbed proteins from various hemoderivatives with pristine and calcium phosphate (CaP)-coated polycaprolactone (PCL) melt electrowritten scaffolds. PCL scaffolds were fabricated by using melt electrospinning writing (MEW). Three hemoderivatives (pure platelet-rich plasma (P-PRP), leucocyte platelet-rich plasma (L-PRP) and injectable platelet-rich fibrin (i-PRF)) and total blood PLASMA (control) were prepared from ovine blood. Hemoderivatives were characterized via SEM/EDX, cross-linking assay, weight loss, pH and protein quantification. The interface between PCL/CaP and hemoderivative was examined via FTIR, XPS and electrophoresis. i-PRF/PCL-CaP (1653 cm-1), PLASMA/PCL-CaP (1652 cm-1) and i-PRF/PCL (1651 cm-1) demonstrated a strong signal at the Amide I region. PLASMA and i-PRF presented similar N1s spectra, with most of the nitrogen involved in N-C=O bonds (≈400 eV). i-PRF resulted in higher adsorption of low molecular weight (LMW) proteins at 60 min, while PLASMA exhibited the lowest adsorption. L-PRP and P-PRP had a similar pattern of protein adsorption. The characteristics of biomaterial interfaces can be customized, thus creating a specific hemoderivative-defined layer on the PCL surface. i-PRF demonstrated a predominant adsorption of LMW proteins. Further investigation of hemoderivative functionalized biomaterials is required to identify the differential protein corona composition, and the resultant immune response and regenerative capacity.

Reduced Fibroblast Activation on Electrospun Polycaprolactone Scaffolds.

In vivo, quiescent fibroblasts reside in three-dimensional connective tissues and are activated in response to tissue injury before proliferating rapidly and becoming migratory and contractile myofibroblasts. When deregulated, chronic activation drives fibrotic disease. Fibroblasts cultured on stiff 2D surfaces display a partially activated phenotype, whilst many 3D environments limit fibroblast activation. Cell mechanotransduction, spreading, polarity, and integrin expression are controlled by material mechanical properties and micro-architecture. Between 3D culture systems, these features are highly variable, and the challenge of controlling individual properties without altering others has led to an inconsistent picture of fibroblast behaviour. Electrospinning offers greater control of mechanical properties and microarchitecture making it a valuable model to study fibroblast activation behaviour in vitro. Here, we present a comprehensive characterisation of the activation traits of human oral fibroblasts grown on a microfibrous scaffold composed of electrospun polycaprolactone. After over 7 days in the culture, we observed a reduction in proliferation rates compared to cells cultured in 2D, with low KI67 expression and no evidence of cellular senescence. A-SMA mRNA levels fell, and the expression of ECM protein-coding genes also decreased. Electrospun fibrous scaffolds, therefore, represent a tuneable platform to investigate the mechanisms of fibroblast activation and their roles in fibrotic disease.

Visualization of porosity and pore size gradients in electrospun scaffolds using laser metrology.

We applied a recently developed method, laser metrology, to characterize the influence of collector rotation on porosity gradients of electrospun polycaprolactone (PCL) widely investigated for use in tissue engineering. The prior- and post-sintering dimensions of PCL scaffolds were compared to derive quantitative, spatially-resolved porosity 'maps' from net shrinkage. Deposited on a rotating mandrel (200 RPM), the central region of deposition reaches the highest porosity, ~92%, surrounded by approximately symmetrical decreases to ~89% at the edges. At 1100 RPM, a uniform porosity of ~88-89% is observed. At 2000 RPM, the lowest porosity, ~87%, is found in the middle of the deposition, rebounding to ~89% at the edges. Using a statistical model of random fiber network, we demonstrated that these relatively small changes in porosity values produce disproportionately large variations in pore size. The model predicts an exponential dependence of pore size on porosity when the scaffold is highly porous (e.g., >80%) and, accordingly, the observed porosity variation is associated with dramatic changes in pore size and ability to accommodate cell infiltration. Within the thickest regions most likely to 'bottleneck' cell infiltration, pore size decreases from ~37 to 23 μm (38%) when rotational speeds increased from 200 to 2000 RPM. This trend is corroborated by electron microscopy. While faster rotational speeds ultimately overcome axial alignment induced by cylindrical electric fields associated with the collector geometry, it does so at the cost of eliminating larger pores favoring cell infiltration. This puts the bio-mechanical advantages associated with collector rotation-induced alignment at odds with biological goals. A more significant decrease in pore size from ~54 to ~19 μm (65%), well below the minimum associated with cellular infiltration, is observed from enhanced collector biases. Finally, similar predictions show that sacrificial fiber approaches are inefficient in achieving cell-permissive pore sizes.

3D-printed vascularized biofunctional scaffold for bone regeneration.

3D-printed biofunctional scaffolds have promising applications in bone tissue regeneration. However, the development of bioinks with rapid internal vascularization capabilities and relatively sustained osteoinductive bioactivity is the primary technical challenge. In this work, we added rat platelet-rich plasma (PRP) to a methacrylated gelatin (GelMA)/methacrylated alginate (AlgMA) system, which was further modified by a nanoclay, laponite (Lap). We found that Lap was effective in retarding the release of multiple growth factors from the PRP-GelMA/AlgMA (PRP-GA) hydrogel and sustained the release for up to 2 weeks. Our in vitro studies showed that the PRP-GA@Lap hydrogel significantly promoted the proliferation, migration, and osteogenic differentiation of rat bone marrow mesenchymal stem cells, accelerated the formation of endothelial cell vascular patterns, and promoted macrophage M2 polarization. Furthermore, we printed hydrogel bioink with polycaprolactone (PCL) layer-by-layer to form active bone repair scaffolds and implanted them in subcutaneous and femoral condyle defects in rats. In vivo experiments showed that the PRP-GA@Lap/PCL scaffolds significantly promoted vascular inward growth and enhanced bone regeneration at the defect site. This work suggests that PRP-based 3D-bioprinted vascularized scaffolds will have great potential for clinical translation in the treatment of bone defects.

3D Electrospun Polycaprolactone Scaffolds to Assess Human Periodontal Ligament Cells Mechanobiological Behaviour.

While periodontal ligament cells are sensitive to their 3D biomechanical environment, only a few 3D in vitro models have been used to investigate the periodontal cells mechanobiological behavior. The objective of the current study was to assess the capability of a 3D fibrous scaffold to transmit a mechanical loading to the periodontal ligament cells. Three-dimensional fibrous polycaprolactone (PCL) scaffolds were synthetized through electrospinning. Scaffolds seeded with human periodontal cells (103 mL-1) were subjected to static (n = 9) or to a sinusoidal axial compressive loading in an in-house bioreactor (n = 9). At the end of the culture, the dynamic loading seemed to have an influence on the cells' morphology, with a lower number of visible cells on the scaffolds surface and a lower expression of actin filament. Furthermore, the dynamic loading presented a tendency to decrease the Alkaline Phosphatase activity and the production of Interleukin-6 while these two biomolecular markers were increased after 21 days of static culture. Together, these results showed that load transmission is occurring in the 3D electrospun PCL fibrous scaffolds, suggesting that it can be used to better understand the periodontal ligament cells mechanobiology. The current study shows a relevant way to investigate periodontal mechanobiology using 3D fibrous scaffolds.

Osteoregenerative Potential of 3D-Printed Poly ε-Caprolactone Tissue Scaffolds In Vitro Using Minimally Manipulative Expansion of Primary Human Bone Marrow Stem Cells.

The repair of orthopedic and maxillofacial defects in modern medicine currently relies heavily on the use of autograft, allograft, void fillers, or other structural material composites. This study examines the in vitro osteo regenerative potential of polycaprolactone (PCL) tissue scaffolding, fabricated via a three-dimensional (3D) additive manufacturing technology, i.e., a pneumatic micro extrusion (PME) process. The objectives of this study were: (i) To examine the innate osteoinductive and osteoconductive potential of 3D-printed PCL tissue scaffolding and (ii) To perform a direct in vitro comparison of 3D-printed PCL scaffolding with allograft Allowash® cancellous bone cubes with regards to cell-scaffold interactions and biocompatibility with three primary human bone marrow (hBM) stem cell lines. This study specifically examined cell survival, cell integration, intra-scaffold cell proliferation, and differentiation of progenitor cells to investigate the potential of 3D-printed PCL scaffolds as an alternative to allograft bone material for the repair of orthopedic injuries. We found that mechanically robust PCL bone scaffolds can be fabricated via the PME process and the resulting material did not elicit detectable cytotoxicity. When the widely used osteogenic model SAOS-2 was cultured in PCL extract medium, no detectable effect was observed on cell viability or proliferation with multiple test groups showing viability ranges of 92.2% to 100% relative to a control group with a standard deviation of ±10%. In addition, we found that the honeycomb infill pattern of the 3D-printed PCL scaffold allowed for superior mesenchymal stem-cell integration, proliferation, and biomass increase. When healthy and active primary hBM cell lines, having documented in vitro growth rates with doubling times of 23.9, 24.67, and 30.94 h, were cultured directly into 3D-printed PCL scaffolds, impressive biomass increase values were observed. It was found that the PCL scaffolding material allowed for biomass increase values of 17.17%, 17.14%, and 18.18%, compared to values of 4.29% for allograph material cultured under identical parameters. It was also found that the honeycomb scaffold infill pattern was superior to the cubic and rectangular matrix structures, and provided a superior microenvironment for osteogenic and hematopoietic progenitor cell activity and auto-differentiation of primary hBM stem cells. Histological and immunohistochemical studies performed in this work confirmed the regenerative potential of PCL matrices in the orthopedic setting by displaying the integration, self-organization, and auto-differentiation of hBM progenitor cells within the matrix. Differentiation products including mineralization, self-organizing "proto-osteon" structures, and in vitro erythropoiesis were observed in conjunction with the documented expression of expected bone marrow differentiative markers including CD-99 (>70%), CD-71 (>60%), and CD-61 (>5%). All of the studies were conducted without the addition of any exogenous chemical or hormonal stimulation and exclusively utilized the abiotic and inert material polycaprolactone; setting this work apart from the vast majority of contemporary investigations into synthetic bone scaffold fabrication In summary, this study demonstrates the unique clinical potential of 3D-printed PCL scaffolds for stem cell expansion and incorporation into advanced microstructures created via PME manufacturing to generate a physiologically inert temporary bony defect graft with significant autograft features for enhanced end-stage healing.

Embedding aligned nanofibrous architectures within 3D-printed polycaprolactone scaffolds for directed cellular infiltration and tissue regeneration

Three-dimensional (3D) printing provides a promising way to fabricate biodegradable scaffolds with designer architectures for the regeneration of various tissues. However, the existing 3D-printed scaffolds commonly suffer from weak cell-scaffold interactions and insufficient cell organizations due to the limited resolution of the 3D-printed features. Here, composite scaffolds with mechanically-robust frameworks and aligned nanofibrous architectures are presented and hybrid manufactured by combining techniques of 3D printing, electrospinning, and unidirectional freeze-casting. It was found that the composite scaffolds provided volume-stable environments and enabled directed cellular infiltration for tissue regeneration. In particular, the nanofibrous architectures with aligned micropores served as artificial extracellular matrix materials and improved the attachment, proliferation, and infiltration of cells. The proposed scaffolds can also support the adipogenic maturation of adipose-derived stem cells (ADSCs) in vitro. Moreover, the composite scaffolds were found to guide directed tissue infiltration and promote nearby neovascularization when implanted into a subcutaneous model of rats, and the addition of ADSCs further enhanced their adipogenic potential. The presented hybrid manufacturing strategy might provide a promising way to produce additional topological cues within 3D-printed scaffolds for better tissue regeneration.

Processing and evaluation of the structure-properties of electrospun PCL/ zirconium nanoparticle scaffolds

Electrospun fibers with large surface area are of particular importance in the tissue engineering field as their highly porous structure favours cell adhesion, proliferation, and migration. Herein, we report a facile approach for the development of polycaprolactone (PCL) scaffolds incorporated with zirconium dioxide (ZrO 2 ) nanoparticles. Zirconium dioxide nanoparticles were incorporated into the fibers at five different concentrations (0.5, 1, 3, 5 and 7 wt. %), based on the amount of polymer. The morphology of the scaffolds was evaluated by scanning electron microscopy which showed interconnected pore structure. The incorporation of nanoparticles was confirmed from EDX studies. The functional groups were analysed using FTIR and crystallinity was analysed by XRD. Mechanical studies demonstrated that the scaffolds containing 1 wt. % Zirconium dioxide nanoparticles exhibited the highest tensile strength. The scaffolds showed antibacterial activity against Gram positive bacterium Staphylococcus lugdunensis and were not active against Gram-negative bacterium Proteus vulgaris, at all time-points. ZrO 2 loaded PCL scaffolds with good mechanical and biological properties will open avenues for the potential applications of the electrospun membranes for tissue engineering.

Polycaprolactone nanofibers colonized by HeLa cells: an alternative for in vitro investigation of fungal infection.

Introduction: In vitro 3D equivalent tissues can be used for studies of fungal infections. Objectives: To develop 3D electrospun nanofibers using polycaprolactone (PCL) colonized by HeLa cells as a possible in vitro model for the investigation of fungal infection. Materials & methods: A PCL solution was synthesized and electrospun. HeLa cells were cultured on the nanostructured PCL scaffolds, forming a 3D structure. Physicochemical, biological and Candida albicans infection assays were performed in this model. Results: The nanostructured PCL scaffolds showed favorable physicochemical characteristics and allowed the colonization of HeLa cells, which showed indications of extracellular matrix production. Conclusions: Fungal infection was evidenced in the 3D nanostructured PCL scaffolds, being viable, economical and compatible to study fungal infections in vitro.

Use of 3D-printed Polycaprolactone + Hyaluronic Acid-Based Scaffold in Orthopedic Practice: Report of Two Cases

3D-printed (3DP) polycaprolactone (PCL)-based scaffolds have gained popularity in the past decade. Despite the wide utility of autografts for bone regeneration in orthopedics practice, 3DP PCL scaffolds may replace this general application. Here we present a 42-year-old patient who had glenohumeral arthritis due to rheumatoid arthritis accompanied by a Walch 3 glenoid defect, and a 19-year-old patient with neglected Galeazzi fracture–dislocation accompanied by a radius non-union. In both patients, a 3DP PCL + hyaluronic acid-based scaffold was used for bone regeneration purposes. The preliminary results showed that the 3DP PCL + hyaluronic acid scaffold provided bone regeneration and may be a promising alternative to autografting for bone regeneration in orthopedic practice.

Effect of Pore Characteristics and Alkali Treatment on the Physicochemical and Biological Properties of a 3D-Printed Polycaprolactone Bone Scaffold.

Polycaprolactone scaffolds were designed and 3D-printed with different pore shapes (cube and triangle) and sizes (500 and 700 μm) and modified with alkaline hydrolysis of different ratios (1, 3, and 5 M). In total, 16 designs were evaluated for their physical, mechanical, and biological properties. The present study mainly focused on the pore size, porosity, pore shapes, surface modification, biomineralization, mechanical properties, and biological characteristics that might influence bone ingrowth in 3D-printed biodegradable scaffolds. The results showed that the surface roughness in treated scaffolds increased compared to untreated polycaprolactone scaffolds (R a = 2.3-10.5 nm and R q = 17- 76 nm), but the structural integrity declined with an increase in the NaOH concentration especially in the scaffolds with small pores and a triangle shape. Overall, the treated polycaprolactone scaffolds particularly with the triangle shape and smaller pore size provided superior performance in mechanical strength similar to that of cancellous bone. Additionally, the in vitro study showed that cell viability increased in the polycaprolactone scaffolds with cubic pore shapes and small pore sizes, whereas mineralization was enhanced in the designs with larger pore sizes. Based on the results obtained, this study demonstrated that the 3D-printed modified polycaprolactone scaffolds exhibit a favorable mechanical property, biomineralization, and better biological properties; therefore, they can be applied in bone tissue engineering.

Biomimetic Growth of Hydroxyapatite in Hybrid Polycaprolactone/Graphene Oxide Ultra-Porous Scaffolds.

This paper reports the preparation and characterization of hybrid scaffolds composed of polycaprolactone (PCL) and different graphene oxide (GO) amounts, intending to incorporate the intrinsic characteristics of their constituents, such as bioactivity and biocidal effect. These materials were fabricated by a solvent-casting/particulate leaching technique showing a bimodal porosity (macro and micro) that was around 90%. The highly interconnected scaffolds were immersed in a simulated body fluid, promoting the growth of a hydroxyapatite (HAp) layer, making them ideal candidates for bone tissue engineering. The growth kinetics of the HAp layer was influenced by the GO content, a remarkable result. Furthermore, as expected, the addition of GO neither significantly improves nor reduces the compressive modulus of PCL scaffolds. The thermal behavior of composites was investigated by differential scanning calorimetry, showing an increase in crystallinity as the addition of GO raised, which implies that GO nanosheets can act as seeds to induce the crystallization of PCL. The improved bioactivity was demonstrated by the deposition of an HAp layer on the surface of the scaffold with GO, especially with a 0.1% GO content.

Covalently Grafted Biomimetic Matrix Reconstructs the Regenerative Microenvironment of the Porous Gradient Polycaprolactone Scaffold to Accelerate Bone Remodeling.

Integrating a biomimetic extracellular matrix to improve the microenvironment of 3D printing scaffolds is an emerging strategy for bone substitute design. Here, a "soft-hard" bone implant (BM-g-DPCL) consisting of a bioactive matrix chemically integrated on a polydopamine (PDA)-coated porous gradient scaffold by polyphenol groups is constructed. The PDA-coated "hard" scaffolds promoted Ca2+ chelation and mineral deposition; the "soft" bioactive matrix is beneficial to the migration, proliferation, and osteogenic differentiation of stem cells in vitro, accelerated endogenous stem cell recruitment, and initiated rapid angiogenesis in vivo. The results of the rabbit cranial defect model (Φ= 10mm) confirmed that BM-g-DPCL promoted the integration between bone tissue and implant and induced the deposition of bone matrix. Proteomics confirmed that cytokine adhesion, biomineralization, rapid vascularization, and extracellular matrix formation are major factors that accelerate bone defect healing. This strategy of highly chemically bonded soft-hard components guided the construction of the bioactive regenerative scaffold.

Non-electric bioelectrical analog strategy by a biophysical-driven nano-micro spatial anisotropic scaffold for regulating stem cell niche and tissue regeneration in a neuronal therapy.

The slow regenerating rate and misdirected axonal growth are primary concerns that disturb the curative outcome of peripheral nerve repair. Biophysical intervention through nerve scaffolds can provide efficient, tunable and sustainable guidance for nerve regrowth. Herein, we fabricate the reduced graphene oxide (rGO)/polycaprolactone (PCL) scaffold characterized with anisotropic microfibers and oriented nanogrooves by electrospinning technique. Adipose-derived stem cells (ADSCs) are seeded on the scaffolds in vitro and the viability, neural differentiation efficiency and neurotrophic potential are investigated. RGO/PCL conduits reprogram the phenotype of seeded cells and efficiently repair 15mm sciatic nerve defect in rats. In summary, biophysical cues on nerve scaffolds are key determinants to stem cell phenotype, and ADSC-seeded rGO/PCL oriented scaffolds are promising, controllable and sustainable approaches to enable peripheral nerve regeneration.

Erratum to "Controlled Vacuum Seeding as a Means of Generating Uniform Cellular Distribution in Electrospun Polycaprolactone (PCL) Scaffolds," ASME J. Biomech. Eng., 2009, 131(7), p. 074521, DOI: 10.1115/1.3173283.

Erratum of figures and tables

Construction of Rapid Extracellular Matrix-Deposited Small-Diameter Vascular Grafts Induced by Hypoxia in a Bioreactor.

Cardiovascular disease has become one of the most globally prevalent diseases, and autologous or vascular graft transplantation has been the main treatment for the end stage of the disease. However, there are no commercialized small-diameter vascular graft (SDVG) products available. The design of SDVGs is promising in the future, and SDVG preparation using an in vitro bioreactor is a favorable method, but it faces the problem of long-term culture of >8 weeks. Herein, we used different oxygen (O2) concentrations and mechanical stimulation to induce greater secretion of extracellular matrix (ECM) from cells in vitro to rapidly prepare SDVGs. Culturing with 2% O2 significantly increased the production of the ECM components and growth factors of human dermal fibroblasts (hDFs). To accelerate the formation of ECM, hDFs were seeded on a polycaprolactone (PCL) scaffold and cultured in a flow culture bioreactor with 2% O2 for only 3 weeks. After orthotopic transplantation in rat abdominal aorta, the cultured SDVGs (PCL-decellularized ECM) showed excellent endothelialization and smooth muscle regeneration. The vascular grafts cultured with hypoxia and mechanical stimulation could accelerate the reconstruction speed and obtain an improved therapeutic effect and thereby provide a new research direction for improving the production and supply of SDVGs.

Improvement of bioactivity with dual bioceramic incorporation to nanofibrous PCL scaffolds

Bone tissue injuries, diseases or related clinical interventions require bone tissue engineering (BTE) approaches for regeneration of large bone defects, especially for compromised situations. Most BTE applications in literature focused on composites of polymers with a single type of bioceramic. However, native bone matrix has various inorganic components. Accordingly, this study aimed to investigate the use of dual bioceramics in BTE scaffolds prepared by wet-electrospinning of Poly-caprolactone (PCL) and bioceramics; hydroxyapatite (HA), carbonated hydroxyapatite (CHA) and akermanite (AKR). After physicochemical characterizations of single and dual bioceramic-discs, it was found that AKR component should be at most 10% and thus, fibrous 3D composite scaffolds (PCL/HA; PCL/CHA; PCL/90HA/10AKR; PCL/90CHA/10AKR) were prepared. Scaffolds were then compared for degradation, swelling and microstructure properties. PCL/CHA, PCL/HA/AKR and PCL/CHA/AKR scaffolds improved apatite formation and presence of CHA, AKR or both in scaffolds increased Ca/P precipitation compared to HA involving scaffolds. Specifically, when two bioceramics were introduced, there was a noticeable increase in mineralization. No significant weight change was detected for scaffolds after two weeks of incubation. Presence of bioceramics in scaffolds significantly reduced swelling of PCL scaffolds with PCL/90HA10AKR and PCL/90CHA10AKR having higher swelling results than PCL/HA and PCL/CHA, respectively. Additionally, scaffolds with dual bioceramics (90HA10AKR, 90CHA10AKR) had higher cell proliferation for bone cells (Saos-2). In terms of osteogenic activity all groups showed increased ALP activity at 14 days compared to 7 days with PCL/CHA scaffolds having the highest value. Results suggests that newly developed PCL/90HA10AKR and PCL/90CHA10AKR composites hold potential for bone tissue engineering applications.

Magnetofection of miR-21 promoted by electromagnetic field and iron oxide nanoparticles via the p38 MAPK pathway contributes to osteogenesis and angiogenesis for intervertebral fusion

BackgroundMagnetofection-mediated gene delivery shows great therapeutic potential through the regulation of the direction and degree of differentiation. Lumbar degenerative disc disease (DDD) is a serious global orthopaedic problem. However, even though intervertebral fusion is the gold standard for the treatment of DDD, its therapeutic effect is unsatisfactory. Here, we described a novel magnetofection system for delivering therapeutic miRNAs to promote osteogenesis and angiogenesis in patients with lumbar DDD.ResultsCo-stimulation with electromagnetic field (EMF) and iron oxide nanoparticles (IONPs) enhanced magnetofection efficiency significantly. Moreover, in vitro, magnetofection of miR-21 into bone marrow mesenchymal stem cells (BMSCs) and human umbilical endothelial cells (HUVECs) influenced their cellular behaviour and promoted osteogenesis and angiogenesis. Then, gene-edited seed cells were planted onto polycaprolactone (PCL) and hydroxyapatite (HA) scaffolds (PCL/HA scaffolds) and evolved into the ideal tissue-engineered bone to promote intervertebral fusion. Finally, our results showed that EMF and polyethyleneimine (PEI)@IONPs were enhancing transfection efficiency by activating the p38 MAPK pathway.ConclusionOur findings illustrate that a magnetofection system for delivering miR-21 into BMSCs and HUVECs promoted osteogenesis and angiogenesis in vitro and in vivo and that magnetofection transfection efficiency improved significantly under the co-stimulation of EMF and IONPs. Moreover, it relied on the activation of p38 MAPK pathway. This magnetofection system could be a promising therapeutic approach for various orthopaedic diseases.Graphical

Fabrication of polycaprolactone/calcium phosphates hybrid scaffolds impregnated with plant extracts using 3D printing for potential bone regeneration

The increase in critical bone diseases and defects in the world's population increases the need for bone substitutes to restore form and function. Organic and inorganic scaffolds with antibacterial properties could provide advantages for bone regeneration. In this study, we obtained scaffolds of polycaprolactone (PCL) charged with calcium phosphates nanoparticles and impregnated with extracts of Colombian plants as an alternative for potential bone regeneration. Calcium phosphate nanoparticles were obtained via auto-combustion synthesis. The nanoparticles were incorporated into the PCL with a chemical dissolution-disperse process. The composite obtained was used to produce a filament to print Triply Periodic Minimal Surface (TPMS) based scaffolds. Such geometry facilitates cellular growth thanks to its interconnected porosity. The scaffolds were impregnated with extracts of Justicia cf colorifera (Acanthaceae), and Billia rosea (Sapindaceae) due to their ancestral medical applications. A physical and biological characterization was conducted. The process to print scaffolds with an enhanced geometry to facilitate the flux of biological fluids was successful. The scaffolds loaded with B. rosea showed strong antibacterial behavior, suggesting the presence of reported terpenoids with antibacterial properties. The approach used in this study evidenced promising prospects for bone defect repair.