An additive manufacturing‐based PCL–alginate–chondrocyte bioprinted scaffold for cartilage tissue engineering

Regenerative medicine is targeted to improve, restore or replace damaged tissues or organs using a combination of cells, materials and growth factors. Both tissue engineering and developmental biology currently deal with the process of tissue self‐assembly and extracellular matrix (ECM) deposition. In this investigation, additive manufacturing (AM) with a multihead deposition system (MHDS) was used to fabricate three‐dimensional (3D) cell‐printed scaffolds using layer‐by‐layer (LBL) deposition of polycaprolactone (PCL) and chondrocyte cell‐encapsulated alginate hydrogel. Appropriate cell dispensing conditions and optimum alginate concentrations for maintaining cell viability were determined. In vitro cell‐based biochemical assays were performed to determine glycosaminoglycans (GAGs), DNA and total collagen contents from different PCL–alginate gel constructs. PCL–alginate gels containing transforming growth factor‐β (TGFβ) showed higher ECM formation. The 3D cell‐printed scaffolds of PCL–alginate gel were implanted in the dorsal subcutaneous spaces of female nude mice. Histochemical [Alcian blue and haematoxylin and eosin (H&E) staining] and immunohistochemical (type II collagen) analyses of the retrieved implants after 4 weeks revealed enhanced cartilage tissue and type II collagen fibril formation in the PCL–alginate gel (+TGFβ) hybrid scaffold. In conclusion, we present an innovative cell‐printed scaffold for cartilage regeneration fabricated by an advanced bioprinting technology. Copyright © 2013 John Wiley & Sons, Ltd.     

Modification of 3D printed PCL scaffolds by PVAc and HA to enhance cytocompatibility and osteogenesis

In the study, a specific material system that contains poly-(ε-caprolactone) (PCL), polyvinyl acetate (PVAc) and hydroxyapatite (HA) was used to fabricate porous scaffolds employing a 3D printing technique for bone regeneration. Four groups of 3D printing scaffolds were fabricated: PCL, PCL/PVAc, PCL/HA and PCL/PVAc/HA for comparision. The morphologies, mechanical properties and biological characteristics of these scaffolds were analyzed using SEM, a material testing machine, in vitro cell culture and in vivo animal experiments. The results showed that these 3D printed scaffolds possessed porous channel structures with a hole size of 375–475 μm and porosity of 74.1–76.1%. The compressive moduli of the scaffolds increased with the addition of HA and decreased with the addition of PVAc. The PCL/PVAc/HA scaffold exhibited higher cell proliferation and bone formation rates than other groups (p < 0.001), which could be attributed to the synergistic effect of PAVc and HA components. Two types of new bone formation patterns in the scaffold were found in this study: one is the new bone formed directly on the grid matrix, and the other is the new bone initially formed in the center of the scaffold channel and then remolded to concentric circles. The osteogenesis pattern of the latter is analogous to the osteon structure of a cortical bone. The 3D printed scaffold based on PCL/PVAc/HA tri-component system is a promising prospect for future individualized bone repair applications.

Effects of PVAc and HA on the surface structure of PCL and the in vivo bone repair activity of scaffolds.     

Cell‐free artificial implants of electrospun fibres in a three‐dimensional gelatin matrix support sciatic nerve regeneration in vivo

Surgical repair of larger peripheral nerve lesions requires the use of autologous nerve grafts. At present, clinical alternatives to avoid nerve transplantation consist of empty tubes, which are only suitable for the repair over short distances and have limited success. We developed a cell‐free, three‐dimensional scaffold for axonal guidance in long‐distance nerve repair. Sub‐micron scale fibres of biodegradable poly‐ε‐caprolactone (PCL) and collagen/PCL (c/PCL) blends were incorporated in a gelatin matrix and inserted in collagen tubes. The conduits were tested by replacing 15‐mm‐long segments of rat sciatic nerves in vivo. Biocompatibility of the implants and nerve regeneration were assessed histologically, with electromyography and with behavioural tests for motor functions. Functional repair was achieved in all animals with autologous transplants, in 12 of 13 rats that received artificial implants with an internal structure and in half of the animals with empty nerve conduits. In rats with implants containing c/PCL fibres, the extent of recovery (compound muscle action potentials, motor functions of the hind limbs) was superior to animals that had received empty implants, but not as good as with autologous nerve transplantation. Schwann cell migration and axonal regeneration were observed in all artificial implants, and muscular atrophy was reduced in comparison with animals that had received no implants. The present design represents a significant step towards cell‐free, artificial nerve bridges that can replace autologous nerve transplants in the clinic. Copyright © 2017 John Wiley & Sons, Ltd.     

Characterization and osteogenic evaluation of mesoporous magnesium–calcium silicate/polycaprolactone/polybutylene succinate composite scaffolds fabricated by rapid prototyping

The properties of scaffolds for bone tissue engineering, including their biocompatibility, highly interconnected porosity, and mechanical integrity, are critical for promoting cell adhesion, proliferation, and osteoinduction. We used various physical and biological assays to obtain in vitro confirmation that the proposed composite scaffolds are potentially suitable for applications to bone tissue engineering. The proposed new composite scaffolds, which we fabricated by a rapid prototyping technique, were composed of mesoporous magnesium–calcium silicate (m_MCS), polycaprolactone (PCL), and polybutylene succinate (PBSu). We systematically evaluated the characteristics of the composite scaffolds, such as the hydrophilicity and bioactivity. We also investigated the proliferation and osteogenic differentiation of human mesenchymal stem cells (MSCs) scaffolded on the m_MCS/PCL/PBSu composite. Our results showed that, compared to the m_MCS/PCL scaffold, the m_MCS/PCL/PBSu scaffold has improved water absorption, in vitro degradability, biocompatibility, and bioactivity in simulated body fluid, while its mechanical strength is reduced. Moreover, the results of the cytotoxicity tests specified in ISO 10993-12 and ISO 10993-5 clearly indicate that the m_MCS/PCL scaffold is not toxic to cells. In addition, we obtained significant increases in initial cell attachment and improvements to the osteogenic MSC differentiation by replacing the m_MCS/PCL scaffold with the m_MCS/PCL/PBSu scaffold. Our results indicate that the m_MCS/PCL/PBSu scaffold achieves enhanced bioactivity, degradability, cytocompatibility, and osteogenesis. As such, this scaffold is a potentially promising candidate for use in stem cell-based bone tissue engineering.

A new composite scaffold consisting of mesoporous magnesium–calcium silicate (m_MCS), polycaprolactone (PCL), and polybutylene succinate (PBSu) was manufactured by a rapid prototyping technique, for stem cell-based bone tissue engineering.     

Correction: Three-dimensional directional nerve guide conduits fabricated by dopamine-functionalized conductive carbon nanofibre-based nanocomposite ink printing

Correction for ’Three-dimensional directional nerve guide conduits fabricated by dopamine-functionalized conductive carbon nanofibre-based nanocomposite ink printing’ by Shadi Houshyar et al., RSC Adv., 2020, 10, 40351–40364, DOI: 10.1039/D0RA06556K.

The authors regret that an incorrect version of Fig. 2 was included in the original article. The correct version of Fig. 2 is presented below.Open in a separate windowFig. 2(a) FTIR spectra of pure PCL and PCL printed with CNF and DA (40 and 100 μg mL⁻¹), where circles emphasize the OH peak (3700 cm⁻¹) of the carboxylated CNF and NH peak (1565 cm⁻¹) of dopamine. (b) Shear stress of the CNF and CNF + DA nanocomposite inks versus shear rate. (c) Viscosity versus shear rate of the prepared nanocomposite inks.The Royal Society of Chemistry apologises for these errors and any consequent inconvenience to authors and readers.     

Gelatin-assisted conglutination of aligned polycaprolactone nanofilms into a multilayered fibre-guiding scaffold for periodontal ligament regeneration

The repair or regeneration of well-aligned periodontal ligaments (PDL) remains a challenging clinical task in reconstructive surgeries and regenerative medicine. Topographical cell guidance has been utilized as a tissue-engineering bionic technique and facilitates the geometric design of composite materials. In this investigation, we manufactured multilayered scaffolds by cementing aligned polycaprolactone (PCL) electrospun films together using gelatin; the fibre-guiding scaffold mimicked the natural structure of periodontal ligaments and was aimed at promoting the growth of functionally oriented ligamentous fibres in vivo. Experiments in vitro demonstrated that this scaffold could provide good attachment and tissue-mimicking microenvironments for “seeding cells”, that is, human periodontal ligament mesenchyme cells (PDLSCs). Histological and immunofluorescence results indicated that a three-dimensional aligned construct could significantly enhance the angulation of new-born PDL-like tissue and facilitate collagen formation and maturation at periodontal fenestration defects compared to an amorphous PCL embedded scaffold. Multilayered fibre-guiding scaffold made of PCL and gelatin was demonstrated to be applicable for oriented neogenesis of periodontium, and it may represent an important potential application for dental stem cell delivery for periodontal regenerative medicine.

The 3D-AL scaffold mimics the physiological structure of periodontal ligaments and could enhance the angulation of regenerated PDL.     

Let-7 microRNAs Regenerate Peripheral Nerve Regeneration by Targeting Nerve Growth Factor

Peripheral nerve injury is a common clinical problem. Nerve growth factor (NGF) promotes peripheral nerve regeneration, but its clinical applications are limited by several constraints. In this study, we found that the time-dependent expression profiles of eight let-7 family members in the injured nerve after sciatic nerve injury were roughly similar to each other. Let-7 microRNAs (miRNAs) significantly reduced cell proliferation and migration of primary Schwann cells (SCs) by directly targeting NGF and suppressing its protein translation. Following sciatic nerve injury, the temporal change in let-7 miRNA expression was negatively correlated with that in NGF expression. Inhibition of let-7 miRNAs increased NGF secretion by primary cultured SCs and enhanced axonal outgrowth from a coculture of primary SCs and dorsal root gangalion neurons. In vivo tests indicated that let-7 inhibition promoted SCs migration and axon outgrowth within a regenerative microenvironment. In addition, the inhibitory effect of let-7 miRNAs on SCs apoptosis might serve as an early stress response to nerve injury, but this effect seemed to be not mediated through a NGF-dependent pathway. Collectively, our results provide a new insight into let-7 miRNA regulation of peripheral nerve regeneration and suggest a potential therapy for repair of peripheral nerve injury.     

Differentiated adipose‐derived stem cells act synergistically with RGD‐modified surfaces to improve neurite outgrowth in a co‐culture model

Peripheral nerve damage is a problem encountered after trauma and during surgery and the development of synthetic polymer conduits may offer a promising alternative to autografts. In order to improve the performance of the polymer to be used for nerve conduits, poly‐ε‐caprolactone (PCL) films were chemically functionalized with RGD moieties, using a chemical reaction previously developed. In vitro cultures of dissociated dorsal root ganglion (DRG) neurons provide a valid model to study different factors affecting axonal growth. In this work, DRG neurons were cultured on RGD‐functionalized PCL films. Adult adipose‐derived stem cells differentiated to Schwann cells (dASCs) were initially cultured on the functionalized PCL films, resulting in improved attachment and proliferation. dASCs were also co‐cultured with DRG neurons on treated and untreated PCL to assess stimulation by dASCs on neurite outgrowth. Neuron response was generally poor on untreated PCL films, but long neurites were observed in the presence of dASCs or RGD moieties. A combination of the two factors enhanced even further neurite outgrowth, acting synergistically. Finally, in order to better understand the extracellular matrix (ECM)–cell interaction, a β1 integrin blocking experiment was carried out. Neurite outgrowth was not affected by the specific antibody blocking, showing that β1 integrin function can be compensated by other molecules present on the cell membrane. Copyright © 2013 John Wiley & Sons, Ltd.     

An electrospun poly(ε-caprolactone) nanocomposite fibrous mat with a high content of hydroxyapatite to promote cell infiltration

Electrospun polymer/inorganic biomimetic nanocomposite scaffolds have emerged for use in a new strategy for bone regeneration. In this study, a poly(ε-caprolactone) (PCL)/hydroxyapatite (HAp) nanocomposite mat with a HAp content as high as 60% was prepared via one-step electrospinning using trifluoroethanol as the solvent, and it has superior dispersibility and spinnability. The structure and physicochemical properties of the scaffolds were studied using scanning electron microscopy and spectroscopic techniques. X-ray diffraction and Fourier transformed infrared spectroscopy confirmed the presence of HAp in the composite PCL fibers. The results of cell culturing suggested that the incorporation of HAp with PCL could regulate the cytoskeleton and the differentiation of cells. More interestingly, the high content of HAp was also found to be conducive to the infiltration of MC-3T3 cells into the mat. The results indicated the potential of PCL/HAp scaffolds as a promising substitute for bone regeneration.

PCL nanofibers with 60% HAp content were fabricated, and the presence of HAp regulated cell morphology to enhance cell infiltration.     

3D printed chitosan/polycaprolactone scaffold for lung tissue engineering: hope to be useful for COVID-19 studies

To prevent or reduce mortality from lung diseases, new biological materials and scaffolds are needed to conduct more accurate research and support lung tissue regeneration. On the other hand, the outbreak of the COVID-19 virus and its targeting of the human lung has caused many deaths worldwide. The main aim of this study was to provide a biologically and mechanically suitable 3D printed scaffold using chitosan/polycaprolactone bioink for lung tissue engineering. Design-Expert software was employed for studying various compositions for 3D printing. The selected scaffolds underwent physiochemical, biological and mechanical studies to evaluate if they are capable of MRC-5 cell line growth, proliferation, and migration. Based on the results, the average diameter of the chitosan/polycaprolactone strands was measured at 360 μm. Chitosan concentration controlled the printability, while changes in polycaprolactone content did not affect printability. The scaffolds showed excellent potential in swelling, degradation, and mechanical behavior, although they can be modified by adjusting the polycaprolactone content. The scaffolds also revealed notable cell adhesion, nontoxicity, low apoptosis, high proliferation, and cell biocompatibility in vitro. To sum up, scaffold 3 (chitosan/polycaprolactone ratio: 4 : 1) revealed better activity for MRC-5 cell culture. Thereby, this scaffold can be a good candidate for lung tissue engineering and may be applicable for more studies on the COVID-19 virus.

The aim of this study was to provide a biologically and mechanically suitable 3D printed scaffold using chitosan/polycaprolactone bioink for lung tissue engineering.     

生物可降解聚合物结合碱性成纤维细胞生长因子与内皮细胞的黏附

背景：可降解材料的应用是体外构建小口径组织工程血管的重要研究部分。如何对可降解材料进行改性,以利于材料本身体外抗凝与促进内皮细胞黏附,是目前血管组织工程研究的热点之一。目的：利用可降解聚己内酯接枝肝素材料体外负载碱性成纤维细胞生长因子,观察其对于内皮细胞黏附的影响。方法：应用聚己内酯可降解材料,将肝素活化后并与聚己内酯的端羟基发生酯化反应从而被锚定在聚己内酯两端。经过电纺丝技术,制备血管支架。同时利用肝素和生长因子间的静电吸附作用,使支架负载碱性成纤维细胞生长因子。采用低密度内皮细胞短期静态种植,观察负载细胞生长因子的可降解聚己内酯材料对内皮细胞生长黏附情况的影响。结果与结论：成功地制备了负载成纤维细胞生长因子的可降解聚己内酯接枝肝素支架。内皮黏附实验证实,该支架利于内皮细胞黏附。提示可降解聚己内酯接枝肝素材料负荷碱性细胞生长因子支架对内皮细胞有很好的体外黏附性。     

A novel therapy strategy for bile duct repair using tissue engineering technique: PCL/PLGA bilayered scaffold with hMSCs

The current clinical treatments for complications caused by hepatobiliary surgery still have some inevitable weakness. The aim of the study was to fabricate a tissue‐engineered bile duct that utilized a novel bilayered polymer scaffold combined with human bone marrow‐derived mesenchymal stem cells (hMSCs) for new treatment of biliary disease. The biocompatibility of polycaprolactone (PCL) (PCL)/poly(lactide‐co‐glycolide) (PLGA) scaffold with hMSCs was first examined, and the hMSC–PCL/PLGA constructs (MPPCs) prepared. The MPPCs and blank scaffolds were then transplanted into 18 pigs for evaluation its efficacy on bile duct repairing, respectively. In vitro, the PCL/PLGA scaffold was verified to support the adhesion, proliferation and matrix deposition of hMSCs. There was no sign of bile duct narrowing and cholestasis in all experimental animals. At 6 months, the MPPCs had a superior repairing effect on the bile duct injury, compared with the blank PCL/PLGA scaffolds. Therefore, the implanted scaffolds could not only support the biliary tract and allow free bile flow but also had direct or indirect positive effects on repair of injured bile duct. Copyright © 2015 John Wiley & Sons, Ltd.     

Minimally Invasive Approach to the Repair of Injured Skeletal Muscle With a Shape-memory Scaffold

Repair of injured skeletal muscle by cell therapies has been limited by poor survival of injected cells. Use of a carrier scaffold delivering cells locally, may enhance in vivo cell survival, and promote skeletal muscle regeneration. Biomaterial scaffolds are often implanted into muscle tissue through invasive surgeries, which can result in trauma that delays healing. Minimally invasive approaches to scaffold implantation are thought to minimize these adverse effects. This hypothesis was addressed in the context of a severe mouse skeletal muscle injury model. A degradable, shape-memory alginate scaffold that was highly porous and compressible was delivered by minimally invasive surgical techniques to injured tibialis anterior muscle. The scaffold controlled was quickly rehydrated in situ with autologous myoblasts and growth factors (either insulin-like growth factor-1 (IGF-1) alone or IGF-1 with vascular endothelial growth factor (VEGF)). The implanted scaffolds delivering myoblasts and IGF-1 significantly reduced scar formation, enhanced cell engraftment, and improved muscle contractile function. The addition of VEGF to the scaffold further improved functional recovery likely through increased angiogenesis. Thus, the delivery of myoblasts and dual local release of VEGF and IGF-1 from degradable scaffolds implanted through a minimally invasive procedure effectively promoted the functional regeneration of injured skeletal muscle.     

Neuronally expressed stem cell factor induces neural stem cell migration to areas of brain injury

Neural stem/progenitor cell (NSPC) migration toward sites of damaged central nervous system (CNS) tissue may represent an adaptive response for the purpose of limiting and/or repairing damage. Little is known of the mechanisms responsible for this migratory response. We constructed a cDNA library of injured mouse forebrain using subtractive suppression hybridization (SSH) to identify genes that were selectively upregulated in the injured hemisphere. We demonstrate that stem cell factor (SCF) mRNA and protein are highly induced in neurons within the zone of injured brain. Additionally, the SCF receptor c-kit is expressed on NSPCs in vitro and in vivo. Finally, we demonstrate that recombinant SCF induces potent NSPC migration in vitro and in vivo through the activation of c-kit on NSPCs. These data suggest that the SCF/c-kit pathway is involved in the migration of NSPCs to sites of brain injury and that SCF may prove useful for inducing progenitor cell recruitment to specific areas of the CNS for cell-based therapeutic strategies.     

Study on antibacterial properties and cytocompatibility of EPL coated 3D printed PCL/HA composite scaffolds

Biomaterial scaffolds play a critical role in bone tissue engineering. Moreover, 3D printing technology has enormous advantage in the manufacture of bioengineering scaffolds for patient-specific bone defect treatments. In order to provide an aseptic environment for bone regeneration, ε-poly-l-lysine (EPL), an antimicrobic cationic polypeptide, was used for surface modification of 3D printed polycaprolactone/hydroxyapatite (PCL/HA) scaffolds which were fabricated by fused deposition modeling (FDM) technology. The scaffold morphology and micro-structure were characterized by scanning electron microscopy (SEM), X-ray diffraction (XRD) and transform infrared spectroscopy (FT-IR). The release profile surface roughness, open porosity, and mechanical properties of the scaffolds were evaluated. Cell adhesion, proliferation, differentiation potential and antibacterial properties were also examined. As a result, 3D printed PCL/HA scaffolds with interconnected pores showed a slightly rough surface and improved mechanical properties due to adding hydroxyapatite (HA) particles. After being modified by EPL, favorable biocompatibility and osteoconductivity of ε-poly-l-lysine/polycaprolactone/hydroxyapatite (EPL/PCL/HA) scaffolds were observed. Moreover, antibacterial activity of the EPL/PCL/HA scaffolds was apparent. As a consequence, the EPL/PCL/HA scaffolds had great potential for bone regeneration and prevention of infections. This would yield a patient-specific bioactive and antibacterial composite scaffold for advanced bone tissue engineering applications.

Biomaterial scaffolds play a critical role in bone tissue engineering.     

Poly‐ε‐caprolactone scaffold and reduced in vitro cell culture: beneficial effect on compaction and improved valvular tissue formation

Tissue‐engineered heart valves (TEHVs), based on polyglycolic acid (PGA) scaffolds coated with poly‐4‐hydroxybutyrate (P4HB), have shown promising in vivo results in terms of tissue formation. However, a major drawback of these TEHVs is compaction and retraction of the leaflets, causing regurgitation. To overcome this problem, the aim of this study was to investigate: (a) the use of the slowly degrading poly‐ε‐caprolactone (PCL) scaffold for prolonged mechanical integrity; and (b) the use of lower passage cells for enhanced tissue formation. Passage 3, 5 and 7 (P3, P5 and P7) human and ovine vascular‐derived cells were seeded onto both PGA–P4HB and PCL scaffold strips. After 4 weeks of culture, compaction, tissue formation, mechanical properties and cell phenotypes were compared. TEHVs were cultured to observe retraction of the leaflets in the native‐like geometry. After culture, tissues based on PGA–P4HB scaffold showed 50–60% compaction, while PCL‐based tissues showed compaction of 0–10%. Tissue formation, stiffness and strength were increased with decreasing passage number; however, this did not influence compaction. Ovine PCL‐based tissues did render less strong tissues compared to PGA–P4HB‐based tissues. No differences in cell phenotype between the scaffold materials, species or cell passage numbers were observed. This study shows that PCL scaffolds may serve as alternative scaffold materials for human TEHVs with minimal compaction and without compromising tissue composition and properties, while further optimization of ovine TEHVs is needed. Reducing cell expansion time will result in faster generation of TEHVs, providing more rapid treatment for patients. Copyright © 2013 John Wiley & Sons, Ltd.     

Low-Intensity Pulsed Ultrasound Promotes Chondrogenic Progenitor Cell Migration via Focal Adhesion Kinase Pathway

Low-intensity pulsed ultrasound (LIPUS) has been studied frequently for its beneficial effects on the repair of injured articular cartilage. We hypothesized that these effects are due to stimulation of chondrogenic progenitor cell (CPC) migration toward injured areas of cartilage through focal adhesion kinase (FAK) activation. CPC chemotaxis in bluntly injured osteochondral explants was examined by confocal microscopy, and migratory activity of cultured CPCs was measured in transwell and monolayer scratch assays. FAK activation by LIPUS was analyzed in cultured CPCs by Western blot. LIPUS effects were compared with the effects of two known chemotactic factors: N-formyl-methionyl-leucyl-phenylalanine (fMLF) and high-mobility group box 1 (HMGB1) protein. LIPUS significantly enhanced CPC migration on explants and in cell culture assays. Phosphorylation of FAK at the kinase domain (Tyr 576/577) was maximized by 5 min of exposure to LIPUS at a dose of 27.5 mW/cm² and frequency of 3.5 MHz. Treatment with fMLF, but not HMBG1, enhanced FAK activation to a degree similar to that of LIPUS, but neither fMLF nor HMGB1 enhanced the LIPUS effect. LIPUS-induced CPC migration was blocked by suppressing FAK phosphorylation with a Src family kinase inhibitor that blocks FAK phosphorylation. Our results imply that LIPUS might be used to promote cartilage healing by inducing the migration of CPCs to injured sites, which could delay or prevent the onset of post-traumatic osteoarthritis.     

The application of POSS nanostructures in cartilage tissue engineering: the chondrocyte response to nanoscale geometry

Despite extensive research into cartilage tissue engineering (CTE), there is still no scaffold ideal for clinical applications. Various synthetic and natural polymers have been investigated in vitro and in vivo, but none have reached widespread clinical use. The authors investigate the potential of POSS–PCU, a synthetic nanocomposite polymer, for use in CTE. POSS–PCU is modified with silsesquioxane nanostructures that improve its biological and physical properties. The ability of POSS–PCU to support the growth of ovine nasoseptal chondrocytes was evaluated against a polymer widely used in CTE, polycaprolactone (PCL). Scaffolds with varied concentrations of the POSS molecule were also synthesized to investigate their effect on chondrocyte growth. Chondrocytes were seeded onto scaffold disks (PCU negative control; POSS–PCU 2%, 4%, 6%, 8%; PCL). Cytocompatibilty was evaluated using cell viability, total DNA, collagen and GAG assays. Chondrocytes cultured on POSS–PCU (2% POSS) scaffolds had significantly higher viability than PCL scaffolds (p < 0.001). Total DNA, collagen and sGAG protein were also greater on POSS–PCU scaffolds compared with PCL (p > 0.05). POSS–PCU (6% and 8% POSS) had improved viability and proliferation over an 18 day culture period compared with 2% and 4% POSS–PCU (p < 0.0001). Increasing the percentage of POSS in the scaffolds increased the size of the pores found in the scaffolds (p < 0.05). POSS–PCU has excellent potential for use in CTE. It supports the growth of chondrocytes in vitro and the POSS modification significantly enhances the growth and proliferation of nasoseptal chondrocytes compared with traditional scaffolds such as PCL. Copyright © 2013 John Wiley & Sons, Ltd.     

Overexpression of GDNF in the Uninjured DRG Exerts Analgesic Effects on Neuropathic Pain Following Segmental Spinal Nerve Ligation in Mice

Glial cell line-derived neurotrophic factor (GDNF), a survival-promoting factor for a subset of nociceptive small-diameter neurons, has been shown to exert analgesic effects on neuropathic pain. However, its detailed mechanisms of action are still unknown. In the present study, we investigated the site-specific analgesic effects of GDNF in the neuropathic pain state using lentiviral vector-mediated GDNF overexpression in mice with left fifth lumbar (L5) spinal nerve ligation (SNL) as a neuropathic pain model. A lentiviral vector expressing both GDNF and enhanced green fluorescent protein (EGFP) was constructed and injected into the left dorsal spinal cord, uninjured fourth lumbar (L4) dorsal root ganglion (DRG), injured L5 DRG, or plantar skin of mice. In SNL mice, injection of the GDNF-EGFP-expressing lentivirus into the dorsal spinal cord or uninjured L4 DRG partially but significantly reduced the mechanical allodynia in association with an increase in GDNF protein expression in each virus injection site, whereas injection into the injured L5 DRG or plantar skin had no effects. These results suggest that GDNF exerts its analgesic effects in the neuropathic pain state by acting on the central terminals of uninjured DRG neurons and/or on the spinal cells targeted by the uninjured DRG neurons.

Perspective

This article shows that GDNF exerts its analgesic effects on neuropathic pain by acting on the central terminals of uninjured DRG neurons and/or on the spinal cells targeted by these neurons. Therefore, research focusing on these GDNF-dependent neurons in the uninjured DRG would provide a new strategy for treating neuropathic pain.