Electrical aligned polyurethane nerve guidance conduit modulates macrophage polarization and facilitates immunoregulatory peripheral nerve regeneration.

Biomaterials can modulate the local immune microenvironments to promote peripheral nerve regeneration. Inspired by the spatial orderly distribution and endogenous electric field of nerve fibers, we aimed to investigate the synergistic effects of electrical and topological cues on immune microenvironments of peripheral nerve regeneration. Nerve guidance conduits (NGCs) with aligned electrospun nanofibers were fabricated using a polyurethane copolymer containing a conductive aniline trimer and degradable L-lysine (PUAT). In vitro experiments showed that the aligned PUAT (A-PUAT) membranes promoted the recruitment of macrophages and induced their polarization towards the pro-healing M2 phenotype, which subsequently facilitated the migration and myelination of Schwann cells. Furthermore, NGCs fabricated from A-PUAT increased the proportion of pro-healing macrophages and improved peripheral nerve regeneration in a rat model of sciatic nerve injury. In conclusion, this study demonstrated the potential application of NGCs in peripheral nerve regeneration from an immunomodulatory perspective and revealed A-PUAT as a clinically-actionable strategy for peripheral nerve injury.

Macrophages in guided bone regeneration: potential roles and future directions.

Guided bone regeneration (GBR) is one of the most widely used and thoroughly documented alveolar bone augmentation surgeries. However, implanting GBR membranes inevitably triggers an immune response, which can lead to inflammation and failure of bone augmentation. It has been shown that GBR membranes may significantly improve in vivo outcomes as potent immunomodulators, rather than solely serving as traditional barriers. Macrophages play crucial roles in immune responses and participate in the entire process of bone injury repair. The significant diversity and high plasticity of macrophages complicate our understanding of the immunomodulatory mechanisms underlying GBR. This review provides a comprehensive summary of recent findings on the potential role of macrophages in GBR for bone defects in situ. Specifically, macrophages can promote osteogenesis or fibrous tissue formation in bone defects and degradation or fibrous encapsulation of membranes. Moreover, GBR membranes can influence the recruitment and polarization of macrophages. Therefore, immunomodulating GBR membranes are primarily developed by improving macrophage recruitment and aggregation as well as regulating macrophage polarization. However, certain challenges remain to be addressed in the future. For example, developing more rational and sophisticated sequential delivery systems for macrophage activation reagents; addressing the interference of bone graft materials and dental implants; and understanding the correlations among membrane degradation, macrophage responses, and bone regeneration.

Osteoinductive micro-nano guided bone regeneration membrane for in situ bone defect repair.

BACKGROUND: Biomaterials used in bone tissue engineering must fulfill the requirements of osteoconduction, osteoinduction, and osseointegration. However, biomaterials with good osteoconductive properties face several challenges, including inadequate vascularization, limited osteoinduction and barrier ability, as well as the potential to trigger immune and inflammatory responses. Therefore, there is an urgent need to develop guided bone regeneration membranes as a crucial component of tissue engineering strategies for repairing bone defects. METHODS: The mZIF-8/PLA membrane was prepared using electrospinning technology and simulated body fluid external mineralization method. Its ability to induce biomimetic mineralization was evaluated through TEM, EDS, XRD, FT-IR, zeta potential, and wettability techniques. The biocompatibility, osteoinduction properties, and osteo-immunomodulatory effects of the mZIF-8/PLA membrane were comprehensively evaluated by examining cell behaviors of surface-seeded BMSCs and macrophages, as well as the regulation of cellular genes and protein levels using PCR and WB. In vivo, the mZIF-8/PLA membrane's potential to promote bone regeneration and angiogenesis was assessed through Micro-CT and immunohistochemical staining. RESULTS: The mineralized deposition enhances hydrophilicity and cell compatibility of mZIF-8/PLA membrane. mZIF-8/PLA membrane promotes up-regulation of osteogenesis and angiogenesis related factors in BMSCs. Moreover, it induces the polarization of macrophages towards the M2 phenotype and modulates the local immune microenvironment. After 4-weeks of implantation, the mZIF-8/PLA membrane successfully bridges critical bone defects and almost completely repairs the defect area after 12-weeks, while significantly improving the strength and vascularization of new bone. CONCLUSIONS: The mZIF-8/PLA membrane with dual osteoconductive and immunomodulatory abilities could pave new research paths for bone tissue engineering.

Fabrication and characterization of a multi-functional GBR membrane of gelatin-chitosan for osteogenesis and angiogenesis.

Guided bone regeneration (GBR) membranes are widely used to treat bone defects. In this study, sequential electrospinning and electrospraying techniques were used to prepare a dual-layer GBR membrane composed of gelatin (Gel) and chitosan (CS) containing simvastatin (Sim)-loaded poly(lactic-co-glycolic acid) (PLGA) microspheres (Sim@PLGA/Gel-CS). As a GBR membrane, Sim@PLGA/Gel-CS could act as a barrier to prevent soft tissue from occupying regions of bone tissue. Furthermore, compared with traditional GBR membranes, Sim@PLGA/Gel-CS played an active role on stimulating osteogenesis and angiogenesis. Determination of the physical, chemical, and biological properties of Sim@PLGA/Gel-CS membranes revealed uniform sizes of the nanofibers and microspheres and appropriate morphologies. Fourier-transform infrared spectroscopy was used to characterize the interactions between Sim@PLGA/Gel-CS molecules and the increase in the number of amide groups in crosslinked membranes. The thermal stability and tensile strength of the membranes increased after N-(3-dimethylaminopropyl)-N9- ethylcarbodiimide/N-hydroxysuccinimide crosslinking. The increased fiber density of the barrier layer decreased fibroblast migration compared with that in the osteogenic layer. Osteogenic function was indicated by the increased alkaline phosphatase activity, calcium deposition, and neovascularization. In conclusion, the multifunctional effects of Sim@PLGA/Gel-CS on the barrier and bone microenvironment were achieved via its dual-layer structure and simvastatin coating. Sim@PLGA/Gel-CS has potential applications in bone tissue regeneration.

Remodeling of the Intra-Conduit Inflammatory Microenvironment to Improve Peripheral Nerve Regeneration with a Neuromechanical Matching Protein-Based Conduit.

Peripheral nerve injury (PNI) remains a challenging area in regenerative medicine. Nerve guide conduit (NGC) transplantation is a common treatment for PNI, but the prognosis of NGC treatment is unsatisfactory due to 1) neuromechanical unmatching and 2) the intra-conduit inflammatory microenvironment (IME) resulting from Schwann cell pyroptosis and inflammatory-polarized macrophages. A neuromechanically matched NGC composed of regenerated silk fibroin (RSF) loaded with poly(3,4-ethylenedioxythiophene): poly(styrene sulfonate) (P:P) and dimethyl fumarate (DMF) are designed, which exhibits a matched elastic modulus (25.1 ± 3.5 MPa) for the peripheral nerve and the highest 80% elongation at break, better than most protein-based conduits. Moreover, the NGC can gradually regulate the intra-conduit IME by releasing DMF and monitoring sciatic nerve movements via piezoresistive sensing. The combination of NGC and electrical stimulation modulates the IME to support PNI regeneration by synergistically inhibiting Schwann cell pyroptosis and reducing inflammatory factor release, shifting macrophage polarization from the inflammatory M1 phenotype to the tissue regenerative M2 phenotype and resulting in functional recovery of neurons. In a rat sciatic nerve crush model, NGC promoted remyelination and functional and structural regeneration. Generally, the DMF/RSF/P:P conduit provides a new potential therapeutic approach to promote nerve repair in future clinical treatments.

Advances in guided bone regeneration membranes: a comprehensive review of materials and techniques.

Guided tissue/bone regeneration (GTR/GBR) is a widely used technique in dentistry to facilitate the regeneration of damaged bone and tissue, which involves guiding materials that eventually degrade, allowing newly created tissue to take its place. This comprehensive review the evolution of biomaterials for guided bone regeneration that showcases a progressive shift from non-resorbable to highly biocompatible and bioactive materials, allowing for more effective and predictable bone regeneration. The evolution of biomaterials for guided bone regeneration GTR/GBR has marked a significant progression in regenerative dentistry and maxillofacial surgery. Biomaterials used in GBR have evolved over time to enhance biocompatibility, bioactivity, and efficacy in promoting bone growth and integration. This review also probes into several promising fabrication techniques like electrospinning and latest 3D printing fabrication techniques, which have shown potential in enhancing tissue and bone regeneration processes. Further, the challenges and future direction of GTR/GBR are explored and discussed.

Development of a PVA/PCL/CS-Based Nanofibrous Membrane for Guided Tissue Regeneration and Controlled Delivery of Doxycycline Hydrochloride in Management of Periodontitis: In Vivo Evaluation in Rats.

Antibiotic administration is an adjacent therapy to guided tissue regeneration (GTR) in the management of periodontitis. This is due to the major role of pathogen biofilm in aggravating periodontal defects. This study aimed to fabricate a GTR membrane for sustained delivery of doxycycline hydrochloride (DOX) while having a space-maintaining function. The membranes were prepared using a polymeric blend of polycaprolactone/polyvinyl alcohol/chitosan by the electrospinning technique. The obtained membranes were characterized in terms of physicochemical and biological properties. Nanofibers showed a mean diameter in the submicron range of < 450 nm while having uniform randomly aligned morphology. The obtained membranes showed high strength and flexibility. A prolonged in vitro release profile during 68 h was observed for manufactured formulations. The prepared membranes showed a cell viability of > 70% at different DOX concentrations. The formulations possessed antimicrobial efficacy against common pathogens responsible for periodontitis. In vivo evaluation also showed prolonged release of DOX for 14 days. The histopathological evaluation confirmed the biocompatibility of the GTR membrane. In conclusion, the developed nanofibrous DOX-loaded GTR membranes may have beneficial characteristics in favour of both sustained antibiotic delivery and periodontal regeneration by space-maintaining function without causing any irritation and tissue damage.

Healing outcomes of open versus closed flap procedures for collagen membrane coverage following immediate dental implant placements with simultaneous guided tissue regeneration.

OBJECTIVE: To compare the outcomes of open healing to complete closure for collagen membrane coverage for immediate implant placements with simultaneous guided bone regeneration (GBR) in two retrospective cohorts. METHODS: The subjects included 118 patients who received Bio-Gide® collagen membrane coverage for immediate implant placements and GBR in 20 anterior and 98 posterior teeth. For 58 patients, gingival flaps were released to achieve full coverage of collagen membrane (CC group). For 60 patients, no efforts were made to release the gingival flaps and collagen membrane was left exposed for open healing (OH group). Antibiotics and analgesics were prescribed for 7 days after surgery. The width of crestal open wounds were measured after surgery (W0), and at 1, 2 and 16 weeks (W16). Changes in bone mass were assessed by cone-beam computed tomography after implant placement and again at W16. Gingival and bone tissues over the implant cover screws were harvested and assessed for 16 patients in the OH group at W16. RESULTS: No wound dehiscence occurred in the CC group from W0 to W16. Both the vertical and horizontal bone dimension changes were not significantly different between the OH and CC group. For the OH group, soft tissue was completely healed at W16 when the initial wound widths were ≤6 mm. For those with initial wound widths ≥ 7 mm, the cover screws were exposed in 5/16 patients at W16 but did not affect the final restorations. Tissue staining showed keratinized mucosa and new bone formation above the dental implant in the OH group. CONCLUSION: Open healing achieved healing outcomes similar to those of complete closure for collagen membrane coverage following immediate implant placements. CLINICAL SIGNIFICANCE: For immediate implant placement requiring bone grafting and collagen membrane coverage, it is unnecessary to release the gingival flaps or use tissue grafts to achieve full coverage of the crestal wounds. Open healing with exposed membrane could achieve similar outcomes with less pain and swelling.

Perspectives on the Novel Multifunctional Nerve Guidance Conduits: From Specific Regenerative Procedures to Motor Function Rebuilding.

Peripheral nerve injury potentially destroys the quality of life by inducing functional movement disorders and sensory capacity loss, which results in severe disability and substantial psychological, social, and financial burdens. Autologous nerve grafting has been commonly used as treatment in the clinic; however, its rare donor availability limits its application. A series of artificial nerve guidance conduits (NGCs) with advanced architectures are also proposed to promote injured peripheral nerve regeneration, which is a complicated process from axon sprouting to targeted muscle reinnervation. Therefore, exploring the interactions between sophisticated NGC complexes and versatile cells during each process including axon sprouting, Schwann cell dedifferentiation, nerve myelination, and muscle reinnervation is necessary. This review highlights the contribution of functional NGCs and the influence of microscale biomaterial architecture on biological processes of nerve repair. Progressive NGCs with chemical molecule induction, heterogenous topographical morphology, electroactive, anisotropic assembly microstructure, and self-powered electroactive and magnetic-sensitive NGCs are also collected, and they are expected to be pioneering features in future multifunctional and effective NGCs.

Electrospun Nanofibrous Conduit Filled with a Collagen-Based Matrix (ColM) for Nerve Regeneration.

Traumatic nerve defects result in dysfunctions of sensory and motor nerves and are usually accompanied by pain. Nerve guidance conduits (NGCs) are widely applied to bridge large-gap nerve defects. However, few NGCs can truly replace autologous nerve grafts to achieve comprehensive neural regeneration and function recovery. Herein, a three-dimensional (3D) sponge-filled nanofibrous NGC (sf@NGC) resembling the structure of native peripheral nerves was developed. The conduit was fabricated by electrospinning a poly(L-lactide-co-glycolide) (PLGA) membrane, whereas the intraluminal filler was obtained by freeze-drying a collagen-based matrix (ColM) resembling the extracellular matrix. The effects of the electrospinning process and of the composition of ColM on the physicochemical performance of sf@NGC were investigated in detail. Furthermore, the biocompatibility of the PLGA sheath and ColM were evaluated. The continuous and homogeneous PLGA nanofiber membrane had high porosity and tensile strength. ColM was shown to exhibit an ECM-like architecture characterized by a multistage pore structure and a high porosity level of over 70%. The PLGA sheath and ColM were shown to possess stagewise degradability and good biocompatibility. In conclusion, sf@NGC may have a favorable potential for the treatment of nerve reconstruction.

Regeneração óssea guiada para aumento horizontal de rebordo utilizando fibrina rica em plaquetas associada a enxertos ósseos: revisão de literatura / Guided bone regenerationfor horizontal bone augmentation using platelet- rich fibrin associated with bone grafts: literature review

Objetivo: revisar a literatura sobre os diferentes tipos de derivados de plaquetas autólogas e o desempenho clínico do uso do sticky bone para aumento ósseo horizontal de rebordo. Materiais e métodos: Para realização dessa revisão foram realizadas buscas nas bases de dados PubMed, Google Scholar e Web of Science, utilizando os seguintes descritores: "platelet-rich fibrin" AND "sticky bone" OR "alveolar bone grafting" AND "sticky bone" OR "guided bone regeneration" AND "sticky bone" AND "alveolar ridge augmentation" OR "Alveolar ridge augmentation" AND "sticky bone". Foram incluídos artigos publicados em inglês, que abordavam conceitos relacionados aos agregados plaquetários e a regeneração óssea guiada para aumento ósseo horizontal de rebordo utilizando fibrina rica em plaquetas associada à enxertos ósseos (sticky bone). Resultados: Após avaliação dos estudos encontrados foram selecionados 11 artigos sobre o uso do sticky bone para aumento horizontal de rebordo. Para compor este trabalho foram selecionados também 14 estudos de revisão e artigos associados ao tema. Por ser de fácil aplicação e obtenção, muitos autores têm estudado as aplicações cirúrgicas do sticky bone e os resultados demonstram que o aumento horizontal do rebordo utilizando essa técnica pode ser realizado de forma previsível. Conclusão: apesar de haver estudos promissores sobre o uso do sticky bone, falta evidência na literatura sobre seu sucesso clínico. Assim, para compreender o potencial regenerativo desta técnica são necessários um maior número de estudos randomizados, com diferentes materiais de enxerto e protocolos padronizados de obtenção do sticky bone.(AU)

Objective: to review the literature on the different types of autologous platelet derivatives and the clinical performance of using sticky bone for horizontal bone ridge augmentation. Materials and methods: In order to conduct this review, it was conducted searches in the PubMed, Google Scholar, and Web of Science databases using the following descriptors: "platelet-rich fibrin" AND "sticky bone" OR "alveolar bone grafting" AND "sticky bone" OR "guided bone regeneration" AND "sticky bone" AND "alveolar ridge augmentation" OR "Alveolar ridge augmentation" AND "sticky bone". It included articles published in English that addressed concepts related to platelet aggregates and guided bone regeneration for horizontal bone augmentation using platelet-rich fibrin associated with bone grafts (sticky bone). Results: After evaluating the studies found, were selected 11 articles on the use of sticky bone for horizontal ridge augmentation. To compose this work, 14 review studies and articles associated with the topic were also selected. Due to its ease of application and availability, many authors have explored the surgical applications of sticky bone, and the results indicate that horizontal ridge augmentation using this technique can be predictably performed. Conclusion: while there are promising studies on the use of sticky bone, the literature lacks evidence regarding its clinical success. Therefore, to fully understand the regenerative potential of this technique, further randomized studies are needed, involving different graft materials and standardized protocols for obtaining sticky bone.(AU)

Tissue-Engineered Core-Shell Silk-Fibroin/Poly-l-Lactic Acid Nerve Guidance Conduit Containing Encapsulated Exosomes of Human Endometrial Stem Cells Promotes Peripheral Nerve Regeneration.

Nerve guide conduits (NGCs) have been shown to be less efficient than nerve autografts in peripheral nerve regeneration. To address this issue, we developed for the first time a novel tissue-engineered nerve guide conduit structure encapsulated with human endometrial stem cell (EnSC) derived exosomes, which promoted nerve regeneration in rat sciatic nerve defects. In this study, we initially indicated the long-term efficacy and safety impacts of newly designed double layered SF/PLLA nerve guide conduits. Then the regeneration effects of SF/PLLA nerve guide conduits containing exosomes derived from human EnSCs were evaluated in rat sciatic nerve defects. The human EnSC derived exosomes were isolated from the supernatant of human EnSC cultures and characterized. Subsequently, the human EnSC derived exosomes were encapsulated in constructed NGCs by fibrin gel. For in vivo studies, entire 10 mm peripheral nerve defects were generated in rat sciatic nerves and restored with NGC encapsulated with human EnSC derived exosomes (Exo-NGC group), nerve guide conduits, and autografts. The efficiency of the NGCs encapsulated with human EnSCs derived exosomes in assisting peripheral nerve regeneration was investigated and compared with other groups. The in vivo results demonstrated that encapsulated human EnSC derived exosomes in NGC (Exo-NGC) significantly benefitted nerve regeneration based on motor function, sensory reaction, and electrophysiological results. Furthermore, immunohistochemistry with histopathology results showed the formation of regenerated nerve fibers, along with blood vessels that newly were developed, as a result of the exosome functions in the Exo-NGC group. These outcomes illustrated that the newly designed core-shell SF/PLLA nerve guide conduit encapsulated with human EnSC derived exosomes enhanced the regeneration process of axons and improved the functional recovery of rat sciatic nerve defects. So, encapsulated human EnSC-derived exosomes in a core-shell SF/PLLA nerve guide conduit are a potential therapeutic cell-free treatment for peripheral nerve defects.

Biodegradable polyurethane-incorporating decellularized spinal cord matrix scaffolds enhance Schwann cell reprogramming to promote peripheral nerve repair.

Decellularized extracellular matrix (dECM) nerve guide conduits (NGCs) are a promising strategy to replace autogenous nerve grafting for the treatment of peripheral nerve system (PNS) injury. However, dECM conduits with mechanical properties that match those of peripheral nerves are yet to be well developed. Herein, we developed polyurethane-based NGCs incorporating decellularized spinal cord (BWPU-DSC NGCs) to repair peripheral nerves. BWPU-DSC NGCs have an inner three-dimensional micro-nanostructure. The mechanical properties of BWPU-DSC NGCs were similar to those of polyurethane NGCs, which were proven to promote peripheral nerve regeneration. An in vitro study indicated that BWPU-DSC NGCs could boost the proliferation and growth of cell processes in Schwann and neuron-like cells. In a rat sciatic nerve transected injury model, BWPU-DSC NGCs exhibited a dramatic increase in nerve repair, similar to that obtained by the current gold standard autograft implantation at only 6 weeks post-implantation, whereas polyurethane NGCs still displayed incomplete nerve repair. Histological analysis revealed that BWPU-DSC NGCs could induce the reprogramming of Schwann cells to promote axon regeneration and remyelination. Moreover, reprogrammed Schwann cells together with BWPU-DSC NGCs had anti-inflammatory effects and altered the activation state of macrophages to M2 phenotypes to enhance PNS regeneration. In this study, we provided a strategy to prepare polyurethane-based dECM NGCs enriched with bioactive molecules to promote PNS regeneration.

Osteo-Immunomodulatory Role of Interleukin-4-Immobilized Plasma Immersion Ion Implantation Membranes for Bone Regeneration.

Barrier membranes for guided tissue regeneration are essential for bone repair and regeneration. The implanted membranes may trigger early inflammatory responses as a foreign material, which can affect the recruitment and differentiation of bone cells during tissue regeneration. The purpose of this study was to determine whether immobilizing interleukin 4 (IL4) on plasma immersion ion implantation (PIII)-activated surfaces may alter the osteo-immunoregulatory characteristics of the membranes and produce pro-osteogenic effects. In order to immobilize IL4, polycaprolactone surfaces were modified using the PIII technology. No discernible alterations were found between the morphology before and after PIII treatment or IL4 immobilization. IL4-immobilized PIII surfaces polarized macrophages to an M2 phenotype and mitigated inflammatory cytokine production under lipopolysaccharide stimulation. Interestingly, the co-culture of macrophages (on IL4-immobilized PIII surfaces) and bone marrow-derived mesenchymal stromal cells enhanced the production of angiogenic and osteogenic factors and triggered autophagy activation. Exosomes produced by PIII + IL4-stimulated macrophages were also found to play a role in osteoblast differentiation. In conclusion, the osteo-immunoregulatory properties of bone materials can be modified by PIII-assisted IL4 immobilization, creating a favorable osteoimmune milieu for bone regeneration.

Additive manufacturing of Schwann cell-laden collagen/alginate nerve guidance conduits by freeform reversible embedding regulate neurogenesis via exosomes secretion towards peripheral nerve regeneration.

Peripheral nerve injury is a common clinical problem that could be debilitating to one's quality of life. The complex nerve guidance conduits (NGCs) with cells in order to improve nerve regeneration. Therefore, we used freeform reversible embedding of suspended hydrogels to fabricate Schwann cells (SCs)-laden collagen/alginate (Col/Alg) NGCs. First, we evaluated Col influence on the characteristics of NGCs. After which, Wharton's jelly mesenchymal stem cells (WJMSC) are seeded onto the inner channel of NGCs and evaluated neural regeneration behaviors. Results indicated the SCs-laden NGCs with 2.5 % Col found the highest proliferation and secretion of neurotrophic protein. Furthermore, co-culture of SCs promoted differentiation of WJMSC as seen from the increased neurogenic-related protein in NGCs. To determine the molecular mechanism between SCs and WJMSC, we demonstrated the neurotrophic factors secreted by SCs act on tropomyosin receptor kinase A (TrkA) receptors of WJMSC to promote nerve regeneration. In addition, our study demonstrated SCs-derived exosomes had a critical role in regulating neural differentiation of WJMSC. Taken together, this study demonstrates the fabrication of SCs-laden Col/Alg NGCs for nerve regeneration and understanding regarding the synergistic regenerative mechanisms of different cells could bring us a step closer for clinical treatment of large nerve defects.

Multifaceted polymeric nerve guidance conduits with distinctive double-layered architecture and plasma-induced inner chemistry gradient for the repair of critical-sized defects.

Despite tissue engineering advances, current nerve guidance conduits (NGCs) are still failing in repairing critical-sized defects. This study aims, therefore, at tackling large nerve gaps (2 cm) by designing NGCs possessing refined physicochemical properties enhancing the activity of Schwann cells (SCs) that support nerve regeneration over long distances. As such, a combinatorial strategy adopting novel plasma-induced surface chemistry and architectural heterogeneity was considered. A mechanically suitable copolymer (Polyactive®) was electrospun to produce nanofibrous NGCs mimicking the extracellular matrix. An innovative seamless double-layered architecture consisting of an inner wall comprised of bundles of aligned fibers with intercalated random fibers and an outer wall fully composed of random fibers was conceived to synergistically provide cell guidance cues and sufficient nutrient inflow. NGCs were subjected to argon plasma treatments using a dielectric barrier discharge (DBD) and a plasma jet (PJ). Surface chemical changes were examined by advanced X-ray photoelectron spectroscopy (XPS) micro-mappings. The DBD homogeneously increased the surface oxygen content from 17 % to 28 % on the inner wall. The PJ created a gradient chemistry throughout the inner wall with an oxygen content gradually increasing from 21 % to 30 %. In vitro studies revealed enhanced primary SC adhesion, elongation and proliferation on plasma-treated NGCs. A cell gradient was observed on the PJ-treated NGCs thus underlining the favorable oxygen gradient in promoting cell chemotaxis. A gradual change from circular to highly elongated SC morphologies mimicking the bands of Büngner was visualized along the gradient. Overall, plasma-treated NGCs are promising candidates paving the way towards critical nerve gap repair.

A Brief Review of In Vitro Models for Injury and Regeneration in the Peripheral Nervous System.

Nerve axonal injury and associated cellular mechanisms leading to peripheral nerve damage are important topics of research necessary for reducing disability and enhancing quality of life. Model systems that mimic the biological changes that occur during human nerve injury are crucial for the identification of cellular responses, screening of novel therapeutic molecules, and design of neural regeneration strategies. In addition to in vivo and mathematical models, in vitro axonal injury models provide a simple, robust, and reductionist platform to partially understand nerve injury pathogenesis and regeneration. In recent years, there have been several advances related to in vitro techniques that focus on the utilization of custom-fabricated cell culture chambers, microfluidic chamber systems, and injury techniques such as laser ablation and axonal stretching. These developments seem to reflect a gradual and natural progression towards understanding molecular and signaling events at an individual axon and neuronal-soma level. In this review, we attempt to categorize and discuss various in vitro models of injury relevant to the peripheral nervous system and highlight their strengths, weaknesses, and opportunities. Such models will help to recreate the post-injury microenvironment and aid in the development of therapeutic strategies that can accelerate nerve repair.

Regenerative Neurology and Regenerative Cardiology: Shared Hurdles and Achievements.

From the first success in cultivation of cells in vitro, it became clear that developing cell and/or tissue specific cultures would open a myriad of new opportunities for medical research. Expertise in various in vitro models has been developing over decades, so nowadays we benefit from highly specific in vitro systems imitating every organ of the human body. Moreover, obtaining sufficient number of standardized cells allows for cell transplantation approach with the goal of improving the regeneration of injured/disease affected tissue. However, different cell types bring different needs and place various types of hurdles on the path of regenerative neurology and regenerative cardiology. In this review, written by European experts gathered in Cost European action dedicated to neurology and cardiology-Bioneca, we present the experience acquired by working on two rather different organs: the brain and the heart. When taken into account that diseases of these two organs, mostly ischemic in their nature (stroke and heart infarction), bring by far the largest burden of the medical systems around Europe, it is not surprising that in vitro models of nervous and heart muscle tissue were in the focus of biomedical research in the last decades. In this review we describe and discuss hurdles which still impair further progress of regenerative neurology and cardiology and we detect those ones which are common to both fields and some, which are field-specific. With the goal to elucidate strategies which might be shared between regenerative neurology and cardiology we discuss methodological solutions which can help each of the fields to accelerate their development.

Metformin-Incorporated Gelatin/Nano-Hydroxyapatite Scaffolds Promotes Bone Regeneration in Critical Size Rat Alveolar Bone Defect Model.

In this study, we fabricated gelatin/nano-hydroxyapatite/metformin scaffold (GHMS) and compared its effectiveness in bone regeneration with extraction-only, Sinbone, and Bio-Oss Collagen® groups in a critical size rat alveolar bone defect model. GHMS was synthesized by co-precipitating calcium hydroxide and orthophosphoric acid within gelatin solution, incorporating metformin, and cross-linked by microbial transglutaminase. The morphology, characterization, and biocompatibility of scaffold were examined. The in vitro effects of GHMS on osteogenic gene and protein expressions were evaluated. In vivo bone formation was assessed in a critical size rat alveolar bone defect model with micro-computed tomography and histological examination by comparing GHMS with extraction-only, Sinbone, and Bio-Oss Collagen®. The synthesized GHMS had a highly interconnected porous structure with a mean pore size of 81.85 ± 13.8 µm. GHMS exhibited good biocompatibility; promoted ALPL, RUNX2, SP7, BGLAP, SPARC and Col1a1 gene expressions; and upregulated the synthesis of osteogenic proteins, including osteonectin, osteocalcin, and collagen type I. In critical size rat alveolar bone defects, GHMS showed superior bone regeneration compared to extraction-only, Sinbone, and Bio-Oss Collagen® groups as manifested by greater alveolar ridge preservation, while more bone formation with a lower percentage of connective tissue and residual scaffold at the defect sites grafted with GHMS in histological staining. The GHMS presented in this study may be used as a potential bone substitute to regenerate alveolar bone. The good biocompatibility, relatively fast degradation, interconnected pores allowing vascularization, and higher bioactivity properties of the components of the GHMS (gelatin, nHA, and metformin) may contribute to direct osteogenesis.

Low-intensity pulsed ultrasound enhances bone marrow-derived stem cells-based periodontal regenerative therapies.

BACKGROUND: Alveolar bone loss is one of the most common consequence for periodontitis, which is a major obstacle in periodontal regeneration. Bone marrow stromal cells (BMSCs) have shown significant promise in the treatment of various disease, which also contribute to the natural bone repair process. Low-intensity pulsed ultrasound (LIPUS) is a therapeutic ultrasound used in our previous studies to promotes alveolar bone regeneration. In addition, LIPUS was found to be a promising method to enhance mesenchymal stromal cell-based therapies. In the current study, we have investigated the effects of LIPUS combined with BMSCs therapies on BMSCs homing and its potential to promote alveolar bone regeneration. METHODS: BMSCs were isolated from rat and characterized by multilineages differentiation assay. Then these cells were labeled with luciferase and green fluorescent protein (GFP) by lentivirus in vitro. Periodontal bone defect was made on the mesial area of the maxillary first molar in rats. A total of 1 × 106 Luc-GFP labeled BMSCs were injected into rat tail vein. Bioluminescence imaging was utilized to track BMSCs in vivo. The rats were sacrificed eight weeks after surgery and the samples were harvested. Micro-computed tomography (Micro-CT) was performed to evaluate alveolar bone regeneration. Paraffin sections were made and subject to hematoxylin-eosin staining, masson staining and immunohistochemistry staining. RESULTS: BMSCs display a fibroblast-like morphology and can differentiate into adipocytes or osteoblasts under appropriate condition. The transfected BMSCs are strongly positive for GFP express. Bioluminescence imaging showed that most of BMSCs were trapped in the lung. A small portion BMSCs were homed to the alveolar bone defect area in BMSCs group, while more cells were observed in BMSCs/LIPUS group compare to other groups on day 3 and 7. Micro-CT results showed that BMSCs/LIPUS group resulted in more new bone formation than other groups. Immunohistochemical results showed higher expression of COL-I and osteopontin in BMSCs/LIPUS group compared with the other groups. CONCLUSIONS: These results suggested that LIPUS can enhance BMSCs-based periodontal alveolar bone regeneration. This study provides new insights into how LIPUS might provide therapeutic benefits by promoting BMSCs homing.