Facilitate Bone Regeneration Research Articles

The construction of a self-assembled coating with chitosan-grafted reduced graphene oxide on porous calcium polyphosphate scaffolds for bone tissue engineering

Bone regeneration in large bone defects remains one of the major challenges in orthopedic surgery. Calcium polyphosphate (CPP) scaffolds possess excellent biocompatibility and exhibits good bone ingrowth. However, the present CPP scaffolds lack enough osteoinductive activity to facilitate bone regeneration at bone defects that exceed the critical size threshold. To endow CPP scaffolds with improved osteoinductive activity for better bone regeneration, in this study, a self-assembled coating with chitosan-grafted reduced graphene oxide (CS-rGO) sheets was successfully constructed onto the surface of CPP scaffolds through strong electrostatic interaction and hydrogen bonds. Our results showed that the obtained CPP/CS-rGO composite scaffolds exhibited highly improved biomineralization and considerable antibacterial activity. More importantly, CPP/CS-rGO composite scaffolds could drive osteogenic differentiation of BMSCs and significantly up-regulate the expression of osteogenesis-related proteins in vitro. Meanwhile, the CS-rGO coating could inhibit aseptic loosening and improve interfacial osseointegration through stimulating bone marrow mesenchymal stem cells (BMSCs) to secrete more osteoprotegerin (OPG) and lesser receptor activator of nuclear factor-κB ligand (RANKL). Overall, the CS-rGO coating adjusts CPP scaffolds’ biological environment interface and endows CPP scaffolds with more bioactivity.

Mussel-inspired multifunctional surface through promoting osteogenesis and inhibiting osteoclastogenesis to facilitate bone regeneration

Osteogenesis and osteoclastogenesis are closely associated during the bone regeneration process. The development of multifunctional bone repair scaffolds with dual therapeutic actions (pro-osteogenesis and anti-osteoclastogenesis) is still a challenging task for bone tissue engineering applications. Herein, through a facile surface coating process, mussel-inspired polydopamine (PDA) is adhered to the surface of a biocompatible porous scaffold followed by the immobilization of a small-molecule activator (LYN-1604 (LYN)) and the subsequent in situ coprecipitation of hydroxyapatite (HA) nanocrystals. PDA, acting as an intermediate bridge, can provide strong LYN immobilization and biomineralization ability, while LYN targets osteoclast precursor cells to inhibit osteoclastic differentiation and functional activity, which endows LYN/HA-coated hybrid scaffolds with robust anti-osteoclastogenesis ability. Due to the synergistic effects of the LYN and HA components, the obtained three-dimensional hybrid scaffolds exhibited the dual effects of osteoclastic inhibition and osteogenic stimulation, thereby promoting bone tissue repair. Systematic characterization experiments confirmed the successful fabrication of LYN/HA-coated hybrid scaffolds, which exhibited an interconnected porous structure with nanoroughened surface topography, favorable hydrophilicity, and improved mechanical properties, as well as the sustained sequential release of LYN and Ca ions. In vitro experiments demonstrated that LYN/HA-coated hybrid scaffolds possessed satisfactory cytocompatibility, effectively promoting cell adhesion, spreading, proliferation, alkaline phosphatase activity, matrix mineralization, and osteogenesis-related gene and protein secretion, as well as stimulating angiogenic differentiation of endothelial cells. In addition to osteogenesis, the engineered scaffolds also significantly reduced osteoclastogenesis, such as tartrate-resistant acid phosphatase activity, F-actin ring staining, and osteoclastogenesis-related gene and protein secretion. More importantly, in a rat calvarial defect model, the newly developed hybrid scaffolds significantly promoted bone repair and regeneration. Microcomputed tomography, histological, and immunohistochemical analyses all revealed that the LYN/HA-coated hybrid scaffolds possessed not only reliable biosafety but also excellent osteogenesis-inducing and osteoclastogenesis-inhibiting effects, resulting in faster and higher-quality bone tissue regeneration. Taken together, this study offers a powerful and promising strategy to construct multifunctional nanocomposite scaffolds by promoting osteo/angiogenesis and suppressing osteoclastogenesis to accelerate bone regeneration.

Angiogenic and Osteogenic Properties of Fibrin in Bone Tissue Engineering

The bone scaffold has become a promising alternative in bone tissue engineering due to the limitation associated with current bone treatments. However, the selection of scaffold material that could accurately mimic the extracellular matrix of native tissue remains challenging. Owing to its biological origin properties, natural materials including fibrin are widely used as scaffold materials as compared to synthetic materials. Fibrin has been recognized as one of the appealing natural biopolymers, which possesses unique characteristic due to its natural formed nano-scaffold, which provide a temporary matrix that facilitates cellular activities of cells. Fibrin has shown remarkable effects over other biomaterials in inducing angiogenesis and osteogenesis in bone regeneration owing to its mechanical and biological properties. In this article, we highlight the significance of fibrin materials in facilitating bone regeneration. We focus on the manipulation of fibrin composition and on the recent developments of fibrin composites in enhancing osteogenesis and angiogenesis for bone healing.

The role of rare earth elements in bone tissue engineering scaffolds - A review

Rare earth elements (REEs) have gained spotlights with particular features to promote bone regeneration. Therefore, the introduction of appropriate REEs into bone tissue engineering (BTE) scaffolds is nowadays a promising strategy in order to achieve optimized bone repair performances. In this review, direct uses of REE compound nanoadditives in BTE scaffolds are first of all reviewed to illustrate the fundamental effects of REE towards bone regeneration. Then the Re-containing biodegradable porous alloys and Re-doped bioceramic-based scaffolds are discussed, respectively. REEs play functional roles in those materials with physical, chemical and biological improvements. Moreover, REEs-related biological effects including osteogenic, angiogenic and anti-bacterial properties that facilitate bone regeneration are also reviewed. This study is aiming to invoke diverse and comprehensive studies to investigate and improve the utilization of REEs in bone tissue engineering.

MiR-153-3p inhibited osteogenic differentiation of human DPSCs through CBFβ signaling.

Dental pulp stem cells (DPSCs) have multilineage differentiation potential and especially show a great foreground in bone regeneration engineering. The mechanism of osteogenic differentiation of DPSCs needs to be explored exactly. As a kind of endogenous and non-coding small RNAs, microRNAs (miRNAs) play an important role in many biological processes including osteogenic differentiation. However, the mechanism of miR-153-3p in osteogenic differentiation of DPSCs is still unknown. Core-binding factors-beta (CBFβ) is a non-DNA-binding factor that combines with the runt-related transcription factor family transcription factors to mediate their DNA-binding affinities, and plays a critical role in regulating osteogenic differentiation. In this study, we explored the mechanisms of miR-153-3p and CBFβ in DPSC osteogenesis. The expression of miR-153-3p and CBFβ was tested under the osteogenic condition, and the influence led by changing the expression of miR-153-3p or CBFβ had also been detected. A luciferase reporter assay confirmed that miR-153-3p directly targeted to CBFβ. The osteogenic markers, alkaline phosphatase (ALP), runt-related transcription factor 2 (Runx2), and bone morphogenetic protein 2 (BMP2), were tested in protein level or mRNA level. ALP and Alizarin red staining were used to detect the osteoblast activity and mineral deposition. In osteogenic condition, the expressions of CBFβ and osteogenic markers were upregulated, whereas that of miR-153-3p was downregulated. miR-153-3p negatively regulated the osteogenic differentiation, and overexpression of CBFβ could offset the negative effect of miR-153-3p. Our findings provided a novel strategy for DPSC application in treatment of bone deficiencies and facilitated bone regeneration.

A COMPARISON OF GBR MEMBRANES

Introduction: Guided bone regeneration has become common practice among surgeons because it gives them the opportunity to place implants in bone defects or absorbed alveolar ridges. This fact raises the question which membrane should one use to achieve the best results. Purpose and Methods: The aim of this study is to present the most recent data around GBR membranes. The existing types of membranes will be compared for their ability to facilitate bone regeneration, their biocompatibility, and their clinical complications. Research for this study was done by evaluating papers of the last decade from the following online databases: PubMed, Medline, and Google Scholar. Finally, selected clinical cases will be presented. Results: Membranes are divided in two main categories resorbable (collagen, synthetic) and non-resorbable (ePTFE, Ti reinforced). It is quite challenging to compare clinical trials because they use different parameters for comparison such as selection criteria of the patients, the type of defect to be treated, the way of executing the treatment, the duration, the materials used and the evaluation criteria. As far as bone regeneration is concerned there seem to be no significant differences in bone level and density of newly formed bone between different types of membranes when used in combination with bone grafts. However, non-resorbable membranes are more likely to cause tissue dehiscence, become exposed to the oral environment and subsequently become infected. Also, there are new types of membranes being studied which aim to play an active role in bone regeneration with embedded active ingredients, but more research needs to be done. Conclusions: According to the most recent protocols the combination of bone grafts and collagen membranes is selected in most cases because it has less clinical complications.

Fabrication of Naturally Derived Double-Network Hydrogels With a Sustained Aspirin Release System for Facilitating Bone Regeneration.

Continuous efforts on pursuit of effective drug delivery systems for engineering hydrogel scaffolds is considered a promising strategy for the bone-related diseases. Here, we developed a kind of acetylsalicylic acid (aspirin, ASA)–based double-network (DN) hydrogel containing the positively charged natural chitosan (CS) and methacrylated gelatin (GelMA) polymers. Combination of physical chain-entanglement, electrostatic interactions, and a chemically cross-linked methacrylated gelatin (GelMA) network led to the formation of a DN hydrogel, which had a suitable porous structure and favorable mechanical properties. After in situ encapsulation of aspirin agents, the resulting hydrogels were investigated as culturing matrices for adipose tissue–derived stromal cells (ADSCs) to evaluate their excellent biocompatibility and biological capacities on modulation of cell proliferation and differentiation. We further found that the long-term sustained ASA in the DN hydrogels could contribute to the anti-inflammation and osteoinductive properties, demonstrating a new strategy for bone tissue regeneration.

A multifunctional strontium/silver-co-substituted hydroxyapatite derived from biogenic source as antibacterial biomaterial

The bioactive properties of hydroxyapatite (HAp) facilitate bone regeneration, however, its physico-chemical and bioactive properties can be further enhanced by ionic substitutions within the crystalline lattice. In this work, substitution with Sr2+ and Ag+ ions was investigated for the improvement of osseointegration and antibacterial activity in potential treatment of traumatic bone injuries. A series of single substituted HAp with Sr2+ or Ag+ ions and Ag/Sr-co-substituted HAp with different degrees of substitution (0, 1, 2.5, and 5 mol%) were obtained by wet precipitation from cuttlefish bone. Rietveld refinement indicated successful Ca2+ substitution by increasing cell parameters due to the larger ionic radii of Sr2+ and Ag+ compared to Ca2+, which was confirmed by elemental mapping showing uniform distribution of substituent cations. Characterization of the zeta-potential of Ag/Sr-co-substituted HAp showed negatively charged populations, at potentials not lower than −15 mV. Increasing the degree of substitution resulted in decreasing zeta-potential. The higher absorption capacity of bovine serum albumin was determined on Ag/Sr-co-substituted HAp powders as compared to non-substituted HAp. All Ag-substituted HAps have shown an antibacterial activity towards Gram-positive Staphylococcus aureus and Gram-negative Escherichia coli, as determined by inhibition zone, viability analysis and scanning electron microscopy. In addition, non-cytotoxicity towards human cells was confirmed by in vitro tests with embryonic kidney 293 and mesenchymal stem cells.

A programmed surface on polyetheretherketone for sequentially dictating osteoimmunomodulation and bone regeneration to achieve ameliorative osseointegration under osteoporotic conditions

Polyetheretherketone (PEEK) is a desirable alternative to conventional biomedical metals for orthopedic implants due to the excellent mechanical properties. However, the inherent bioinertness of PEEK contributes to inferior osseointegration of PEEK implants, especially under pathological conditions of osteoporosis. Herein, a programmed surface is designed and fabricated on PEEK to dictate osteoimmunomodulation and bone regeneration sequentially. A degradable hybrid coating consisting of poly(lactide-co-glycolide) and alendronate (ALN) loaded nano-hydroxyapatite is deposited on PEEK and then interleukin-4 (IL-4) is grafted onto the outer surface of the hybrid coating with the aid of N2 plasma immersion ion implantation and subsequent immersion in IL-4 solution. Dominant release of IL-4 together with ALN and Ca2+ during the first few days synergistically mitigates the early acute inflammatory reactions and creates an osteoimmunomodulatory microenvironment that facilitates bone regeneration. Afterwards, slow and sustained delivery of ALN and Ca2+ in the following weeks boosts osteogenesis and suppresses osteoclastogenesis simultaneously, consequently ameliorating bone-implant osseointegration even under osteoporotic conditions. By taking into account the different phases in bone repair, this strategy of constructing advanced bone implants with sequential functions provides customizable and clinically viable therapy to osteoporotic patients.

Inhibition of miRNA-100 facilitates bone regeneration defects of mesenchymal stem cells in osteoporotic mice through the protein kinase B pathway

ABSTRACT Osteoporotic patients suffer from bone microstructure damage and are prone to fracture and bone defect. Due to the damage of bone healing ability, the bone repair of osteoporotic patients is usually slow. Here we aimed to explore the function and potential molecular mechanism of miR-100 in osteogenic differentiation ability of bone marrow stem cells (BMSCs). Ovariectomy was performed on mice to induce osteoporosis. BMSCs were extracted from normal and ovariectomized (OVX) mice to examine the effect of microRNA (miR)-100 on BMSC osteogenic differentiation. Hematoxylin and eosin (H&E) staining and safranin O-fast green staining assays were performed on femur tissues to reveal pathological changes. The osteogenic differentiation of BMSCs were determined by Alkaline Phosphatase and Alizarin red staining assays. The results showed that miR-100 expression was significantly upregulated in bone tissues and BMSCs from osteoporotic mice. MiR-100 knockdown partially improved osteogenic function of OVX mice-derived BMSCs. Next, mechanistic target of rapamycin kinase (MTOR) was identified as the target downstream miR-100. MiR-100 deficiency can activate the protein kinase B (AKT)/mTOR pathway. MiR-100 controlled the osteogenic function of BMSCs by the AKT/mTOR pathway. Collectively, our findings demonstrate that inhibition of miR-100 facilitates bone regeneration defects of BMSCs in osteoporotic mice through AKT pathway, indicating that miR-100 might be an effective target for the treatment of osteoporotic mandibular injury and bone defect diseases.

Independent and complementary bio-functional effects of CuO and Ga2O3 incorporated as therapeutic agents in silica- and phosphate-based bioactive glasses

The incorporation of therapeutic-capable ions into bioactive glasses (BGs), either based on silica (SBGs) or phosphate (PBGs), is currently envisaged as a proficient path for facilitating bone regeneration. In conjunction with this view, the single and complementary structural and bio-functional roles of CuO and Ga2O3 (in the 2–5 mol% range) were assessed, by deriving a series of SBG and PBG formulations starting from the parent glass systems, FastOs®BG – 38.5SiO2—36.1CaO—5.6P2O5—19.2MgO—0.6CaF2, and 50.0P2O5—35.0CaO—10.0Na2O—5.0 Fe2O3 (mol%), respectively, using the process of melt-quenching. The inter-linked physico-chemistry – biological response of BGs was assessed in search of bio-functional triggers. Further light was shed on the structural role – as network former or modifier – of Cu and Ga, immersed in SBG and PBG matrices. The preliminary biological performance was surveyed in vitro by quantification of Cu and Ga ion release under homeostatic conditions, cytocompatibility assays (in fibroblast cell cultures) and antibacterial tests (against Staphylococcus aureus). The similar (Cu) and dissimilar (Ga) structural roles in the SBG and PBG vitreous networks governed their release. Namely, Cu ions were leached in similar concentrations (ranging from 10–35 ppm and 50–110 ppm at BG doses of 5 and 50 mg/mL, respectively) for both type of BGs, while the release of Ga ions was 1–2 orders of magnitude lower in the case of SBGs (i.e., 0.2–6 ppm) compared to PBGs (i.e., 9–135 ppm). This was attributed to the network modifier role of Cu in both types of BGs, and conversely, to the network former (SBGs) and network modifier (PBGs) roles of Ga. All glasses were cytocompatible at a dose of 5 mg/mL, while at the same concentration the antimicrobial efficiency was found to be accentuated by the coupled release of Cu and Ga ions from SBG. By collective assessment, the most prominent candidate material for the further development of implant coatings and bone graft substitutes was delineated as the 38.5SiO2—34.1CaO—5.6P2O5—16.2MgO—0.6CaF2—2.0CuO—3.0Ga2O3 (mol%) SBG system, which yielded moderate Cu and Ga ion release, excellent cytocompatibility and marked antibacterial efficacy.

A polydopamine-assisted strontium-substituted apatite coating for titanium promotes osteogenesis and angiogenesis via FAK/MAPK and PI3K/AKT signaling pathways

Early osteointegration is essential for biomedical implants. Surface modifications can significantly compensate for an implant's lack of biocompatibility and osteo-differentiation. They can also be designed to promote angiogenesis in order to assist osteogenesis and ultimately facilitate bone regeneration. In this study, a polydopamine-assisted strontium-substituted apatite coating (Ti@PDA + SrHA) was fabricated on a multifunctional titanium implant to induce both angiogenic and osteogenic abilities for rapid osseointegration. Polydopamine and Sr-substituted hydroxyapatite were coated on the implant through biomineralization. The in vitro results showed that Ti@PDA + SrHA improved cell adhesion and increased the proliferation of rat bone marrow-derived mesenchymal stem cells (rBMSCs) and human umbilical vein endothelial cells (HUVECs). Ti@PDA + SrHA upregulated the expression of ALP activity and osteogenic genes in rBMSCs and elevated angiogenic genes in both rBMSCs and HUVECs. Mechanically, the FAK/MAPK signaling pathway was activated in rBMSCs, and the PI3K/AKT signaling pathway was activated in both rBMSCs and HUVECs. Consistent with these findings, Ti@PDA + SrHA accelerated new bone formation and rapid osseointegration in the femoral condyle implantation study with good stability. Overall, we fabricated a multifunctional biocompatible implant with better angiogenic and osteogenic performance compared to the non-coated implant.

Front Cover: Biomaterials and nanomedicine for bone regeneration: Progress and future prospects (EXP2 2/2021)

In article 20210011, Claudia Corbo, Wei Tao, and co-workers present an overview of common causes of bone defects, the unmet clinical needs, and the utility of biomaterials and nanotechnology to facilitate bone regeneration. The authors propose several key approaches to improve bone repair strategies that guide the future research direction of bone repair to improve treatment.

Polarized M2 macrophages induced by mechanical stretching modulate bone regeneration of the craniofacial suture for midfacial hypoplasia treatment.

The underlying mechanism of the trans-sutural distraction osteogenesis (TSDO) technique as an effective treatment that improves the symptoms of midfacial hypoplasia syndromes is not clearly understood. Increasing findings in the orthopedics field indicate that macrophages are mechanically sensitive and their phenotypes can respond to mechanical cues. However, how macrophages respond to mechanical stretching and consequently influence osteoblast differentiation of suture-derived stem cells (SuSCs) remains unclear, particularly during the TSDO process. In the present study, we established a TSDO rat model to determine whether and how macrophages were polarized in response to stretching and consequently affected bone regeneration of the suture frontal edge. Notably, after performing immunofluorescence, RNA-sequencing, and micro-computed tomography, it was demonstrated that macrophages are first recruited by various chemokines factors and polarized to the M2 phenotype upon optimal stretching. The latter in turn regulates SuSC activity and facilitates bone regeneration in sutures. Moreover, when the activated M2 macrophages were suppressed by pharmacological manipulation, new bone microarchitecture could rarely be detected under mechanical stretching and the expansion of the sutures was clear. Additionally, macrophages achieved M2 polarization in response to the optimal mechanical stretching (10%, 0.5Hz) and strongly facilitated SuSC osteogenic differentiation and human umbilical vein endothelial cell angiogenesis using an indirect co-culture system in vitro. Collectively, this study revealed the mechanical stimulation-immune response-bone regeneration axis and clarified at least in part how sutures achieve bone regeneration in response to mechanical force.

Enhancement of Bone Regeneration Through the Converse Piezoelectric Effect, A Novel Approach for Applying Mechanical Stimulation.

Serious bone injuries have devastating effects on the lives of patients including limiting working ability and high cost. Orthopedic implants can aid in healing injuries to an extent that exceeds the natural regenerative capabilities of bone to repair fractures or large bone defects. Autografts and allografts are the standard implants used, but disadvantages such as donor site complications, a limited quantity of transplantable bone, and high costs have led to an increased demand for synthetic bone graft substitutes. However, replicating the complex physiological properties of biological bone, much less recapitulating its complex tissue functions, is challenging. Extensive efforts to design biocompatible implants that mimic the natural healing processes in bone have led to the investigation of piezoelectric smart materials because the bone has natural piezoelectric properties. Piezoelectric materials facilitate bone regeneration either by accumulating electric charge in response to mechanical stress, which mimics bioelectric signals through the direct piezoelectric effect or by providing mechanical stimulation in response to electrical stimulation through the converse piezoelectric effect. Although both effects are beneficial, the converse piezoelectric effect can address bone atrophy from stress shielding and immobility by improving the mechanical response of a healing defect. Mechanical stimulation has a positive impact on bone regeneration by activating cellular pathways that increase bone formation and decrease bone resorption. This review will highlight the potential of the converse piezoelectric effect to enhance bone regeneration by discussing the activation of beneficial cellular pathways, the properties of piezoelectric biomaterials, and the potential for the more effective administration of the converse piezoelectric effect using wireless control.

Current Biomaterial-Based Bone Tissue Engineering and Translational Medicine.

Bone defects cause significant socio-economic costs worldwide, while the clinical “gold standard” of bone repair, the autologous bone graft, has limitations including limited graft supply, secondary injury, chronic pain and infection. Therefore, to reduce surgical complexity and speed up bone healing, innovative therapies are needed. Bone tissue engineering (BTE), a new cross-disciplinary science arisen in the 21st century, creates artificial environments specially constructed to facilitate bone regeneration and growth. By combining stem cells, scaffolds and growth factors, BTE fabricates biological substitutes to restore the functions of injured bone. Although BTE has made many valuable achievements, there remain some unsolved challenges. In this review, the latest research and application of stem cells, scaffolds, and growth factors in BTE are summarized with the aim of providing references for the clinical application of BTE.

Effectiveness of treating segmental bone defects with a synergistic co-delivery approach with platelet-rich fibrin and tricalcium phosphate

Several studies have applied tricalcium phosphate (TCP) or autografts in bone tissue engineering to enhance the clinical regeneration of bone. Unfortunately, there are several drawbacks related to the use of autografts, including a risk of infection, blood loss, limited quantities, and donor-site morbidities. Platelet-rich fibrin (PRF) is a natural extracellular matrix (ECM) biomaterial that possesses bioactive factors, which can generally be used in regenerative medicine. The goal of the present investigation was to develop osteoconductive TCP incorporated with bioactive PRF for bio-synergistic bone regeneration and examine the potential biological mechanisms and applications. Our in vitro results showed that PRF plus TCP had excellent biosafety and was favorable for initiating osteoblast cell attachment, slow release of bioactive factors, cell proliferation, cell migration, and ECM formation that potentially impacted bone repair. In a rabbit femoral segmental bone defect model, regeneration of bone was considerably augmented in defects locally implanted by PRF plus TCP according to radiographic and histologic examinations. Notably, the outcomes of this investigation suggest that the combination of PRF and TCP possesses novel synergistic and bio-inspired functions that facilitate bone regeneration.

Programmed surface on poly(aryl-ether-ether-ketone) initiating immune mediation and fulfilling bone regeneration sequentially

The immune responses are involved in every stage after implantation but the reported immune-regulated materials only work at the beginning without fully considering the different phases of bone healing. Here, poly(aryl-ether-ether-ketone) (PEEK) is coated with a programmed surface, which rapidly releases interleukin-10 (IL-10) in the first week and slowly delivers dexamethasone (DEX) up to 4 weeks. Owing to the synergistic effects of IL-10 and DEX, an aptly weak inflammation is triggered within the first week, followed by significant M2 polarization of macrophages and upregulation of the autophagy-related factors. The suitable immunomodulatory activities pave the way for osteogenesis and the steady release of DEX facilitates bone regeneration thereafter. The sequential immune-mediated process is also validated by an 8-week implementation on a rat model. This is the first attempt to construct implants by taking advantage of both immune-mediated modulation and sequential regulation spanning all bone regeneration phases, which provides insights into the fabrication of advanced biomaterials for tissue engineering and immunological therapeutics.

The Crosstalk between Mesenchymal Stem Cells and Macrophages in Bone Regeneration: A Systematic Review.

Bone regeneration is a complex and well-coordinated process that involves crosstalk between immune cells and resident cells in the injury site. Transplantation of mesenchymal stem cells (MSCs) is a promising strategy to enhance bone regeneration. Growing evidence suggests that macrophages have a significant impact on osteogenesis during bone regeneration. However, the precise mechanisms by which macrophage subtypes influence bone regeneration and how MSCs communicate with macrophages have not yet been fully elucidated. In this systematic literature review, we gathered evidence regarding the crosstalk between MSCs and macrophages during bone regeneration. According to the PRISMA protocol, we extracted literature from PubMed and Embase databases by using “mesenchymal stem cells” and “macrophages” and “bone regeneration” as keywords. Thirty-three studies were selected for this review. MSCs isolated from both bone marrow and adipose tissue and both primary macrophages and macrophage cell lines were used in the selected studies. In conclusion, anti-inflammatory macrophages (M2) have significantly more potential to strengthen bone regeneration compared with naïve (M0) and classically activated macrophages (M1). Transplantation of MSCs induced M1-to-M2 transition and transformed the skeletal microenvironment to facilitate bone regeneration in bone fracture and bone defect models. This review highlights the complexity between MSCs and macrophages, providing more insight into the polarized macrophage behavior in this evolving field of osteoimmunology. The results may serve as a useful reference for definite success in MSC-based therapy based on the critical interaction with macrophages.

Surface engineering of titania nanotubes incorporated with double-layered extracellular vesicles to modulate inflammation and osteogenesis.

Titania nanotubes (TNT) generated on titanium implant are emerged as important modification technique to facilitate bone regeneration. Mesenchymal stem cells (MSCs)-derived exosomes are membrane bound extracellular vesicles (EVs), which play an important role in tissue regeneration. The objective of this study was to generate an EVs hybrid TNT aiming at regulating inflammation, MSCs recruitment and osteogenesis. We isolated EVs from MSCs (MSCs EVs) and 3-day osteogenically differentiated MSCs (3d EVs). MSC EVs and 3d EVs exhibited round morphology under TEM, which also showed robust internalization by human bone marrow derived MSCs (hBMSCs). Next, we fabricated 3d EVs/MSC EVs hybrid TNT. When inflammatory macrophages were co-cultured with EVs hybrid TNT, the gene and protein expression of inflammatory cytokine were significantly reduced. Macrophage morphology was also examined by confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM). Further migratory ability study using hBMSCs indicated significant enhancement of MSCs migration in EVs hybrid TNT. In addition, we further demonstrated significant increase of osteogenic differentiation of hBMSCs in EVs hybrid TNT. This study suggests that EVs hybrid TNT may serve as a viable therapeutic approach to enhance osteogenesis and bone regeneration.