Proliferation Of Marrow Stromal Cells Research Articles

Digital light processing of customized elastic scaffolds by efficient thiol-yne crosslinking

Digital light processing (DLP) based 3D printing technology has the advantages of fast printing rate and high spatial resolution in the manufacture of tissue engineering scaffolds. However, the limited availability of printable materials for DLP technology, especially the lack of biodegradable elastomeric materials hinders the development of customized soft tissue scaffolds. Herein, liquid poly(4-methyl-ε-caprolactone) dialkynylate (PMCLDY) was prepared as a DLP printable ink based on efficient thiol-yne photocrosslinking. With the aid of photorheology and in situ FT-IR and tensile tests, an ink formulation for DLP printing was optimized as the P50 ink (PMCLDY to diluent PEGDY mass ratio of 1), which possessed fast cure rates (<15 s), low viscosity (<2 Pa·s), moderate mechanical properties (failure strain > 40 %) and maximum toughness (0.153 MJ·m−3). A commercial DLP printer equipped with the P50 ink realized 3D scaffolds models with microstructure and macroscopic shape precisely. It was demonstrated that the 3D printed scaffold with biodegradability could withstand dynamic compression cycles. Furthermore, the cytocompatibility experiments showed that the scaffold could effectively support the growth and proliferation of rat bone marrow stromal cells (BMSCs). These results demonstrate that the P50 ink is a promising material for manufacturing elastic scaffolds by DLP printing for tissue engineering applications.

Graphene Oxide Functionalized Gelatin Methacryloyl Microgel for Enhanced Biomimetic Mineralization and in situ Bone Repair.

The formation of bone-like apatite (Ap) on natural polymers through biomimetic mineralization using simulated body fluid (SBF) can improve osteoconductivity and biocompatibility, while lowering immunological rejection. Nonetheless, the coating efficiency of the bone-like Ap layer on natural polymers requires improvement. Carbonyls (-COOH) and hydroxyls (-OH) are abundant in graphene oxide (GO), which may offer more active sites for biomimetic mineralization and promote the proliferation of rat bone marrow stromal cells (BMSCs). In this study, gelatin methacryloyl (GelMA) microgels were infused with GO (0, 0.5, 1, and 2 mg/mL) and embedded into microgels in SBF for 1, 7, and 14 days. Systematic in vitro and in vivo experiments were performed to evaluate the structure of the microgel and its effect on cell proliferation and ability to repair bone defects in rats. The resulting GO-GelMA-Ap microgels displayed a porous, interconnected structure with uniformly coated surfaces in bone-like Ap, and the Ca/P ratio of the 1 mg/mL GO-GelMA-Ap group was comparable to that of natural bone tissue. Moreover, the 1 mg/mL GO-GelMA-Ap group exhibited a greater Ap abundance, enhanced proliferation of BMSCs in vitro and increased bone formation in vivo compared to the GelMA-Ap group. Overall, this study offers a novel method for incorporating GO into microgels for bone tissue engineering to promote biomimetic mineralization.

Injectable, pro-osteogenic and antioxidant composite microspheres composed of cerium-containing mesoporous bioactive glass and chitosan for bone regeneration applications

Repair of bone defects caused by excess reactive oxygen species (ROS) remains a clinical challenge. It is essential to inhibit the overaccumulation of ROS to accelerate the bone healing process. Herein, we developed an injectable composite phenylboronic acid-modified chitosan microspheres (CSP@BG) containing cerium-doped mesoporous bioactive glass nanoparticles (Ce-MBGNs) using a W/O emulsification method and investigated the structure, antioxidant, and osteogenic activity of microspheres. We systematically evaluated the microscopic morphology and chemical structure of the microspheres using FT-IR, XRD, SEM and TGA. When the content of incorporated Ce-MBGNs ranged from 0.83 to 14.37 wt%, the microspheres retained their morphology and their particle size varied from ∼10 to ∼23 μm. In vitro tests indicated that the CSP@BG microspheres exhibited desirable antioxidant activity, scavenged a variety of ROS such as H2O2 and ·OH, and demonstrated significant DPPH radical scavenging efficiency with up to 72.8% DPPH was scavenged. In addition, the results of CCK8 assay, ALP and ARS staining, and RT-PCR assay demonstrated that the CSP@BG microspheres could significantly promote the proliferation and osteogenic differentiation of human bone marrow stromal cells by tuning the amount of Ce-MBGNs added. In conclusion, this study systematically investigated the effects of Ce-MBGNs content on the morphological characteristic, antioxidant properties, and osteogenic activity of CSP@BG microspheres. Our results demonstrate the potential of pro-osteogenic and antioxidant composite microspheres composed of Ce-MBGNs and chitosan for bone regeneration applications.

SOX9 Inhibits the Progression of Osteonecrosis of the Femoral Head via the Activation of the Wnt/Beta-Catenin Pathway

Background: In this study, we aimed to explore whether the SRY-box transcription factor 9 (SOX9) can play protective roles against the occurrence and development of osteonecrosis of the femoral head (ONFH) by regulating the proliferation, apoptosis, and osteogenic differentiation of human bone marrow stromal cells (hBMSCs) via the Wnt/β-catenin pathway. Methods: We used 1600 mg of glucocorticoid (GC) to induce hBMSCs to establish an ONFH cell model and performed various experiments. Reverse transcription-quantitative polymerase chain reaction and western blotting assays were used to determine the expression levels of SOX9 and osteoblast markers, such as the RUNX family transcription factor 2 (RUNX2), alkaline phosphatase (ALP), osterix, Wnt3a, and β-catenin. An ALP detection kit was used to measure the ALP activity. Flow cytometry and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays were performed to determine the cell viability. Results: GC treatment decreased the expression levels of RUNX2, ALP, and osterix, suppressed ALP activity, and inhibited SOX9 expression. SOX9 overexpression promoted GC-induced cell proliferation and decreased cell apoptosis. Additionally, hBMSCs were transfected with SOX9-small interfering RNA in GC treatment, and SOX9 knockdown was found to suppress the osteogenic differentiation of cells and decrease their viability. Conclusion: Our results revealed that SOX9 is related to the Wnt/β-catenin pathway in ONFH. Moreover, SOX9 participated in ONFH development by activating the Wnt/β-catenin pathway.

A 2-Stage Root Analog Implant with Compact Structure, Uniform Roughness, and High Accuracy

Immediate implant placement has the advantages of shortening the operation time, reducing the treatment cycle and cost. At present, this technology has been used widely, but the indications of immediate implantation are still limited. Here, a novel type of root analog implant (RAI) was manufactured by selective laser melting technology to address the limitation. Under optimized condition, RAIs were printed with the internal density of 99.73% and the uniform surface roughness of 11 μm (Sa). Besides, the deviation between RAI specimen and design models is controlled within 0.15 mm after optimizing scanning parameters. The substrate printed could promote human bone marrow stromal cell proliferation, spreading, and osteogenic differentiation. The bone–implant contact (BIC, 75% ± 7%) and bone volume/total volume (BV/TV, 74% ± 7%) of RAIs were significantly higher than that of conventional implants (BIC, 66% ± 5%; BV/TV, 62% ± 5%) in in vivo experiments. Further, customized abutments were designed for the RAIs, improving the masticatory ability of the beagle dogs after crown restoration. This study aims to design a personalized 2-stage RAI with compact structure and uniform roughness, in order to achieve better fracture resistance, initial osseointegration efficiency, and dispersed stress in immediate implantation. It provides a certain guiding value for standardizing the manufacture and clinical application of RAI in immediate implantation.

Evaluation of mechanical and biological properties of akermanite/poly-ether-ether- ketone composite fabricated by high- temperature laser powder bed fusion.

High-temperature laser bed powder fusion (HT-LPBF) technology is an ideal method for processing poly-ether-ether-ketone (PEEK) implants with personalized bionic structures, but the biological inertia of PEEK limits its medical applications. In this study, we evaluated the mechanical and biological properties of a novel akermanite (AKM)/PEEK composite for HT-LPBF. The results showed that tiny AKM particles are evenly attached to the surface of the PEEK particle. The delayed peak crystallization temperature and stable sintering window ensure the processing feasibility of the AKM/PEEK composites. The tensile strength and Young's modulus are in the range of 30.83-98.73 MPa and 2.27-3.71 GPa, respectively, which can match the properties of cancellous bones and meet their implanting requirement. The CCK-8 experiments demonstrated the biocompatibility of the composites and the good proliferation of bone marrow stromal cells. The dense hydroxyapatite network layer and petal-like hydroxyapatite demonstrates biological activity, indicating that the composite has a good potential in the orthopedics fields.

Chordin-Like 1 Regulates Bone Marrow Stem Cell Proliferation and Excretion of Exosome and Myocardial Damage Under High Sugar

This study assesses the role of CHRDL1 (Chordin-like 1) i in the bone marrow stromal cells (BMSC) proliferation and excretion of exosome and myocardial damage under high sugar. BMSC from rats with high CHRDL1 expression was established. The exosome in the supernatant of BMSC with high CHRDL1 level was collected. H9C2 cells were assigned into control set, high sugar set and exo-CHRDL1-BMSC set followed by analysis of CHRDL1 level, BMSC proliferation and apoptosis, expression of Beclin-1, Atg5, Bcl-2 and Bax, and ROS and SOD activity. Cell proliferation was prompted and apoptotic activity was reduced in exo-CHRDL1-BMSC set with reduced ROS activity and increased SOD activity as well as upregulated Bcl-2 and downregulated Bax. In addition, exo-CHRDL1-BMSC set presented increased CHRDL1 secretion and upregulated Beclin-1 and Atg5 expression. In conclusion, proliferation of BMSC under high sugar is prompted and apoptosis is reduced by CHRDL1 through regulating the autophagy.

Chemically Crosslinked Amphiphilic Degradable Shape Memory Polymer Nanocomposites with Readily Tuned Physical, Mechanical, and Biological Properties.

Facile surgical delivery and stable fixation of synthetic scaffolds play roles just as critically as degradability and bioactivity in ensuring successful scaffold-guided tissue regeneration. Properly engineered shape memory polymers (SMPs) may meet these challenges. Polyhedral oligomeric silsesquioxanes (POSSs) can be covalently integrated with urethane-crosslinked polylactide (PLA) to give high-strength, degradable SMPs around physiological temperatures. To explore their potential for guided bone regeneration, here we tune their hydrophilicity, degradability, cytocompatibility, and osteoconductivity/osteoinductivity by crosslinking star-branched POSS-PLA with hydrophilic polyethylene glycol diisocyanates of different lengths and up to 60 wt % hydroxyapatite (HA). The composites exhibit high compliance, toughness, up to gigapascal storage moduli, and excellent shape recovery (>95%) at safe triggering temperatures. Water swelling ratios and hydrolytic degradation rates positively correlated with the hydrophilic crosslinker lengths, while the negative impact of degradation on the proliferation and osteogenesis of bone marrow stromal cells was mitigated with HA incorporation. Macroporous composites tailored for a rat femoral segmental defect were fabricated, and their ability to stably retain and sustainedly release recombinant osteogenic bone morphogenetic protein-2 and support cell attachment and osteogenesis was demonstrated. These properties combined make these amphiphilic osteoconductive degradable SMPs promising candidates as next-generation synthetic bone grafts.

Improving the osseointegration and soft tissue sealing of zirconia ceramics by the incorporation of akermanite via sol infiltration for dental implants.

Zirconia ceramics are promising dental implant materials due to their high-grade biocompatibility, high mechanical strength, and distinctive aesthetic appearance. Nevertheless, zirconia ceramics are bio-inert with a lack of osseointegration and soft tissue sealing, which limits dental implant applications. As such, the fabrication of zirconia ceramics with high mechanical strength, excellent osseointegration and soft tissue sealing performance remains a great challenge in the dental restoration field. In this article, a novel zirconia ceramic with akermanite (AKT) modification by the negative pressure infiltration method is presented. The effects of AKT sol infiltration at different times on the morphology, phase composition, mechanical properties, bioactivity, osseointegration and soft tissue sealing of the modified zirconia ceramics have been systematically investigated. The modified zirconia ceramics feature excellent mechanical properties and significantly improved surface roughness, hydrophilia, and apatite mineralization ability as compared with unmodified zirconia ceramics. Furthermore, cell-culture experiment results indicated that the surface modification of zirconia ceramics could promote adhesion, spreading, migration, proliferation and osteogenic differentiation of mouse bone marrow stromal stem cells (mBMSCs), as well as the early adhesion, spreading, proliferation and fibroblast differentiation of human gingival fibroblasts (HGFs) in vitro. The prepared bioactive zirconia distinctively enhanced the alkaline phosphate (ALP) activity, osteogenesis-related gene expression of mBMSCs and fibroblast-related-gene expression of HGFs. The in vivo evaluation confirmed that 15-TZP ceramics could promote bone-implant osseointegration to the greatest extent as compared with pure zirconia ceramics. To conclude, our research has shown that AKT-modified zirconia ceramics can achieve bone integration and soft tissue sealing, indicating that they have a lot of potential for application as a novel dental implant material in the clinical setting.

Adhesion and proliferation of bone marrow stromal cells on acellular spinal cord scaffolds

Objectives This study aimed to produce an acellular spinal cord scaffold-bone marrow stromal cell (ASCS-BMSC) complex in which the growth of BMSCs transplanted into the spinal cord of rats could be simulated in vitro, facilitating the observation and evaluation of the growth of BMSCs on the ASCS for the first time. Methods Freeze-thaw, chemical extraction and mechanical shaking approaches were used to remove the cellular components and prepare a rat ASCS containing only the extracellular matrix (ECM) structure from the rat spinal cord. BMSCs were embedded into ASCSs and freeze-dried agarose scaffolds (FASs), and cell migration and proliferation were observed via fluorescence microscopy and the MTT assay. Results Compared with the normal rat spinal cord, the ASCS had no cell structure and retained ECM components such as type IV collagen, fibronectin and laminin, showing a three-dimensional network structure with good voids. The growth and proliferation of BMSCs on the ASCS was good, as shown by the MTT assay. Scanning electron microscopy showed that BMSCs covered 65% of the ASCS surface, and the mitochondria of BMSCs were developed and adhered to collagen fibres, as demonstrated by transmission electron microscopy. HE staining showed that BMSCs could grow inside the ASCS, and immunohistochemical staining showed that BMSCs still expressed CD44 and CD90 on the ASCS and had stem cell characteristics. Conclusions The results of the experiment indicate that the ASCS has the ability to improve cell adhesion and proliferation. Thus, the ASCS-BMSC combination may be used to treat spinal cord injury.

MiR-145 inhibits the differentiation and proliferation of bone marrow stromal mesenchymal stem cells by GABARAPL1 in steroid-induced femoral head necrosis

Steroid-induced osteonecrosis of femoral head (SANFH) involves impaired differentiation of bone marrow mesenchymal stem cells (BMSC), the mechanism of which is regulated by multiple microRNAs. Studies have shown that miR-145 is a key regulatory molecule of BMSC cells, but its mechanism in steroid-induced femur head necrosis remains unclear. The present study mainly explored the specific mechanism of miR-145 involved in SANFH. In this study dexamethasone, a typical glucocorticoid, was used to induce osteogenic differentiation of BMSC cells. Western blot, qPCR, CCK8 and flow cytometry were used to investigate the effects of miR-145 on the proliferation and differentiation of BMSC. The relationship between miR-145 and GABA Type A Receptor Associated Protein Like 1(GABARAPL1) was identified using dual luciferase reports and the effects of the two molecules on BMSC were investigated in vitro. The results showed that miR-145 was up-regulated in SANFH patients, while GABARAPL1 was down-regulated. Inhibition of miR-145 can improve apoptosis and promote proliferation and activation of BMSC. GABARAPL1 is a downstream target gene of miR-145 and is negatively regulated by miR-145. In conclusion, miR-145 regulates the proliferation and differentiation of glucocorticoid-induced BMSC cells through GABARAPL1 and pharmacologically inhibit targeting miR-145 may provide new aspect for the treatment of SANFH.

Fabrication of BMP-2-peptide-Deferoxamine- and QK-peptide-functionalized nanoscaffolds and their application for bone defect treatment.

The microenvironment in the healing process of large bone defects requires suitable conditions to promote osteogenesis and angiogenesis. Coaxial electrospinning is a mature method in bone tissue engineering (BTE) and allows functional modification. Appropriate modification methods can be used to improve the bioactivity of scaffolds for BTE. In this study, coaxial electrospinning with QK peptide (a Vascular endothelial growth factor mimetic peptide) and BMP-2 peptide-DFO (BD) was performed to produce double-modified PQBD scaffolds with vascularizing and osteogenic features. The morphology of coaxially electrospun scaffolds was verified by scanning electron microscopy (SEM) and transmission electron microscopy. Laser scanning confocal microscopy and Fourier transform infrared spectroscopy confirmed that BD covalently bound to the surface of the P and PQ scaffolds. In vitro, the PQBD scaffold promoted the adhesion and proliferation of bone marrow stromal cells (BMSCs). Both QK peptide and BD showed sustainable release and preservation of biological activity, enhancing the osteogenic differentiation of BMSCs and the migration of human umbilical vein endothelial cells and promoting angiogenesis. The combined ability of these factors to promote osteogenesis and angiogenesis is superior to that of each alone. In vivo, the PQBD scaffold was implanted into the bone defect, and after 8weeks, the defect area was almost completely covered by new bone tissue. Histology showed more mature bone tissue and more blood vessels. PQBD scaffolds promote both angiogenesis and osteogenesis, offering a promising approach to enhance bone regeneration in the treatment of large bone defects.

Apoptotic Body-Rich Media from Tenocytes Enhance Proliferation and Migration of Tenocytes and Bone Marrow Stromal Cells.

The intrinsic healing following tendon injury is ideal, in which tendon progenitor cells proliferate and migrate to the injury site to directly bridge or regenerate tendon tissue. However, the mechanism determining why and how those cells are attracted to the injury site for tendon healing is not understood. Since the tenocytes near the injury site go through apoptosis or necrosis following injury, we hypothesized that secretions from injured tenocytes might have biological effects on cell proliferation and migration to enhance tendon healing. Tenocyte apoptosis was induced by 24 h cell starvation. Apoptotic body-rich media (T-ABRM) and apoptotic body-depleted media (T-ABDM) were collected from culture media after centrifuging. Tenocytes and bone marrow-derived stem cells (BMDSCs) were isolated and cultured with the following four media: (1) T-ABRM, (2) T-ABDM, (3) GDF-5, or (4) basal medium with 2% fetal calf serum (FCS). The cell activities and functions were evaluated. Both T-ABRM and T-ABDM treatments significantly stimulated the cell proliferation, migration, and extracellular matrix synthesis for both tenocytes and BMDSCs compared to the control groups (GDF-5 and basal medium). However, cell proliferation, migration, and extracellular matrix production of T-ABRM-treated cells were significantly higher than the T-ABDM, which indicates the apoptotic bodies are critical for cell activities. Our study revealed the possible mechanism of the intrinsic healing of the tendon in which apoptotic bodies, in the process of apoptosis, following tendon injury promote tenocyte and stromal cell proliferation, migration, and production. Future studies should analyze the components of the apoptotic bodies that play this role, and, thus, the targeting of therapeutics can be developed.

Adhesive Gelatin-Catechol Complex Reinforced Poly(Acrylic Acid) Hydrogel with Enhanced Toughness and Cell Affinity for Cartilage Regeneration.

The repair of cartilage damage caused by trauma, wear, or degenerative deformation remains a major challenge in modern medicine. Therefore, it is essential to develop a mechanically compatible and bioactive scaffold for cartilage tissue regeneration. In this study, a mussel-inspired, tough, adhesive polydopamine/gelatin-poly(acrylic acid) (PDA/Gel-PAA) composite hydrogel was developed for cartilage regeneration. The hydrogel achieved a high compressive strength of up to 0.67 MPa and a toughness of 420 J/m2 because of the unique chemical-physical cross-linking structure by introducing the PDA/Gel complex into the PAA network. PAA chains with rich carboxyl groups mimic the negatively charged glycosaminoglycans (GAGs) in the natural cartilage extracellular matrix (ECM), leading to strong water retention in the hydrogel. The incorporation of the PDA/Gel complex with catechol groups on PDA and arginine-glycine-aspartic acid (RGD) sequences on gelatin chains provided abundant adhesive motifs to improve the cell affinity and tissue adhesiveness of PAA, thereby facilitating the adhesion and proliferation of bone marrow stromal cells (BMSCs). In addition, transforming growth factor-β3 (TGFβ3) was stably immobilized and released from the PDA/Gel-PAA hydrogel. Thus, adhesive hydrogels can provide a suitable microenvironment to promote cell migration in the defect area and induce chronogenesis for cartilage regeneration.

Rhodiola rosea polysaccharides promote the proliferation of bone marrow haematopoietic progenitor cells and stromal cells in mice with aplastic anaemia

Context The effects of Rhodiola rosea L. (Crassulaceae) polysaccharides (RRPs) on haematopoiesis are poorly understood. Objective To determine the effects of RRPs on haematopoiesis in mice with aplastic anaemia. Materials and methods Aplastic anaemia was induced in Kunming mice by 60Coγ (2.0 Gy) irradiation and cyclophosphamide administration (50 mg/kg/day for 3 consecutive days; intraperitoneal injection). The in vivo effects of RRPs (10, 20, and 40 mg/kg; intraperitoneal injection) on haematopoiesis were analyzed using peripheral blood tests, histopathological examination of haematopoietic tissues, culture of haematopoietic progenitors and bone marrow stromal cells (BMSCs), and Western blotting of Fas and Fas ligand (FasL). The in vitro effects of RRPs on bone-marrow haematopoietic progenitors and BMSCs were also evaluated. Results Compared to anaemic controls, high-dose RRPs (40 mg/kg) significantly increased red blood cells (8.21 ± 0.57835 versus 6.13 ± 1.34623 × 1012/L), white blood cells (5.11 ± 1.6141 versus l.54 ± 1.1539 × 109/L), and BMSCs (10.33 ± 1.5542 versus 5.87 ± 3.1567 × 1012/L) in mice with aplastic anaemia (all p < 0.01). High-dose RRPs significantly increased the formation of colony-forming unit-granulocyte macrophage (CFU-GM), burst-forming unit-erythroid (BFU-E), and colony-forming unit-erythroid (CFU-E; p < 0.01). Fas and FasL protein expression in BMSCs decreased after RRPs administration. Especially at the high dose, RRPs (150 μg/mL) significantly promoted in vitro CFUs-E, BFUs-E, and CFUs-GM formation. RRPs (150–300 μg/mL) also promoted BMSC proliferation. Discussion and conclusions RRPs helped to promote haematopoietic recovery in mice with aplastic anaemia, facilitating haematopoietic tissue recovery. This study indicated some mechanisms of the haematopoietic regulatory effects of RRPs. Our findings provide a laboratory basis for clinical research on RRPs.

Degradation-Kinetics-Controllable and Tissue-Regeneration-Matchable Photocross-linked Alginate Hydrogels for Bone Repair.

Photocross-linked alginate hydrogels, due to their biodegradability, biocompatibility, strong control for gelling kinetics in space and time, and admirable adaptability for in situ polymerization with a minimally invasive approach in surgical procedures, have created great expectations in bone regeneration. However, hydrogels with suitable degradation kinetics that can match the tissue regeneration process have not been designed, which limits their further application in bone tissue engineering. Herein, we finely developed an oxidation strategy for alginate to obtain hydrogels with more suitable degradation rates and comprehensively explored their physical and biological performances in vitro and in vivo to further advance the clinical application for the hydrogels in bone repair. The physical properties of the gels can be tuned via tailoring the degree of alginate oxidation. In particular, in vivo degradation studies showed that the degradation rates of the gels were significantly increased by oxidizing alginate. The activity, proliferation, initial adhesion, and osteogenic differentiation of rat and rabbit bone marrow stromal cells (BMSCs) cultured with/in the hydrogels were explored, and the results demonstrated that the gels possessed excellent biocompatibility and that the encapsulated BMSCs were capable of osteogenic differentiation. Furthermore, in vivo implantation of rabbit BMSC-loaded gels into tibial plateau defects of rabbits demonstrated the feasibility of hydrogels with appropriate degradation rates for bone repair. This study indicated that hydrogels with increasingly controllable and matchable degradation kinetics and satisfactory bioproperties demonstrate great clinical potential in bone tissue engineering and regenerative medicine and could also provide references for drug/growth-factor delivery therapeutic strategies for diseases requiring specific drug/growth-factor durations of action.

Preparation of BMP-2/PDA-BCP Bioceramic Scaffold by DLP 3D Printing and its Ability for Inducing Continuous Bone Formation.

Digital light processing (DLP)-based 3D printing is suitable to fabricate bone scaffolds with small size and high precision. However, the published literature mainly deals with the fabrication procedure and parameters of DLP printed bioceramic scaffold, but lacks the subsequent systematic biological evaluations for bone regeneration application. In this work, a biphasic calcium phosphate (BCP) macroporous scaffold was constructed by DLP-based 3D printing technique. Furthermore, bone morphogenetic protein-2 (BMP-2) was facilely incorporated into this scaffold through a facile polydopamine (PDA) modification process. The resultant scaffold presents an interconnected porous structure with pore size of ∼570 μm, compressive strength (∼3.6 MPa), and the self-assembly Ca-P/PDA nanocoating exhibited excellent sustained-release property for BMP-2. Notably, this BMP-2/PDA-BCP scaffold presents favorable effects on the adhesion, proliferation, osteogenic differentiation, and mineralization of bone marrow stromal cells (BMSCs). Furthermore, in vivo experiments conducted on rats demonstrated that the scaffolds could induce cell layer aggregation adjacent to the scaffolds and continuous new bone generation within the scaffold. Collectively, this work demonstrated that the BMP-2/PDA-BCP scaffold is of immense potential to treat small craniofacial bone defects in demand of high accuracy.

An ECM-Mimetic Hydrogel to Promote the Therapeutic Efficacy of Osteoblast-Derived Extracellular Vesicles for Bone Regeneration.

The use of extracellular vesicles (EVs) is emerging as a promising acellular approach for bone regeneration, overcoming translational hurdles associated with cell-based therapies. Despite their potential, EVs short half-life following systemic administration hinders their therapeutic efficacy. EVs have been reported to bind to extracellular matrix (ECM) proteins and play an essential role in matrix mineralisation. Chitosan and collagen type I are naturally-derived pro-osteogenic biomaterials, which have been demonstrated to control EV release kinetics. Therefore, this study aimed to develop an injectable ECM-mimetic hydrogel capable of controlling the release of osteoblast-derived EVs to promote bone repair. Pure chitosan hydrogels significantly enhanced compressive modulus (2.48-fold) and osteogenic differentiation (3.07-fold), whilst reducing gelation times (2.09-fold) and proliferation (2.7-fold) compared to pure collagen gels (p ≤ 0.001). EV release was strongly associated with collagen concentration (R2 > 0.94), where a significantly increased EV release profile was observed from chitosan containing gels using the CD63 ELISA (p ≤ 0.001). Hydrogel-released EVs enhanced human bone marrow stromal cells (hBMSCs) proliferation (1.12-fold), migration (2.55-fold), and mineralisation (3.25-fold) compared to untreated cells (p ≤ 0.001). Importantly, EV-functionalised chitosan-collagen composites significantly promoted hBMSCs extracellular matrix mineralisation when compared to the EV-free gels in a dose-dependent manner (p ≤ 0.001). Taken together, these findings demonstrate the development of a pro-osteogenic thermosensitive chitosan-collagen hydrogel capable of enhancing the therapeutic efficacy of osteoblast-derived EVs as a novel acellular tool for bone augmentation strategy.

Magnesium oxide regulates the degradation behaviors and improves the osteogenesis of poly(lactide-co-glycolide) composite scaffolds

Poly (lactic-co-glycolic acid) (PLGA) is a star biodegradable polymer widely studied and applied in the biomedical field. Improving the acidic microenvironment caused by its degradation products and regulating its degradation behavior are still urgent scientific and technological problems to be solved. In this study, to regulate the degradation behaviors of PLGA and improve its bioactivity, hydroxyapatite (HA) and magnesium oxide (MgO) were incorporated into PLGA substrate in different proportions and a series of 3D-printing PLGA/HA/MgO (PHM) composite porous scaffolds were prepared. Then the physicochemical properties, degradation behaviors, in vitro and in vivo biological performance of fabricated scaffolds were systematically studied. The in vitro experimental results showed that PHM composite scaffolds exhibited hydrophilic and mechanical properties similar to natural bone, the presence of MgO could effectively neutralize the acidic environment, and promote bone marrow stromal cells (BMSCs) adhesion, proliferation, and differentiation. The in vivo osteogenic experiments demonstrated that the PHM5 (25 wt% HA, 5 wt% MgO) scaffold could reduce the inflammatory response and promote new bone formation. This study provides a new idea for the regulation of the degradation behavior and the improvement of osteogenic properties of PLGA-based biomaterials. The prepared 3D-printed PHM5 scaffold shows great application potential in the bone repair field.

3D printed biocompatible graphene oxide, attapulgite, and collagen composite scaffolds for bone regeneration.

Tissue-engineered bone material is one of the effective methods to repair bone defects, but the application is restricted in clinical because of the lack of excellent scaffolds that can induce bone regeneration as well as the difficulty in making scaffolds with personalized structures. 3D printing is an emerging technology that can fabricate bespoke 3D scaffolds with precise structure. However, it is challenging to develop the scaffold materials with excellent printability, osteogenesis ability, and mechanical strength. In this study, graphene oxide (GO), attapulgite (ATP), type I collagen (Col I) and polyvinyl alcohol were used as raw materials to prepare composite scaffolds via 3D bioprinting. The composite materials showed excellent printability. The microcosmic architecture and properties was characterized by scanning electron microscopy, Fourier transform infrared and thermal gravimetric analyzer, respectively. To verify the biocompatibility of the scaffolds, the viability, proliferation and osteogenic differentiation of Bone Marrow Stromal Cells (BMSCs) on the scaffolds were assessed by CCK-8, Live/Dead staining and Real-time PCR in vitro. The composited scaffolds were then implanted into the skull defects on rat for bone regeneration. Hematoxylin-eosin staining, Masson staining and immunohistochemistry staining were carried out in vivo to evaluate the regeneration of bone tissue.The results showed that GO/ATP/COL scaffolds have been demonstrated to possess controlled porosity, water absorption, biodegradability and good apatite-mineralization ability. The scaffold consisting of 0.5% GO/ATP/COL have excellent biocompatibility and was able to promote the growth, proliferation and osteogenic differentiation of mouse BMSCs in vitro. Furthermore, the 0.5% GO/ATP/COL scaffolds were also able to promote bone regeneration of in rat skull defects. Our results illustrated that the 3D printed GO/ATP/COL composite scaffolds have good mechanical properties, excellent cytocompatibility for enhanced mouse BMSCs adhesion, proliferation, and osteogenic differentiation. All these advantages made it potential as a promising biomaterial for osteogenic reconstruction.