Increased BMSC Exosomal miR-140-3p Alleviates Bone Degradation and Promotes Bone Restoration by Targeting Plxnb1 in Diabetic Rats

Abstract Background: Diabetes mellitus (DM) is considered to be an important factor for bone degeneration disorders such as bone defect nonunion, which is characterized by physical disability and tremendous economy cost to families and society. Exosomal miRNAs of BMSCs have been reported to participate in osteoblastogenesis and modulating bone formation. However, their impacts on the development of bone degeneration in DM are not yet known. The role of miRNAs in BMSCs exosomes on regulating hyperglycemia bone degeneration was investigated in the present study. Results: The osteogenic potential in bone defect repair of exosomes derived from diabetes mellitus BMSCs derived exosomes (DM-Exos) were revealed to be lower than that in normal BMSCs derived exosomes (N-Exos) in vitro and in vivo. Here, we demonstrate that miR-140-3p level was significantly altered in exosomes derived from BMSCs, ADSCs and serum from DM rats. In in vitro experiments, upregulated miR-140-3p exosomes promoted DM BMSCs differentiation into osteoblasts. The effects were exerted by miR-140-3p targeting plxnb1, plexin B1 is the receptor of semaphoring 4D(Sema4D) that inhibited osteocytes differentiation, thereby promoting bone formation. In DM rats with bone defect, miR-140-3p upregulated exosomes were transplanted into injured bone and accelerated bone regeneration. Besides, miR-140-3p in the exosomes was transferred into BMSCs and osteoblasts and promoted bone regeneration by targeting the plexin B1/RohA/ROCK signaling pathway. Conclusions: Normal-Exos and miR-140-3p overexpressed-Exos accelerated diabetic wound healing by promoting the osteoblastogenesis function of BMSCs through inhibition plexin B1 expression which is the receptor of Sema4D and the plexin B1/RhoA/ROCK pathway compared with diabetes mellitus-Exos. This offers a new insight and a new therapy for treating diabetic bone unhealing.

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Enhanced Bone Regeneration and Bone Defect Repair Using Magnesium/Lithium-Co-Modified, Porous, Hydroxyapatite Composite Scaffolds

Science of Advanced Materials ◽

10.1166/sam.2020.3681 ◽

2020 ◽

Vol 12 (1) ◽

pp. 124-136

Author(s):

Hai-Yan Zhao ◽

Releken-Yeersheng ◽

Ya-Yi Xia ◽

Xing-Wen Han ◽

Chao Zhang ◽

...

Keyword(s):

Bone Regeneration ◽

Bone Defect ◽

Biological Degradation ◽

Porous Hydroxyapatite ◽

Bone Defect Repair ◽

Hydroxyapatite Composite ◽

Defect Repair ◽

Osteogenic Effect

Background: Hydroxyapatite (HA) has been frequently used in clinic, but it is hard to be degraded, and insufficient in osteogenesis and angiogenesis. This study aimed to modify HA by doping magnesium/lithium (Mg/Li) and assess the Mg/LiHA scaffold's bone regeneration and bone defect repair effects. Materials and Methods: The biomaterial was identified using XRD, FTIR and SEM. The porosity, cell mediated degradation behavior and mechanical property were investigated. Meanwhile, cell proliferation and adhesion were also exploited. Finally, osteogenic effect of Mg/LiHA scaffold in vitro, and bone defect repair effect in vivo were researched. Results: The results suggested that low-content of Mg/Li incorporation did not influence on the structure of HA. The cells mediated degradation experiments indicated that Mg/Li doped HA could improve the biological degradation and release the Mg2+ and Li+ sustainedly. The compressive strength of Mg/LiHA scaffolds with 63% porosity reached to 3.9 MPa. Cells proliferation and adhesion experiments demonstrated that Mg/LiHA scaffolds were beneficial to cell growth and attachment. Furthermore, Mg/LiHA scaffolds increased ALP expression, calcium phosphate deposition and VEGF expression in vitro. The bone defect repair in vivo was enhanced by using Mg/LiHA scaffolds. Conclusion: Mg/Li-co-substituted HA could enhance bone regeneration and bone defect repair, and may be recommended to further research on bone defect repair.

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Construction of hollow polydopamine nanoparticle based drug sustainable release system and its application in bone regeneration

International Journal of Oral Science ◽

10.1038/s41368-021-00132-6 ◽

2021 ◽

Vol 13 (1) ◽

Author(s):

Lu Wang ◽

Shuwei Liu ◽

Chunxia Ren ◽

Siyuan Xiang ◽

Daowei Li ◽

...

Keyword(s):

Bone Regeneration ◽

Bone Defect ◽

Load Capacity ◽

Good Prospect ◽

Alizarin Red ◽

Drug Load ◽

Load Rate ◽

Low Toxicity

AbstractNanomaterial-based drug sustainable release systems have been tentatively applied to bone regeneration. They, however, still face disadvantages of high toxicity, low biocompatibility, and low drug-load capacity. In view of the low toxicity and high biocompatibility of polymer nanomaterials and the excellent load capacity of hollow nanomaterials with high specific surface area, we evaluated the hollow polydopamine nanoparticles (HPDA NPs), in order to find an optimal system to effectively deliver the osteogenic drugs to improve treatment of bone defect. Data demonstrated that the HPDA NPs synthesized herein could efficiently load four types of osteogenic drugs and the drugs can effectively release from the HPDA NPs for a relatively longer time in vitro and in vivo with low toxicity and high biocompatibility. Results of qRT-PCR, ALP, and alizarin red S staining showed that drugs released from the HPDA NPs could promote osteogenic differentiation and proliferation of rat bone marrow mesenchymal stem cells (rBMSCs) in vitro. Image data from micro-CT and H&E staining showed that all four osteogenic drugs released from the HPDA NPs effectively promoted bone regeneration in the defect of tooth extraction fossa in vivo, especially tacrolimus. These results suggest that the HPDA NPs, the biodegradable hollow polymer nanoparticles with high drug load rate and sustainable release ability, have good prospect to treat the bone defect in future clinical practice.

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The Bone Regeneration Capacity of BMP-2 + MMP-10 Loaded Scaffolds Depends on the Tissue Status

Pharmaceutics ◽

10.3390/pharmaceutics13070979 ◽

2021 ◽

Vol 13 (7) ◽

pp. 979

Author(s):

Patricia Garcia-Garcia ◽

Ricardo Reyes ◽

José Antonio Rodriguez ◽

Tomas Martín ◽

Carmen Evora ◽

...

Keyword(s):

Bone Formation ◽

Bone Regeneration ◽

Growth Promotion ◽

Tissue Growth ◽

Bone Morphogenetic Protein 2 ◽

Morphogenetic Protein ◽

Therapeutic Molecules ◽

Positive Effect

Biomaterials-mediated bone formation in osteoporosis (OP) is challenging as it requires tissue growth promotion and adequate mineralization. Based on our previous findings, the development of scaffolds combining bone morphogenetic protein 2 (BMP-2) and matrix metalloproteinase 10 (MMP-10) shows promise for OP management. To test our hypothesis, scaffolds containing BMP-2 + MMP-10 at variable ratios or BMP-2 + Alendronate (ALD) were prepared. Systems were characterized and tested in vitro on healthy and OP mesenchymal stem cells and in vivo bone formation was studied on healthy and OP animals. Therapeutic molecules were efficiently encapsulated into PLGA microspheres and embedded into chitosan foams. The use of PLGA (poly(lactic-co-glycolic acid)) microspheres as therapeutic molecule reservoirs allowed them to achieve an in vitro and in vivo controlled release. A beneficial effect on the alkaline phosphatase activity of non-OP cells was observed for both combinations when compared with BMP-2 alone. This effect was not detected on OP cells where all treatments promoted a similar increase in ALP activity compared with control. The in vivo results indicated a positive effect of the BMP-2 + MMP-10 combination at both of the doses tested on tissue repair for OP mice while it had the opposite effect on non-OP animals. This fact can be explained by the scaffold’s slow-release rate and degradation that could be beneficial for delayed bone regeneration conditions but had the reverse effect on healthy animals. Therefore, the development of adequate scaffolds for bone regeneration requires consideration of the tissue catabolic/anabolic balance to obtain biomaterials with degradation/release behaviors suited for the existing tissue status.

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Matrix Vesicles: Role in Bone Mineralization and Potential Use as Therapeutics

Pharmaceuticals ◽

10.3390/ph14040289 ◽

2021 ◽

Vol 14 (4) ◽

pp. 289

Author(s):

Sana Ansari ◽

Bregje W. M. de de Wildt ◽

Michelle A. M. Vis ◽

Carolina E. de de Korte ◽

Keita Ito ◽

...

Keyword(s):

Bone Formation ◽

Bone Regeneration ◽

Bone Cells ◽

Bone Mineralization ◽

Recipient Cell ◽

Organic Matrix ◽

Matrix Vesicles ◽

Matrix Mineralization

Bone is a complex organ maintained by three main cell types: osteoblasts, osteoclasts, and osteocytes. During bone formation, osteoblasts deposit a mineralized organic matrix. Evidence shows that bone cells release extracellular vesicles (EVs): nano-sized bilayer vesicles, which are involved in intercellular communication by delivering their cargoes through protein–ligand interactions or fusion to the plasma membrane of the recipient cell. Osteoblasts shed a subset of EVs known as matrix vesicles (MtVs), which contain phosphatases, calcium, and inorganic phosphate. These vesicles are believed to have a major role in matrix mineralization, and they feature bone-targeting and osteo-inductive properties. Understanding their contribution in bone formation and mineralization could help to target bone pathologies or bone regeneration using novel approaches such as stimulating MtV secretion in vivo, or the administration of in vitro or biomimetically produced MtVs. This review attempts to discuss the role of MtVs in biomineralization and their potential application for bone pathologies and bone regeneration.

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Alendronate Can Improve Bone Alterations in Experimental Diabetes by Preventing Antiosteogenic, Antichondrogenic, and Proadipocytic Effects of AGEs on Bone Marrow Progenitor Cells

BioMed Research International ◽

10.1155/2016/5891925 ◽

2016 ◽

Vol 2016 ◽

pp. 1-13 ◽

Cited By ~ 1

Author(s):

Sara Rocío Chuguransky ◽

Ana María Cortizo ◽

Antonio Desmond McCarthy

Keyword(s):

Bone Marrow ◽

Experimental Diabetes ◽

Bone Microarchitecture ◽

Bone Matrix ◽

Diabetic Rats ◽

Long Bone ◽

Osteogenic Potential ◽

Bone Marrow Progenitor

Bisphosphonates such as alendronate are antiosteoporotic drugs that inhibit the activity of bone-resorbing osteoclasts and secondarily promote osteoblastic function. Diabetes increases bone-matrix-associated advanced glycation end products (AGEs) that impair bone marrow progenitor cell (BMPC) osteogenic potential and decrease bone quality. Here we investigated the in vitro effect of alendronate and/or AGEs on the osteoblastogenic, adipogenic, and chondrogenic potential of BMPC isolated from nondiabetic untreated rats. We also evaluated the in vivo effect of alendronate (administered orally to rats with insulin-deficient Diabetes) on long-bone microarchitecture and BMPC multilineage potential. In vitro, the osteogenesis (Runx2, alkaline phosphatase, type 1 collagen, and mineralization) and chondrogenesis (glycosaminoglycan production) of BMPC were both decreased by AGEs, while coincubation with alendronate prevented these effects. The adipogenesis of BMPC (PPARγ, intracellular triglycerides, and lipase) was increased by AGEs, and this was prevented by coincubation with alendronate. In vivo, experimental Diabetes (a) decreased femoral trabecular bone area, osteocyte density, and osteoclastic TRAP activity; (b) increased bone marrow adiposity; and (c) deregulated BMPC phenotypic potential (increasing adipogenesis and decreasing osteogenesis and chondrogenesis). Orally administered alendronate prevented all these Diabetes-induced effects on bone. Thus, alendronate could improve bone alterations in diabetic rats by preventing the antiosteogenic, antichondrogenic, and proadipocytic effects of AGEs on BMPC.

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Osteogenic Potential of Multipotent Adult Progenitor Cells for Calvaria Bone Regeneration

Advances in Medicine ◽

10.1155/2016/2803081 ◽

2016 ◽

Vol 2016 ◽

pp. 1-11

Author(s):

Dong Joon Lee ◽

Yonsil Park ◽

Wei-Shou Hu ◽

Ching-Chang Ko

Keyword(s):

Bone Regeneration ◽

Progenitor Cells ◽

Adult Stem Cells ◽

Single Cells ◽

Osteogenic Potential ◽

Type I ◽

Differentiation Potential ◽

Multipotent Adult Progenitor Cells

Osteogenic cells derived from rat multipotent adult progenitor cells (rMAPCs) were investigated for their potential use in bone regeneration. rMAPCs are adult stem cells derived from bone marrow that have a high proliferation capacity and the differentiation potential to multiple lineages. They may also offer immunomodulatory properties favorable for applications for regenerative medicine. rMAPCs were cultivated as single cells or as 3D aggregates in osteogenic media for up to 38 days, and their differentiation to bone lineage was then assessed by immunostaining of osteocalcin and collagen type I and by mineralization assays. The capability of rMAPCs in facilitating bone regeneration was evaluatedin vivoby the direct implantation of multipotent adult progenitor cell (MAPC) aggregates in rat calvarial defects. Bone regeneration was examined radiographically, histologically, and histomorphometrically. Results showed that rMAPCs successfully differentiated into osteogenic lineage by demonstrating mineralized extracellular matrix formationin vitroand induced new bone formation by the effect of rMAPC aggregatesin vivo. These outcomes confirm that rMAPCs have a good osteogenic potential and provide insights into rMAPCs as a novel adult stem cell source for bone regeneration.

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Effect of Silicate Incorporation in Alpha-Tricalcium Phosphate on Behaviors of Osteoblast-Like Cells

Key Engineering Materials ◽

10.4028/www.scientific.net/kem.720.90 ◽

2016 ◽

Vol 720 ◽

pp. 90-94

Author(s):

Masanobu Kamitakahara ◽

Takashi Shirato ◽

Taishi Yokoi ◽

Hideaki Matsubara ◽

Yasuaki Shibata ◽

...

Keyword(s):

Bone Formation ◽

Bone Regeneration ◽

Tricalcium Phosphate ◽

Proliferation And Differentiation ◽

Porous Silicate

Silicate-containing alpha-tricalcium phosphate (α-TCP) ceramics are expected to be useful scaffolds for bone regeneration because α-TCP shows high biodegradability and silicate ions are expected to promote the bone formation. We previously revealed that the porous silicate-containing α-TCP granules provided earlier bone formation and showed lower biodegradability than the porous silicate-free α-TCP granules in vivo. In order to reveal the mechanism of the bone formation promoted by silicate incorporation, the proliferation and differentiation of osteoblast-like cells on the silicate-containing and silicate-free α-TCP ceramics were examined in vitro. The silicate incorporation in α-TCP promoted the differentiation of osteoblast-like cells, and it might be one of the factors to promote bone formation In Vivo.

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Effects of P-15 Peptide Coated Hydroxyapatite on Tibial Defect Repair In Vivo in Normal and Osteoporotic Rats

BioMed Research International ◽

10.1155/2015/253858 ◽

2015 ◽

Vol 2015 ◽

pp. 1-14 ◽

Cited By ~ 4

Author(s):

Rasmus Hestehave Pedersen ◽

Marina Rasmussen ◽

Søren Overgaard ◽

Ming Ding

Keyword(s):

Bone Formation ◽

Bone Regeneration ◽

Volume Fraction ◽

Structure Type ◽

Osteoporotic Bone ◽

Bone Volume Fraction ◽

Defect Repair ◽

Paired Design ◽

Tibia Defect

This study assessed the efficacy of anorganic bone mineral coated with P-15 peptide (ABM/P-15) on tibia defect repair longitudinally in both normal and osteoporotic rats in vivo. A paired design was used. 24 Norwegian brown rats were divided into normal and osteoporotic groups. 48 cylindrical defects were created in proximal tibias bilaterally. Defects were filled with ABM/P-15 or left empty. Osteoporotic status was assessed by microarchitectural analysis. Microarchitectural properties of proximal tibial defects were evaluated at 4 time points. 21 days after surgery, tibias were harvested for histology and histomorphometry. Significantly increased bone volume fraction, surface density, and connectivity were seen in all groups at days 14 and 21 compared with day 0. Moreover, the structure type of ABM/P-15 group was changed toward typical plate-like structure. Microarchitectural properties of ABM/P-15 treated newly formed bones at 21 days were similar in normal and osteoporotic rats. Histologically, significant bone formation was seen in all groups. Interestingly, significantly increased bone formation was seen in osteoporotic rats treated with ABM/P-15 indicating optimized healing potential. Empty defects showed lower healing potential in osteoporotic bone. In conclusion, ABM/P-15 accelerated bone regeneration in osteoporotic rats but did not enhance bone regeneration in normal rats.

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The potential of stromal cell-derived factor-1 delivery using a collagen membrane for bone regeneration

Journal of Biomaterials Applications ◽

10.1177/0885328216686727 ◽

2017 ◽

Vol 31 (7) ◽

pp. 1049-1061 ◽

Cited By ~ 10

Author(s):

Tadahiro Takayama ◽

Jisen Dai ◽

Keita Tachi ◽

Ryutaro Shohara ◽

Hironori Kasai ◽

...

Keyword(s):

Cell Proliferation ◽

Growth Factor ◽

Bone Formation ◽

Bone Regeneration ◽

Stromal Cell ◽

In Vitro Studies ◽

New Bone Formation ◽

Stromal Cell Derived Factor

Stromal cell-derived factor-1 (SDF-1) is a cytokine that is important in stem and progenitor cell recruitment in tissue repair after injury. Regenerative procedures using collagen membranes (CMs) are presently well established in periodontal and implant dentistry. The objective of this study is to test the subsequent effects of the released SDF-1 from a CM on bone regeneration compared to platelet-derived growth factor (PDGF) in vitro and in vivo. For in vitro studies, cell proliferation, alkaline phosphatase activity, and osteoblastic differentiation marker genes were assessed after MC3T3-E1 mouse preosteoblasts were cultured with CMs containing factors. In vivo effects were investigated by placement of CMs containing SDF-1 or PDGF using a rat mandibular bone defect model. At 4 weeks after the surgery, the new bone formation was measured using micro-computed tomography (µCT) and histological analysis. The results of in vitro studies revealed that CM delivery of SDF-1 significantly induced cell proliferation, ALP activity, and gene expression of all osteogenic markers compared to the CM alone or control, similar to PDGF. Quantitative and qualitative µCT analysis for volume of new bone formation and the percentage of new bone area showed that SDF-1-treated groups significantly increased and accelerated bone regeneration compared to control and CM alone. The enhancement of bone formation in SDF-1-treated animals was dose-dependent and with levels similar to those measured with PDGF. These results suggest that a CM with SDF-1 may be a great candidate for growth factor delivery that could be a substitute for PDGF in clinical procedures where bone regeneration is necessary.

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Lipoteichoic Acid Accelerates Bone Healing by Enhancing Osteoblast Differentiation and Inhibiting Osteoclast Activation in a Mouse Model of Femoral Defects

International Journal of Molecular Sciences ◽

10.3390/ijms21155550 ◽

2020 ◽

Vol 21 (15) ◽

pp. 5550

Author(s):

Chih-Chien Hu ◽

Chih-Hsiang Chang ◽

Yi-min Hsiao ◽

Yuhan Chang ◽

Ying-Yu Wu ◽

...

Keyword(s):

Mouse Model ◽

Bone Formation ◽

Bone Regeneration ◽

Bone Healing ◽

Lipoteichoic Acid ◽

Calcium Deposition ◽

Micro Computed Tomography ◽

Osteoclast Activation

Lipoteichoic acid (LTA) is a cell wall component of Gram-positive bacteria. Limited data suggest that LTA is beneficial for bone regeneration in vitro. Thus, we used a mouse model of femoral defects to explore the effects of LTA on bone healing in vivo. Micro-computed tomography analysis and double-fluorochrome labeling were utilized to examine whether LTA can accelerate dynamic bone formation in vivo. The effects of LTA on osteoblastogenesis and osteoclastogenesis were also studied in vitro. LTA treatment induced prompt bone bridge formation, rapid endochondral ossification, and accelerated healing of fractures in mice with femoral bone defects. In vitro, LTA directly enhanced indicators of osteogenic factor-induced MC3T3-E1 cell differentiation, including alkaline phosphatase activity, calcium deposition and osteopontin expression. LTA also inhibited osteoclast activation induced by receptor activator of nuclear factor-kappa B ligand. We identified six molecules that may be associated with LTA-accelerated bone healing: monocyte chemoattractant protein 1, chemokine (C-X-C motif) ligand 1, cystatin C, growth/differentiation factor 15, endostatin and neutrophil gelatinase-associated lipocalin. Finally, double-fluorochrome, dynamic-labeling data indicated that LTA significantly enhanced bone-formation rates in vivo. In conclusion, our findings suggest that LTA has promising bone-regeneration properties.

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