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

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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Ursolic acid loaded-mesoporous hydroxylapatite/ chitosan therapeutic scaffolds promote the bone regeneration

10.21203/rs.3.rs-84118/v1 ◽

2020 ◽

Author(s):

Xijiao Yu ◽

Yuxuan Wang ◽

Xiao-Liang Liu ◽

Degang Yu ◽

Shanyong Zhang

Keyword(s):

Bone Regeneration ◽

Ursolic Acid ◽

Control Release ◽

The Novel ◽

Alizarin Red ◽

Biomaterial Scaffolds ◽

Scaffold Materials ◽

Related Proteins

Abstract Background: Mesoporous hydroxylapatite (MHAP) could play an important role in bone regeneration, and UA (Ursolic acid) also promote the osteogenic differentiation. Accordingly, we developed the UA loaded MHAP scaffolds to cure bone defects. In vitro, we synthesize biomaterial scaffolds. By SEM, XRD, EDS and FTIR, we test the performance of the hybrid scaffolds. By drug release, ALP staining, Alizarin red staining, and Western blotting, we test the osteo-inductive properties of scaffold materials. In vivo, We verify bone regeneration through a rat skull defect model.Results: The MHAP is a rod-shaped structure with a length of 100~300nm and a diameter of 40~60nm. The critical structure gives the micro scaffold a property of control release due to the pore sizes of 1.6~4.3 nm in hydroxyapatite and the hydrogen bonding between the scaffolds and UA drugs. The released UA drugs could notably promote the expression of osteogenic-related genes (COL1, ALP, OPG) and osteogenic-related proteins (BMP-2, RUNX2 and COL1). Both the images of μCT and the results of double fluorochrome labelling demonstrated that therapeutic scaffolds promoted the bone regeneration. We obtained the similar results through immunohistochemistry. Conclusions: The MHAP-CS-UA scaffolds have good osteo-inductivity and bone regeneration. And they will be the novel and promising candidates to cure the bone disease.

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Activation of Mesenchymal Stem Cells Promotes New Bone Formation Within Dentigerous Cyst

10.21203/rs.3.rs-63524/v1 ◽

2020 ◽

Author(s):

Yejia Yu ◽

Mengyu Li ◽

Yuqiong Zhou ◽

Yueqi Shi ◽

Wenjie Zhang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Regeneration ◽

Bone Defect ◽

Dentigerous Cyst ◽

Healing Process ◽

Cell Counting ◽

Cranial Bone ◽

Alizarin Red

Abstract Background: Dentigerous cyst (DC) is a bone destructive disease and remains a challenge for clinicians. Marsupialization enables bone to regenerate with capsules maintaining, making it a preferred therapeutic means for DC adjacent to vital anatomical structures. Given that capsules of DC derive from odontogenic epithelium remnants at embryonic stage, we investigated whether there were mesenchymal stem cells (MSCs) located in DC capsules and the role that they played in the bone regeneration after marsupialization.Methods: Samples obtained before and after marsupialization were used for histological detection and cell culture. The stemness of cells isolated from fresh tissues were analyzed by morphology, surface marker and multi-differentiation assays. Comparison of proliferation ability between Am-DCSCs and Bm-DCSCs were evaluated by Cell Counting Kit-8 (CCK-8), fibroblast colony-forming units (CFU-F) and 5’‐ethynyl‐2’‐deoxyuridine (EdU) assay. Their osteogenic capacity in vitro was detected by Alkaline phosphatase (ALP) and Alizarin Red staining (ARS), combined with Real-time polymerase chain reaction (RT-PCR) and immunofluorescence (IF) staining. Subcutaneous ectopic osteogenesis as well as cranial bone defect model in nude mice were performed to detect their bone regeneration and bone defect repair ability.Results: Bone tissue and strong ALP activity were detected in the capsule of DC after marsupialization. Two types of MSCs were isolated from fibrous capsules of DC both before (Bm-DCSCs) and after (Am-DCSCs) marsupialization. These fibroblast-like, colony forming cells expressed MSC markers (CD44+, CD90+, CD31-, CD34-, CD45-), and they could differentiate into osteoblast-, adipocyte- and chondrocyte-like cells under induction. Notably, Am-DCSCs performed better in cell proliferation and self-renewal. Moreover, Am-DCSCs showed greater osteogenic capacity both in vitro and in vivo compared with Bm-DCSCs. Conclusions: There are MSCs residing in capsules of DC, and the cell viability as well as osteogenic capacity of them are largely enhanced after marsupialization. Our findings suggested that MSCs might play a crucial role in the healing process of DC after marsupialization, thus providing new insight into the treatment for DC by promoting the osteogenic differentiation of MSCs inside capsules.

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

10.21203/rs.3.rs-1073550/v1 ◽

2021 ◽

Author(s):

Ning Wang ◽

Xuanchen Liu ◽

Zhen Tang ◽

Xinghui Wei ◽

Hui Dong ◽

...

Keyword(s):

Diabetes Mellitus ◽

Bone Formation ◽

Bone Regeneration ◽

Bone Defect ◽

Diabetic Rats ◽

Osteogenic Potential ◽

Bone Degradation ◽

Defect Repair

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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Activation of mesenchymal stem cells promotes new bone formation within dentigerous cyst

Stem Cell Research & Therapy ◽

10.1186/s13287-020-01999-8 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Yejia Yu ◽

Mengyu Li ◽

Yuqiong Zhou ◽

Yueqi Shi ◽

Wenjie Zhang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Regeneration ◽

Bone Defect ◽

Dentigerous Cyst ◽

Healing Process ◽

Cell Counting ◽

Cranial Bone ◽

Alizarin Red

Abstract Background Dentigerous cyst (DC) is a bone destructive disease and remains a challenge for clinicians. Marsupialization enables the bone to regenerate with capsule maintaining, making it a preferred therapeutic means for DC adjacent to vital anatomical structures. Given that capsules of DC are derived from odontogenic epithelium remnants at the embryonic stage, we investigated whether there were mesenchymal stem cells (MSCs) located in DC capsules and the role that they played in the bone regeneration after marsupialization. Methods Samples obtained before and after marsupialization were used for histological detection and cell culture. The stemness of cells isolated from fresh tissues was analyzed by morphology, surface marker, and multi-differentiation assays. Comparison of proliferation ability between MSCs isolated from DC capsules before (Bm-DCSCs) and after (Am-DCSCs) marsupialization was evaluated by Cell Counting Kit-8 (CCK-8), fibroblast colony-forming units (CFU-F), and 5′-ethynyl-2′-deoxyuridine (EdU) assay. Their osteogenic capacity in vitro was detected by alkaline phosphatase (ALP) and Alizarin Red staining (ARS), combined with real-time polymerase chain reaction (RT-PCR) and immunofluorescence (IF) staining. Subcutaneous ectopic osteogenesis as well as cranial bone defect model in nude mice was performed to detect their bone regeneration and bone defect repairability. Results Bone tissue and strong ALP activity were detected in the capsule of DC after marsupialization. Two types of MSCs were isolated from fibrous capsules of DC both before (Bm-DCSCs) and after (Am-DCSCs) marsupialization. These fibroblast-like, colony-forming cells expressed MSC markers (CD44+, CD90+, CD31−, CD34−, CD45−), and they could differentiate into osteoblast-, adipocyte-, and chondrocyte-like cells under induction. Notably, Am-DCSCs performed better in cell proliferation and self-renewal. Moreover, Am-DCSCs showed a greater osteogenic capacity both in vitro and in vivo compared with Bm-DCSCs. Conclusions There are MSCs residing in capsules of DC, and the cell viability as well as the osteogenic capacity of them is largely enhanced after marsupialization. Our findings suggested that MSCs might play a crucial role in the healing process of DC after marsupialization, thus providing new insight into the treatment for DC by promoting the osteogenic differentiation of MSCs inside capsules.

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Effects of strontium ions with potential antibacterial activity on in vivo bone regeneration

10.21203/rs.3.rs-18054/v2 ◽

2020 ◽

Author(s):

Nafiseh Baheiraei ◽

Hossein Eyni ◽

Bita bakhshi ◽

Raziyeh Najafloo

Keyword(s):

Antibacterial Activity ◽

Bone Formation ◽

Bone Regeneration ◽

Bone Repair ◽

Early Stage ◽

Antibacterial Activities ◽

Calvarial Bone ◽

Alizarin Red

Abstract Background: Bioactive glasses (BGs) have attracted added attention in the structure of the scaffolds for bone repair applications. Different metal ions could be doped in BGs to induce specific biological responses. Among these ions, strontium (Sr) is considered as an effective and safe doping element with promising effects on bone formation and regeneration.Methods: In this experiment, we evaluated the antibacterial activities of the gelatin-BG (Gel-BG) and Gel-BG/Sr scaffolds in vitro. The osteogenic properties of the prepared scaffolds were also assessed in rabbit calvarial bone defects for 12 weeks. Alizarin Red, Hematoxylin & Eosin (H&E) and Masson’s Trichrome staining were performed to assess bone regeneration and the obtained results were compared with those without Sr. Also, histomorphometric data were obtained to evaluate the new bone, residual graft, and connective tissue.Results: Both scaffolds showed in vivo bone formation during 12 weeks with the newly formed bone area in Gel-BG/Sr scaffold was higher than that in Gel-BG scaffolds after the whole period. Based on the histological results, Gel-BG/Sr exhibited acceleration of early-stage bone formation in vivo. The results of antibacterial investigation showed that although both Gel-BG/Sr and Gel-BG effectively inhibited the growth of Escherichia coli (E. coli) but, only Gel-BG/Sr structure could lead to a 3 log reduction in Staphylococcus aureus (S. aureus). Conclusions: Our results confirmed that Sr doped BG is a favorable candidate for bone tissue engineering with superior antibacterial activity and bone regeneration capacity compared with similar counterparts having no Sr ion.

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Activation of mesenchymal stem cells promotes new bone formation within dentigerous cyst

10.21203/rs.3.rs-63524/v2 ◽

2020 ◽

Author(s):

Yejia Yu ◽

Mengyu Li ◽

Yuqiong Zhou ◽

Yueqi Shi ◽

Wenjie Zhang ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Bone Regeneration ◽

Bone Defect ◽

Dentigerous Cyst ◽

Healing Process ◽

Cell Counting ◽

Cranial Bone ◽

Alizarin Red

Abstract Background: Dentigerous cyst (DC) is a bone destructive disease and remains a challenge for clinicians. Marsupialization enables bone to regenerate with capsules maintaining, making it a preferred therapeutic means for DC adjacent to vital anatomical structures. Given that capsules of DC derive from odontogenic epithelium remnants at embryonic stage, we investigated whether there were mesenchymal stem cells (MSCs) located in DC capsules and the role that they played in the bone regeneration after marsupialization. Methods: Samples obtained before and after marsupialization were used for histological detection and cell culture. The stemness of cells isolated from fresh tissues were analyzed by morphology, surface marker and multi-differentiation assays. Comparison of proliferation ability between Am-DCSCs and Bm-DCSCs were evaluated by Cell Counting Kit-8 (CCK-8), fibroblast colony-forming units (CFU-F) and 5’‐ethynyl‐2’‐deoxyuridine (EdU) assay. Their osteogenic capacity in vitro was detected by Alkaline phosphatase (ALP) and Alizarin Red staining (ARS), combined with Real-time polymerase chain reaction (RT-PCR) and immunofluorescence (IF) staining. Subcutaneous ectopic osteogenesis as well as cranial bone defect model in nude mice were performed to detect their bone regeneration and bone defect repair ability. Results: Bone tissue and strong ALP activity were detected in the capsule of DC after marsupialization. Two types of MSCs were isolated from fibrous capsules of DC both before (Bm-DCSCs) and after (Am-DCSCs) marsupialization. These fibroblast-like, colony forming cells expressed MSC markers (CD44+, CD90+, CD31-, CD34-, CD45-), and they could differentiate into osteoblast-, adipocyte- and chondrocyte-like cells under induction. Notably, Am-DCSCs performed better in cell proliferation and self-renewal. Moreover, Am-DCSCs showed greater osteogenic capacity both in vitro and in vivo compared with Bm-DCSCs. Conclusions: There are MSCs residing in capsules of DC, and the cell viability as well as osteogenic capacity of them are largely enhanced after marsupialization. Our findings suggested that MSCs might play a crucial role in the healing process of DC after marsupialization, thus providing new insight into the treatment for DC by promoting the osteogenic differentiation of MSCs inside capsules.

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Effects of strontium ions with potential antibacterial activity on in vivo bone regeneration

10.21203/rs.3.rs-18054/v1 ◽

2020 ◽

Author(s):

Nafiseh Baheiraei ◽

Hossein Eyni ◽

Bita bakhshi ◽

Raziyeh Najafloo

Keyword(s):

Antibacterial Activity ◽

Bone Formation ◽

Bone Regeneration ◽

Bone Repair ◽

Early Stage ◽

Antibacterial Activities ◽

Calvarial Bone ◽

Alizarin Red

Abstract Background Bioactive glasses (BGs) have attracted added attention in the structure of the scaffolds for bone repair applications. Different metal ions could be doped in BGs to induce specific biological responses. Among these ions, strontium (Sr) is considered as an effective and safe doping element with promising effects on bone formation and regeneration. Methods In this experiment, we evaluated the antibacterial activities of the gelatin-BG (Gel-BG) and Gel-BG/Sr scaffolds in vitro. The osteogenic properties of the prepared scaffolds were also assessed in rabbit calvarial bone defects for 12 weeks. Alizarin Red, Hematoxylin & Eosin (H&E) and Masson’s Trichrome staining were performed to assess bone regeneration and the obtained results were compared with those without Sr. Also, histomorphometric data were obtained to evaluate the new bone, residual graft, and connective tissue. Results Both scaffolds showed in vivo bone formation during 12 weeks with the newly formed bone area in Gel-BG/Sr scaffold was higher than that in Gel-BG scaffolds after the whole period. Based on the histological results, Gel-BG/Sr exhibited acceleration of early-stage bone formation in vivo. The results of antibacterial investigation showed that although both Gel-BG/Sr and Gel-BG effectively inhibited the growth of Escherichia coli (E. coli) but, only Gel-BG/Sr structure could lead to a 3 log reduction in Staphylococcus aureus (S. aureus). Conclusions: Our results confirmed that Sr doped BG is a favorable candidate for bone tissue engineering with superior antibacterial activity and bone regeneration capacity compared with similar counterparts having no Sr ion.

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The Co-Culture of ASCs and EPCs Promotes Vascularized Bone Regeneration in Critical-Sized Bone Defects of Cranial Bone in Rats

10.21203/rs.3.rs-24037/v1 ◽

2020 ◽

Author(s):

yuanjia he ◽

Shuang Lin ◽

Qiang Ao ◽

Xiaoning He

Keyword(s):

Bone Regeneration ◽

Bone Tissue ◽

Bone Defect ◽

Culture System ◽

Donor Site ◽

Bone Defects ◽

Cranial Bone ◽

Vascularized Bone

Abstract Background: The repair of critical-sized bone defect represents a challenging problem in bone tissue engineering. To address the most important problem in bone defect repair, namely insufficient blood supply, this study aimed to find a method that can promote the formation of vascularized bone tissue.Method The phenotypes of ASCs and EPCs were identified respectively, and ASCs/EPCs were co-cultured in vitro to detect the expression of osteogenic and angiogenic genes. Furthermore, the co-culture system combined with scaffold material was used to repair the critical-sized bone defects of the cranial bone in rats.Results The co-culture of ASCs/EPCs could increase osteogenesis and angiogenesis-related gene expression in vitro. The results of in vivo animal experiments demonstrated that the ASCs/EPCs group could promote bone regeneration and vascularization in the meantime and, then significantly accelerate the repair of critical-sized bone defects.Conclusion It is feasible to replace traditional single seed cells with ASCs/EPCs co-culture system for vascularized bone regeneration. This system could ultimately enable clinicians to better repair the defect of craniofacial bone and avoid donor site morbidity.

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The Co-Culture of ASCs and EPCs Promotes Vascularized Bone Regeneration in Critical-Sized Bone Defects of Cranial Bone in Rats

10.21203/rs.3.rs-24037/v2 ◽

2020 ◽

Author(s):

yuanjia he ◽

Shuang Lin ◽

Qiang Ao ◽

Xiaoning He

Keyword(s):

Bone Regeneration ◽

Bone Tissue ◽

Bone Defect ◽

Culture System ◽

Donor Site ◽

Bone Defects ◽

Cranial Bone ◽

Vascularized Bone

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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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