The Impact of Nano-Crystal Hydroxyapatites on the Regeneration of Bone Defects

Abstract Calcium hydroxyapatite is a widely used material for replacing bone defects. However, the effectiveness of nano-crystalline calcium hydroxyapatite produced from eggshells in the replacement of bone defects has not been investigated yet. The study aimed to evaluate the effectiveness of using nano-crystalline calcium hydroxyapatite made from eggshell for the healing of bone defect of the femur in rats. Forty-eight (n=48) rats underwent a surgical procedure to simulate femoral defect. The animals were sub-divided into 4 groups (each with n=12) depending on the methods of bone defect replacement: I control group (CG) (without bone defect replacement); II intervention group (the bone defect was replaced by PRP (PRP); III intervention group (the bone defect was replaced by nano-crystalline hydroxyapatite obtained from eggshell) (HA) and IV interventional group (the bone defect was replaced by a combination of hydroxyapatite and PRP) (HA+PRP). The degree of effectiveness of studied methods was assessed using radiological (on the 14th day), histological (on the 61st day), and biomechanical analysis (on the 61st day). According to radiographic data, the CG group had the lowest level of bone regeneration after 14 days (4.2 ±1.7%). In the HA + PRP group, the level of bone regeneration was 22.1±7.1 %, which was higher in comparison with the rates of consolidation of bone defects in the HA group (20.7± 9.3) (p = 0.023). According to the histo-morphometry data, the rates of bone tissue regeneration in the PRP group (19.8 ±4.2%) were higher in comparison with the CG group (12.7 ± 7.3%), (p>0.05). In the HA+PRP group, bone regeneration rates (48.9±9.4 %) were significantly higher (p=0.001) than in the HA group (35.1±9.8%). According to the results of biomechanical assessment under the maximum stress (121.0722), the maximum bending deformation of the contralateral bone without defect was 0.028746, which was higher than the indicators of the HA+PRP group, where at the maximum stress (90.67979) the bending deformation was 0.024953 (p>0.05). Compared to CG, PRP, and HA, biomechanical bone strength was significantly higher in the HA + PRP group (p≤0.01). At the maximum stress (51.81391), the maximum bending strain in the CG group was 0.03869, which was lower than in the PRP group, where the maximum stress and bending strain were 59.45824 and 0.055171, respectively (p>0.05). However, the bone strength of the HA group was statistically significantly higher compared to the CG and PRP groups (p<0.01). The results demonstrated the effectiveness of the use of nanocrystalline calcium hydroxyapatite obtained from eggshell in the healing of a bone defect. The best results were observed in the group of the combined use of nano-crystalline calcium hydroxyapatite and PRP.

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3D-printed Poly-Lactic Co-Glycolic Acid (PLGA) scaffolds in non-critical bone defects impede bone regeneration in rabbit tibia bone

Bio-Medical Materials and Engineering ◽

10.3233/bme-216017 ◽

2021 ◽

pp. 1-7

Author(s):

Jin Xi Lim ◽

Min He ◽

Alphonsus Khin Sze Chong

Keyword(s):

Computed Tomography ◽

Bone Regeneration ◽

Bone Defect ◽

Volume Ratio ◽

Bone Defects ◽

Control Group ◽

Micro Computed Tomography ◽

Rabbit Tibia ◽

3D Printed ◽

Critical Bone Defects

BACKGROUND: An increasing number of bone graft materials are commercially available and vary in their composition, mechanism of action, costs, and indications. OBJECTIVE: A commercially available PLGA scaffold produced using 3D printing technology has been used to promote the preservation of the alveolar socket after tooth extraction. We examined its influence on bone regeneration in long bones of New Zealand White rabbits. METHODS: 5.0-mm-diameter circular defects were created on the tibia bones of eight rabbits. Two groups were studied: (1) control group, in which the bone defects were left empty; (2) scaffold group, in which the PLGA scaffolds were implanted into the bone defect. Radiography was performed every two weeks postoperatively. After sacrifice, bone specimens were isolated and examined by micro-computed tomography and histology. RESULTS: Scaffolds were not degraded by eight weeks after surgery. Micro-computed tomography and histology showed that in the region of bone defects that was occupied by scaffolds, bone regeneration was compromised and the total bone volume/total volume ratio (BV/TV) was significantly lower. CONCLUSION: The implantation of this scaffold impedes bone regeneration in a non-critical bone defect. Implantation of bone scaffolds, if unnecessary, lead to a slower rate of fracture healing.

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Repairing Rabbit Radius Bone Defects with Simvastatin Compound Biological Bone

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2021.2710 ◽

2021 ◽

Vol 11 (7) ◽

pp. 1263-1270

Author(s):

Zhong-Yu Liu ◽

Jin-Li Zhang ◽

Yang Zhang ◽

Shi-Lian Kan ◽

Jun Liang ◽

...

Keyword(s):

Bone Defect ◽

Maximum Stress ◽

Biomechanical Testing ◽

Bone Defects ◽

Bending Test ◽

Bone Material ◽

X Ray ◽

Group A ◽

Group B ◽

Group D

Objective: This study aimed to investigate the feasibility of repairing rabbit radius bone defects with simvastatin compound biological bone. Methods: Simvastatin biological bone material was prepared, and osteoblasts were cultured. A total of 42 New Zealand white rabbits were randomly divided into four groups, and a bone defect with a length of 15 mm was created at the middle part of the radial shaft of both limbs in each rabbit, thereby establishing a bone defect model. The grafts in group A were biological bones of osteoblasts combined with simvastatin; the grafts in group B were biological bones of simvastatin; the grafts in group C were biological compound bones of osteoblasts; and the grafts in group D were simple biological bones. In each group, four animals were randomly sacrificed at the sixth and twelfth week after surgery, and specimens were collected for gross observation, X-ray examination, histological observation, and biomechanical testing. In each group, two animals were randomly sacrificed at the twelfth week after surgery; a three-point bending test was performed using a biomechanical testing machine, and the results were compared with those of a normal radius. Results: The X-ray and histological examinations at 6 and 12 weeks after surgery revealed that the osteogenesis ability of the simvastatin biological bone and osteoblast-simvastatin biological bone was better than that of the osteoblast biological bone and simple biological bone, which was superior in group A and group B to group C and group D. The results of the biomechanical examination revealed that the maximum stress of the normal radius was significantly higher than that of the experimental groups. Among the experimental groups, the difference between group A and group B was not statistically significant, and the maximum stress was higher in groups A and B than in groups C and D. Conclusion: Simvastatin biological bone material can promote the repair of rabbit radius defects and increase the quality of bone healing.

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Guided Bone Regeneration in Calvarial Bone Defects Using Polytetrafluoroethylene Membranes

The Cleft Palate-Craniofacial Journal ◽

10.1597/1545-1569_1995_032_0311_gbricb_2.3.co_2 ◽

1995 ◽

Vol 32 (4) ◽

pp. 311-317 ◽

Cited By ~ 18

Author(s):

Carles Bosch ◽

Birte Melsen ◽

Karin Vargervik

Keyword(s):

Bone Regeneration ◽

Bone Defect ◽

Dura Mater ◽

Bone Defects ◽

Guided Bone Regeneration ◽

Parietal Bone ◽

Bony Defect ◽

Calvarial Bone ◽

Double Membrane ◽

Single Membrane

Guided bone regeneration is defined as controlled stimulation of new bone formation in a bony defect, either by osteogenesis, osteoinduction, or osteoconduction, re-establishing both structural and functional characteristics. Bony defects may be found as a result of congenital anomalies, trauma, neoplasms, or infectious conditions. Such conditions are often associated with severe functional and esthetic problems. Corrective treatment is often complicated by limitations in tissue adaptations. The aim of the investigation was to compare histologically the amount of bone formed in an experimentally created parietal bone defect protected with one or two polytetrafluoroethylene membranes with a contralateral control defect. A bony defect was created bilaterally in the parietal bone lateral to the sagittal suture in 29 6-month-old male Wistar rats. The animals were divided into two groups: (1) In the double membrane group (n=9), the left experimental bone defect was protected by an outer polytetrafluoroethylene membrane under the periosteum and parietal muscles and an inner membrane between the dura mater and the parietal bone. (2) In the single membrane group (n=20), only the outer membrane was placed. The right defect was not covered with any membrane and served as control. The animals were killed after 30 days. None of the control defects demonstrated complete or partial bone regeneration. In the single membrane group, the experimental site did not regenerate in 15 animals, partially in four, and completely in one. In the double membrane group, six of the experimental defects had complete closure with bone, two had partial closure, and one no closure. The use of two membranes protecting the bone edges of the parietal defect from the overlying tissues and underlying brain enhanced bone regeneration in experimental calvarial bone defects. The biologic role of the dura mater may not be of critical importance in new bone regeneration in these calvarial bone defects.

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The Development of Gelatin/Hyaluronate Copolymer Mixed with Calcium Sulfate, Hydroxyapatite, and Stromal-Cell-Derived Factor-1 for Bone Regeneration Enhancement

Polymers ◽

10.3390/polym11091454 ◽

2019 ◽

Vol 11 (9) ◽

pp. 1454 ◽

Cited By ~ 6

Author(s):

Yun-Liang Chang ◽

Chia-Ying Hsieh ◽

Chao-Yuan Yeh ◽

Feng-Huei Lin

Keyword(s):

Bone Regeneration ◽

Bone Defect ◽

Calcium Sulfate ◽

Stromal Cell ◽

Femoral Condyle ◽

Bone Defects ◽

Blood Analysis ◽

Control Group ◽

Defect Model ◽

Stromal Cell Derived Factor

In clinical practice, bone defects still remain a challenge. In recent years, apart from the osteoconductivity that most bone void fillers already provide, osteoinductivity has also been emphasized to promote bone healing. Stromal-cell-derived factor-1 (SDF-1) has been shown to have the ability to recruit mesenchymal stem cells (MSCs), which play an important role in the bone regeneration process. In this study, we developed a gelatin–hyaluronate (Gel-HA) copolymer mixed with calcium sulfate (CS), hydroxyapatite (HAP), and SDF-1 in order to enhance bone regeneration in a bone defect model. The composites were tested in vitro for biocompatibility and their ability to recruit MSCs after material characterization. For the in vivo test, a rat femoral condyle bone defect model was used. Micro computed tomography (Micro-CT), two-photon excitation microscopy, and histology analysis were performed to assess bone regeneration. As expected, enhanced bone regeneration was well observed in the group filled with Gel-HA/CS/HAP/SDF-1 composites compared with the control group in our animal model. Furthermore, detailed blood analysis of rats showed no obvious systemic toxicity or side effects after material implantation. In conclusion, the Gel-HA/CS/HAP/SDF-1 composite may be a safe and applicable material to enhance bone regeneration in bone defects.

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Fabrication of Stromal Cell-Derived Factor-1 Contained in Gelatin/Hyaluronate Copo006Cymer Mixed with Hydroxyapatite for Use in Traumatic Bone Defects

Micromachines ◽

10.3390/mi12070822 ◽

2021 ◽

Vol 12 (7) ◽

pp. 822

Author(s):

Yun-Liang Chang ◽

Chia-Ying Hsieh ◽

Chao-Yuan Yeh ◽

Chih-Hao Chang ◽

Feng-Huei Lin

Keyword(s):

Bone Regeneration ◽

Bone Defect ◽

Stromal Cell ◽

Second Harmonic ◽

Bone Defects ◽

In Vivo Studies ◽

Control Group ◽

Defect Model ◽

Stromal Cell Derived Factor ◽

Bone Void

Bone defects of orthopedic trauma remain a challenge in clinical practice. Regarding bone void fillers, besides the well-known osteoconductivity of most bone substitutes, osteoinductivity has also been gaining attention in recent years. It is known that stromal cell-derived factor-1 (SDF-1) can recruit mesenchymal stem cells (MSCs) in certain circumstances, which may also play an important role in bone regeneration. In this study, we fabricated a gelatin/hyaluronate (Gel/HA) copolymer mixed with hydroxyapatite (HAP) and SDF-1 to try and enhance bone regeneration in a bone defect model. After material characterization, these Gel/HA–HAP and Gel/HA–HAP–SDF-1 composites were tested for their biocompatibility and ability to recruit MSCs in vitro. A femoral condyle bone defect model of rats was used for in vivo studies. For the assessment of bone healing, micro-CT analysis, second harmonic generation (SHG) imaging, and histology studies were performed. As a result, the Gel/HA–HAP composites showed no systemic toxicity to rats. Gel/HA–HAP composite groups both showed better bone generation compared with the control group in an animal study, and the composite with the SDF-1 group even showed a trend of faster bone growth compared with the composite without SDF-1 group. In conclusion, in the management of traumatic bone defects, Gel/HA–HAP–SDF-1 composites can be a feasible material for use as bone void fillers.

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Effect of Gellan Gum/Tuna Skin Film in Guided Bone Regeneration in Artificial Bone Defect in Rabbit Calvaria

Materials ◽

10.3390/ma13061318 ◽

2020 ◽

Vol 13 (6) ◽

pp. 1318

Author(s):

Seunggon Jung ◽

Hee-Kyun Oh ◽

Myung-Sun Kim ◽

Ki-Young Lee ◽

Hongju Park ◽

...

Keyword(s):

Bone Regeneration ◽

Bone Defect ◽

Bone Defects ◽

Guided Bone Regeneration ◽

Gellan Gum ◽

Collagen Membrane ◽

New Bone Formation ◽

Average Cell ◽

Beta Tricalcium Phosphate

It is necessary to prevent the invasion of soft tissue into bone defects for successful outcomes in guided bone regeneration (GBR). For this reason, many materials are used as protective barriers to bone defects. In this study, a gellan gum/tuna skin gelatin (GEL/TSG) film was prepared, and its effectiveness in bone regeneration was evaluated. The film exhibited average cell viability in vitro. Experimental bone defects were prepared in rabbit calvaria, and a bone graft procedure with beta-tricalcium phosphate was done. The film was used as a membrane of GBR and compared with results using a commercial collagen membrane. Grafted material did not show dispersion outside of bone defects and the film did not collapse into the bone defect. New bone formation was comparable to that using the collagen membrane. These results suggest that the GEL/TSG film could be used as a membrane for GBR.

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Exosomal miR-335 derived from mature dendritic cells enhanced mesenchymal stem cell-mediated bone regeneration of bone defects in athymic rats

Molecular Medicine ◽

10.1186/s10020-021-00268-5 ◽

2021 ◽

Vol 27 (1) ◽

Author(s):

Zhongliu Cao ◽

Yanfeng Wu ◽

Lingling Yu ◽

Lingfeng Zou ◽

Liu Yang ◽

...

Keyword(s):

Dendritic Cells ◽

Bone Regeneration ◽

Osteogenic Differentiation ◽

Bone Defect ◽

Animal Experiments ◽

Bone Defects ◽

Induction Medium ◽

Hippo Signaling ◽

Femoral Bone ◽

The Matrix

Abstract Background Transplantation of bone marrow-derived mesenchymal stem cells (BM-MSCs) embedded in a bio-compatible matrix has been demonstrated as a promising strategy for the treatment of bone defects. This study was designed to explore the effect and mechanism of exosomes derived from mature dendritic cells (mDC-Exo) on the BM-MSCs-mediated bone regeneration using the matrix support in an athymic rat model of femoral bone defect. Methods The BM-MSCs were isolated from rats and incubated with osteoblast induction medium, exosomes derived from immature DCs (imDC-Exo), mDC-Exo, and miR-335-deficient mDC-Exo. BM-MSCs treated without or with mDC-Exo were embedded in a bio-compatible matrix (Orthoss®) and then implanted into the femoral bone defect of athymic rats. Results mDC-Exo promoted the proliferation and osteogenic differentiation of BM-MSCs by transferring miR-335. Mechanistically, exosomal miR-335 inhibited Hippo signaling by targeting large tongue suppressor kinase 1 (LATS1) and thus promoted the proliferation and osteogenic differentiation of BM-MSCs. Animal experiments showed that mDC-Exo enhanced BM-MSCs-mediated bone regeneration after bone defect, and this effect was abrogated when miR-335 expression was inhibited in mDC-Exo. Conclusion mDC-Exo promoted osteogenic differentiation of BM-MSCs and enhanced BM-MSCs-mediated bone regeneration after femoral bone defect in athymic rats by transferring miR-335.

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The combinatory effect of sinusoidal electromagnetic field and VEGF promotes osteogenesis and angiogenesis of mesenchymal stem cell-laden PCL/HA implants in a rat subcritical cranial defect

Stem Cell Research & Therapy ◽

10.1186/s13287-019-1464-x ◽

2019 ◽

Vol 10 (1) ◽

Cited By ~ 1

Author(s):

Jingyuan Chen ◽

Chang Tu ◽

Xiangyu Tang ◽

Hao Li ◽

Jiyuan Yan ◽

...

Keyword(s):

Tissue Engineering ◽

Electromagnetic Field ◽

Bone Formation ◽

Bone Regeneration ◽

Bone Defect ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Bone Defects ◽

Histological Evaluation ◽

Cranial Defect

Abstract Background Restoration of massive bone defects remains a huge challenge for orthopedic surgeons. Insufficient vascularization and slow bone regeneration limited the application of tissue engineering in bone defect. The effect of electromagnetic field (EMF) on bone defect has been reported for many years. However, sinusoidal EMF (SEMF) combined with tissue engineering in bone regeneration remains poorly investigated. Methods In the present study, we investigated the effect of SEMF and vascular endothelial growth factor (VEGF) on osteogenic and vasculogenic differentiation of rat bone marrow-derived mesenchymal stem cells (rBMSCs). Furthermore, pretreated rBMSC- laden polycaprolactone-hydroxyapatite (PCL/HA) scaffold was constructed and implanted into the subcritical cranial defect of rats. The bone formation and vascularization were evaluated 4 and 12 weeks after implantation. Results It was shown that SEMF and VEGF could enhance the protein and mRNA expression levels of osteoblast- and endothelial cell-related markers, respectively. The combinatory effect of SEMF and VEGF slightly promoted the angiogenic differentiation of rBMSCs. The proteins of Wnt1, low-density lipoprotein receptor-related protein 6 (LRP-6), and β-catenin increased in all inducted groups, especially in SEMF + VEGF group. The results indicated that Wnt/β-catenin pathway might participate in the osteogenic and angiogenic differentiation of rBMSCs. Histological evaluation and reconstructed 3D graphs revealed that tissue-engineered constructs significantly promoted the new bone formation and angiogenesis compared to other groups. Conclusion The combinatory effect of SEMF and VEGF raised an efficient approach to enhance the osteogenesis and vascularization of tissue-engineered constructs, which provided a useful guide for regeneration of bone defects.

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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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