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13 pages, 4932 KiB

Open AccessArticle

Dose-Dependent Effects of a Novel Selective EP₄ Prostaglandin Receptor Agonist on Treatment of Critical Size Femoral Bone Defects in a Rat Model

by Corina Vater, Elisabeth Mehnert, Henriette Bretschneider, Julia Bolte, Lisa Findeisen, Lucas-Maximilian Matuszewski and Stefan Zwingenberger

Biomedicines 2021, 9(11), 1712; https://doi.org/10.3390/biomedicines9111712 - 18 Nov 2021

Cited by 2 | Viewed by 1868

Abstract

Difficulties in treating pseudarthrosis and critical bone defects are still evident in physicians’ clinical routines. Bone morphogenetic protein 2 (BMP-2) has shown promising osteoinductive results but also considerable side effects, not unexpected given that it is a morphogen. Thus, the bone regenerative potential [...] Read more.

Difficulties in treating pseudarthrosis and critical bone defects are still evident in physicians’ clinical routines. Bone morphogenetic protein 2 (BMP-2) has shown promising osteoinductive results but also considerable side effects, not unexpected given that it is a morphogen. Thus, the bone regenerative potential of the novel selective, non-morphogenic EP₄ prostaglandin receptor agonist KMN-159 was investigated in this study. Therefore, mineralized collagen type-1 matrices were loaded with different amounts of BMP-2 or KMN-159 and implanted into a 5 mm critical-sized femoral defect in rats. After 12 weeks of observation, micro-computed tomography scans were performed to analyze the newly formed bone volume (BV) and bone mineral density (BMD). Histological analysis was performed to evaluate the degree of defect healing and the number of vessels, osteoclasts, and osteoblasts. Data were evaluated using Kruskal-Wallis followed by Dunn’s post hoc test. As expected, animals treated with BMP-2, the positive control for this model, showed a high amount of newly formed BV as well as bone healing. For KMN-159, a dose-dependent effect on bone regeneration could be observed up to a dose optimum, demonstrating that this non-morphogenic mechanism of action can stimulate bone formation in this model system. Full article

(This article belongs to the Special Issue Bone Tissue Regeneration: Biology and Strategies)

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17 pages, 5171 KiB

Open AccessArticle

Rapid Fabrication of MgNH₄PO₄·H₂O/SrHPO₄ Porous Composite Scaffolds with Improved Radiopacity via 3D Printing Process

by Xiaofeng Cao, Wufei Ge, Yihu Wang, Ming Ma, Ying Wang, Bing Zhang, Jianing Wang and Yanchuan Guo

Biomedicines 2021, 9(9), 1138; https://doi.org/10.3390/biomedicines9091138 - 02 Sep 2021

Cited by 4 | Viewed by 2227

Abstract

Although bone repair scaffolds are required to possess high radiopacity to be distinguished from natural bone tissues in clinical applications, the intrinsic radiopacity of them is usually insufficient. For improving the radiopacity, combining X-ray contrast agents with bone repair scaffolds is an effective [...] Read more.

Although bone repair scaffolds are required to possess high radiopacity to be distinguished from natural bone tissues in clinical applications, the intrinsic radiopacity of them is usually insufficient. For improving the radiopacity, combining X-ray contrast agents with bone repair scaffolds is an effective method. In the present research, MgNH₄PO₄·H₂O/SrHPO₄ 3D porous composite scaffolds with improved radiopacity were fabricated via the 3D printing technique. Here, SrHPO₄ was firstly used as a radiopaque agent to improve the radiopacity of magnesium phosphate scaffolds. X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive spectroscopy (EDS) were used to characterize the phases, morphologies, and element compositions of the 3D porous composite scaffolds. The radiography image showed that greater SrHPO₄ contents corresponded to higher radiopacity. When the SrHPO₄ content reached 9.34%, the radiopacity of the composite scaffolds was equal to that of a 6.8 mm Al ladder. The porosity and in vitro degradation of the porous composite scaffolds were studied in detail. The results show that magnesium phosphate scaffolds with various Sr contents could sustainably degrade and release the Mg, Sr, and P elements during the experiment period of 28 days. In addition, the cytotoxicity on MC3T3-E1 osteoblast precursor cells was evaluated, and the results show that the porous composite scaffolds with a SrHPO₄ content of 9.34% possessed superior cytocompatibility compared to that of the pure MgNH₄PO₄·H₂O scaffolds when the extract concentration was 0.1 g/mL. Cell adhesion experiments showed that all of the scaffolds could support MC3T3-E1 cellular attachment well. This research indicates that MgNH₄PO₄·H₂O/SrHPO₄ porous composite scaffolds have potential applications in the bone repair fields. Full article

(This article belongs to the Special Issue Bone Tissue Regeneration: Biology and Strategies)

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15 pages, 2937 KiB

Open AccessArticle

Comparison of Autologous Blood Clots with Fibrin Sealant as Scaffolds for Promoting Human Muscle-Derived Stem Cell-Mediated Bone Regeneration

by Xueqin Gao, Haizi Cheng, Xuying Sun, Aiping Lu, Joseph J. Ruzbarsky, Bing Wang and Johnny Huard

Biomedicines 2021, 9(8), 983; https://doi.org/10.3390/biomedicines9080983 - 09 Aug 2021

Cited by 7 | Viewed by 1883

Abstract

Background. Fibrin sealant has been used as a scaffold to deliver genetically modified human muscle-derived stem cells (hMDSCs) for bone regeneration. Alternatively, autologous blood clots are safe, economic scaffolds. This study compared autologous blood clot (BC) with fibrin sealant (FS) as a [...] Read more.

Background. Fibrin sealant has been used as a scaffold to deliver genetically modified human muscle-derived stem cells (hMDSCs) for bone regeneration. Alternatively, autologous blood clots are safe, economic scaffolds. This study compared autologous blood clot (BC) with fibrin sealant (FS) as a scaffold to deliver lenti-BMP2/GFP-transduced hMDSCs for bone regeneration. Methods. In vitro osteogenic differentiation was performed using 3D pellet culture and evaluated using microCT and Von Kossa staining. The lenti-GFP transduced cells were then mixed with human blood for evaluation of osteogenic differentiation. Furthermore, a murine critical- sized calvarial defect model was utilized to compare BC and FS scaffolds for lenti-BMP2/GFP-transduced hMDSCs mediated bone regeneration and evaluated with micro-CT and histology. Results. Lenti-BMP2/GFP transduced hMDSCs formed significantly larger mineralized pellets than non-transduced hMDSCs. hMDSCs within the human blood clot migrated out and differentiated into ALP⁺ osteoblasts. In vivo, BC resulted in significantly less new bone formation within a critical-sized calvarial bone defect than FS scaffold, despite no difference observed for GFP⁺ donor cells, osteoclasts, and osteoblasts in the newly formed bone. Conclusions. Human lenti-BMP2/GFP-transduced hMDSCs can efficiently undergo osteogenic differentiation in vitro. Unexpectedly, the newly regenerated bone in BC group was significantly less than the FS group. The autologous blood clot scaffold is less efficacious for delivering stem cells for bone regeneration than fibrin sealant. Full article

(This article belongs to the Special Issue Bone Tissue Regeneration: Biology and Strategies)

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15 pages, 17550 KiB

Open AccessEditor’s ChoiceArticle

3D-Printed Ginsenoside Rb1-Loaded Mesoporous Calcium Silicate/Calcium Sulfate Scaffolds for Inflammation Inhibition and Bone Regeneration

by Cheng-Yu Chen, Ming-You Shie, Alvin Kai-Xing Lee, Yun-Ting Chou, Chun Chiang and Chun-Pin Lin

Biomedicines 2021, 9(8), 907; https://doi.org/10.3390/biomedicines9080907 - 28 Jul 2021

Cited by 17 | Viewed by 2763

Abstract

Bone defects are commonly found in the elderly and athletic population due to systemic diseases such as osteoporosis and trauma. Bone scaffolds have since been developed to enhance bone regeneration by acting as a biological extracellular scaffold for cells. The main advantage of [...] Read more.

Bone defects are commonly found in the elderly and athletic population due to systemic diseases such as osteoporosis and trauma. Bone scaffolds have since been developed to enhance bone regeneration by acting as a biological extracellular scaffold for cells. The main advantage of a bone scaffold lies in its ability to provide various degrees of structural support and growth factors for cellular activities. Therefore, we designed a 3D porous scaffold that can not only provide sufficient mechanical properties but also carry drugs and promote cell viability. Ginsenoside Rb1 (GR) is an extract from panax ginseng, which has been used for bone regeneration and repair since ancient Chinese history. In this study, we fabricated scaffolds using various concentrations of GR with mesoporous calcium silicate/calcium sulfate (MSCS) and investigated the scaffold’s physical and chemical characteristic properties. PrestoBlue, F-actin staining, and ELISA were used to demonstrate the effect of the GR-contained MSCS scaffold on cell proliferation, morphology, and expression of the specific osteogenic-related protein of human dental pulp stem cells (hDPSCs). According to our data, hDPSCs cultivated in GR-contained MSCS scaffold had preferable abilities of proliferation and higher expression of the osteogenic-related protein and could effectively inhibit inflammation. Finally, in vivo performance was assessed using histological results that revealed the GR-contained MSCS scaffolds were able to further achieve more effective hard tissue regeneration than has been the case in the past. Taken together, this study demonstrated that a GR-containing MSCS 3D scaffold could be used as a potential alternative for future bone tissue engineering studies and has good potential for clinical use. Full article

(This article belongs to the Special Issue Bone Tissue Regeneration: Biology and Strategies)

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25 pages, 8058 KiB

Open AccessArticle

Lumbar Interbody Fusion Conducted on a Porcine Model with a Bioresorbable Ceramic/Biopolymer Hybrid Implant Enriched with Hyperstable Fibroblast Growth Factor 2

by Milan Krticka, Ladislav Planka, Lucy Vojtova, Vladimir Nekuda, Premysl Stastny, Radek Sedlacek, Adam Brinek, Michaela Kavkova, Eduard Gopfert, Vera Hedvicakova, Michala Rampichova, Leos Kren, Kvetoslava Liskova, Daniel Ira, Jana Dorazilová, Tomas Suchy, Tomas Zikmund, Jozef Kaiser, David Stary, Martin Faldyna and Martin Trunec Show full author list Hide full author list

Biomedicines 2021, 9(7), 733; https://doi.org/10.3390/biomedicines9070733 - 25 Jun 2021

Cited by 5 | Viewed by 2959

Abstract

Many growth factors have been studied as additives accelerating lumbar fusion rates in different animal models. However, their low hydrolytic and thermal stability both in vitro and in vivo limits their workability and use. In the proposed work, a stabilized vasculogenic and prohealing [...] Read more.

Many growth factors have been studied as additives accelerating lumbar fusion rates in different animal models. However, their low hydrolytic and thermal stability both in vitro and in vivo limits their workability and use. In the proposed work, a stabilized vasculogenic and prohealing fibroblast growth factor-2 (FGF2-STAB^®) exhibiting a functional half-life in vitro at 37 °C more than 20 days was applied for lumbar fusion in combination with a bioresorbable scaffold on porcine models. An experimental animal study was designed to investigate the intervertebral fusion efficiency and safety of a bioresorbable ceramic/biopolymer hybrid implant enriched with FGF2-STAB^® in comparison with a tricortical bone autograft used as a gold standard. Twenty-four experimental pigs underwent L2/3 discectomy with implantation of either the tricortical iliac crest bone autograft or the bioresorbable hybrid implant (BHI) followed by lateral intervertebral fixation. The quality of spinal fusion was assessed by micro-computed tomography (micro-CT), biomechanical testing, and histological examination at both 8 and 16 weeks after the surgery. While 8 weeks after implantation, micro-CT analysis demonstrated similar fusion quality in both groups, in contrast, spines with BHI involving inorganic hydroxyapatite and tricalcium phosphate along with organic collagen, oxidized cellulose, and FGF2- STAB^® showed a significant increase in a fusion quality in comparison to the autograft group 16 weeks post-surgery (p = 0.023). Biomechanical testing revealed significantly higher stiffness of spines treated with the bioresorbable hybrid implant group compared to the autograft group (p < 0.05). Whilst histomorphological evaluation showed significant progression of new bone formation in the BHI group besides non-union and fibrocartilage tissue formed in the autograft group. Significant osteoinductive effects of BHI based on bioceramics, collagen, oxidized cellulose, and FGF2-STAB^® could improve outcomes in spinal fusion surgery and bone tissue regeneration. Full article

(This article belongs to the Special Issue Bone Tissue Regeneration: Biology and Strategies)

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19 pages, 7656 KiB

Open AccessArticle

Hydroxycoumarin Scopoletin Inhibits Bone Loss through Enhancing Induction of Bone Turnover Markers in a Mouse Model of Type 2 Diabetes

by Eun-Jung Lee, Woojin Na, Min-Kyung Kang, Yun-Ho Kim, Dong-Yeon Kim, Hyeongjoo Oh, Soo-Il Kim, Su-Yeon Oh, Sohyun Park, Kyungho Park and Young-Hee Kang

Biomedicines 2021, 9(6), 648; https://doi.org/10.3390/biomedicines9060648 - 07 Jun 2021

Cited by 9 | Viewed by 2920

Abstract

Diabetes induces bone deterioration, which leads to increased risk of fracture, osteopenia, and osteoporosis. Thus, diabetes-associated bone fragility has been recognized as a diabetic complication. However, the pathophysiological effects of hyperglycemia on bone turnover remain unclear. Literature evidence demonstrates that anti-diabetic medications increase [...] Read more.

Diabetes induces bone deterioration, which leads to increased risk of fracture, osteopenia, and osteoporosis. Thus, diabetes-associated bone fragility has been recognized as a diabetic complication. However, the pathophysiological effects of hyperglycemia on bone turnover remain unclear. Literature evidence demonstrates that anti-diabetic medications increase the risk of fractures in individuals with type 2 diabetes. Scopoletin is a naturally occurring hydroxycoumarin potentially exhibiting anti-inflammatory and antioxidant activities and ameliorating insulin resistance as an anti-diabetic agent. However, little is known regarding the effects of scopoletin on the impairment of bone remodeling that is caused by diabetes. The aim of this study was to identify that scopoletin was capable of inhibiting the impairment of bone remodeling and turnover in a mouse model of type 2 diabetes. Submicromolar scopoletin accelerated the formation TRAP-positive multinucleated osteoclasts (40.0 vs. 105.1%) and actin ring structures impaired by 33 mM glucose. Further, 1–20 μM scopoletin enhanced bone resorption and the induction of matrix-degrading enzymes in diabetic osteoclasts. The oral administration of 10 mg/kg scopoletin elevated serum RANKL/OPG ratio and osteocalcin level reduced in db/db mice along with an increase in BMD by ~6–14%; however, it was not effective in lowering blood glucose and hemoglobin glycation. In addition, the supplementation of scopoletin elevated the formation of trabecular bones and collagen fibers in femoral epiphysis and metaphysis with a thicker epiphyseal plate and cortical bones. Furthermore, 1–20 μM scopoletin enhanced ALP activity (4.39 vs. 7.02 nmol p-nitrophenyl phosphate/min/mg protein) and deposits of mineralized bone nodules in cultured osteoblasts reduced by 33 mM glucose. The treatment of diabetic osteoblasts with scopoletin stimulated the cellular induction of BMP-2 and osteopontin and Runx2 transcription. Accordingly, the administration of scopoletin protected mice from type 2 diabetes-associated bone loss through boosting bone remodeling via the robust induction of bone turnover markers of both osteoclasts and osteoblasts. These findings suggest that scopoletin could be a potential osteoprotective agent for the treatment of diabetes-associated bone loss and fractures. Full article

(This article belongs to the Special Issue Bone Tissue Regeneration: Biology and Strategies)

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22 pages, 9485 KiB

Open AccessArticle

Identification of Sclerostin as a Putative New Myokine Involved in the Muscle-to-Bone Crosstalk

by Maria Sara Magarò, Jessika Bertacchini, Francesca Florio, Manuela Zavatti, Francesco Potì, Francesco Cavani, Emanuela Amore, Ilaria De Santis, Alessandro Bevilacqua, Luca Reggiani Bonetti, Pietro Torricelli, Delphine B. Maurel, Stefano Biressi and Carla Palumbo

Biomedicines 2021, 9(1), 71; https://doi.org/10.3390/biomedicines9010071 - 12 Jan 2021

Cited by 24 | Viewed by 3938

Abstract

Bone and muscle have been recognized as endocrine organs since they produce and secrete “hormone-like factors” that can mutually influence each other and other tissues, giving rise to a “bone–muscle crosstalk”. In our study, we made use of myogenic (C2C12 cells) and osteogenic [...] Read more.

Bone and muscle have been recognized as endocrine organs since they produce and secrete “hormone-like factors” that can mutually influence each other and other tissues, giving rise to a “bone–muscle crosstalk”. In our study, we made use of myogenic (C2C12 cells) and osteogenic (2T3 cells) cell lines to investigate the effects of muscle cell-produced factors on the maturation process of osteoblasts. We found that the myogenic medium has inhibitory effects on bone cell differentiation and we identified sclerostin as one of the myokines produced by muscle cells. Sclerostin is a secreted glycoprotein reportedly expressed by bone/cartilage cells and is considered a negative regulator of bone growth due to its role as an antagonist of the Wnt/β-catenin pathway. Given the inhibitory role of sclerostin in bone, we analyzed its expression by muscle cells and how it affects bone formation and homeostasis. Firstly, we characterized and quantified sclerostin synthesis by a myoblast cell line (C2C12) and by murine primary muscle cells by Western blotting, real-time PCR, immunofluorescence, and ELISA assay. Next, we investigated in vivo production of sclerostin in distinct muscle groups with different metabolic and mechanical loading characteristics. This analysis was done in mice of different ages (6 weeks, 5 and 18 months after birth) and revealed that sclerostin expression is dynamically modulated in a muscle-specific way during the lifespan. Finally, we transiently expressed sclerostin in the hind limb muscles of young mice (2 weeks of age) via in vivo electro-transfer of a plasmid containing the SOST gene in order to investigate the effects of muscle-specific overproduction of the protein. Our data disclosed an inhibitory role of the muscular sclerostin on the bones adjacent to the electroporated muscles. This observation suggests that sclerostin released by skeletal muscle might synergistically interact with osseous sclerostin and potentiate negative regulation of osteogenesis possibly by acting in a paracrine/local fashion. Our data point out a role for muscle as a new source of sclerostin. Full article

(This article belongs to the Special Issue Bone Tissue Regeneration: Biology and Strategies)

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19 pages, 2735 KiB

Open AccessArticle

Enhancing Mechanical Properties and Biological Performances of Injectable Bioactive Glass by Gelatin and Chitosan for Bone Small Defect Repair

by Mehri Sohrabi, Bijan Eftekhari Yekta, Hamidreza Rezaie, Mohammad Reza Naimi-Jamal, Ajay Kumar, Andrea Cochis, Marta Miola and Lia Rimondini

Biomedicines 2020, 8(12), 616; https://doi.org/10.3390/biomedicines8120616 - 15 Dec 2020

Cited by 21 | Viewed by 3367

Abstract

Bioactive glass (BG) represents a promising biomaterial for bone healing; here injectable BG pastes biological properties were improved by the addition of gelatin or chitosan, as well as mechanical resistance was enhanced by adding 10 or 20 wt% 3-Glycidyloxypropyl trimethoxysilane (GPTMS) cross-linker. Composite [...] Read more.

Bioactive glass (BG) represents a promising biomaterial for bone healing; here injectable BG pastes biological properties were improved by the addition of gelatin or chitosan, as well as mechanical resistance was enhanced by adding 10 or 20 wt% 3-Glycidyloxypropyl trimethoxysilane (GPTMS) cross-linker. Composite pastes exhibited bioactivity as apatite formation was observed by Scanning Electron Microscopy (SEM) and X-Ray Diffraction (XRD) after 14 days immersion in simulated body fluid (SBF); moreover, polymers did not enhance degradability as weight loss was >10% after 30 days in physiological conditions. BG-gelatin-20 wt% GPTMS composites demonstrated the highest compressive strength (4.8 ± 0.5 MPa) in comparison with the bulk control paste made of 100% BG in water (1.9 ± 0.1 MPa). Cytocompatibility was demonstrated towards human mesenchymal stem cells (hMSC), osteoblasts progenitors, and endothelial cells. The presence of 20 wt% GPTMS conferred antibacterial properties thus inhibiting the joint pathogens Staphylococcus aureus and Staphylococcus epidermidis infection. Finally, hMSC osteogenesis was successfully supported in a 3D model as demonstrated by alkaline phosphatase release and osteogenic genes expression. Full article

(This article belongs to the Special Issue Bone Tissue Regeneration: Biology and Strategies)

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16 pages, 3493 KiB

Open AccessArticle

Shockwave Therapy Modulates the Expression of BMP2 for Prevention of Bone and Cartilage Loss in the Lower Limbs of Postmenopausal Osteoporosis Rat Model

by Shan-Ling Hsu, Wen-Yi Chou, Chieh-Cheng Hsu, Jih-Yang Ko, Shun-Wun Jhan, Ching-Jen Wang, Meng-Shiou Lee, Tsai-Chin Hsu and Jai-Hong Cheng

Biomedicines 2020, 8(12), 614; https://doi.org/10.3390/biomedicines8120614 - 15 Dec 2020

Cited by 6 | Viewed by 2235

Abstract

Osteoporosis (OP) causes bone loss and weakness, increasing the risk of bone fracture. In this study, rats were divided into Sham, OP, SW(F) (0.25 mJ/mm² with 1600 impulses to the left medial femur), and SW(T) (0.25 mJ/mm² with 1600 impulses to [...] Read more.

Osteoporosis (OP) causes bone loss and weakness, increasing the risk of bone fracture. In this study, rats were divided into Sham, OP, SW(F) (0.25 mJ/mm² with 1600 impulses to the left medial femur), and SW(T) (0.25 mJ/mm² with 1600 impulses to the left medial tibia). The bone strength results following SW(T) were better than SW(F) in the modulus, extension at peak load, handleability, and strain at break. SW(T) had the best prevention for bone loss in both lower limbs of ovariectomized (OVX) rats. The cartilage cellular matrixes of both knees were improved in SW(T) and SW(F) compared to that of OP. Serum bone morphogenetic protein 2 (BMP2) in rats undergoing SW(T) or SW(F) was significantly improved compared to that in Sham and OP. The expressions of BMP2, BMP4, and SMAD family member 4 (Smad4) in addition to the Wnt family member 3A (Wnt3a) and Cyclin D1 signaling key factors were significantly induced in the cartilage of both knees by shockwave (SW). SW(T) presented the best efficacy to induce serum BMP2 to prevent bone loss from both lower limbs. Here, we display the protective effects of SW therapy to induce BMP2, BMP4, Smad4, Wnt3a, and Cyclin D1 signaling factors for cartilage loss in both knees of OVX rats. Full article

(This article belongs to the Special Issue Bone Tissue Regeneration: Biology and Strategies)

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Review

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20 pages, 2406 KiB

Open AccessReview

Tissue Engineering Through 3D Bioprinting to Recreate and Study Bone Disease

by Adriene Pavek, Christopher Nartker, Maamoon Saleh, Matthew Kirkham, Sana Khajeh Pour, Ali Aghazadeh-Habashi and Jared J. Barrott

Biomedicines 2021, 9(5), 551; https://doi.org/10.3390/biomedicines9050551 - 14 May 2021

Cited by 12 | Viewed by 4325

Abstract

The applications of 3D bioprinting are becoming more commonplace. Since the advent of tissue engineering, bone has received much attention for the ability to engineer normal bone for tissue engraftment or replacement. While there are still debates on what materials comprise the most [...] Read more.

The applications of 3D bioprinting are becoming more commonplace. Since the advent of tissue engineering, bone has received much attention for the ability to engineer normal bone for tissue engraftment or replacement. While there are still debates on what materials comprise the most durable and natural replacement of normal tissue, little attention is given to recreating diseased states within the bone. With a better understanding of the cellular pathophysiology associated with the more common bone diseases, these diseases can be scaled down to a more throughput way to test therapies that can reverse the cellular pathophysiology. In this review, we will discuss the potential of 3D bioprinting of bone tissue in the following disease states: osteoporosis, Paget’s disease, heterotopic ossification, osteosarcoma, osteogenesis imperfecta, and rickets disease. The development of these 3D bioprinted models will allow for the advancement of novel therapy testing resulting in possible relief to these chronic diseases. Full article

(This article belongs to the Special Issue Bone Tissue Regeneration: Biology and Strategies)

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