Augmentation of bone healing of nonunion fracture using stem cell based tissue engineering in a dog: a case report

A 4-year-old, intact male crossbreed dog, weighing 27 kg, was referred for the treatment of a nonunion fracture. The radiographs revealed displacement of the radius and ulna bone fracture fragment and a sclerotic fracture end of the radius. Autologous adipose derived stem cells (ADSCs) were isolated and expanded ex vivo in a culture. The ADSCs (3.2 × 107 cells) were seeded on a composition scaffold made from hydroxyapatite (HA) and chitosan (CH) fibers. The seeded scaffold with ADSCs was placed on the fracture site and the bone fracture was stabilized. A sample of seeded scaffold with ADSCs was taken to evaluate the extent of cell attachment and morphology on the scaffold using scanning electron microscopy (SEM). SEM showed that ADSCs adhered to the scaffold well and many bone nodules formed from the bone matrix secreted by ADSCs. Three months after surgery, the nonunion had successfully healed with no complications. The application of a composition scaffold of HA and CH containing ADSCs can be used to treat a nonunion fracture by augmenting bone healing and may decrease the risk of surgical failure of nonunion fractures.

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Novel Fracture Site Targeting Drug Improves Fracture Healing with Pain Relief: Preclinical Evaluation of MAK123

Proceedings of IMPRS ◽

10.18060/25901 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Abduallah Elsayed ◽

Jeffery Nielsen ◽

Natalie Taylor ◽

Mohammed Juboori ◽

Caio de Andrade Staut ◽

...

Keyword(s):

Bone Healing ◽

Femoral Fracture ◽

Ex Vivo ◽

Demineralized Bone Matrix ◽

Bone Matrix ◽

Fracture Site ◽

X Rays ◽

Opioid Use ◽

Fracture Callus ◽

Surgically Induced

Non-union bone fracture occurs in 5-10% of fracture injuries. Interventions include surgery with local implantation of autograft, allograft, demineralized bone matrix, and/or bone morphogenetic proteins. These types of fracture injuries are also accompanied by acute and chronic pain states. In most instances, opioids are provided to injured patients during and after surgery. With the opioid crisis, identifying new analgesic therapies that could reduce or eliminate opioid use, while also improving bone healing is important. Here we show the ability of a novel compound, MAK123, to both enhance bone healing and reduce pain behavior in a surgically induced femoral fracture mouse model. Briefly, 20 male C57BL/6 mice underwent a surgically induced femoral fracture and then were treated with 0, 2, 6, or 20 mg/kg, 3X/week for the 3 week study duration. Weekly X-rays were used to examine healing progression. Prior to euthanasia, mice underwent behavioral testing to measure evoked pain behaviors. Upon euthanasia, ex vivo µCT imaging and analysis was completed to assess fracture callus size and composition. While all doses of MAK123 tested resulted in improved healing, the 6mg/kg dose resulted in accelerated bone healing and a significant increase in mineralized callus volume (p<0.05). Similarly, while all doses of MAK123 reduced evoked responses to tactile stimulus as demonstrated by increased paw withdrawal thresholds, 6 mg/kg of MAK123 resulted in a more robust and significant improvement (p<0.05). We postulate that optimization of the dosing schedule/concentration could further improve both bone healing and behavioral measures thought to represent pain in rodents. That said, these promising pre-clinical data warrant further evaluation as MAK123 may prove to be a unique tool for orthopaedic surgery usage whereby it could both improve bone healing and reduce clinical pain, improving overall patient outcomes.

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MAK122: A Novel Drug Utilizing Innovative Fracture Site Targeting Technology to Improve Bone Healing

Proceedings of IMPRS ◽

10.18060/25922 ◽

2021 ◽

Vol 4 (1) ◽

Author(s):

Nicholas Hux ◽

Jeffery Nielson ◽

Caio De Andrade Staut ◽

Vincent Alentado ◽

Abduallah Elsayed ◽

...

Keyword(s):

Fracture Healing ◽

Bone Healing ◽

Ex Vivo ◽

Healing Process ◽

Fracture Site ◽

Fracture Repair ◽

X Rays ◽

Invasive Treatment ◽

Post Surgery ◽

Novel Drug

Megakaryocytes play a pivotal role in the bone fracture healing process through enhancing osteoblast proliferation, osteoclastogenesis, and angiogenesis. Current fracture repair therapies require direct implantation during surgery (BMP-2, grafts etc.), which has limitations. In order to address this, a novel drug, compound MAK122, was created with targeting technology that directs its actions to the fracture site without needing to be implanted during surgery, limiting undesirable offsite effects, increasing the quantity of drug at the fracture site, and allowing for non-invasive treatment following assessment of the natural healing process. Therefore, this study examined the ability of MAK122 to stimulate megakaryocytes and subsequent bone healing. To accomplish this, male mice on a C57BL/6 background underwent a surgically induced femoral fracture. Following surgery, the mice were injected daily for the first 7 days with either saline (vehicle) or MAK122. Mice were then euthanized 2, 3 and 4 weeks post-surgery. Fracture healing was assessed by standard and novel methodologies. Biweekly X-rays were evaluated and bone union was scored showing that MAK122 accelerated bone healing compared to controls. Ex vivo µCT analysis demonstrated that MAK122 increased callus volume and the percentage of mineralized callus tissue compared to vehicle treatment. Biomechanical testing showed that MAK122 treatment resulted in stronger repairs as compared to vehicle treated controls with nearly a 2-fold increase in twist to failure and toughness parameters. Additionally, histological assessment demonstrated accelerated remodeling in MAK122 treated femurs compared to those treated with saline. Taken together, these pre-clinical data suggest that MAK122 is capable of promoting an environment in which megakaryocytes can favorably influence bone remodeling mechanisms, expediting fracture repair in murine models. Though further pharmacokinetic, pharmacodynamic, and toxicology studies are required, MAK122 displays potential to serve as a state-of-the-art therapy for improving fracture healing in humans.

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Bone-Healing Patterns Affected by Loading, Fracture Fragment Stability, Fracture Type, and Fracture Site Compression

Clinical Orthopaedics and Related Research ◽

10.1097/00003086-199308000-00003 ◽

1993 ◽

Vol &NA; (293) ◽

pp. 8???17 ◽

Cited By ~ 4

Author(s):

HANNU T. ARO ◽

EDMUND Y. S. CHAO

Keyword(s):

Bone Healing ◽

Fracture Site ◽

Fracture Type ◽

Fracture Fragment

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Simultaneous Treatment of Photobiomodulation and Demineralized Bone Matrix With Adipose-Derived Stem Cells Improve Bone Healing in an osteoporotic bone defect

Journal of lasers in medical sciences ◽

10.34172/jlms.2021.41 ◽

2021 ◽

Vol 12 (1) ◽

pp. e41-e41

Author(s):

Rouhallah Gazor ◽

Mehrdad Asgari ◽

Mohammad-Amin Abdollajhifar ◽

Pejman Kiani ◽

Fatemeh Zare ◽

...

Keyword(s):

Stem Cells ◽

Bone Healing ◽

Demineralized Bone Matrix ◽

Bone Matrix ◽

Adipose Derived Stem Cells ◽

Simultaneous Treatment ◽

Whitney Test ◽

Mann Whitney Test ◽

Ovx Rats ◽

Demineralized Bone

Introduction: The ability of simultaneous treatment of critical-sized femoral defects (CSFDs) with photobiomodulation (PBM) and demineralized bone matrix (DBM) with or without seeded adipose-derived stem cells (ASCs) to induce bone reconstruction in ovariectomized induced osteoporotic (OVX) rats was investigated. Methods: The OVX rats with CSFD were arbitrarily separated into 6 groups: control, scaffold (S, DBM), S + PBM, S + alendronate (ALN), S + ASCs, and S + PBM + ASCs. Each group was assessed by cone beam computed tomography (CBCT) and histological examinations. Results: In the fourth week, CBCT and histological analyses revealed that the largest volume of new bone formed in the S + PBM and S + PBM + ASC groups. The S + PBM treatment relative to the S and S + ALN treatments remarkably reduced the CSFD (Mann-Whitney test, P = 0.009 and P = 0.01). Furthermore, S + PBM + ASCs treatment compared to the S and S + ALN treatments significantly decreased CSFD (Mann Whitney test, P = 0.01). In the eighth week, CBCT analysis showed that extremely enhanced bone regeneration occurred in the CSFD of the S + PBM group. Moreover, the CSFD in the S + PBM group was substantially smaller than S, S + ALN and S + ASCs groups (Mann Whitney test, P = 0.01, P = 0.02 and P = 0.009). Histological observations showed more new bone formation in the treated CSFD of S + PBM + ASCs and S + PBM groups. Conclusion: The PBM plus DBM with or without ASCs significantly enhanced bone healing in the CSFD in OVX rats compared to control, DBM alone, and ALN plus DBM groups. The PBM plus DBM with or without ASCs significantly decreased the CSFD area compared to either the solo DBM or ALN plus DBM treatments.

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Paracrine Effects of Recombinant Human Adiponectin Promote Bone Regeneration

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.762335 ◽

2021 ◽

Vol 9 ◽

Author(s):

Yanping Gong ◽

Yang Wang ◽

Yiqing Zhang ◽

Liangchen Wang ◽

Lijuan Wan ◽

...

Keyword(s):

Fracture Healing ◽

Bone Regeneration ◽

Bone Healing ◽

Bone Fracture ◽

Underlying Mechanism ◽

Fracture Site ◽

High Yield ◽

Sprague Dawley ◽

Paracrine Effects ◽

Non Union

Bone regeneration is a delicate physiological process. Non-union and delayed fracture healing remains a great challenge in clinical practice nowadays. Bone and fat hold a close relationship to remain balanced through hormones and cytokines. Adiponectin is a well-known protein to maintain the hemostasis, which may be an interesting target for fracture healing. Herein, we provided a facile and efficient method to obtain high-purity and high-yield recombinant human adiponectin (ADPN). The biocompatibility and the pharmaceutical behaviors were evaluated in Sprague–Dawley rats. The paracrine effects of adiponectin on bone fracture healing were investigated with a rat tibia fracture model via intrabone injection. Significantly accelerated bone healing was observed in the medulla injection group, indicating the paracrine effects of adiponectin could be potentially utilized for clinical treatments. The underlying mechanism was primarily assessed, and the expression of osteogenic markers, including bone morphogenic protein 2, alkaline phosphatase, and osteocalcin, along with adiponectin receptor 1 (AdipoR1), was markedly increased at the fracture site. The increased bone healing of ADPN treatment may result from both enhanced osteogenic proliferation as well as differentiation. Cell experiments confirmed that the expression of osteogenesis markers increased significantly in ADPN treatment groups, while it decreased when the expression of AdipoR1 was knocked down by siRNA. Our study provided a feasible and efficacious way for bone fracture treatment with local administration of ADPN, which could be rapidly translated into the clinics.

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Analysis of the effects of spaceflight and local administration of thrombopoietin to a femoral defect injury on distal skeletal sites

npj Microgravity ◽

10.1038/s41526-021-00140-0 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Ariane Zamarioli ◽

Zachery R. Campbell ◽

Kevin A. Maupin ◽

Paul J. Childress ◽

Joao P. B. Ximenez ◽

...

Keyword(s):

Bone Loss ◽

Mechanical Loading ◽

Bone Healing ◽

Space Flight ◽

Bone Fracture ◽

Local Administration ◽

Systemic Effects ◽

Human Presence ◽

Femoral Defect ◽

Healing Agent

AbstractWith increased human presence in space, bone loss and fractures will occur. Thrombopoietin (TPO) is a recently patented bone healing agent. Here, we investigated the systemic effects of TPO on mice subjected to spaceflight and sustaining a bone fracture. Forty, 9-week-old, male, C57BL/6 J were divided into 4 groups: (1) Saline+Earth; (2) TPO + Earth; (3) Saline+Flight; and (4) TPO + Flight (n = 10/group). Saline- and TPO-treated mice underwent a femoral defect surgery, and 20 mice were housed in space (“Flight”) and 20 mice on Earth for approximately 4 weeks. With the exception of the calvarium and incisor, positive changes were observed in TPO-treated, spaceflight bones, suggesting TPO may improve osteogenesis in the absence of mechanical loading. Thus, TPO, may serve as a new bone healing agent, and may also improve some skeletal properties of astronauts, which might be extrapolated for patients on Earth with restraint mobilization and/or are incapable of bearing weight on their bones.

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Impaired Alignment of Bone Matrix Microstructure Associated with Disorganized Osteoblast Arrangement in Malignant Melanoma Metastasis

Biomolecules ◽

10.3390/biom11020131 ◽

2021 ◽

Vol 11 (2) ◽

pp. 131

Author(s):

Aira Matsugaki ◽

Yumi Kimura ◽

Ryota Watanabe ◽

Fumihito Nakamura ◽

Ryo Takehana ◽

...

Keyword(s):

Malignant Melanoma ◽

Cell Activation ◽

Ex Vivo ◽

Bone Matrix ◽

Melanoma Metastasis ◽

Mineral Density ◽

High Fracture ◽

Matrix Microstructure ◽

Metastasis Model

Malignant melanoma favors spreading to bone, resulting in a weakened bone with a high fracture risk. Here, we revealed the disorganized alignment of apatite crystals in the bone matrix associated with the homing of cancer cells by developing an artificially controlled ex vivo melanoma bone metastasis model. The ex vivo metastasis model reflects the progressive melanoma cell activation in vivo, resulting in decreased bone mineral density and expression of MMP1-positive cells. Moreover, less organized intercellular connections were observed in the neighboring osteoblasts in metastasized bone, indicating the abnormal and randomized organization of bone matrix secreted by disconnected osteoblasts. Our study revealed that the deteriorated microstructure associated with disorganized osteoblast arrangement was a determinant of malignant melanoma-related bone dysfunction.

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ZNF521 Represses Osteoblastic Differentiation in Human Adipose-Derived Stem Cells

International Journal of Molecular Sciences ◽

10.3390/ijms19124095 ◽

2018 ◽

Vol 19 (12) ◽

pp. 4095 ◽

Cited By ~ 8

Author(s):

Emanuela Chiarella ◽

Annamaria Aloisio ◽

Stefania Scicchitano ◽

Valeria Lucchino ◽

Ylenia Montalcini ◽

...

Keyword(s):

Stem Cells ◽

Zinc Finger Protein ◽

Lentiviral Vector ◽

Adipogenic Differentiation ◽

Bone Matrix ◽

Osteoblastic Differentiation ◽

Adipose Derived Stem Cells ◽

Matrix Mineralization ◽

Calcium Deposits ◽

Vector Transduction

Human adipose-derived stem cells (hADSCs) are multipotent mesenchymal cells that can differentiate into adipocytes, chondrocytes, and osteocytes. During osteoblastogenesis, the osteoprogenitor cells differentiate into mature osteoblasts and synthesize bone matrix components. Zinc finger protein 521 (ZNF521/Zfp521) is a transcription co-factor implicated in the regulation of hematopoietic, neural, and mesenchymal stem cells, where it has been shown to inhibit adipogenic differentiation. The present study is aimed at determining the effects of ZNF521 on the osteoblastic differentiation of hADSCs to clarify whether it can influence their osteogenic commitment. The enforced expression or silencing of ZNF521 in hADSCs was achieved by lentiviral vector transduction. Cells were cultured in a commercial osteogenic medium for up to 20 days. The ZNF521 enforced expression significantly reduced osteoblast development as assessed by the morphological and molecular criteria, resulting in reduced levels of collagen I, alkaline phosphatase, osterix, osteopontin, and calcium deposits. Conversely, ZNF521 silencing, in response to osteoblastic stimuli, induced a significant increase in early molecular markers of osteogenesis and, at later stages, a remarkable enhancement of matrix mineralization. Together with our previous findings, these results show that ZNF521 inhibits both adipocytic and osteoblastic maturation in hADSCs and suggest that its expression may contribute to maintaining the immature properties of hADSCs.

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Mechanical flexural ex vivo study of osteotomized swine femurs stabilized with two types of polyamide 12 rods

Ciência Rural ◽

10.1590/0103-8478cr20160757 ◽

2017 ◽

Vol 47 (8) ◽

Author(s):

Juliana Scarpa da Silveira Almeida ◽

Débora de Oliveira Garcia ◽

Renato Camargo Bortholin ◽

Carlos Amaral Razzino ◽

Cristiane dos Santos Honsho ◽

...

Keyword(s):

Bone Fractures ◽

Ex Vivo ◽

Polymeric Materials ◽

Fracture Site ◽

Medullary Canal ◽

Modulus Of Rupture ◽

Long Bone ◽

Bone Implant ◽

Polyamide 12 ◽

Implant System

ABSTRACT: Long bone fractures are commonly in surgery routine and several bone imobilization techniques are currently available. Technological progress has enabled to use low cost materials in surgical procedures. Thus, the aim of this study was to evaluate the applicability of polyamide 12 rods, solid and hollow in swine femurs, comparing them through flexion strength. This study had as second aim to fix the locking errors, commom place in interlocking nails, once polyamide 12 allows perforation in any direction by orthopaedic screw. Six groups were used: G1 - eight whole swine femurs; G2 - eight whole swine femurs with drilled medullary canal; G3 - two solid polyamide 12 rods; G4 - two hollow polyamide 12 rods; G5 - eight osteotomized drilled swine femurs with a solid polyamide 12 rod implanted in the medullary canal and locked by four 316L stainless steel screws; and G6 - similar to G5 but using hollow rods instead of solid ones. No significant differences were observed for the modulus of rupture between solid and hollow rods, demonstrating that both rods had similar performances. These results led to the speculation that the addition of other polymers to the hollow rods could increase their strength and thus the bone-implant system. Furthermore, the comparison between G1, G5 and G6 could be analyzed using the finite element method in future. New polymeric materials may be developed based on the data from this study, strengthening the bone-implant system and making possible screws to be placed in any direction, nullifying the detrimental forces on the fracture site.

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