Systemic modulation of skeletal mineralization by magnesium implant promoting fracture healing: Radiological exploration enhanced with PCA-based machine learning in a rat femoral model

The clinical application of magnesium(Mg)and its alloys for bone fractures has been well supported by in vitro and in vivo trials.However,there were studies indicating negative effects of high dose Mg intake and sustained local release of Mg ions on bone metabolism or repair,which should not be ignored when developing Mg-based implants.Thus,it remains necessary to assess the biological effects of Mg implants in animal models relevant to clinical treatment modalities.The primary purpose of this study was to validate the beneficial effects of intramedullary Mg implants on the healing outcome of femoral fractures in a modified rat model.In addition,the mineralization parameters at multiple anatomical sites were evaluated,to investigate their association with healing outcome and potential clinical applications.Compared to the control group without Mg implantation,postoperative imaging at week 12 demonstrated better healing outcomes in the Mg group,with more stable unions in 3D analysis and high-mineralized bridging in 2D evaluation.The bone tissue mineral density(TMD)was higher in the Mg group at the non-operated femur and lumbar vertebra,while no differences between groups were identified regarding the bone tissue volume(TV),TMD and bone mineral content(BMC)in humerus.In the surgical femur,the Mg group presented higher TMD,but lower TV and BMC in the distal metaphyseal region,as well as reduced BMC at the osteotomy site.Principal component analysis(PCA)-based machine learning revealed that by selecting clinically relevant parameters,radiological markers could be constructed for differentiation of healing outcomes,with better performance than 2D scoring.The study provides insights and preclinical evidence for the rational investigation of bioactive materials,the identification of potential adverse effects,and the promotion of diagnostic capabilities for fracture healing.

Peri-implant gas accumulation in response to magnesium-based musculoskeletal biomaterials:Reframing current evidence for preclinical research and clinical evaluation

Historically,the rapid degradation and massive gas release from magnesium(Mg)implants resulted in severe emphysema and mechanical failure.With the advent of new alloys and surface treatment methods,optimized Mg implants have re-entered clinics since last decade with reliable performance.However,the optimization aims at slowing down the degradation process,rather than exemption of the gas release.This study involved a systematic evaluation of current preclinical and clinical evidence,regarding the physical signs,symptoms,radiological features,pathological findings and complications potentially associated with peri±implant gas accumulation(PIGA)after musculoskeletal Mg implantation.The literature search identified 196 potentially relevant publications,and 51 papers were enrolled for further analysis,including 22 preclinical tests and 29 clinical studies published from 2005 to 2023.Various Mg-based materials have been evaluated in animal research,and the application of pure Mg and Mg alloys have been reported in clinical follow-ups involving multiple anatomical sites and musculoskeletal disorders.Soft tissue and intraosseous PIGA are common in both animal tests and clinical follow-ups,and potentially associated with certain adverse events.Radiological examinations especially micro-CT and clinical CT scans provide valuable information for quantitative and longitudinal analysis.While according to simulation tests involving Mg implantation and chemical processing,tissue fixation could lead to an increase in the volume of gas cavity,thus the results obtained from ex vivo imaging or histopathological evaluations should be interpreted with caution.There still lacks standardized procedures or consensus for both preclinical and clinical evaluation of PIGA.However,by providing focused insights into the topic,this evidence-based study will facilitate future animal tests and clinical evaluations,and support developing biocompatible Mg implants for the treatment of musculoskeletal disorders.

Preclinical in vivo research of magnesium-based implants for fracture treatment: A systematic review of animal model selection and study design

Background:Degradable magnesium implants are promising for clinical fracture treatment,providing less stress-shielding mechanical support and superior bone-strengthening benefits to traditional materials.The quality of preclinical research is essential for developing Mg implants;however,there are considerable variations in the model selection and study design in published studies,posing challenges for safe and effective clinical translation of lab discoveries.Objective:The purpose of this systematic review is to investigate the current progress on in vivo research of Mg implants for fracture treatment,focusing on model selection,Mg materials,implant design,methodological and analyzing techniques,aiming to provide comprehensive guidance for future preclinical research.Materials and methods:Pub Med and Embase online databases were searched to identify researches investigating Mg implants in animal models of fracture from 1960 to December 2019,using a combination of keywords:magnesium and fracture.Eligible studies were included without language restriction.Data extraction was conducted for qualitative analysis.Meta-analysis was not performed due to extensive heterogeneities among studies.Results:Twenty studies published from 2014 to 2019 were included.Publication information,animal model,methodological quality,implants preparation,and evaluation techniques were extracted.The methodological assessment revealed low to high risk of bias among studies.Both rodent and non-rodent species were selected,and the anatomical sites for inducing fractures included both cranial-facial and limb bones.Pure Mg and alloys with or without surface modification were evaluated,covering implant designs of both intra-medullary and extra-medullary fixation.Radiological and histological evaluations were commonly conducted.Conclusions:Published in vivo evidence confirmed the role of Mg-based implants in promoting fracture healing.However,considering heterogeneity in animal selection,implant preparation,and evaluation methods,there still lacks a standardized reference model.By analyzing the information extracted from included studies,the systematic review may facilitate planning and conducting preclinical research with translational perspective.

Effects of proteins on magnesium degradation-static vs.dynamic conditions

The interaction between organic molecules and biomaterial surfaces determines the fate of biomaterials during their service life,which is also the research hotspots in the field of biomaterials.To understand the mechanism of protein interaction with magnesium(Mg)degradation,alloying elements,immersion time,protein concentration and surface conditions have been previously considered for the effect of proteins on Mg degradation.However,fluid flow,as one of the critical factors,drew little attention in this case.In the present study,the effect of bovine serum albumin(BSA)and fetal bovine serum(FBS)on Mg degradation was compared under static and dynamic conditions.The results revealed that both BSA and FBS slightly decreased the degradation rate of Mg in Hanks’balanced salt solution(HBSS)under static immersion due to the protein adsorption and the formation of a Ca/P-rich top layer on Mg surface,whereas under dynamic flow condition the degradation of Mg was significantly accelerated in the presence of BSA or FBS.The reasons seemed to stem from the weakened protein adsorption on Mg surface in this case and the dynamically enhanced interaction between proteins and ions/products in solutions,which largely weaken the combination of the top Ca/P-rich layer with the inner corrosion product layer.These results highlight the importance of testing conditions for Mg characterization in vitro and the synergistic effect between different parameters on Mg degradation.

Treatment of osteoarthritis by implantation of Mg- and WE43-cylinders - A preclinical study on bone and cartilage changes and their influence on pain sensation in rabbits

With its main features of cartilage degeneration, subchondral bone sclerosis and osteophyte formation, osteoarthritisrepresents a multifactorial disease with no effective treatment options. As biomechanical shift in thetrabecular network may be a driver for further cartilage degeneration, bone enhancement could possibly delayOA progression. Magnesium is known to be osteoconductive and already showed positive effects in OA models.We aimed to use magnesium cylinders to enhance subchondral bone quality, condition of cartilage and painsensation compared to sole drilling in vivo. After eight weeks of implantation in rabbits, significant increase insubchondral bone volume and trabecular thickness with constant bone mineral density was found indicatingfavored biomechanics. As representative for pain, a higher number of CD271+ vessels were present in controlsamples without magnesium. However, this result could not be confirmed by sensitive, objective lamenessevaluation using a pressure sensing mat and no positive effect could be shown on either cartilage degenerationevaluated by OARSI score nor the presence of regenerative cells in CD271-stained samples. The presented resultsshow a relevant impact of implanted magnesium on key structures in OA pain with missing clinical relevanceregarding pain. Further studies with shifted focus should examine additional structures as joint capsule orosteophytes.

Multi-modal investigation of the bone micro- and ultrastructure, and elemental distribution in the presence of Mg-xGd screws at mid-term healing stages

Magnesium(Mg)–based alloys are becoming attractive materials for medical applications as temporary bone implants for support of fracture healing,e.g.as a suture anchor.Due to their mechanical properties and biocompatibility,they may replace titanium or stainless-steel implants,commonly used in orthopedic field.Nevertheless,patient safety has to be assured by finding a long-term balance between metal degradation,osseointegration,bone ultrastructure adaptation and element distribution in organs.In order to determine the implant behavior and its influence on bone and tissues,we investigated two Mg alloys with gadolinium contents of 5 and 10 wt percent in comparison to permanent materials titanium and polyether ether ketone.The implants were present in rat tibia for 10,20 and 32 weeks before sacrifice of the animal.Synchrotron radiation-based micro computed tomography enables the distinction of features like residual metal,degradation layer and bone structure.Additionally,X-ray diffraction and X-ray fluorescence yield information on parameters describing the bone ultrastructure and elemental composition at the bone-to-implant interface.Finally,with element specific mass spectrometry,the elements and their accumulation in the main organs and tissues are traced.The results show that Mg-xGd implants degrade in vivo under the formation of a stable degradation layer with bone remodeling similar to that of Ti after 10 weeks.No accumulation of Mg and Gd was observed in selected organs,except for the interfacial bone after 8 months of healing.Thus,we confirm that Mg-5Gd and Mg-10Gd are suitable material choices for bone implants.

In vitro and in vivo degradation behavior of Mg-0.45Zn-0.45Ca(ZX00)screws for orthopedic applications

Magnesium (Mg) alloys have become a potential material for orthopedic implants due to their unnecessary implant removal, biocompatibility, and mechanical integrity until fracture healing. This study examined the in vitro and in vivo degradation of an Mg fixation screw composed of Mg-0.45Zn-0.45Ca (ZX00, in wt.%). With ZX00 human-sized implants, in vitro immersion tests up to 28 days under physiological conditions, along with electrochemical measurements were performed for the first time. In addition, ZX00 screws were implanted in the diaphysis of sheep for 6, 12, and 24 weeks to assess the degradation and biocompatibility of the screws in vivo. Using scanning electron microscopy (SEM) coupled with energy dispersive X-ray spectroscopy (EDX), micro-computed tomography (μCT), X-ray photoelectron spectroscopy (XPS), and histology, the surface and cross-sectional morphologies of the corrosion layers formed, as well as the bone-corrosion-layer-implant interfaces, were analyzed. Our findings from in vivo testing demonstrated that ZX00 alloy promotes bone healing and the formation of new bone in direct contact with the corrosion products. In addition, the same elemental composition of corrosion products was observed for in vitro and in vivo experiments;however, their elemental distribution and thicknesses differ depending on the implant location. Our findings suggest that the corrosion resistance was microstructure-dependent. The head zone was the least corrosion-resistant, indicating that the production procedure could impact the corrosion performance of the implant. In spite of this, the formation of new bone and no adverse effects on the surrounding tissues demonstrated that the ZX00 is a suitable Mg-based alloy for temporary bone implants.