Could trace elements be linked to fracture healing acceleration in TBI-related skeletal multitrauma?

doi:10.21203/rs.3.rs-7623864/v1

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Could trace elements be linked to fracture healing acceleration in TBI-related skeletal multitrauma?

https://doi.org/10.21203/rs.3.rs-7623864/v1

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Dealing with trauma cases, physicians may have noticed that patients with concomitant TBI has shorter terms in bone callus formation. Such phenomenon was firstly reviewed by Morley back in 2005. Identification of the most significant factors affecting the acceleration of bone union may give opportunities for nonunion treatment. Materials and methods. To reproduce the isolated fracture and TBI-related polytrauma, 90 white female rats were used. According to the timing of the samples harvest(7, 14, 21 days), the animals were divided into six equal subgroups. Hip fracture was modeled by blunt guillotine injury, while for TBI modified control cortical impact was used. Results. At day 14 the concentration of both blood and bone copper increased in the polytrauma animals(p = 0.002) and in the sham(p = 0.004). There was a spike of 43% in the average blood copper in sham group on day 21. Meanwhile, bone copper decreased by 17%(p = 0.013). The difference in bone zinc concentration on day 21 was 25% in favor of the sham group (p < 0.001), while blood zinc at 21 days was amounted to 39% in favor of the polytrauma rats. On the 14th day the average concentration of blood magnesium reached equal concentrations for both groups (p = 0.47). At the time of day 21, bone magnesium of both groups was almost the same in favor of the multitrauma (p = 0.2). At the time of 14 days, the bone calcium of polytrauma animals increased by 1.4 times (p < 0.001). After reaching this peak on day 14, the calcium returned to a value equal to sham group one week later(p = 0.12). Conclusions. The critical period of acceleration could be day 14, which corresponds to the peak values of bone zinc, magnesium, and calcium. The third week was accompanied by a decrease in the bone trace elements with a simultaneous increase in the concentration of blood cadmium.

multitrauma

trace elements

traumatic brain injury

fracture healing

experimental animal models

While polytrauma is a leading cause of disability in young persons, among all cases of multiple injuries, the most common (approx. 19%) is combined head and limb injury[1, 2]. Dealing with trauma cases, physicians may have noticed that patients with concomitant TBI has shorter terms in bone callus formation. In 1991, Mueller et al. described the phenomenon of systemic increase of bone density after damage to the cortical layer of any bone of the axial skeleton in animal model [3]. However, only by 2005, Morley approved a similar phenomenon in patients with TBI-related polytrauma with increased mineral density, although the mechanism of acceleration remained unclear[4, 5]. This advantage was proved by separate studies by Morgan [6] and Tsitsilonis [7], where the healing stages in rats were shorter in the case of concomitant TBI.

The cause of acceleration is still unclear. In one of the latest reviews Hofman evaluated humoral and biochemical markers that may be responsible for accretion, no consistent results were obtained [8]. Other articles points on the possible influence of leptin, cytokine, growth factors, and neuropeptide Y. The dependence of plasma levels of bone-dependent trace elements: protocollagen, osteocalcin, and consolidation time was established by Stoffel [9]. There are several publications confirming a systemic decrease in bone density in patients with polytrauma with a more pronounced decrease in ipsilateral brain injury [10-14]. Unfortunately, Einhorn's retrospective and experimental studies of humoral factors affecting bone union also failed to reveal reliable results [15]. Reliable data is proving that the location of the brain injury has a crucial role in the acceleration phenomenon [16]. Identification of the most significant factors affecting the acceleration of bone union, in addition to deepening the understanding of the biology of the process, can open new courses for treating nonunion [17].

Data on the level of bone-related trace elements in polytrauma are very limited [18]. It is known that such trace elements as calcium, phosphorus, magnesium, fluorine, zinc, copper, and cadmium have a significant impact on bone metabolism, which served as the basis for the study of these trace elements in this paper.

All animal procedures were approved by the meeting of the local bioethics committee (protocol #124/21) and performed following ARRIVE v2.0 guidelines [19]. To reproduce the isolated hip fracture and TBI-related polytrauma, 90 sexually mature white female rats with an average weight of 207 (95% CI=197.2-216.8) g were used. Until reaching the required body weight and age, the animals were kept in standard cages under natural light conditions with access to food and water ad libitum for 4 animals each[20, 21. After the surgical interventions, the animals were allocated to cages of two. At the time of surgery, the rats had reached a least 12 weeks of age. Surgical interventions were performed in all animals during the same period of daylight hours. All procedures were carried out in compliance with the requirements of the European Convention for the Protection of Vertebrate Animals Used for Experimental and Other Scientific Purposes.

Group assignment. Adult animals were randomly divided into two groups: polytrauma group (n=45) and the isolated femur fracture (sham) group (n=45). According to the timing of the samples harvest(7, 14, 21 days), the animals were divided into six equal subgroups (n=15).

Isolated fracture model. All subsequent manipulations were performed under intraperitoneal anaesthesia using a combination of ketamine (65 mg/kg), xylazine (13 mg/kg), and acepromazine (1.25 mg/kg) [22]. For perioperative infection prevention cefazolin (50 mg/kg) was used under the data on the absence of any effect of cefazolin on fracture healing in rats [23]. To model a closed diaphyseal fracture of the femur in both groups, after skin incision and soft tissue separation, an entrance to the marrow canal was formed using a G18 needle [24]. A 0.8 mm Kirschner wire was inserted into the femoral canal, followed by a blunt guillotine blow with a kinetic energy of 1.6 J with a 1.5 mm limit to the edge of the table to reduce soft tissue injury. The signs of a fracture were confirmed palpably. In the absence of signs of fracture, the impact was repeated with an increase in kinetic energy to 2.15 J by increasing the fall height from 15 cm to 20 cm. The next step was to suture the knee joint wound with full-layer sutures (Ethilon 3-0), followed by dressing with a 10% povidone-iodine solution (Betadine). For post-procedural pain relief, a solution of ketoprofen (5 mg/kg) was injected subcutaneously twice within 4 hours.

TBI-related polytrauma model. The model was achieved by closed left-side TBI followed by closed right-side femur fracture by a method described above. The reproducibility of TBI was achieved according to the algorithm proposed by Flierl with slight modifications [25]. Under aseptic conditions, after skin incision in the area of the sagittal suture of the skull and visualization of the bregma, a blunt trauma with a kinetic energy of 42 mJ was applied in the projection of the motor cortex, which is sufficient to simulate mild to moderate TBI [25]. In our experiment, the injury was achieved by using a self-modeled device with a 4 mm wide blunt impactor and possible values for the kinetic force of 21, 42, and 63 mJ, which corresponds to different severity levels of TBI(mild-moderate-severe). The severity of traumatic brain injury was determined by using the neurological severity score (NSS) in small rodents, which was adapted to use the most representative time frames of its application - 1 hour and 4 hours after trauma[26]. NSS is a scoring system for assessing the behavior of rodents according to the main criteria, such as the ability to find an exit from a circle with a diameter of 30 cm, the absence of hemiparesis, the seeking behavior, and others[25]. A higher score indicates a more complex neurological injury. Next, the head wound was sutured with a continuous suture (Prolene 3-0), followed by wound dressing with 10% povidone-iodine solution (Betadine).

Material harvesting. One TBI-related polytrauma animal died before the sampling stage, and another one was excluded from the experiment due to the detection of a large intra-abdominal tumor at the harvesting. The concentration of copper, zinc, cadmium, magnesium and calcium in whole blood and femurs were determined using atomic absorption spectroscopy. In brief, the method is based on atomizing a solution of mineralizate in an air-acetylene flame and measuring the resonant absorption of element atoms by atomic spectrophotometer(Shimadzu AA-7000).

The statistical analysis of the results was performed using Microsoft Excel 365 with the additional add-on Xrealstats. The normality of the distribution was determined by the d'Agostino-Pearson and Shapiro-Wilk tests for α=0.05 in six subgroups [27]. Since the distribution of all 6 subgroups was normal (p>0.05 for normality tests), the data were interpreted using the mean and standard deviation (Mean±SD) accompanied by a 95% Confidence Interval(95% CI). The significance of intergroup data differences was determined using a two-sided Student's t_α test for small independent samples, with p<0.05 considered as significant [28].

Copper. On day 7(Fig. 1), the average concentration of whole blood copper in the polytrauma group was 10% higher than in the sham, but without a sufficiently significant difference between the groups (p = 0.07). In contrast, the concentration of bone copper of the poly group was significantly 1.7 times lower compared to the sham animals(p < 0.001). The average value of copper in the blood in both groups was close to equal on day 14. For the same term, the concentration of bone copper increased statistically significantly in both groups: by 36% (p = 0.002) in the polytrauma animals and by 28% in the sham group (p = 0.004). There was a spike of 43% in the average blood copper in sham group on day 21 (p < 0.0001), while in the main group this value did not change (p = 0.5), which led to a significant difference of 1.4 times between the groups (p < 0.001). Meanwhile, the average bone copper in sham group decreased by 17% (p = 0.013), while in polytrauma group, it did not change (p = 0.92) at all.

Zinc. The average blood zinc concentration(Fig. 2) in polytrauma group on day 7 was significantly lower by 29% compared to the sham (p = 0.002). For the bone zinc, we gained a significantly lower concentration by 14% on day 7(p = 0.01). While on day 14 the average value of blood zinc of sham group did not change compared to day 7 (p = 0.19), in multitrauma group it increased by 131% (p < 0.001) and by 55% in bones(p < 0.001). The difference in blood zinc at 21 days was statistically significant and amounted to 39% in favor of the poly group (p < 0.001). At the same time, the average value in the bones group significantly decreased by 1.7 times (p < 0.001), while in the sham group decreased only by 13% (p = 0.013). The difference in bone zinc concentration on day 21 was 25% in favor of the sham group (p < 0.001).

Cadmium. The average cadmium concentration both in blood and femurs between the sham and experimental groups differed significantly following day 7 in favor of the sham group (4.4 times for blood and 1.4 times for bones, p < 0.001). On day 14, the concentration of cadmium in the blood of the sham group significantly decreased by 6.2 times (p < 0.001), in polytrauma animals it increased almost twice (p < 0.001). At the same time, bone cadmium in sham group did not change within the statistical error (p = 0.34), and there was a significant increase in the polytrauma group by 25% (p < 0.001). In both the sham group and poly group, a significant increase (12.5 and 4.1 times, respectively, p < 0.001) in blood cadmium on day 21 was observed. The bone cadmium at same term statistically significant decreased by 19% only in polytrauma animals (p < 0.001).

Calcium. The average whole blood calcium of both sham and polytrauma group showed a tendency to statistically significant increase during all observed periods (p < 0.001). On the one hand, the sham group blood calcium increased by 12.6% on day 14 and by another 9% on day 21. On the other hand, the bone calcium showed an inverse tendency to decrease. The blood calcium concentration in case of polytrauma showed similar to isolated fracture increase by 7% on day 14 and by 13.5% in the next 7 days. Still, as of day 21 the average blood calcium in the sham group was significantly higher by 26.5% (p < 0.001). At the time of 14 days, the bone calcium in case of TBI-related polytrauma significantly increased by 1.4 times (p < 0.001). After reaching the peak value, on day 21 it’s returned to a value equal to that of the sham group (p = 0.12).

Copper. Copper acts as a coenzyme and affects the synthesis of hormones and neurotransmitters[29], which may take main role in neuroacceleration of bone repair. The musculoskeletal system is the main depot of copper[30]. Copper is a coenzyme in oxidoreductase processes, including collagen formation and normal calcium and phosphorus deposition [30]. Since copper is a coenzyme of lysyl oxidase, a decrease in its level disrupts the formation and cross-linking of collagen molecules, which leads to bone structure disorders and fragility[31]. A study conducted by Li indicates the inhibition of osteoclasts by copper molecules, i.e. a decrease in bone resorption [32]. Moreover, Qi et al. indicate the induction of osteoblast enzymes by copper molecules[33]. At the macrostructural level, the relationship between copper levels, age, and bone strength has been demonstrated in several studies [30; 34]. In our study, serum copper levels increased sharply at 21 days due to release from the bone depot in the group with isolated femoral injury, while bone copper concentration showed an inverse correlation. In general, the bone concentration of copper in case of TBI-related polytrauma was lower than in case of isolated fracture. This phenomenon may be related to the redistribution of the trace element between other bones of the skeleton, since the main copper depot in the body is the ribs and shin bones [30], and the phenomenon of acceleration has systemic nature [8].

Zinc. The function of zinc in osteogenesis is also limited to coenzyme function[30], although bones contain only up to 20% of the total zinc mass. Zinc acts as a coenzyme and regulates gene expression, especially during anabolic processes [35]. The zinc molecule is responsible for the formation of many hormones and hormone-like compounds, including growth factors [30]. This may be the reason for the data on the stimulation of osteoblasts and inhibition of osteoclasts by increased concentrations of the zinc transport protein (ZIP1) [36; 37]. A decrease in bone zinc concentration leads not only to impaired bone reparation but also to a decrease in bone mineral density [38] Again, the increase in bone zinc on day 14 in the poly group with a sharp decrease on day 21, may indicate that the most critical and active period of acceleration is day 14. A similar conclusion was reached by other authors in the histological examination of the bone callus [10, 39]

Cadmium. Although cadmium is better known for its toxicity, its role is to regulate bone resorption [35]. Increased cadmium concentrations reduce bone mineralisation, worsening their physical and mechanical properties [40, 41], causing an osteotoxic effect by inducing osteoblast apoptosis and disrupting the formation of their cytoskeleton [30, 42]. On the other hand, by inducing osteoclast function, cadmium may be one of the markers of the phases of reparative osteogenesis[43]. There are reports of osteoclast activation starting from week 2 in cranio-skeletal polytrauma [16]. The massive release of cadmium into the blood in both groups on day 21 may be caused by osteoclasts activity and, accordingly, bone resorption with the release of trapped cadmium. At the same time, the bone cadmium in polytrauma decreased, which may indicate a more active resorption at the third week.

Magnesium. The effect of magnesium on fusion remains controversial. Some authors point to the falsity of the paradigm about the osteogenic effect of the trace element, pointing to its suppression in the process of hydroxyapatite formation [35]. Magnesium is involved as a coenzyme in more than 300 enzyme systems and energy-dependent cellular functions [30]. It is also an inducer of osteoblast function as a coenzyme in phosphatase enzymes [44, 45]. Since bone tissue is the main dynamic depot of magnesium, changes in its concentration can be a marker of bone repair and mineralisation activity [46]. In our study, the highest concentration of magnesium in both the blood and bone was observed on day 14, which further confirms the statement about the increased activity of bone repair during this period.

Calcium. Calcium plays a major role in mineralisation and is responsible for hard callus formation [30]. The process of remodelling and repair is calcium-dependent due to its direct effect on osteon cells [47]. This trace element is critical for the functioning of the human body, affecting the function of both intra- and extracellular processes [48]. More than 99% of calcium mass is stored in the form of hydroxyapatite [49] Due to its direct involvement in bone remodelling, calcium is one of the markers of bone resorption and osteogenesis[49]. We found a peak increase in calcium concentration on day 14 in the polytrauma group compared to a gradual decrease in the sham group against the background of an increase in serum calcium. This trend may be a marker of significantly more active remodelling of the injured bone in the presence of a cranial component of polytrauma.

The severity of anabolic processes at the level of soft bone regenerate can be indicated by the average concentrations of trace elements that are part of coenzymes - zinc, magnesium and copper. According to the results obtained, it was on day 14 that the bone concentration of copper, zinc and magnesium increased in the case of TBI-related multitrauma. Whereas in the group of animals without TBI, the concentration of these trace elements increased slightly or, on the contrary, decreased during the 14th day. An increase in the activity of anabolic processes in polytrauma after 2 weeks was also observed in Locher's study [5], observing the filling of the osteotomy cavity. In addition, the femur in the polytrauma group showed greater resistance to torsional loads after 3 weeks [50]. The decrease in bone concentrations of copper, zinc and magnesium found in our study against the background of decreased bone mineralisation with calcium in the poly group at 21 days may be a direct cause of the decrease in bone resistance to torsional loads.

Study limitations. It is necessary to take into account the physiological differences in the process and, especially, in the timing of bone regeneration in rodents and humans. The sample size for certain study periods is small, which requires further research using a larger sample size for area of interest. At the stage of study design, we faced a lack of previous studies on the analysis of trace elements in polytrauma, especially in humans. Exactly why representativeness may be one of the study limitations. Great care should be taken when applying the results of this study in the clinic.

The presence of TBI in polytrauma directly affects the concentration of trace elements in both blood and bone tissue.

In the animals of the polytrauma group, a significantly lower concentration of copper and calcium in the blood and bone tissue was observed, but a higher concentration of zinc. The average concentration of copper and cadmium in the bone tissue was lower for polytrauma.

The critical period of acceleration is considered to be day 14, which corresponds to the peak values of bone zinc, magnesium, and calcium. The third week was accompanied by a sharp decrease in the bone trace elements with a simultaneous increase in the concentration of cadmium in the blood, indicating the predominance of bone resorption.

Further clinical, experimental, and histological studies are needed to identify possible factors that accelerate bone healing in TBI-related polytrauma.

Could trace elements be linked to fracture healing acceleration in TBI-related skeletal multitrauma?

Declaration of Conflicting Interest: The Authors declare that there is no conflict of interest

Funding: This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.

Ethical approval: Ethical approval for this study was obtained from Ivano-Frankivsk local committee of bioethics (protocol #124/21 from 29/11/2021)

Data Availability: The datasets generated during and/or analysed during the current study are available in the “Experimental dataset” repository on URL https://shorturl.at/0RQp0 or Experimental dataset.

Contributorship: Bihun Roman researched literature and conceived the study, involved in protocol development, gaining ethical approval, and performed data analysis, wrote, and translate the manuscript draft. Sulyma Vadym was involved in protocol development, provided mentoring and draft review as well as approve published version. Sribniak Andrii was involved in gathering data, involved in protocol development. The draft of the manuscript was written by Ruslana Bihun. All authors reviewed and edited the manuscript and approved the final version of the manuscript.

Acknowledgements: We would like to thank the stuff of Bioelementology center of Ivano-Frankivsk national medical university for their assistance in conducting this research. We also would like to thank Andrii Sachko for help in conducting research and gathering material for laboratory data.

Gunawan B, Dumastoro R, Kamal A (2018) Trauma and Injury Severity Score in Predicting Mortality of Polytrauma Patients. eJournal Kedokteran Indonesia 5. 10.23886/ejki.5.8148
Turculeţ CŞ, Georgescu TF, Iordache F et al (2021) Polytrauma: The European Paradigm. Chirurgia (Bucur) 116(6):664–668. 10.21614/chirurgia.116.6.664
Mueller M, Schilling T, Minne HW et al (1991) A systemic acceleratory phenomenon (SAP) accompanies the regional acceleratory phenomenon (RAP) during healing of a bone defect in the rat. J Bone Min Res 6(4):401–410. 10.1002/jbmr.5650060412
Morley J, Marsh S, Drakoulakis E et al (2005) Does traumatic brain injury result in accelerated fracture healing? Injury 36(3):363–368. 10.1016/j.injury.2004.08.028
Locher RJ, Lünnemann T, Garbe A et al (2015) Traumatic brain injury and bone healing: radiographic and biomechanical analyses of bone formation and stability in a combined murine trauma model. J Musculoskelet Neuronal Interact 15(4):309–315
Morgan EF, De Giacomo A, Gerstenfeld LC (2014) Overview of skeletal repair (fracture healing and its assessment). Methods Mol Biol 1130:13–31. 10.1007/978-1-62703-989-5_2
Tsitsilonis S, Seemann R, Misch M et al (2015) The effect of traumatic brain injury on bone healing: an experimental study in a novel in vivo animal model. Injury 46(4):661–665. 10.1016/j.injury.2015.01.044
Hofman M, Koopmans G, Kobbe P et al (2015) Improved fracture healing in patients with concomitant traumatic brain injury: proven or not? Mediators Inflamm 2015:204842. 10.1155/2015/204842
Stoffel K, Engler H, Kuster M et al (2007) Changes in biochemical markers after lower limb fractures. Clin Chem 53(1):131–134. 10.1373/clinchem.2006.076976
Brady RD, Shultz SR, Sun M et al (2016) Experimental Traumatic Brain Injury Induces Bone Loss in Rats. J Neurotrauma 33(23):2154–2160. 10.1089/neu.2014.3836
Smith É, Comiskey C, Carroll Á (2016) Prevalence of and risk factors for osteoporosis in adults with acquired brain injury. Ir J Med Sci 185(2):473–481. 10.1007/s11845-016-1399-5
Oppl B, Michitsch G, Misof B et al (2014) Low bone mineral density and fragility fractures in permanent vegetative state patients. J Bone Miner Res. ;29(5):1096 – 100. 10.1002/jbmr.2122. PMID: 24470043
Beaupre GS, Lew HL (2006) Bone-density changes after stroke. Am J Phys Med Rehabil 85(5):464–472. 10.1097/01.phm.0000214275.69286.7a
Smith EM, Comiskey CM, Carroll AM (2009) A study of bone mineral density in adults with disability. Arch Phys Med Rehabil 90(7):1127–1135. 10.1016/j.apmr.2008.09.578
Einhorn TA, Gerstenfeld LC (2015) Fracture healing: mechanisms and interventions. Nat Rev Rheumatol 11(1):45–54. 10.1038/nrrheum.2014.164
Morioka K, Marmor Y, Sacramento JA et al (2019) Differential fracture response to traumatic brain injury suggests dominance of neuroinflammatory response in polytrauma. Sci Rep 9(1):12199. 10.1038/s41598-019-48126-z
Einhorn TA, Gerstenfeld LC (2015) Fracture healing: mechanisms and interventions. Nat Rev Rheumatol 11(1):45–54. 10.1038/nrrheum.2014.164
Ciosek Ż, Kot K, Kosik-Bogacka D et al (2021) The Effects of Calcium, Magnesium, Phosphorus, Fluoride, and Lead on Bone Tissue. Biomolecules 11(4):506. 10.3390/biom11040506
Percie du Sert N, Ahluwalia A, Alam S et al (2020) Reporting animal research: Explanation and elaboration for the ARRIVE guidelines 2.0. PLoS Biol 18(7):e3000411. 10.1371/journal.pbio.3000411
National Research Council (US) (2011) Committee for the Update of the Guide for the Care and Use of Laboratory Animals. Guide for the Care and Use of Laboratory Animals, 8th edn. National Academies Press (US), Washington (DC)
Makowska IJ, Weary DM (2021) A Good Life for Laboratory Rodents? ILAR J. ;60(3):373–388. 10.1093/ilar/ilaa001. PMID: 32311030
Ahmadi-Noorbakhsh S, Farajli Abbasi M, Ghasemi M et al (2022) Anesthesia and analgesia for common research models of adult mice. Lab Anim Res 38(1):40. 10.1186/s42826-022-00150-3
Natividad-Pedreño M, Nuñez-Chia A, Cobo-Valenzuela N et al (2016) Effect of cefazolin and cefuroxime on fracture healing in rats. Injury 47(Suppl 3):S3–S6. 10.1016/S0020-1383(16)30599-X
Aurégan JC, Coyle RM, Danoff JR et al (2013) The rat model of femur fracture for bone and mineral research: An improved description of expected comminution, quantity of soft callus and incidence of complications. Bone Joint Res 2(8):149–154. 10.1302/2046-3758.28.2000171
Flierl MA, Stahel PF, Beauchamp KM et al (2009) Mouse closed head injury model induced by a weight-drop device. Nat Protoc 4(9):1328–1337. 10.1038/nprot.2009.148
Tsenter J, Beni-Adani L, Assaf Y et al (2008) Dynamic changes in the recovery after traumatic brain injury in mice: effect of injury severity on T2-weighted MRI abnormalities, and motor and cognitive functions. J Neurotrauma 25(4):324–333. 10.1089/neu.2007.0452
Lang TA, Altman DG (2015) Basic statistical reporting for articles published in biomedical journals: the Statistical Analyses and Methods in the Published Literature or the SAMPL Guidelines. Int J Nurs Stud 52(1):5–9. 10.1016/j.ijnurstu.2014.09.006
Tan YD (2022) Two-sample tα -test for testing hypotheses in small-sample experiments. Int J Biostat 19(1):1–19. 10.1515/ijb-2021-0047
Vetlényi E, Rácz G (2020) A réz élettani funkciója, a rézfelhalmozódás és a rézhiány kóroktani szerepe [The physiological function of copper, the etiological role of copper excess and deficiency]. Orv Hetil 161(35):1488–1496 Hungarian. 10.1556/650.2020.31854
Ciosek Ż, Kot K, Rotter I, Iron (2023) Zinc, Copper, Cadmium, Mercury, and Bone Tissue. Int J Environ Res Public Health 20(3):2197. 10.3390/ijerph20032197
Kabata-Pendias A, Mukherjee AB (2007) Trace elements from soil to human. Springer-, Berlin. http://dx.doi.org/10.1007/978-3-540-32714-1
Li BB, Yu SF (2007) [In vitro study of the effects of copper ion on osteoclastic resorption in various dental mineralized tissues]. Zhonghua Kou Qiang Yi Xue Za Zhi 42(2):110–113 Chinese
Qi Y, Wang H, Chen X et al (2021) The role of TGF-β1/Smad3 signaling pathway and oxidative stress in the inhibition of osteoblast mineralization by copper chloride. Environ Toxicol Pharmacol 84:103613. 10.1016/j.etap.2021.103613
Kuo HW, Kuo SM, Chou CH et al (2000) Determination of 14 elements in Taiwanese bones. Sci Total Environ 255(1–3):45–54. 10.1016/s0048-9697(00)00448-4
Hara T, Takeda TA, Takagishi T et al (2017) Physiological roles of zinc transporters: molecular and genetic importance in zinc homeostasis. J Physiol Sci 67(2):283–301. 10.1007/s12576-017-0521-4Epub 2017 Jan 27. PMID: 28130681; PMCID: PMC10717645
Lioumi M, Ferguson CA, Sharpe PT et al (1999) Isolation and characterization of human and mouse ZIRTL, a member of the IRT1 family of transporters, mapping within the epidermal differentiation complex. Genomics 62(2):272–280. 10.1006/geno.1999.5993
Seo HJ, Cho YE, Kim T et al (2010) Zinc may increase bone formation through stimulating cell proliferation, alkaline phosphatase activity and collagen synthesis in osteoblastic MC3T3-E1 cells. Nutr Res Pract 4(5):356–361. 10.4162/nrp.2010.4.5.356
Helliwell TR, Kelly SA, Walsh HP et al (1996) Elemental analysis of femoral bone from patients with fractured neck of femur or osteoarthrosis. Bone 18(2):151–157. 10.1016/8756-3282(95)00440-8
Collier CD, Hausman BS, Zulqadar SH et al (2020) Characterization of a reproducible model of fracture healing in mice using an open femoral osteotomy. Bone Rep 12:100250. 10.1016/j.bonr.2020.100250
Engström A, Michaëlsson K, Vahter M et al (2012) Associations between dietary cadmium exposure and bone mineral density and risk of osteoporosis and fractures among women. Bone 50(6):1372–1378. 10.1016/j.bone.2012.03.018
Wallin M, Barregard L, Sallsten G et al (2016) Low-Level Cadmium Exposure Is Associated With Decreased Bone Mineral Density and Increased Risk of Incident Fractures in Elderly Men: The MrOS Sweden Study. J Bone Min Res 31(4):732–741. 10.1002/jbmr.2743
Bhattacharyya MH (2009) Cadmium osteotoxicity in experimental animals: mechanisms and relationship to human exposures. Toxicol Appl Pharmacol 238(3):258–265. 10.1016/j.taap.2009.05.015
Al-Ghafari A, Elmorsy E, Fikry E et al (2019) The heavy metals lead and cadmium are cytotoxic to human bone osteoblasts via induction of redox stress. PLoS ONE 14(11):e0225341. 10.1371/journal.pone.0225341
Leidi M, Dellera F, Mariotti M et al (2011) High magnesium inhibits human osteoblast differentiation in vitro. Magnes Res 24(1):1–6. 10.1684/mrh.2011.0271
Rondanelli M, Faliva MA, Tartara A et al (2021) An update on magnesium and bone health. Biometals 34(4):715–736. 10.1007/s10534-021-00305-0
Kosik-Bogacka DI, Lanocha-Arendarczyk N, Kot K et al (2018) Calcium, magnesium, zinc and lead concentrations in the structures forming knee joint in patients with osteoarthritis. J Trace Elem Med Biol 50:409–414. 10.1016/j.jtemb.2018.08.007
Ciosek Ż, Kot K, Kosik-Bogacka D et al (2021) The Effects of Calcium, Magnesium, Phosphorus, Fluoride, and Lead on Bone Tissue. Biomolecules 11(4):506. 10.3390/biom11040506
Beto JA (2015) The role of calcium in human aging. Clin Nutr Res 4(1):1–8. 10.7762/cnr.2015.4.1.1
Fischer V, Haffner-Luntzer M, Amling M et al (2018) Calcium and vitamin D in bone fracture healing and post-traumatic bone turnover. Eur Cell Mater 35:365–385. 10.22203/eCM.v035a25
Kesavan C, Rundle C, Mohan S (2022) Repeated mild traumatic brain injury impairs fracture healing in male mice. BMC Res Notes 15(1):25. 10.1186/s13104-022-05906-7

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Reviewers agreed at journal
23 Sep, 2025
Reviewers invited by journal
17 Sep, 2025
Editor assigned by journal
15 Sep, 2025
Submission checks completed at journal
15 Sep, 2025
First submitted to journal
15 Sep, 2025

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Could trace elements be linked to fracture healing acceleration in TBI-related skeletal multitrauma?

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Status:

Version 1