Composition for promoting rapid bone rehabilitation after limb waste reloading and composite nutritional supplement and application thereof

Abstract

The invention discloses a composition for promoting rapid rehabilitation of bones after limb waste reloading and a composite nutritional supplement and application thereof, and belongs to the technical field of bone injury and osteoporosis rehabilitation. The composition disclosed by the invention consists of glutamine, glucose and branched chain amino acid. The composition is a nutrition regulation scheme based on cell energy metabolism and can be further prepared into a nutritional supplement, and the components in the composition are low in cost, easy to obtain, safe to use and easy to widely popularize and popularize clinically. The composition not only has a positive effect on rapid recovery of limb waste reloaded bones, but also has a positive effect on recovery of muscle atrophy after reloading.

Description

Technical Field

[0001] This invention belongs to the field of bone injury and osteoporosis rehabilitation technology, and relates to a composition that promotes rapid bone recovery after disuse reloading of limbs, its compound nutritional supplement, and its application. Background Technology

[0002] The skeletal system is an extremely complex and highly sensitive biological structure. It not only provides support and protection for the body but also adapts to changes in the external mechanical environment through continuous self-adjustment and remodeling. Under normal circumstances, bones can sense the mechanical stimuli brought about by body activity and adapt to these changes by increasing bone density and improving bone structure to enhance their load-bearing capacity. However, a lack of mechanical stimulation leads to bone degeneration and bone loss, causing disuse bone loss. First, patients who are bedridden for extended periods (such as those with spinal cord injuries, traumatic brain injuries, or severe illnesses) experience a rapid decline in bone density due to reduced mechanical stimulation of the bones caused by prolonged lack of normal weight-bearing activity. Data shows that weight-bearing bone loss in middle-aged and elderly people over 50 years old who are bedridden for extended periods is at a rate of >3% per month. Second, fixation methods for fractured limbs, such as plaster casts, splints, external fixators, and internal fixation surgery, restrict patient movement, resulting in a lack of appropriate mechanical stimulation of the bones and subsequent bone loss. Data shows that long bone disuse after a fracture for 3 months leads to more than 40% muscle and bone loss, increasing the risk of refracture by nearly 5 times. Secondly, another common and significant scenario for disuse bone loss is the weightlessness experienced by astronauts during spaceflight. Weightlessness-induced bone loss is the leading risk factor affecting astronauts' health. During spaceflight, astronauts lose approximately 1% to 2% of their bone mass per month, equivalent to the annual bone loss of a postmenopausal woman. Despite disuse bone loss being a serious health problem, current technological means to effectively inhibit this phenomenon remain limited. Studies show that even after long-term bedridden patients or astronauts resume normal weight-bearing activities, disuse bone loss continues, and the time required for bone recovery after reloading is approximately 3 to 4 times longer than during unloading, leaving them at a long-term high risk of fracture. Furthermore, even after prolonged recovery, patients' bone mass and quality may not fully recover to pre-disuse levels, indicating that normal weight-bearing activity cannot fully stimulate bone recovery in patients with disuse bone loss, and their skeletal biomechanical sensitivity is significantly reduced. Currently, numerous studies focus on bone loss caused by disuse / unloading, but there are few reports on interventions for bone recovery after reloading. Therefore, improving the biomechanical sensitivity of bones after disuse is of great significance for accelerating the rapid rehabilitation of limbs after disuse.

[0003] Adult bone tissue primarily contains three cell types: osteoblasts, osteoclasts, and osteocytes. Osteocytes, derived from osteoblasts, account for 95% of all bone cells in adults and are the most abundant and longest-lived bone cells, with a lifespan of up to 50 years. Osteocytes are deeply embedded in the mineralized bone matrix and interconnected via synapses, forming a vast network within the lacunar infundibulocochlear system (LCS). Previously considered inert, non-functional cells within the mineralized bone matrix, osteocytes have been increasingly recognized as key mechanosensors in bone, potentially communicating with osteoclasts and osteoblasts on the bone surface via synapses to regulate bone remodeling and remodeling processes. Osteocytes regulate bone formation and resorption by releasing cytokines such as receptor activator of nuclear factor κB ligand (RANKL), sclerostin, and osteoprotegerin (OPG). Cytokines such as RANKL and Sclerostin released by osteocytes provide new potential drug targets for the treatment of osteoporosis, making osteocytes a novel interventional cell for the prevention and treatment of osteoporosis. However, the mechanisms by which osteocytes sense and respond to mechanical signals during unloading and reloading processes and regulate bone resorption and bone formation remain unclear.

[0004] Calcium ions are important and ubiquitous intracellular second messengers, regulating various cellular processes and participating in early signaling responses of osteoblasts to external physical stimuli. Intracellularly, calcium ion concentration is not constant but dynamically changes in an oscillatory manner, known as calcium oscillation. Compared to a constant or continuously rising calcium ion concentration, this oscillatory calcium signaling pattern can more efficiently and precisely promote intracellular gene transcription. Previous studies have demonstrated that macroscopic mechanical stimulation of bone tissue can induce interstitial fluid flow in the lacunar-tubular microenvironment, triggering calcium oscillations in osteoblasts characterized by strong, repetitive calcium spikes. Furthermore, it was found that the calcium kinetic response of osteoblasts to mechanical stimulation is more significant than that of osteoblasts (Jing D, et al. Bone. 2013, 53:531-40; Jing D, et al. FASEB J. 2014, 28:1582-92). Mechanically induced osteocyte calcium oscillations regulate the secretion of various osteocyte-related cytokines (such as OPG, RANKL, and Sclerostin), thereby modulating the activity and function of osteoclasts and osteoblasts.

[0005] Furthermore, the applicant's research group's latest findings (Liu X, et al. J Clin Invest, 2023, 133:e164508) revealed that mice with hindlimb immobilization for 4 weeks showed significantly lower increases in bone mass and mechanical strength after stress reloading compared to normal mice, indicating a close correlation between impaired bone recovery and reduced bone stress sensitivity after reloading of disused bone tissue. Simultaneously, the study also found that the in-situ calcium signaling response of osteocytes in the tibia of mice with hindlimb immobilization for 4 weeks was significantly weakened after cyclic stress reloading, suggesting that the reduced stress sensitivity of disused bone tissue to mechanical reloading may be significantly related to the reduced calcium signaling response of osteocytes to mechanical reloading. Inhibition of adenine triphosphate (ATP) synthesis in mitochondria and blocking of the ATP-related P2 receptor (P2R) signaling pathway both significantly inhibited stress-induced osteocyte calcium oscillation response, enhancing stress-induced osteocyte calcium oscillation highly dependent on ATP. Further transcriptome sequencing revealed that prolonged disuse significantly altered the energy metabolism of osteocytes, manifested as decreased oxidative phosphorylation and significantly increased glycolysis, leading to a significant reduction in ATP synthesis. Interestingly, osteocytes in long-term disused bone tissue showed a highly significant and specific increase in glutaminase 2 (GLS2) and pyruvate dehydrogenase kinase 1 (PDK1). Therefore, the above research suggests that regulating glutamine and glucose metabolism in osteocytes to restore ATP energy production and improve osteocyte calcium oscillation dynamics may be an effective technique for accelerating rapid bone quality recovery after long-term bone tissue reloading.

[0006] In general, existing technologies for disuse bone injury mainly focus on how to inhibit and counteract bone loss caused by disuse during limb disuse. However, there are currently no effective technologies for promoting rapid bone recovery after stress reloading, especially for improving the stress response of disused bone tissue after mechanical reloading. Summary of the Invention

[0007] In order to overcome the shortcomings of existing technologies, such as numerous obstacles and slow progress in bone recovery after long-term disuse bone tissue reloading, the purpose of this invention is to provide a composition that promotes rapid bone recovery after limb disuse reloading, its compound nutritional supplement, and its application.

[0008] To achieve the above objectives, the present invention employs the following technical solution: This invention discloses a composition for promoting rapid bone recovery after disuse reloading of limbs, which consists of glutamine, glucose and branched-chain amino acids.

[0009] Preferably, the molar ratio of glutamine, glucose, and branched-chain amino acids in the composition is 2-16:4-16:0.05-2. All molar concentrations of each component are expressed in mmol / L.

[0010] More preferably, the molar ratio of glutamine, glucose and branched-chain amino acids in the composition is 10:10:1.

[0011] Preferably, the branched-chain amino acids include leucine, isoleucine, and valine, and the molar ratio of leucine, isoleucine, and valine is 2:1:1.

[0012] The present invention also discloses the use of the above-described composition for promoting rapid bone recovery after disuse reloading of limbs in the preparation of a medicament for promoting rapid bone recovery after disuse reloading of limbs.

[0013] The present invention also discloses the use of the above-mentioned composition for promoting rapid bone recovery after disuse reloading of limbs in the preparation of a medicament for promoting the recovery of muscle atrophy after disuse reloading of limbs.

[0014] The present invention also discloses a compound nutritional supplement that promotes rapid bone recovery after disuse reloading of limbs, characterized in that it is composed of the above-mentioned composition for promoting rapid bone recovery after disuse reloading of limbs with or without the addition of other nutritional components.

[0015] Preferably, the compound nutritional supplement is administered orally or intravenously.

[0016] Preferably, the compound nutritional supplement is administered once daily, with each administration consisting of 10 grams of glutamine, 10 grams of glucose, and 1 gram of branched-chain amino acids.

[0017] Preferably, other nutrients include one or more of the following: bone metabolism-related nutrients, energy metabolism aids, tissue repair and immune modulators, and food-grade excipients.

[0018] Compared with the prior art, the present invention has the following beneficial effects: This invention takes regulating cellular energy metabolism to enhance osteocyte stress response as its innovative entry point, constructing a new technical solution based on nutritional regulation. It discloses a composition that promotes rapid bone recovery after disuse reloading, providing a novel approach to the rehabilitation of disuse bone injuries. Through cell experiments, animal experiments, and multi-dimensional clinical experiments, this invention has identified a composition consisting of glutamine, glucose, and branched-chain amino acids. The nutrients in this composition are inexpensive and readily available. Supplements made from this composition significantly enhance the number and intensity of calcium signal peaks in the disuse osteocyte network under fluid shear stress stimulation. By regulating the ATP synthesis efficiency of osteocytes, it improves the calcium oscillation dynamics, fundamentally improving the stress response of disuse bone tissue to mechanical reloading. Clinical trials have confirmed that this composition is significantly more effective than the control group and the glutamine-only group in restoring bone density, effectively solving the problem of existing technologies failing to promote rapid recovery of disuse bone tissue after reloading.

[0019] Furthermore, through experimental verification, this invention has clarified the optimal dosage of each component, avoiding efficacy reduction or safety risks caused by excessive dosage. This invention discloses an optimized ratio of glutamine, glucose, and branched-chain amino acids of 10:10:1 (4mM:4mM:0.4mM at the cellular level, 600mg / kg:600mg / kg:60mg / kg at the animal level, and 5-10g:5-10g:0.5-1g daily for clinical adults).

[0020] Through experimental verification, this invention shows that the nutritional supplement can not only actively promote the rapid recovery of bone after the reloading of disused bone tissue, but also have a positive effect on the recovery of muscle atrophy that accompanies the reloading. It breaks through the limitations of single bone rehabilitation technology, realizes the synergistic improvement of bone and muscle-related injuries, and improves the overall rehabilitation effect. Attached Figure Description

[0021] Figure 1 The results show the optimal glutamine concentration screening based on in vitro osteocyte calcium signaling dynamics; where A represents the effect of different glutamine concentrations on the calcium signal response ability of osteocytes in waste bone tissue under fluid shear force, and B is a statistical graph of the number and intensity of calcium signal peaks; (compared with the control group: *** P <0.001; compared with the obsolete group: # P <0.05, ## P <0.01, ### P <0.001) Figure 2 The results of screening for the optimal glucose concentration based on in vitro osteocyte calcium signaling kinetics (compared with the obsolete group): ** P <0.01); where A is the effect of different glucose concentrations on the calcium signal response ability of osteocytes in in vitro waste bone tissue under the action of fluid shear force, and B is a statistical graph of the number and intensity of calcium signal peaks; Figure 3 The results of screening for optimal branched-chain amino acid concentrations based on in vitro osteocyte calcium signaling dynamics (compared to the obsolete group): * P <0.05, *** P <0.001); where A is the effect of different concentrations of branched-chain amino acids on the calcium signal response ability of osteocytes in in vitro waste bone tissue under the action of fluid shear force, and B is a statistical graph of the number and intensity of calcium signal peaks; Figure 4The results of screening for the optimal glutamine concentration based on an animal model of discarded bone tissue (compared with the control group): *** P <0.001; compared with the obsolete group: # P <0.05, ## P <0.01); where A is the experimental flowchart, B is the effect of different concentrations of glutamine on the bone microstructure after mechanical reloading of waste bone tissue in vivo, C is the effect of different concentrations of glutamine on the trabecular morphology of waste bone tissue after mechanical reloading in vivo, and D is the effect of different concentrations of glutamine on the mechanical response of waste bone tissue after mechanical reloading in vivo. Figure 5 The results of screening for optimal branched-chain amino acid concentrations based on a waste bone tissue animal model (compared to the waste group): ** P <0.01, *** P <0.001); where A is the experimental flowchart, B is the effect of different concentrations of branched-chain amino acids on the bone microstructure after mechanical reloading of waste bone tissue in vivo, C is the effect of different concentrations of branched-chain amino acids on the trabecular morphology of waste bone tissue after mechanical reloading in vivo, and D is the effect of different concentrations of branched-chain amino acids on the bone mechanical response capacity after mechanical reloading of waste bone tissue in vivo. Detailed Implementation

[0022] To enable those skilled in the art to better understand the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

[0023] It should be noted that the terms "first," "second," etc., in the specification, claims, and accompanying drawings of this invention are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged where appropriate so that the embodiments of the invention described herein can be implemented in orders other than those illustrated or described herein. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover a non-exclusive inclusion; for example, a process, method, system, product, or apparatus that comprises a series of steps or units is not necessarily limited to those steps or units explicitly listed, but may include other steps or units not explicitly listed or inherent to such processes, methods, products, or apparatus.

[0024] The present invention will now be described in further detail with reference to the accompanying drawings: The composition and its compound nutritional supplement disclosed in this invention for promoting rapid bone recovery after disuse reloading of limbs are based on research on promoting rapid bone recovery of disused bone tissue after stress reloading. (1) Previous studies have found that bone recovery impairment after reloading of disused bone tissue is closely related to the decrease in bone stress sensitivity; (2) Previous studies have found that the decrease in bone stress sensitivity is mediated by the decrease in the calcium signal response ability of osteocytes to mechanical reloading; (3) Stress-induced osteocyte calcium oscillation is highly dependent on ATP, and transcriptome sequencing has revealed that long-term disuse has significantly altered the energy metabolism function of osteocytes, manifested as weakened oxidative phosphorylation and significantly enhanced glycolysis, resulting in a significant decrease in ATP synthesis; (4) Transcriptome sequencing also revealed that osteocytes of long-term disused bone tissue showed a significant specific increase in GLS2 and PDK1.

[0025] Based on the aforementioned innovative discoveries, a solid theoretical foundation has been provided for the effective composition proposed in this invention that promotes rapid bone recovery after reloading of disused bone tissue based on the regulation of bone cell energy metabolism.

[0026] The nutritional supplement promotes rapid bone recovery after reloading of disused bone tissue, primarily by increasing the stress sensitivity of the bone.

[0027] The nutritional supplement enhances the stress sensitivity of disused bone tissue after reloading primarily by increasing the ATP synthesis efficiency of bone cells, thereby improving the calcium oscillation dynamics of bone cells under stress.

[0028] The nutritional supplement is a composition of glutamine, glucose, and branched-chain amino acids. While individual glutamine, glucose, and branched-chain amino acids can improve the speed of bone recovery after reloading to a certain extent, the intervention effect of the composition is far superior to that of a single component. The optimal ratio of the nutritional formula in the composition to promote rapid bone recovery after reloading is glutamine:glucose:branched-chain amino acids at a concentration ratio of 10:10:1. Other ratios can also produce certain effects, but the glutamine:glucose:branched-chain amino acid ratio of 10:10:1 is the most effective.

[0029] In the aforementioned nutritional supplement, the optimal ratio of leucine, valine, and isoleucine among the branched-chain amino acids is 2:1:1. Other ratios may also produce certain effects, but the above-mentioned concentration ratio is optimal.

[0030] In addition to its positive effect on rapid bone recovery after limb disuse reloading, this technical solution also has a positive effect on the recovery of muscle atrophy after reloading.

[0031] Example 1 Mouse-derived MLO-Y4 bone cell line was cultured in α-MEM medium containing 5% fetal bovine serum and 5% calf serum. A 2D vortex mixer was used to simulate microgravity during cell culture. MLO-Y4 cells were cultured at 3.5 × 10⁻⁶ cells / day before in vitro culture. 5 Seeds were planted at a density of 1 cell / flask at T-25 cm⁻¹. 2 Cell culture flasks. Cells are placed in a 37°C, 5% CO2 cell culture incubator overnight. When the cells reach 40% confluence, preheated complete culture medium is added to fill the flasks, and all air bubbles are removed. The prepared cell culture flasks are placed on a rotary table and rotated around a horizontal axis at 24 revolutions per minute for 48 hours, generating a microgravity equivalent to near-Earth orbit (approximately 1 × 10⁻⁶). 2 G). During the experiment, the rotary instrument was placed in a 37 °C, 5% CO2 cell culture incubator, while the control group was cultured normally in the same incubator. Subsequently, the cells were cultured at 3.5 × 10⁻⁶ cells / year. 5 Cells were seeded at a density of 1 cell / slide onto slides coated with type I collagen from mouse tails. The cells were incubated for 20 min with a 10 μM Calbryte-520 AM calcium fluorescent probe, rinsed repeatedly, and allowed to stand for 10 min. The cell slides were then carefully mounted into a laminar flow chamber. Calcium fluorescence imaging was performed using a laser confocal microscope with a 10× Olympus objective, an excitation wavelength of 488 nm, and an absorption wavelength of 520 nm. The total fluorescence acquisition time was 10 min, including a 1 min baseline control (no fluid stimulation) and 9 min of oscillating fluid shear stimulation. One frame of calcium fluorescence image was acquired every 3 s using the imaging system.

[0032] Parameter optimization analysis was performed using stress-induced calcium kinetic response in discarded osteocytes, and the results are as follows: Figure 1-3As shown: First, by incubating osteocytes from waste bone tissue in vitro with 1mM, 2mM, 4mM, 8mM, and 16mM glutamine, it was found that glutamine at doses of 2mM, 4mM, 8mM, and 16mM significantly increased the number and intensity of calcium signal peaks in the osteocyte network under fluid shear stress stimulation, with the 4mM concentration showing the most significant effect (reaching its peak at 4mM and decreasing at 8mM). Second, based on 4mM glutamine, waste bone cells were further incubated with different doses of glucose (concentrations of 1mM, 2mM, 4mM, 8mM, and 16mM). The results showed that 4mM, 8mM, and 16mM glucose significantly enhanced the calcium signal response of the osteocyte network under fluid shear stress stimulation, manifested as a significant increase in the number and intensity of calcium peaks; however, there was no significant difference in the calcium signal response induced by the three concentration gradients of 4mM, 8mM, and 16mM. Therefore, a lower concentration of 4 mM glucose was selected as the optimized glucose dose to induce a significant signal response in the osteocyte network. Subsequently, using 4 mM glutamine and 4 mM glucose as substrates, the osteocyte network was further incubated with different doses of branched-chain amino acids (concentrations of 0.05 mM, 0.1 mM, 0.2 mM, 0.4 mM, 1.0 mM, and 2.0 mM, respectively). The results showed that 0.4 mM branched-chain amino acids had a significantly higher promoting effect on the ability of disused osteocytes to respond to fluid shear force than other dose groups, as evidenced by a significant increase in the number and intensity of calcium spikes.

[0033] Example 2 A rat hind limb disuse model was established using the tail suspension method. A section of copper wire was bent and secured to both sides of the rat's tail with medical tape. The medical tape was wrapped approximately 2 cm from the base of the tail and then 2-3 cm downwards from there. When wrapping the tape, the tail was first wrapped once, and then the copper wire was wrapped inside. A certain gap was left between the copper wire and the tail on both sides for ventilation. After securing the tail with the copper wire, paperclips were used to create "rings" and "chains" that allowed the rat to move freely in a specially designed tail-suspended cage, and these were connected to the copper wire. Finally, the rat was suspended with its body at a 30° angle to the bottom of the cage. The rat's hind limb suspension and health status were observed daily. During the tail suspension process, a supersaturated benzoin solution prepared with 95% alcohol was applied to the tail to prevent ulceration at the tape-wrapped area. This condition was maintained for 4 weeks to establish a hind limb disuse bone loss model. A self-made small animal long bone stress loading system was used to conduct physiological cyclic loading experiments on live animals. The entire system consists of four main parts: a stress loading device, an actuator control module, a real-time data acquisition and processing module, and a human-computer interaction module. The right tibia and knee joint of the rat was fixed in a movable clamp, and the carpal joint was fixed in a fixed clamp. The loading parameters were as follows: loading frequency 4 Hz, ramp loading, average peak pressure ensuring a strain of approximately 1500 με at the point of maximum physiological curvature of the tibia, 2000 cycles per day, for two consecutive weeks.

[0034] By subjecting rats to tail-suspended hindlimb unloading for 4 weeks with subsequent 2 weeks of stress loading and nutritional supplementation, the results are as follows: Figure 4-5 As shown: First, cyclic compression loads were applied daily to the tibias of rats that had been tail-suspended for 4 weeks for 2 consecutive weeks. During this period, glutamine was administered daily by gavage at doses of 300 mg / kg, 600 mg / kg, and 1000 mg / kg. All of these concentrations of glutamine improved the mechanical response of disused bone tissue to mechanical reloading to some extent, with 600 mg / kg showing the most significant effect. Second, the effects of glutamine synergistically with glucose and branched-chain amino acids on bone rehabilitation after disused bone tissue reloading were compared. Glutamine (600 mg / kg) synergistically with glucose (600 mg / kg) showed a slightly better effect on bone mass recovery after disused bone tissue reloading than glutamine alone; glutamine and glucose synergistically with branched-chain amino acids showed a better pre-intervention effect, with a concentration of 60 mg / kg of branched-chain amino acids showing the most significant effect.

[0035] Example 3 The specific procedure of this embodiment is as follows: Twenty-four patients, all postmenopausal women aged 49-61 years, who had been bedridden for 8 weeks due to spinal cord injury, were selected. All patients had no heart, liver, kidney, or other endocrine and metabolic diseases, and had not taken any hormone medications within the past 3 months. Before treatment, proximal femoral bone mineral density was measured using a dual-energy X-ray absorptiometry (DXA) system from Lunar Pharmaceuticals, USA. The 24 patients were divided into a control group, a single glutamine group, and a mixed supplement group according to age, bone mineral density, and serum osteocalcin level, with 8 patients in each group. The single glutamine group received 10 grams of glutamine daily for 6 months after mechanical reloading following bed rest, while the mixed supplement group received 10 grams of glutamine, 10 grams of glucose, and 1 gram of branched-chain amino acids daily.

[0036] After a 6-month trial, bone mineral density (BMD) of the proximal femur was measured in all three groups using dual-energy X-ray absorptiometry (DXA). The results are shown in Table 1: Six months after the intervention, comparison of baseline BMD values ​​before bed rest with imaging bone mineral density scans revealed that the BMD in the glutamine monotherapy group was significantly higher than that in the control group, while the BMD in the mixed supplement group was significantly higher than both the control group and the glutamine monotherapy group.

[0037] Table 1: Changes in femoral bone mineral density (%) after 8 weeks of bed rest and 6 months of stress reloading compared to before bed rest (%) n =8)

[0038] Compared with the control group:* P <0.05 Compared with the glutamine group: # P <0.05 In summary, this invention addresses the challenging problem of slow and difficult bone recovery after stress reloading in long-term disused bone tissue. Starting with the regulation of calcium kinetics mediated by osteocyte energy metabolism, it provides an oral nutritional supplement based on a composition of glutamine, glucose, and branched-chain amino acids. This supplement has been proven to significantly improve the stress response of disused bone tissue after mechanical reloading, thus providing a new approach and strategy for rapid bone rehabilitation after reloading in disused limb populations (such as long-term bedridden patients, patients with fractures and other traumatic bone diseases, and astronauts). The disclosed solution, starting from the innovative perspective of improving osteocyte calcium oscillation dynamics by regulating the efficiency of ATP synthesis, and supported and validated by multi-dimensional data from cells, animals, and clinical trials, proposes an innovative composition that promotes rapid bone recovery after stress reloading in disused bone tissue. This composition is a nutritional regulation scheme based on cellular energy metabolism, which can be further prepared into a nutritional supplement. The components of the composition are inexpensive, readily available, and safe to use, facilitating widespread clinical application and promotion. In addition to its positive effect on rapid bone recovery after limb disuse reloading, this technical solution also has a positive effect on the recovery of muscle atrophy after reloading. The solution proposed in this application is applicable to the following groups: 1) patients with fractures, nonunion, delayed bone healing, bone atrophy, bone defects, and stress fractures; 2) patients recovering from long-term bed rest due to illness (such as spinal cord injury, traumatic brain injury, or critically ill patients); 3) people who have been exposed to microgravity environments for a long time (such as astronauts).

[0039] The above content is only for illustrating the technical concept of the present invention and should not be construed as limiting the scope of protection of the present invention. Any modifications made to the technical solution based on the technical concept proposed in this invention shall fall within the scope of protection of the claims of this invention.

Claims

1. A composition for promoting rapid bone rehabilitation after disuse reloading of limbs, characterized in that, It is composed of glutamine, glucose and branched-chain amino acids.

2. The composition for promoting rapid bone rehabilitation after disuse reloading of limbs according to claim 1, characterized in that, The molar ratio of glutamine, glucose and branched-chain amino acids in this composition is (2-16):(4-16):(0.05-2).

3. The composition for promoting rapid bone rehabilitation after disuse reloading of limbs according to claim 2, characterized in that, The molar ratio of glutamine, glucose, and branched-chain amino acids in this composition is 10:10:

1.

4. The composition for promoting rapid bone rehabilitation after disuse reloading of limbs according to any one of claims 1-3, characterized in that, The branched-chain amino acids include leucine, isoleucine, and valine, and the molar ratio of leucine, isoleucine, and valine is 2:1:

1.

5. The use of the composition according to any one of claims 1-4 for promoting rapid bone recovery after disuse reloading in the preparation of a medicament for promoting rapid bone recovery after disuse reloading.

6. The use of the composition according to any one of claims 1-4 for promoting rapid bone recovery after disuse reloading in the preparation of a medicament for promoting the recovery of muscle atrophy after disuse reloading.

7. A compound nutritional supplement that promotes rapid bone recovery after disuse reloading of limbs, characterized in that, The composition comprising, with or without the addition of other nutrients, the composition for promoting rapid bone recovery after disuse reloading of limbs as described in any one of claims 1-4.

8. The compound nutritional supplement for promoting rapid bone recovery after disuse reloading of limbs according to claim 7, characterized in that, This compound nutritional supplement can be administered orally or intravenously.

9. The compound nutritional supplement for promoting rapid bone recovery after disuse reloading of limbs according to claim 7, characterized in that, The compound nutritional supplement is to be administered once a day, with each dose consisting of 10 grams of glutamine, 10 grams of glucose, and 1 gram of branched-chain amino acids.

10. The compound nutritional supplement for promoting rapid bone recovery after disuse reloading of limbs according to claim 7, characterized in that, Other nutrients include one or more of the following: bone metabolism-related nutrients, energy metabolism aids, tissue repair and immune modulators, and food-grade excipients.

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