Objective: To assess the effects of axial vibrations on gene expression and lumbar intervertebral disc degeneration in vivo. Methods: A modified bipedal rat model was established using a brachial plexus rhizotomy appr...Objective: To assess the effects of axial vibrations on gene expression and lumbar intervertebral disc degeneration in vivo. Methods: A modified bipedal rat model was established using a brachial plexus rhizotomy approach to imitate human upright posture. The experimental animals were randomly divided into three groups: control, vertical vibration, and whole-body vibration. Gene expression in degeneration of the intervertebral discs was assessed by reverse transcription-quantitative polymerase chain reaction. Results: The expression of aggrecan, Col1α1, Col2α1, and decorin were shown to be up-regulated in 14-week-old rats in the vertical vibration and whole-body vibration groups, whereas biglycan and versican expression was down-regulated in 14-week-old rats of the two experimental groups. Furthermore, biglycan and versican expression levels were shown to be lower in the whole-body vibration group than in the vertical vibration group(P<0.05). Conclusions: This in-vivo study demonstrated that vibrations can influence the expression of anabolic genes. Furthermore, whole-body vibrations seem to have a greater effect in this regard than vertical vibrations. A new method is expected to relieve the low back pain of the patients through our research.展开更多
KLD-12 peptide with a sequence of AcN-KLDLKLDLKLDL-CNH2 was synthesized and its biocompatibility was assessed in animals. Rabbit MSCs were cultured in the hydrogel for 2 weeks. Live cells were counted by using Calcein...KLD-12 peptide with a sequence of AcN-KLDLKLDLKLDL-CNH2 was synthesized and its biocompatibility was assessed in animals. Rabbit MSCs were cultured in the hydrogel for 2 weeks. Live cells were counted by using Calcein-AM/P1 fluorescence staining. MTT was employed to assess the viability of MSCs cultured in KLD-12 peptide solution of 0.01%, 0.03%, and 0.05%. Hemolysis test, skin irritation test and implantation test were conducted to evaluate its biocompatibility with host tissues. Our results demonstrated that the MSCs in hydrogel grew well and maintained round shape. Cell survival rate was 92.15% （mean： 92.15%±1.17%） at the 7th day and there was no difference in survival rate between day 7 and day 14. Cell proliferation test showed that the A value of the KLD-12 solutions was not significantly different from that of control groups （complete culture media） （P〉0.05） at the 24th and 48th h. The hemolysis rate of KLD-12 solution was 0.112%. Skin irritation test showed that the skin injected with KLD-12 solution remained normal and the score of skin irritation was 0. The histological examination with HE staining exhibited that the skin layers were clear and there was no infiltration with neutrophilic granulocytes and lymphocytes. It is concluded that KLD-12 peptide hydrogel bad a good biocompatibility with host rabbit and MSCs, and KLD-12 pep- tide hydrogel can provide an appropriate microenvironment for MSCs.展开更多
Objective:To explore the survival and migration of bone mesenchymal stem cells transplantated in intervertebral disc of rabbits and expression of the exogenic genes.Methods.Thirty-two rabbits were used,A randomized bl...Objective:To explore the survival and migration of bone mesenchymal stem cells transplantated in intervertebral disc of rabbits and expression of the exogenic genes.Methods.Thirty-two rabbits were used,A randomized block design was used and discs in the same rabbit were one block,the lumbar discs from L2-3 to L5-6 were randomly divided into blank group,saline group,cell transplantation groupⅠand cell transplantation groupⅡ.The fluorescence microscopy was used to determine the fluorescence of the maker protein GFP and DNA-PCR was used to analyze the copies of DNA of neomycin-resistant gene at 1,3,6,months after transplantation.Results:There was fluorescence in cell transplantation groupⅠandⅡand none in blank group,saline group at 1,3,6 months after transplantation.In cell transplantation groups,the fluorescent distribution was more scatter with time,but no significant difference between cell groupsⅠandⅡ.The test of neomycin resistant gene expressed in cell transplantation groupⅠandⅡand quantitative analysis showed that there was no significant difference between the cell groupsⅠandⅡ(P〉0.05).Conclusion:The transplanted bone mesenchymal stem cells can survive,migrate and the transfer genes can express efficiently,it suggests that the BMSC therapy may be effective to prevent and treat intervertebral disc degeneration.展开更多
This study examined the correlation between osteoporosis and the degeneration of intervertebral discs. Sprague-Dawley rats were maintained up to 22 or 28 months. The femoral bone, tibial bone and lumbar vertebra were ...This study examined the correlation between osteoporosis and the degeneration of intervertebral discs. Sprague-Dawley rats were maintained up to 22 or 28 months. The femoral bone, tibial bone and lumbar vertebra were histologically studied and the expression of collagen type Ⅱ and Ⅹ in intervertebral discs was immunohistochemiscally determined. Several indices for the degeneration of intervertebral discs and osteoporosis and the correlation among them were then analyzed. Close correlations were found among the indices for the degeneration of intervertebral discs, including the relative area of the vascular bud, the ratio of the uncalcified and the calcified layers, the expression of collagen type Ⅱ and Ⅹ. The correlation with collagen type Ⅹ was negative. There existed positive correlations among the indices for osteoporosis, including the thickness ratio of cortical bone, the relative area of bone trabecula, the density of femoral and vertebral body bones, and the maximum stress and strain on bone. Analysis on the relationship of osteoporosis and the disease on disc showed that the indices of osteoporosis were negatively correlated with the indices of the degeneration of intervertebral discs but the expression of collagen type Ⅹ was positively correlated, with the density of vertebral body bones having the strongest dependence on collagen type Ⅹ. The maximum stress and strain bore no correlation with the degeneration of intervertebral discs. These results suggest that osteoporosis was negatively correlated with the degeneration of intervertebral discs.展开更多
Summary: To investigate therapeutic efficiency of Ad/CMV-hTGF-β1 gene for rabbit intervertebral disc degeneration model. 60 Japanese white rabbits were selected to form the L5-L6 Anterior-Lateral-Anulus-Fibrosus-Inci...Summary: To investigate therapeutic efficiency of Ad/CMV-hTGF-β1 gene for rabbit intervertebral disc degeneration model. 60 Japanese white rabbits were selected to form the L5-L6 Anterior-Lateral-Anulus-Fibrosus-Incision-Induced model in order to simulate human intervertebral disc degeneration. 36 rabbits, whose corresponding intervertebral discs were injected with 20 μl (10×106 pfu)of Ad/CMV-hTGF-β1 gene, constituted the therapy group, 12 were injected with 20 μl (10×106 pfu)of Ad/CMV-LacZ gene as comparison group, while 12 were only injected with equivalent capacity of saline for empty comparison group. 3 weeks after injection, examples were taken for investigation of HE staining, MRI, Western Blotting and immunohistochemical research TGF-β1. Wide distribution of TGF-β1 was detected by immunohistochemical research in the degenerated annulus fibrosus after injection. Western Blotting research showed significant increase of TGF-β1 content in intervertebral discs treated with TGF-β1 gene than comparison groups. MRI signal transformed from low to comparatively high and that intervertebral disc pathological degree improved. Ad/CMV-hTGF-β1 gene transfection is a potential method to increase TGF-β1 content and reverse intervertebral disc degeneration.展开更多
Objective:Underwater shock can produce extremely high accelerations, resulting in severe human injuries on shipboard, and human thoraco lumbar spines are prone to suffer from injuries by ship shock motion. To observe ...Objective:Underwater shock can produce extremely high accelerations, resulting in severe human injuries on shipboard, and human thoraco lumbar spines are prone to suffer from injuries by ship shock motion. To observe the viscoelasticity of thoracolumbar of young fresh cadavers, and to provide biomechanical parameters for both research and clinical practice. Materials and Methods:5 fresh young male cadavers (aged 22 to 31 years) were provided, and 15 thoracolumbar spinal anatomies of 5 samples were harvested within 1 hour of death. WE-10A universal testing machine was used for creep and relaxation tests.Results:Stress relaxation and creep deformation equations are derived from the biomechanics model and the measured and simulated curves are compared. The creep in vertebral bodies and intervertebral discs exhibited significantly changes in the first 5 min and 10 min, respectively. The stress rapidly decreased in the first 2 min, and then gradually went balance during the relaxation process. Conclusion:The change in creep rate is significant at early stage,and gradually slows down.This indicates that the differences between internal pressure and local pressure are decreased until balance. The simulated curve derived from equation coincides with the experimental data to a large degree, which states that the equation is rational and reliable.展开更多
The aim of the present study was to compare the relationship of morphologically defined non-bulging/herni-ated, bulging and herniated intervertebral lumbar discs with quantitative apparent diffusion coefficient （ADC...The aim of the present study was to compare the relationship of morphologically defined non-bulging/herni-ated, bulging and herniated intervertebral lumbar discs with quantitative apparent diffusion coefficient （ADC）. Thirty-two healthy volunteers and 28 patients with back pain or sciatica were examined by MRI. All intervertebral lumbar discs from L1 to S1 were classified according to morphological abnormality and degenerated grades. The ADC values of nucleus pulposus （NP） were measured and recorded. The significant differences about mean ADC values of NP were found between non-bulging/herniated discs and bulging discs as well as herniated discs （P 0.05）, whereas there were no significant differences in ADC values between bulging and herniated discs （P 0.05）. Moreover, statistically significant relationship was found in the mean ADC values of NP between ＂non-bulging/herniated and non-degenerated discs＂ and ＂non-bulging/herniated degenerated discs＂ as well as herniated discs （P 0.05）. Linear regression analysis between ADC value and disc level revealed an inverse correlation （r = -0.18）. The ADC map of the NP is a potentially useful tool for the quantitative assessment of componential and molecular alterations accompanied with lumbar disc abnormalities.展开更多
Intervertebral disc(IVD)degeneration is a leading cause of back pain and precursor to more severe conditions,including disc herniation and spinal stenosis.While traditional growth factor therapies(e.g.,TGFβ)are effec...Intervertebral disc(IVD)degeneration is a leading cause of back pain and precursor to more severe conditions,including disc herniation and spinal stenosis.While traditional growth factor therapies(e.g.,TGFβ)are effective at transiently reversing degenerated disc by stimulation of matrix synthesis,it is increasingly accepted that bioscaffolds are required for sustained,complete IVD regeneration.Current scaffolds(e.g.,metal/polymer composites,non-mammalian biopolymers)can be improved in one or more IVD regeneration demands:biodegradability,noninvasive injection,recapitulated healthy IVD biomechanics,predictable crosslinking,and matrix repair induction.To meet these demands,tetrazine-norbornene bioorthogonal ligation was combined with gelatin to create an injectable bioorthogonal hydrogel(BIOGEL).The liquid hydrogel precursors remain free-flowing across a wide range of temperatures and crosslink into a robust hydrogel after 5-10 min,allowing a human operator to easily inject the therapeutic constructs into degenerated IVD.Moreover,BIOGEL encapsulation of TGFβpotentiated histological repair(e.g.,tissue architecture and matrix synthesis)and functional recovery(e.g.,high water retention by promoting the matrix synthesis and reduced pain)in an in vivo rat IVD degeneration/nucleotomy model.This BIOGEL procedure readily integrates into existing nucleotomy procedures,indicating that clinical adoption should proceed with minimal difficulty.Since bioorthogonal crosslinking is essentially non-reactive towards biomolecules,our developed material platform can be extended to other payloads and degenerative injuries.展开更多
In previous studies of disc degeneration,the structural and mechanical properties of the endplate were often neglected.In this paper,the station legislation was used to construct an animal model of minor trauma disc d...In previous studies of disc degeneration,the structural and mechanical properties of the endplate were often neglected.In this paper,the station legislation was used to construct an animal model of minor trauma disc degeneration,and the mechanism of disc degeneration was further investigated by observing the changes of mesoscopic structure and developing the mechanical properties of endplate bone.Twenty-eight 6-month-old Japanese white rabbits were divided into two groups:control group and experimental group.An animal model of intervertebral disc degeneration was established by upright experiment in the experimental group.The bone mesoscopic structures in different areas of each endplate were observed by histological and imaging methods,and the mechanical properties of the endplates were measured by indentation method.The two groups of data were compared by one-way ANOVA.After the experimental animals stood for 17 weeks,The experimental group showed the characteristics of early disc degeneration.The microstructure of the degenerative group showed that the end plate mineralization degree was higher,the bone mass was larger,and the number and thickness of bone trabeculae were larger.The results of indentation test showed that the mechanical properties of the degeneration group were enhanced,and the lower endplate was obviously enhanced.We successfully established a model of human disc degeneration with non invasive trauma and more consistent with the process of human disc degeneration through the standing experiment.In the experimental group,the internal structure of the endplate was dense and pore distance was reduced.The change of bone mesoscopic structure further affects the endplate,resulting in the enhancement of the mechanical properties of the endplate after intervertebral disc degeneration.The reduction of the pore distance and the narrowing of the internal channel structure of the endplate also hinder the nutrition supply of the intervertebral disc,which may be the key reason affecting the degeneration of the intervertebral disc.A biomechanical method for investigating the mechanism of intervertebral disc degeneration can be provided in this paper.展开更多
Aging is a pivotal risk factor for intervertebral disc degeneration(IVDD)and chronic low back pain(LBP).The restoration of aging nucleus pulposus cells(NPCs)to a youthful epigenetic state is crucial for IVDD treatment...Aging is a pivotal risk factor for intervertebral disc degeneration(IVDD)and chronic low back pain(LBP).The restoration of aging nucleus pulposus cells(NPCs)to a youthful epigenetic state is crucial for IVDD treatment,but remains a formidable challenge.Here,we proposed a strategy to partially reprogram and reinstate youthful epigenetics of senescent NPCs by delivering a plasmid carrier that expressed pluripotency-associated genes(Oct4,Klf4 and Sox2)in Cavin2-modified exosomes(OKS@M-Exo)for treatment of IVDD and alleviating LBP.The functional OKS@M-Exo efficaciously alleviated senescence markers(p16^(INK4a),p21^(CIP1)and p53),reduced DNA damage and H4K20me3 expression,as well as restored proliferation ability and metabolic balance in senescent NPCs,as validated through in vitro experiments.In a rat model of IVDD,OKS@M-Exo maintained intervertebral disc height,nucleus pulposus hydration and tissue structure,effectively ameliorated IVDD via decreasing the senescence markers.Additionally,OKS@MExo reduced nociceptive behavior and downregulated nociception markers,indicating its efficiency in alleviating LBP.The transcriptome sequencing analysis also demonstrated that OKS@M-Exo could decrease the expression of age-related pathways and restore cell proliferation.Collectively,reprogramming by the OKS@M-Exo to restore youthful epigenetics of senescent NPCs may hold promise as a therapeutic platform to treat IVDD.展开更多
Intervertebral disc degeneration is a major risk factor contributing to chronic low back and neck pain.While the etiological factors for disc degeneration vary,age is still one of the most important risk factors.Recen...Intervertebral disc degeneration is a major risk factor contributing to chronic low back and neck pain.While the etiological factors for disc degeneration vary,age is still one of the most important risk factors.Recent studies have shown the promising role of SIRT6 in mammalian aging and skeletal tissue health,however its role in the intervertebral disc health remains unexplored.We investigated the contribution of SIRT6 to disc health by studying the age-dependent spinal phenotype of mice with conditional deletion of Sirt6 in the disc(AcanCreERT2;Sirt6fl/fl).Histological studies showed a degenerative phenotype in knockout mice compared to Sirt6fl/fl control mice at 12 months,which became pronounced at 24 months.RNA-Seq analysis of NP and AF tissues,in vitro quantitative histone analysis,and RNA-seq with ATAC-seq multiomic studies revealed that SIRT6-loss resulted in changes in acetylation and methylation status of specific Histone 3 lysine residues and affected DNA accessibility and transcriptomic landscape.A decrease in autophagy and an increase in DNA damage were also noted in Sirt6-deficient cells.Further mechanistic insights revealed that loss of SIRT6 increased senescence and SASP burden in the disc characterized by increased p21,p19,γH2AX,IL-6,IL-1β,and TGF-βabundance.Taken together,our study highlights the contribution of SIRT6 in modulating DNA damage,autophagy,and cell senescence and its importance in maintaining disc health during aging,thereby underscoring it as a potential therapeutic target to treat intervertebral disc degeneration.展开更多
Intervertebral disc degeneration(IDD)is a progressive and dynamic process in which the senescence-associated secretory phenotype(SASP)of nucleus pulposus cells(NPC)plays a significant role.While impaired chaperone-med...Intervertebral disc degeneration(IDD)is a progressive and dynamic process in which the senescence-associated secretory phenotype(SASP)of nucleus pulposus cells(NPC)plays a significant role.While impaired chaperone-mediated autophagy(CMA)has been associated with inflammation and cellular senescence,its specific involvement in the self-perpetuating feedback loop of NPC senescence remains poorly understood.Through LAMP2A knockout in NPC,we identified a significant upregulation of DYRK1A,a core mediator of premature senescence in Down syndrome.Subsequent validation established DYRK1A as the critical driver of premature senescence in CMA-deficient NPC.Combinatorial transcription factor analysis revealed that under IL1B stimulation or CMA inhibition,elevated DYRK1A promoted FOXC1 phosphorylation and nuclear translocation,initiating transcriptional activation of cell cycle arrest.Intriguingly,CMA impairment concurrently enhanced glutamine metabolic flux in senescent NPC,thereby augmenting their survival fitness.Transcriptomic profiling demonstrated that CMA reactivation in senescent NPC facilitated fate transition from senescence to apoptosis,mediated by decreased glutamine flux via GLUL degradation.Therefore,CMA exerts protective effects against IDD by maintaining equilibrium between premature senescence and senolysis.This study elucidates CMA’s regulatory role in SASP-mediated senescence amplification circuits,providing novel therapeutic insights for IDD and other age-related pathologies.展开更多
Intervertebral disc degeneration(IDD)results from an imbalance within the intervertebral disc,leading to alterations in extracellular matrix composition,loss of nucleus pulposus cells,increased oxidative stress,and in...Intervertebral disc degeneration(IDD)results from an imbalance within the intervertebral disc,leading to alterations in extracellular matrix composition,loss of nucleus pulposus cells,increased oxidative stress,and inflammatory cascade.While IDD naturally progresses with age,some factors such as mechanical trauma,lifestyle choices,and genetic abnormalities can elevate the risk of symptomatic disease progression.Current treatments,including pharmacological and surgical interventions,fail to halt disease progression or restore IDD function.Although biological therapies have been evaluated,their effectiveness in reversing long-term disc degeneration remains inconsistent.Mesenchymal stem cellbased therapies have demonstrated potential for IDD regeneration but are hindered by biological limitations,ethical issues,etc.To date,mesenchymal stem cell-derived extracellular vesicles(EVs)have emerged as promising therapeutic agents for regeneration and anti-inflammation.Their therapeutic effects are attributed to several mechanisms,such as the induction of regenerative phenotype,apoptosis mitigation,and immunomodulation.In addition,the abundance of microRNAs within EVs play a crucial role in modulating the disc degeneration.Due to the problems in clinical use,however,the efficiency of the EVs should be overcome further by optimizing cell culture conditions,engineering them to deliver drugs and targeting molecules,etc.展开更多
Intervertebral disc degeneration(IVDD)is the primary contributor to a range of spinal diseases.Dynamin-related protein 1(Drp1)-mediated mitochondrial fission has recently been identified as a new cause of nucleus pulp...Intervertebral disc degeneration(IVDD)is the primary contributor to a range of spinal diseases.Dynamin-related protein 1(Drp1)-mediated mitochondrial fission has recently been identified as a new cause of nucleus pulposus cell(NPC)death and IVDD,but the underlying mechanisms remain unclear.Although the effects of Drp1 phosphorylation in IVDD have been studied,it is currently unknown if small ubiquitin-like modifications(SUMOylation)of Drp1 regulate IVDD.This study aimed to investigate the functions and mechanisms of mitochondria-anchored protein ligase(MAPL),a mitochondrial SUMO E3 ligase,during IVDD progression.The expression of genes related to SUMOylation and mitochondrial dynamics in TNF-α-stimulated NPCs was analysed via RNA sequencing.展开更多
Intervertebral disc degeneration is a leading cause of lower back pain and is characterized by pathological processes such as nucleus pulposus cell apoptosis,extracellular matrix imbalance,and annulus fibrosus rupture...Intervertebral disc degeneration is a leading cause of lower back pain and is characterized by pathological processes such as nucleus pulposus cell apoptosis,extracellular matrix imbalance,and annulus fibrosus rupture.These pathological changes result in disc height loss and functional decline,potentially leading to disc herniation.This comprehensive review aimed to address the current challenges in intervertebral disc degeneration treatment by evaluating the regenerative potential of stem cell-based therapies,with a particular focus on emerging technologies such as exosomes and gene vector systems.Through mechanisms such as differentiation,paracrine effects,and immunomodulation,stem cells facilitate extracellular matrix repair and reduce nucleus pulposus cell apoptosis.Despite recent advancements,clinical applications are hindered by challenges such as hypoxic disc environments and immune rejection.By analyzing recent preclinical and clinical findings,this review provided insights into optimizing stem cell therapy to overcome these obstacles and highlighted future directions in the field.展开更多
In healthy intervertebral discs(IVDs),nerves and blood vessels are present only in the outer annulus fibrosus,while in degenerative IVDs,a large amount of nerve and blood vessel tissue grows inward.Evidence supports t...In healthy intervertebral discs(IVDs),nerves and blood vessels are present only in the outer annulus fibrosus,while in degenerative IVDs,a large amount of nerve and blood vessel tissue grows inward.Evidence supports that neurogenic inflammation produced by neuropeptides such as substance P and calcitonin gene related peptide released by the nociceptive nerve fibers innervating the IVDs plays a crucial role in the process of IVD degeneration.Recently,non-neuronal cells,including IVD cells and infiltrating immune cells,have emerged as important players in neurogenic inflammation.IVD cells and infiltrating immune cells express functional receptors for neuropeptides through which they receive signals from the nervous system.In return,IVD cells and immune cells produce neuropeptides and nerve growth factor,which stimulate nerve fibers.This communication generates a positive bidirectional feedback loop that can enhance the inflammatory response of the IVD.Recently emerging transient receptor potential channels have been recognized as contributors to neurogenic inflammation in the degenerative IVDs.These findings suggest that neurogenic inflammation involves complex pathophysiological interactions between sensory nerves and multiple cell types in the degenerative IVDs.Clarifying the mechanism of neurogenic inflammation in IVD degeneration may provide in-depth understanding of the pathology of discogenic low back pain.展开更多
Lumbar intervertebral disc degeneration is thought to be the main cause of low back pain,although the mechanisms by which it occurs and leads to pain remain unclear.In healthy adult discs,vessels and nerves are presen...Lumbar intervertebral disc degeneration is thought to be the main cause of low back pain,although the mechanisms by which it occurs and leads to pain remain unclear.In healthy adult discs,vessels and nerves are present only in the outer layer of the annulus fibrosus and in the bony endplate.Animal models,and histological and biomechanical studies have shown that annulus tear or endplate injury is the initiating factor for painful disc degeneration.Injury to the disc triggers a local inflammatory repair response that activates nociceptors and promotes the synthesis of neuropeptides such as substance P and calcitonin generelated peptide,by dorsal root ganglion neurons.These neuropeptides are transported to injured discs and act as pro-inflammatory molecules,promoting the production of an“inflammatory soup”by inducing vasodilatation and plasma extravasation as well as by promoting the release of chemical mediators from disc cells and infiltrating immune cells,causing neurogenic inflammation that leads to progressive disc degeneration and discogenic pain.展开更多
Effective treatment of intervertebral disc degeneration with biomaterials remains a challenge,owing to the difficulty in simultaneously overcoming oxidative stress and its associated cascades in the nucleus pulposus m...Effective treatment of intervertebral disc degeneration with biomaterials remains a challenge,owing to the difficulty in simultaneously overcoming oxidative stress and its associated cascades in the nucleus pulposus microenvironment,which includes cellular senescence,apoptosis,inflammation,and extracellular matrix(ECM)degradation.To address these issues,a multifunctional hydrogel(HG-QNT)loaded with transforming growth factorβ1(TGFβ1)and quercetin-based nanoparticles(QUNPs)is developed through borate ester bonding and Schiffbase reaction-induced crosslinking.Specifically,QUNPs fabricated via coordination and hydrophobic interactions endow the hydrogel with extraordinary antioxidative properties.Benefiting from the multi-dynamic crosslinking,the hydrogel achieves self-healing,mechanical stability,and pH-responsive release of QUNPs and TGFβ1.The HG-QNT hydrogel is demonstrated to enhance the proliferation of encapsulated nucleus pulposus cells,thereby providing an ideal platform for cell transplantation.The cooperative antioxidation of QUNPs and the hydrogel carrier renders HG-QNT effective in mitigating oxidative stress,resulting in the suppression of cellular senescence,mitochondrial dysfunction,apoptosis,excessive inflammation,and abnormal catabolism.Afterwards,TGFβ1 and QUNPs act in synergy with the hydrogel to restore the anabolic/catabolic balance by enhancing ECM synthesis.Overall,the strategy orchestrating multiple modulation by HG-QNT hydrogel shows great potential for application in intervertebral disc regeneration.展开更多
Background:Renshen Guben oral liquid(RGSB)has the ability to consolidate constitution,restore vitality,and delay the ageing process and the onset of geriatric diseases.RGSB also has potential therapeutic effects on in...Background:Renshen Guben oral liquid(RGSB)has the ability to consolidate constitution,restore vitality,and delay the ageing process and the onset of geriatric diseases.RGSB also has potential therapeutic effects on intervertebral disc degeneration(IVDD).This study investigated the therapeutic effects of RGSB on IVDD and explored the underlying mechanisms.Methods:Single-cell sequencing and network pharmacology analyses were conducted in combination to determine the mechanisms of IVDD.Thirty-two C57BL/6J mice were randomly assigned to four groups:the sham operation group,IVDD group,Ibuprofen(IB)group,and RGSB group.All groups except for the sham operation group underwent needle puncture of the caudal IVDs.Behavioral tests,ELISA,PCR,TUNEL staining,and immunohistochemical staining were performed to investigate the therapeutic mechanisms associated with IVDD.Results:Bioinformatics analysis revealed that apoptosis is involved in the development of IVDD and that RSGB might alleviate IVDD by inhibiting apoptosis.After needle puncture,the mice exhibited from thermal allodynia,a decrease in disc height,an increase in the Pfirrmann grade,and destruction of IVDs.Surgery to induce IVDD increased the levels of Matrix Metalloproteinase-3(MMP3),Caspase-6,and Cyt-c in nucleus pulposus cells and the periphery;serum levels of interleukin-1β(IL‐1β)and TNF‐α;and the mRNA levels of IL-6,TNF-α,and IL-1β.Moreover,IVDD decreased the IL-10 mRNA level and the expression of Col-I and BCL2.RSGB reversed these changes.Conclusion:RSGB alleviated IVDD by relieving inflammation and apoptosis and alleviating neuroinflammation.展开更多
基金supported by the National Natural Science Foundation of China(Grant No.81401768,81301646)the Natural Science Foundation of Jiangsu Province(Grant No.BK20140289)+1 种基金the Specialized Research Fund for the Doctoral Program of Higher Education of China(Grant No.20123201120018)China Postdoctoral Science Foundation on the 53rd general program(Grant No.2013M531404)
文摘Objective: To assess the effects of axial vibrations on gene expression and lumbar intervertebral disc degeneration in vivo. Methods: A modified bipedal rat model was established using a brachial plexus rhizotomy approach to imitate human upright posture. The experimental animals were randomly divided into three groups: control, vertical vibration, and whole-body vibration. Gene expression in degeneration of the intervertebral discs was assessed by reverse transcription-quantitative polymerase chain reaction. Results: The expression of aggrecan, Col1α1, Col2α1, and decorin were shown to be up-regulated in 14-week-old rats in the vertical vibration and whole-body vibration groups, whereas biglycan and versican expression was down-regulated in 14-week-old rats of the two experimental groups. Furthermore, biglycan and versican expression levels were shown to be lower in the whole-body vibration group than in the vertical vibration group(P<0.05). Conclusions: This in-vivo study demonstrated that vibrations can influence the expression of anabolic genes. Furthermore, whole-body vibrations seem to have a greater effect in this regard than vertical vibrations. A new method is expected to relieve the low back pain of the patients through our research.
基金supported by a grant from the National High Technology Research and Development Program of China (Program 863) (No. 2006AA02A124)
文摘KLD-12 peptide with a sequence of AcN-KLDLKLDLKLDL-CNH2 was synthesized and its biocompatibility was assessed in animals. Rabbit MSCs were cultured in the hydrogel for 2 weeks. Live cells were counted by using Calcein-AM/P1 fluorescence staining. MTT was employed to assess the viability of MSCs cultured in KLD-12 peptide solution of 0.01%, 0.03%, and 0.05%. Hemolysis test, skin irritation test and implantation test were conducted to evaluate its biocompatibility with host tissues. Our results demonstrated that the MSCs in hydrogel grew well and maintained round shape. Cell survival rate was 92.15% （mean： 92.15%±1.17%） at the 7th day and there was no difference in survival rate between day 7 and day 14. Cell proliferation test showed that the A value of the KLD-12 solutions was not significantly different from that of control groups （complete culture media） （P〉0.05） at the 24th and 48th h. The hemolysis rate of KLD-12 solution was 0.112%. Skin irritation test showed that the skin injected with KLD-12 solution remained normal and the score of skin irritation was 0. The histological examination with HE staining exhibited that the skin layers were clear and there was no infiltration with neutrophilic granulocytes and lymphocytes. It is concluded that KLD-12 peptide hydrogel bad a good biocompatibility with host rabbit and MSCs, and KLD-12 pep- tide hydrogel can provide an appropriate microenvironment for MSCs.
基金The Study of Differentiation of Bone Mesenchymal Stem Cells Transplanted in Intervertebral Disc and Expression of ExogenousGene(30400163)
文摘Objective:To explore the survival and migration of bone mesenchymal stem cells transplantated in intervertebral disc of rabbits and expression of the exogenic genes.Methods.Thirty-two rabbits were used,A randomized block design was used and discs in the same rabbit were one block,the lumbar discs from L2-3 to L5-6 were randomly divided into blank group,saline group,cell transplantation groupⅠand cell transplantation groupⅡ.The fluorescence microscopy was used to determine the fluorescence of the maker protein GFP and DNA-PCR was used to analyze the copies of DNA of neomycin-resistant gene at 1,3,6,months after transplantation.Results:There was fluorescence in cell transplantation groupⅠandⅡand none in blank group,saline group at 1,3,6 months after transplantation.In cell transplantation groups,the fluorescent distribution was more scatter with time,but no significant difference between cell groupsⅠandⅡ.The test of neomycin resistant gene expressed in cell transplantation groupⅠandⅡand quantitative analysis showed that there was no significant difference between the cell groupsⅠandⅡ(P〉0.05).Conclusion:The transplanted bone mesenchymal stem cells can survive,migrate and the transfer genes can express efficiently,it suggests that the BMSC therapy may be effective to prevent and treat intervertebral disc degeneration.
基金supported by grants from the National Natu-ral Science Foundation of China(No.81171761,30973063)the Fundamental Research Funds for the Central Uni-versity(No.08143045)
文摘This study examined the correlation between osteoporosis and the degeneration of intervertebral discs. Sprague-Dawley rats were maintained up to 22 or 28 months. The femoral bone, tibial bone and lumbar vertebra were histologically studied and the expression of collagen type Ⅱ and Ⅹ in intervertebral discs was immunohistochemiscally determined. Several indices for the degeneration of intervertebral discs and osteoporosis and the correlation among them were then analyzed. Close correlations were found among the indices for the degeneration of intervertebral discs, including the relative area of the vascular bud, the ratio of the uncalcified and the calcified layers, the expression of collagen type Ⅱ and Ⅹ. The correlation with collagen type Ⅹ was negative. There existed positive correlations among the indices for osteoporosis, including the thickness ratio of cortical bone, the relative area of bone trabecula, the density of femoral and vertebral body bones, and the maximum stress and strain on bone. Analysis on the relationship of osteoporosis and the disease on disc showed that the indices of osteoporosis were negatively correlated with the indices of the degeneration of intervertebral discs but the expression of collagen type Ⅹ was positively correlated, with the density of vertebral body bones having the strongest dependence on collagen type Ⅹ. The maximum stress and strain bore no correlation with the degeneration of intervertebral discs. These results suggest that osteoporosis was negatively correlated with the degeneration of intervertebral discs.
文摘Summary: To investigate therapeutic efficiency of Ad/CMV-hTGF-β1 gene for rabbit intervertebral disc degeneration model. 60 Japanese white rabbits were selected to form the L5-L6 Anterior-Lateral-Anulus-Fibrosus-Incision-Induced model in order to simulate human intervertebral disc degeneration. 36 rabbits, whose corresponding intervertebral discs were injected with 20 μl (10×106 pfu)of Ad/CMV-hTGF-β1 gene, constituted the therapy group, 12 were injected with 20 μl (10×106 pfu)of Ad/CMV-LacZ gene as comparison group, while 12 were only injected with equivalent capacity of saline for empty comparison group. 3 weeks after injection, examples were taken for investigation of HE staining, MRI, Western Blotting and immunohistochemical research TGF-β1. Wide distribution of TGF-β1 was detected by immunohistochemical research in the degenerated annulus fibrosus after injection. Western Blotting research showed significant increase of TGF-β1 content in intervertebral discs treated with TGF-β1 gene than comparison groups. MRI signal transformed from low to comparatively high and that intervertebral disc pathological degree improved. Ad/CMV-hTGF-β1 gene transfection is a potential method to increase TGF-β1 content and reverse intervertebral disc degeneration.
文摘Objective:Underwater shock can produce extremely high accelerations, resulting in severe human injuries on shipboard, and human thoraco lumbar spines are prone to suffer from injuries by ship shock motion. To observe the viscoelasticity of thoracolumbar of young fresh cadavers, and to provide biomechanical parameters for both research and clinical practice. Materials and Methods:5 fresh young male cadavers (aged 22 to 31 years) were provided, and 15 thoracolumbar spinal anatomies of 5 samples were harvested within 1 hour of death. WE-10A universal testing machine was used for creep and relaxation tests.Results:Stress relaxation and creep deformation equations are derived from the biomechanics model and the measured and simulated curves are compared. The creep in vertebral bodies and intervertebral discs exhibited significantly changes in the first 5 min and 10 min, respectively. The stress rapidly decreased in the first 2 min, and then gradually went balance during the relaxation process. Conclusion:The change in creep rate is significant at early stage,and gradually slows down.This indicates that the differences between internal pressure and local pressure are decreased until balance. The simulated curve derived from equation coincides with the experimental data to a large degree, which states that the equation is rational and reliable.
基金supported by the National Natural Foundation of China (No.30970797)Shaanxi Science and Technology Plan Projects (No. 2008k09-1)
文摘The aim of the present study was to compare the relationship of morphologically defined non-bulging/herni-ated, bulging and herniated intervertebral lumbar discs with quantitative apparent diffusion coefficient （ADC）. Thirty-two healthy volunteers and 28 patients with back pain or sciatica were examined by MRI. All intervertebral lumbar discs from L1 to S1 were classified according to morphological abnormality and degenerated grades. The ADC values of nucleus pulposus （NP） were measured and recorded. The significant differences about mean ADC values of NP were found between non-bulging/herniated discs and bulging discs as well as herniated discs （P 0.05）, whereas there were no significant differences in ADC values between bulging and herniated discs （P 0.05）. Moreover, statistically significant relationship was found in the mean ADC values of NP between ＂non-bulging/herniated and non-degenerated discs＂ and ＂non-bulging/herniated degenerated discs＂ as well as herniated discs （P 0.05）. Linear regression analysis between ADC value and disc level revealed an inverse correlation （r = -0.18）. The ADC map of the NP is a potentially useful tool for the quantitative assessment of componential and molecular alterations accompanied with lumbar disc abnormalities.
文摘Intervertebral disc(IVD)degeneration is a leading cause of back pain and precursor to more severe conditions,including disc herniation and spinal stenosis.While traditional growth factor therapies(e.g.,TGFβ)are effective at transiently reversing degenerated disc by stimulation of matrix synthesis,it is increasingly accepted that bioscaffolds are required for sustained,complete IVD regeneration.Current scaffolds(e.g.,metal/polymer composites,non-mammalian biopolymers)can be improved in one or more IVD regeneration demands:biodegradability,noninvasive injection,recapitulated healthy IVD biomechanics,predictable crosslinking,and matrix repair induction.To meet these demands,tetrazine-norbornene bioorthogonal ligation was combined with gelatin to create an injectable bioorthogonal hydrogel(BIOGEL).The liquid hydrogel precursors remain free-flowing across a wide range of temperatures and crosslink into a robust hydrogel after 5-10 min,allowing a human operator to easily inject the therapeutic constructs into degenerated IVD.Moreover,BIOGEL encapsulation of TGFβpotentiated histological repair(e.g.,tissue architecture and matrix synthesis)and functional recovery(e.g.,high water retention by promoting the matrix synthesis and reduced pain)in an in vivo rat IVD degeneration/nucleotomy model.This BIOGEL procedure readily integrates into existing nucleotomy procedures,indicating that clinical adoption should proceed with minimal difficulty.Since bioorthogonal crosslinking is essentially non-reactive towards biomolecules,our developed material platform can be extended to other payloads and degenerative injuries.
基金the National Natural Science Foundation of China(11472185,11972243)for fundingFund Program for the Scientific Activities of Selected Returned Overseas Professionals in Shanxi Province(2020040)for funding。
文摘In previous studies of disc degeneration,the structural and mechanical properties of the endplate were often neglected.In this paper,the station legislation was used to construct an animal model of minor trauma disc degeneration,and the mechanism of disc degeneration was further investigated by observing the changes of mesoscopic structure and developing the mechanical properties of endplate bone.Twenty-eight 6-month-old Japanese white rabbits were divided into two groups:control group and experimental group.An animal model of intervertebral disc degeneration was established by upright experiment in the experimental group.The bone mesoscopic structures in different areas of each endplate were observed by histological and imaging methods,and the mechanical properties of the endplates were measured by indentation method.The two groups of data were compared by one-way ANOVA.After the experimental animals stood for 17 weeks,The experimental group showed the characteristics of early disc degeneration.The microstructure of the degenerative group showed that the end plate mineralization degree was higher,the bone mass was larger,and the number and thickness of bone trabeculae were larger.The results of indentation test showed that the mechanical properties of the degeneration group were enhanced,and the lower endplate was obviously enhanced.We successfully established a model of human disc degeneration with non invasive trauma and more consistent with the process of human disc degeneration through the standing experiment.In the experimental group,the internal structure of the endplate was dense and pore distance was reduced.The change of bone mesoscopic structure further affects the endplate,resulting in the enhancement of the mechanical properties of the endplate after intervertebral disc degeneration.The reduction of the pore distance and the narrowing of the internal channel structure of the endplate also hinder the nutrition supply of the intervertebral disc,which may be the key reason affecting the degeneration of the intervertebral disc.A biomechanical method for investigating the mechanism of intervertebral disc degeneration can be provided in this paper.
基金supported by the Ministry of Science and Technology of China(2020YFA0908900)National Natural Science Foundation of China(21935011 and 82072490)+1 种基金Shenzhen Science and Technology Innovation Commission(KQTD20200820113012029 and KJZD20230923114612025)Guangdong Provincial Key Laboratory of Advanced Biomaterials(2022B1212010003).
文摘Aging is a pivotal risk factor for intervertebral disc degeneration(IVDD)and chronic low back pain(LBP).The restoration of aging nucleus pulposus cells(NPCs)to a youthful epigenetic state is crucial for IVDD treatment,but remains a formidable challenge.Here,we proposed a strategy to partially reprogram and reinstate youthful epigenetics of senescent NPCs by delivering a plasmid carrier that expressed pluripotency-associated genes(Oct4,Klf4 and Sox2)in Cavin2-modified exosomes(OKS@M-Exo)for treatment of IVDD and alleviating LBP.The functional OKS@M-Exo efficaciously alleviated senescence markers(p16^(INK4a),p21^(CIP1)and p53),reduced DNA damage and H4K20me3 expression,as well as restored proliferation ability and metabolic balance in senescent NPCs,as validated through in vitro experiments.In a rat model of IVDD,OKS@M-Exo maintained intervertebral disc height,nucleus pulposus hydration and tissue structure,effectively ameliorated IVDD via decreasing the senescence markers.Additionally,OKS@MExo reduced nociceptive behavior and downregulated nociception markers,indicating its efficiency in alleviating LBP.The transcriptome sequencing analysis also demonstrated that OKS@M-Exo could decrease the expression of age-related pathways and restore cell proliferation.Collectively,reprogramming by the OKS@M-Exo to restore youthful epigenetics of senescent NPCs may hold promise as a therapeutic platform to treat IVDD.
基金supported by the Michael Michelson Gift FundNIA grants R01AG073349 (M.V.R.), R01AG044034 (R.F.L.), and R01AG078609 (J.C.)
文摘Intervertebral disc degeneration is a major risk factor contributing to chronic low back and neck pain.While the etiological factors for disc degeneration vary,age is still one of the most important risk factors.Recent studies have shown the promising role of SIRT6 in mammalian aging and skeletal tissue health,however its role in the intervertebral disc health remains unexplored.We investigated the contribution of SIRT6 to disc health by studying the age-dependent spinal phenotype of mice with conditional deletion of Sirt6 in the disc(AcanCreERT2;Sirt6fl/fl).Histological studies showed a degenerative phenotype in knockout mice compared to Sirt6fl/fl control mice at 12 months,which became pronounced at 24 months.RNA-Seq analysis of NP and AF tissues,in vitro quantitative histone analysis,and RNA-seq with ATAC-seq multiomic studies revealed that SIRT6-loss resulted in changes in acetylation and methylation status of specific Histone 3 lysine residues and affected DNA accessibility and transcriptomic landscape.A decrease in autophagy and an increase in DNA damage were also noted in Sirt6-deficient cells.Further mechanistic insights revealed that loss of SIRT6 increased senescence and SASP burden in the disc characterized by increased p21,p19,γH2AX,IL-6,IL-1β,and TGF-βabundance.Taken together,our study highlights the contribution of SIRT6 in modulating DNA damage,autophagy,and cell senescence and its importance in maintaining disc health during aging,thereby underscoring it as a potential therapeutic target to treat intervertebral disc degeneration.
基金supported by the National Natural Science Foundation of China (NSFC) (No.82172497)
文摘Intervertebral disc degeneration(IDD)is a progressive and dynamic process in which the senescence-associated secretory phenotype(SASP)of nucleus pulposus cells(NPC)plays a significant role.While impaired chaperone-mediated autophagy(CMA)has been associated with inflammation and cellular senescence,its specific involvement in the self-perpetuating feedback loop of NPC senescence remains poorly understood.Through LAMP2A knockout in NPC,we identified a significant upregulation of DYRK1A,a core mediator of premature senescence in Down syndrome.Subsequent validation established DYRK1A as the critical driver of premature senescence in CMA-deficient NPC.Combinatorial transcription factor analysis revealed that under IL1B stimulation or CMA inhibition,elevated DYRK1A promoted FOXC1 phosphorylation and nuclear translocation,initiating transcriptional activation of cell cycle arrest.Intriguingly,CMA impairment concurrently enhanced glutamine metabolic flux in senescent NPC,thereby augmenting their survival fitness.Transcriptomic profiling demonstrated that CMA reactivation in senescent NPC facilitated fate transition from senescence to apoptosis,mediated by decreased glutamine flux via GLUL degradation.Therefore,CMA exerts protective effects against IDD by maintaining equilibrium between premature senescence and senolysis.This study elucidates CMA’s regulatory role in SASP-mediated senescence amplification circuits,providing novel therapeutic insights for IDD and other age-related pathologies.
基金Supported by 2024 Yeungnam University Grant,No.224A480005.
文摘Intervertebral disc degeneration(IDD)results from an imbalance within the intervertebral disc,leading to alterations in extracellular matrix composition,loss of nucleus pulposus cells,increased oxidative stress,and inflammatory cascade.While IDD naturally progresses with age,some factors such as mechanical trauma,lifestyle choices,and genetic abnormalities can elevate the risk of symptomatic disease progression.Current treatments,including pharmacological and surgical interventions,fail to halt disease progression or restore IDD function.Although biological therapies have been evaluated,their effectiveness in reversing long-term disc degeneration remains inconsistent.Mesenchymal stem cellbased therapies have demonstrated potential for IDD regeneration but are hindered by biological limitations,ethical issues,etc.To date,mesenchymal stem cell-derived extracellular vesicles(EVs)have emerged as promising therapeutic agents for regeneration and anti-inflammation.Their therapeutic effects are attributed to several mechanisms,such as the induction of regenerative phenotype,apoptosis mitigation,and immunomodulation.In addition,the abundance of microRNAs within EVs play a crucial role in modulating the disc degeneration.Due to the problems in clinical use,however,the efficiency of the EVs should be overcome further by optimizing cell culture conditions,engineering them to deliver drugs and targeting molecules,etc.
基金supported by National Natural Science Foundation of China(82272549,82472505,and 82472498)National key Research and Development plan,Ministry of Science and Technology of the People’s Republic of China(2022YFC2407203)+2 种基金the Young Health Talents of Shanghai Municipal Health Commission,China(2022YQ011)China Medical Education Association(3030537245)The Youth Talent Project of Huashan Hospital(30302164006).
文摘Intervertebral disc degeneration(IVDD)is the primary contributor to a range of spinal diseases.Dynamin-related protein 1(Drp1)-mediated mitochondrial fission has recently been identified as a new cause of nucleus pulposus cell(NPC)death and IVDD,but the underlying mechanisms remain unclear.Although the effects of Drp1 phosphorylation in IVDD have been studied,it is currently unknown if small ubiquitin-like modifications(SUMOylation)of Drp1 regulate IVDD.This study aimed to investigate the functions and mechanisms of mitochondria-anchored protein ligase(MAPL),a mitochondrial SUMO E3 ligase,during IVDD progression.The expression of genes related to SUMOylation and mitochondrial dynamics in TNF-α-stimulated NPCs was analysed via RNA sequencing.
基金Supported by Henan Province Key Research and Development Program,No.231111311000Henan Provincial Science and Technology Research Project,No.232102310411+2 种基金Henan Province Medical Science and Technology Key Project,No.LHGJ20220566 and No.LHGJ20240365Henan Province Medical Education Research Project,No.WJLX2023079Zhengzhou Medical and Health Technology Innovation Guidance Program,No.2024YLZDJH022.
文摘Intervertebral disc degeneration is a leading cause of lower back pain and is characterized by pathological processes such as nucleus pulposus cell apoptosis,extracellular matrix imbalance,and annulus fibrosus rupture.These pathological changes result in disc height loss and functional decline,potentially leading to disc herniation.This comprehensive review aimed to address the current challenges in intervertebral disc degeneration treatment by evaluating the regenerative potential of stem cell-based therapies,with a particular focus on emerging technologies such as exosomes and gene vector systems.Through mechanisms such as differentiation,paracrine effects,and immunomodulation,stem cells facilitate extracellular matrix repair and reduce nucleus pulposus cell apoptosis.Despite recent advancements,clinical applications are hindered by challenges such as hypoxic disc environments and immune rejection.By analyzing recent preclinical and clinical findings,this review provided insights into optimizing stem cell therapy to overcome these obstacles and highlighted future directions in the field.
文摘In healthy intervertebral discs(IVDs),nerves and blood vessels are present only in the outer annulus fibrosus,while in degenerative IVDs,a large amount of nerve and blood vessel tissue grows inward.Evidence supports that neurogenic inflammation produced by neuropeptides such as substance P and calcitonin gene related peptide released by the nociceptive nerve fibers innervating the IVDs plays a crucial role in the process of IVD degeneration.Recently,non-neuronal cells,including IVD cells and infiltrating immune cells,have emerged as important players in neurogenic inflammation.IVD cells and infiltrating immune cells express functional receptors for neuropeptides through which they receive signals from the nervous system.In return,IVD cells and immune cells produce neuropeptides and nerve growth factor,which stimulate nerve fibers.This communication generates a positive bidirectional feedback loop that can enhance the inflammatory response of the IVD.Recently emerging transient receptor potential channels have been recognized as contributors to neurogenic inflammation in the degenerative IVDs.These findings suggest that neurogenic inflammation involves complex pathophysiological interactions between sensory nerves and multiple cell types in the degenerative IVDs.Clarifying the mechanism of neurogenic inflammation in IVD degeneration may provide in-depth understanding of the pathology of discogenic low back pain.
文摘Lumbar intervertebral disc degeneration is thought to be the main cause of low back pain,although the mechanisms by which it occurs and leads to pain remain unclear.In healthy adult discs,vessels and nerves are present only in the outer layer of the annulus fibrosus and in the bony endplate.Animal models,and histological and biomechanical studies have shown that annulus tear or endplate injury is the initiating factor for painful disc degeneration.Injury to the disc triggers a local inflammatory repair response that activates nociceptors and promotes the synthesis of neuropeptides such as substance P and calcitonin generelated peptide,by dorsal root ganglion neurons.These neuropeptides are transported to injured discs and act as pro-inflammatory molecules,promoting the production of an“inflammatory soup”by inducing vasodilatation and plasma extravasation as well as by promoting the release of chemical mediators from disc cells and infiltrating immune cells,causing neurogenic inflammation that leads to progressive disc degeneration and discogenic pain.
基金supported by the National Natural Science Foun-dation of China(Grant No.52073103,52272276,51873069,and 52373128)Beijing Municipal Health Commission(BMHC-2021-6,BJRITO-RDP-2024)Beijing Municipal Public Welfare Devel-opment and Reform Pilot Project for Medical Research Institutes(JYY2023-11,JYY2023-8).
文摘Effective treatment of intervertebral disc degeneration with biomaterials remains a challenge,owing to the difficulty in simultaneously overcoming oxidative stress and its associated cascades in the nucleus pulposus microenvironment,which includes cellular senescence,apoptosis,inflammation,and extracellular matrix(ECM)degradation.To address these issues,a multifunctional hydrogel(HG-QNT)loaded with transforming growth factorβ1(TGFβ1)and quercetin-based nanoparticles(QUNPs)is developed through borate ester bonding and Schiffbase reaction-induced crosslinking.Specifically,QUNPs fabricated via coordination and hydrophobic interactions endow the hydrogel with extraordinary antioxidative properties.Benefiting from the multi-dynamic crosslinking,the hydrogel achieves self-healing,mechanical stability,and pH-responsive release of QUNPs and TGFβ1.The HG-QNT hydrogel is demonstrated to enhance the proliferation of encapsulated nucleus pulposus cells,thereby providing an ideal platform for cell transplantation.The cooperative antioxidation of QUNPs and the hydrogel carrier renders HG-QNT effective in mitigating oxidative stress,resulting in the suppression of cellular senescence,mitochondrial dysfunction,apoptosis,excessive inflammation,and abnormal catabolism.Afterwards,TGFβ1 and QUNPs act in synergy with the hydrogel to restore the anabolic/catabolic balance by enhancing ECM synthesis.Overall,the strategy orchestrating multiple modulation by HG-QNT hydrogel shows great potential for application in intervertebral disc regeneration.
基金supported by the National Natural Science Foundation of China(grant No.82405081)Guangdong Basic and Applied Basic Research Foundation(grant No.2025A1515011267)the National Postdoctoral Research Program(certificate number:GZC20230979)+4 种基金the China Postdoctoral Science Foundation(certificate numbers:2023TQ0135 and 2024M751136)Guangzhou Basic and Applied Basic Research(2025A04J3397)the Guangzhou Key Laboratory of Formula-Patterns of Traditional Chinese Medicine(No.202102010014)the Project of Administration of Traditional Chinese Medicine of Guangdong Province of China(No.20232179)the Fundamental Research Funds for the Central Universities(No.21624362).
文摘Background:Renshen Guben oral liquid(RGSB)has the ability to consolidate constitution,restore vitality,and delay the ageing process and the onset of geriatric diseases.RGSB also has potential therapeutic effects on intervertebral disc degeneration(IVDD).This study investigated the therapeutic effects of RGSB on IVDD and explored the underlying mechanisms.Methods:Single-cell sequencing and network pharmacology analyses were conducted in combination to determine the mechanisms of IVDD.Thirty-two C57BL/6J mice were randomly assigned to four groups:the sham operation group,IVDD group,Ibuprofen(IB)group,and RGSB group.All groups except for the sham operation group underwent needle puncture of the caudal IVDs.Behavioral tests,ELISA,PCR,TUNEL staining,and immunohistochemical staining were performed to investigate the therapeutic mechanisms associated with IVDD.Results:Bioinformatics analysis revealed that apoptosis is involved in the development of IVDD and that RSGB might alleviate IVDD by inhibiting apoptosis.After needle puncture,the mice exhibited from thermal allodynia,a decrease in disc height,an increase in the Pfirrmann grade,and destruction of IVDs.Surgery to induce IVDD increased the levels of Matrix Metalloproteinase-3(MMP3),Caspase-6,and Cyt-c in nucleus pulposus cells and the periphery;serum levels of interleukin-1β(IL‐1β)and TNF‐α;and the mRNA levels of IL-6,TNF-α,and IL-1β.Moreover,IVDD decreased the IL-10 mRNA level and the expression of Col-I and BCL2.RSGB reversed these changes.Conclusion:RSGB alleviated IVDD by relieving inflammation and apoptosis and alleviating neuroinflammation.
