Collagen Degradation Products Research Articles

A Novel High-Throughput Immunoaffinity LC-MS/MS Assay for P-III-NP and Other Fragments of Type III Procollagen in Human Serum.

The amino-terminal propeptide of type III procollagen (P-III-NP) is used with IGF-I to detect the illicit use of growth hormone and to monitor growth hormone therapy. However, the only currently available assays for P-III-NP are immunoassays, which are not well harmonized. In addition, other fragments of type III procollagen may better evaluate collagen turnover. We aimed to develop a high-throughput assay using immunoaffinity enrichment coupled to ultra-high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) to quantify peptides belonging to three different regions of type III procollagen in human serum simultaneously. To facilitate higher throughput, we transferred the assay from microcentrifuge tubes to a 96-well plate format with partially automated pipetting. The method was linear (Pearson's R ≥ 0.994) over an estimated concentration range of 1.35-13.3 nM, 0.04-2.28 nM, and 0.26-5.1 nM for each surrogate peptide of P-III-NP, collagen degradation products, and the carboxyl-terminal propeptide, respectively. Intra-day and inter-day imprecision were both < 13.6%, and the results of robustness testing were also encouraging. The method was successfully applied to capillary blood samples obtained using Tasso+ microsampling devices. Modest correlation of P-III-NP concentration was observed between our new method and a WADA-approved immunoassay (N = 40, Pearson's R = 0.789) with a significant bias of -87.8%. Our method simultaneously quantifies four peptides belonging to three regions of type III procollagen in human serum. High bias between assays highlights the need for common higher-order calibrators or reference materials to help improve the comparability of results across laboratories.

A Comprehensive Review of Advanced Biomarkers for Chronic Kidney Disease in Older Adults: Current Insights and Future Directions.

Chronic kidney disease (CKD) is a growing health concern, particularly in older adults, due to its high prevalence and association with increased morbidity and mortality. Early detection and effective management are crucial to slowing disease progression and reducing complications such as cardiovascular events and end-stage renal disease (ESRD). Traditional biomarkers, including serum creatinine and estimated glomerular filtration rate (eGFR), often have limitations in older populations, where age-related physiological changes can obscure early signs of kidney dysfunction. Advanced biomarkers offer a more precise and comprehensive understanding of kidney health, providing insights into pathological processes such as inflammation, fibrosis, oxidative stress, and tubular injury. These biomarkers have the potential to enhance early diagnosis, predict disease progression, and inform personalized treatment approaches, particularly in the elderly. This review explores the current landscape of advanced biomarkers for CKD in older adults, highlighting their clinical utility and limitations. Key biomarkers, including those related to inflammation (C-reactive protein, interleukin-6), fibrosis (transforming growth factor-beta, collagen degradation products), oxidative stress (F2-isoprostanes, malondialdehyde), and tubular injury (kidney injury molecule-1, neutrophil gelatinase-associated lipocalin), are examined in the context of CKD. Emerging technologies, such as multi-omics and machine learning, are also discussed as they offer new opportunities for biomarker discovery and integration into clinical practice. While challenges remain, including the need for longitudinal studies and better standardization, advanced biomarkers hold promise for transforming CKD management in older adults, paving the way for earlier detection, better risk stratification, and more targeted therapeutic interventions.

Identification of Risk Factors for Primary Osteoporosis: The Role of Cervical Ligament Ossification.

BACKGROUND Long-term clinical practice has suggested a possible association between ossification of cervical ligament (OCL) and primary osteoporosis (POP). However, there is a lack of relevant research data. This study aimed to clarify the potential relationship between OCL and POP, and propose new strategies for preventing the onset of POP. MATERIAL AND METHODS The study involved 107 patients. The patients' diagnosis included OCL (ossification of the posterior longitudinal ligament, ossification of the ligamentum flavum, and ossification of the nuchal ligament) and POP. Bone mineral density (BMD), types of OCL, types of ossification of posterior longitudinal ligament, age, sex, serum calcium, serum phosphorus, alkaline phosphatase, type I collagen amino-terminal extension peptide, type I collagen degradation products, osteocalcin N-terminal molecular fragments, 25-hydroxyvitamin D, and history of taking steroid drugs were collected. SPSS24.0 and GraphPad Prism 8 were used to obtain the risk factors for POP. RESULTS One-way analysis of variance found that OCL, ossification of posterior longitudinal ligament, alkaline phosphatase, and osteocalcin N-terminal molecular fragments had statistical significance on BMD of the femoral neck (P<0.05). The independent sample t test showed that patient sex had statistical significant effect on BMD (femoral neck) (P=0.036). Incorporating the above factors into multiple linear regression analysis, it was found that OCL, alkaline phosphatase, and osteocalcin N-terminal molecular fragments were risk factors affecting BMD of femoral neck (P<0.05). CONCLUSIONS OCL, osteocalcin N-terminal molecular fragments, and alkaline phosphatase are risk factors for POP.

Vitamin D supplementation reduces systemic modulation of oxidative stress and collagen degradation in keratoconus patients

Aims/Purpose: To assess whether vitamin D (Vit D) supplementation modulates systemic biomarkers involved in collagen degradation and oxidative stress in adolescent patients affected with keratoconus (KC).Methods: This study included forty adolescent patients (age range: 16–19 years) with KC and Vit D insufficiency (&lt;30 ng/mL). The patients were supplemented with vitamin D for 6 months and followed up for additional 6 months (total study duration: 12 months). Blood samples were collected at 1, 6, and 12 months. The levels of Vit D or its receptor (VDR), collagen catabolism (type I collagen degradation product; ICTP), and oxidative stress (Peroxisome proliferator‐activated receptor gamma; PPAR‐γ) were quantified by ELISA or RT‐PCR.Results: We found that Vit D levels significantly increased at 6 months after supplementation. Moreover, at month 6 the expression of VDR was upregulated (p &lt; 0.01), while collagen degradation (ICTP; p &lt; 0.01) and oxidative stress (PPAR‐γ; p &lt; 0.05) were significantly reduced.Conclusions: Our findings show that Vit D supplementation reduces oxidative stress and collagen catabolism, which have a key role in KC pathophysiology. These findings provide a potential mechanism by which Vit D could impact KC progression and provide preliminary evidence that KC may be a systemic disease.

STUDY OF SERUM URIC ACID, CALCIUM AND ALPAND URINARY HYDROXYPROLINE CREATININE RATIO IN POSTMENOPAUSAL WOMEN

During menopause women succumb to physiological and biochemical changes along with middle age health hazards. In view of sudden fall in Estrogens during menopause Osteoporosis of aging is fortied by Estrogen deciency. The present study was done to evaluate bone health through biochemical parameters in Guntur Urban area between September 2021 and January 2022. 50 Premenopausal women and 60 postmenopausal women were selected for the study after screening verbally . Serum Calcium, Phosphorus, Alk, Phosphatase and Uric acid along with 24Hrs urinary excretion of Creatinine, Calcium and Hydroxyproline were estimated in both groups. We could not nd wide difference in the mean values of serum Ca , P, ALP&amp; UAbut Urine 24 Hrs Ca was lower in postmenopausal women (104.55±23.86) compared to premenopausal group (124.42 ±17.78 ). Urine Hydroxyproline(HOP) showed higher values in postmenopausal women (25.79±1.44 ) than in Premenopausal women.(18.00 ± 2.55). All the analytes showed statistically signicant values between the groups and within the groups with P value value of &lt;0.0001. 24Hrs urine Hydroxyproline , which is collagen degradation product correlated with 24hrs urine Ca and Creatinine.With P value &lt; 0.0001 Serum ALP , Ca &amp; Uric acid as shown by ANOVA, both within groups and between groups was statistically signicant P &lt;0.0001. We conclude that collagen degradation product like HOPcan be evaluated and considered as marker for assessment of bone health in postmenopausal women.

The Mortality Risk and Pulmonary Fibrosis Investigated by Time-Resolved Fluorescence Spectroscopy from Plasma in COVID-19 Patients.

A method of rapidly pointing out the risk of developing persistent pulmonary fibrosis from a sample of blood is extraordinarily needed for diagnosis, prediction of death, and post-infection prognosis assessment. Collagen scar formation has been found to play an important role in the lung remodeling following SARS-CoV-2 infection. For this reason, the concentration of collagen degradation products in plasma may reflect the process of lung remodeling and determine the extent of fibrosis. According to our previously published results of an in vitro study, an increase in the concentration of type III collagen degradation products in plasma resulted in a decrease in the fluorescence lifetime of plasma at a wavelength of 450 nm. The aim of this study was to use time-resolved fluorescence spectroscopy to assess pulmonary fibrosis, and to find out if the lifetime of plasma fluorescence is shortened in patients with COVID-19. The presented study is thus far the only one to explore the fluorescence lifetime of plasma in patients with COVID-19 and pulmonary fibrosis. The time-resolved spectrometer Life Spec II with the sub-nanosecond pulsed 360 nm EPLED® diode was used in order to measure the fluorescence lifetime of plasma. The survival analysis showed that COVID-19 mortality was associated with a decreased mean fluorescence lifetime of plasma. The AUC of mean fluorescence lifetime in predicting death was 0.853 (95% CI 0.735–0.972, p < 0.001) with a cut-off value of 7 ns, and with 62% sensitivity and 100% specificity. We observed a significant decrease in the mean fluorescence lifetime in COVID-19 non-survivors (p < 0.001), in bacterial pneumonia patients without COVID-19 (p < 0.001), and in patients diagnosed with idiopathic pulmonary fibrosis (p < 0.001), relative to healthy subjects. Furthermore, these results suggest that the development of pulmonary fibrosis may be a real and serious problem in former COVID-19 patients in the future. A reduction in the mean fluorescence lifetime of plasma was observed in many patients 6 months after discharge. On the basis of these data, it can be concluded that a decrease in the mean fluorescence lifetime of plasma at 450 nm may be a risk factor for mortality, and probably also for pulmonary fibrosis in hospitalized COVID-19 patients.

A Targeted Liquid Chromatography-Tandem Mass Spectrometry Method for Simultaneous Quantification of Peptides from the Carboxyl-terminal Region of Type III Procollagen, Biomarkers of Collagen Turnover.

The development of analytical approaches to help reduce the risk of growth hormone (GH) doping is important to fair competition and the health of athletes. However, the reliable detection of GH use remains challenging. The identification of novel biomarkers of GH administration could lead to a better understanding of the physiological response to GH, more sensitive detection of the illicit use of GH in sport, and better management of patients treated for GH disorders. We developed a targeted liquid chromatography-tandem mass spectrometry method to simultaneously quantify the carboxyl-terminal propeptide of type III procollagen (P-III-CP) and type III collagen degradation products in human serum. Following proteolysis, we instituted a simple acid precipitation step to reduce digested sample complexity before peptide immunoenrichment, which improved the recovery of one target peptide from serum. We evaluated the concentration of each biomarker at different age ranges and after GH administration in healthy participants. The assay was linear over an estimated concentration range of 0.3 to1.0 nM and 0.1 to 0.4 nM for each surrogate peptide of P-III-CP and collagen fragments, respectively. Intra-day and inter-day coefficients of variation were ≤15%. Biomarker concentrations appeared to vary with age and to reflect age-specific collagen turnover. Moreover, their concentrations changed after GH administration. Our method quantifies the proteins belonging to the family of P-III-CP and type III collagen degradation products in human serum, which could be used to detect GH administration in athletes and better understand diseases involving GH therapy or altered type III collagen turnover.

Clinical evaluation of muscle functions in neurofibromatosis type 1.

Muscle weakness, fatigue and speech problems can occur in neurofibromatosis type 1 (NF1). The pathogenesis of these symptoms is unclear, likely multifactorial. We examined motor function in limb and speech muscles in NF1 patients. We evaluated NF1 and control groups aged 4-18 years for muscle strength, tone and mobility using standard manual testing, joint motion and Beighton score measurements. Speech and language functions were assessed by speech articulation and resonance. As a marker of muscle tissue turnover, we determined collagen degradation products in urine before and after submaximal exercise. NF1 patients had reduced strength in proximal limb muscles compared to control subjects. Speech articulation problems and hypernasality were more common in NF1 (47% and 38%, respectively). Collagen products excreted in urine correlated with gluteal and biceps muscle strength. Muscle dysfunction can be detected in some children with NF1 and may explain certain clinical features including fatigue, speech and articulation problems. If confirmed by further research, these findings may be relevant to the management of this condition.

Longitudinal serological assessment of type VI collagen turnover is related to progression in a real-world cohort of idiopathic pulmonary fibrosis

BackgroundRemodeling of the extracellular matrix (ECM) is a central mechanism in the progression of idiopathic pulmonary fibrosis (IPF), and remodeling of type VI collagen has been suggested to be associated with disease progression. Biomarkers that reflect and predict the progression of IPF would provide valuable information for clinicians when treating IPF patients.MethodsTwo serological biomarkers reflecting formation (PRO-C6) and degradation (C6M) of type VI collagen were evaluated in a real-world cohort of 178 newly diagnoses IPF patients. All patients were treatment naïve at the baseline visit. Blood samples and clinical data were collected from baseline, six months, and 12 months visit. The biomarkers were measured by competitive ELISA using monoclonal antibodies.ResultsPatients with progressive disease had higher (P = 0.0099) serum levels of PRO-C6 compared to those with stable disease over 12 months with an average difference across all timepoints of 12% (95% CI 3–22), whereas C6M levels tended (P = 0.061) to be higher in patients with progressive disease compared with stable patients over 12 months with an average difference across all timepoints of 12% (95% CI − 0.005–27). Patients who did not receive antifibrotic medicine had a greater increase of C6M (P = 0.043) compared to treated patients from baseline over 12 months with an average difference across all timepoints of 12% (95% CI − 0.07–47). There were no differences in biomarker levels between patients receiving pirfenidone or nintedanib.ConclusionsType VI collagen formation was related to progressive disease in patients with IPF in a real-world cohort and antifibrotic therapy seemed to affect the degradation of type VI collagen. Type VI collagen formation and degradation products might be potential biomarkers for disease progression in IPF.

Development of a Pure Small Molecule αVβ3 Antagonist

# Contributed equallyαVβ3 has been implicated in the pathophysiology of important disorders, including osteoporosis, sickle cell disease, tumor angiogenesis, metastasis, virus invasion, glomerulonephritis, dermal and hepatic fibrosis, acute myelogenous leukemia, supravalvular aortic stenosis, bowel strictures, and T-cell lymphoma, but no αVβ3 antagonists have been approved for human therapy. The RGD-based αVβ3 antagonist MK-0429 (MK429) had favorable results in osteoporosis patients in Phase 2, but was not advanced, and the RGD-based antagonist cilengitide was studied in patients with malignancies, but did not demonstrate efficacy. Since RGD-based antagonists can under certain circumstances activate αVβ3, prime it to bind ligand, induce apoptosis, increase tumor angiogenesis and growth, and increase hepatic fibrosis, we sought to develop a pure αVβ3 antagonist that does not induce high affinity ligand binding or exposure of the epitope for mAb AP5, both of which depend on the swing-out of the β3 PSI domain. We built on Arnaout's group's studies of a variant fibronectin fragment with high affinity for αVβ3 (hFN10) that did not induce the β3 swing-out motion because a Trp substitution on the ligand formed a π-π interaction with β3 Y122 on the β1-α1 loop, thus preventing the latter's movement toward the MIDAS, which is the trigger for β3 swing-out. To develop a small-molecule that mimics the effect of hFN10, we used inferences from a predicted docking pose of MK429 into the hFN10 binding site of the αVβ3 crystal structure to rationally design a compound with an aromatic ring that directly engages β3 Y122. Based on the results of molecular dynamics simulations, the aromatic group of this compound, TDI-4161, is only 4.4 Å from Y122 (and forms a π-π interaction), which compares with 4.7 Å for hFN10 and 7.0 Å for MK429 (which does not form a π-π interaction). We analyzed compounds in 2 assays: 1. Adhesion of HEK293 cells expressing human αVβ3 (HEK-αVβ3) to fibrinogen (reported as the concentration producing 50% inhibition; IC50). 2. Exposure of the epitope for mAb AP5 (the concentration producing 50% exposure; EC50). The racemate of MK429 (rMK429) has an IC50=0.0036 µM and an EC50=0.019 µM, and cilengitide has an IC50=0.050 µM and an EC50=0.034 µM, indicating that both are potent αVβ3 antagonists and potent inducers of the αVβ3 active conformation. In sharp contrast, TDI-4161 has an IC50=0.022 µM and an EC50>10 µM, indicating that it does not induce the β3 conformational change even at 400X the IC50. TDI-4161 inhibited the binding of: purified αVβ3 to adenovirus 2 penton base (IC50 = 0.028 µM) and purified αVβ5 to vitronectin (IC50 =2.09 µM), but not purified αVβ6 to TGF-β1 Latency Associated Peptide even at 10 µM. TDI-4161inhibited adhesion of mouse endothelial cells containing murine αVβ3 to fibrinogen (IC50 = 0.437 µM). We tested TDI-4161's ability to 'prime“ αVβ3 by: incubating it with HEK-αVβ3 at 10 µM, fixing the cells with paraformaldehyde, washing, incubating with fluorescent fibrinogen, and measuring bound fibrinogen by flow cytometry. The peptide RGDS (100 µM) increased fibrinogen binding from 8.7 ± 1.5 to 24.3 ± 1.4 GMFI (p <0.001; n=3), but TDI-4161 did not (7.0 ± 2.6 GMFI; p<0.001 vs RGDS). After oral administration of TDI-4161 to mice at 100 mg/kg, the Cmax was 7.5 µM at the Tmax of 0.94 hours; the T1/2 was 3.6 hours. Once-daily oral dosing of TDI-4161 for 28 days at 10, 30, and 100 mg/kg was well tolerated in male CD-1 mice (n=3). Day 1 and day 28 plasma levels 1 hour after dosing were similar. We tested the effect of TDI-4161 on osteoclast function by adding it at 10 µM (0.1% DMSO) to bone marrow-derived murine macrophages and cultured them either on plastic or bovine bone slices with RANKL and a source of M-CSF. After 5 days, cells incubated with TDI-4161 demonstrated an osteoclast phenotype similar to that of β3 null mice; osteoclast numbers were better preserved than with rMK429, but less well than with the DMSO control. Supernatants from bone slices were assayed for cross-linked collagen degradation products (CrossLaps). When compared to the DMSO control (32.7 nM), rMK429 (10 µM) produced 82% inhibition (5.9 nM) and TDI-4161 produced 74% inhibition (8.3 nM). In summary, using inferences from structural data derived from hFN10, we have synthesized an orally bioavailable pure αVβ3 antagonist that reacts with both human and murine αVβ3, does not prime αVβ3 to bind fibrinogen, and inhibits osteoclast bone resorption. DisclosuresFilizola:Icahn School of Medicine at Mount Sinai: Patents & Royalties. Foley:Tri-Institutional Therapeutics Discovery Institute: Patents & Royalties.

Evaluation of Intra-Articular Amikacin Administration in an Equine Non-inflammatory Joint Model to Identify Effective Bactericidal Concentrations While Minimizing Cytotoxicity.

Septic arthritis causes significant morbidity and mortality in veterinary and human clinical practice and is increasingly complicated by multidrug-resistant infections. Intra-articular (IA) antibiotic administration achieves high local drug concentrations but is considered off-label usage, and appropriate doses have not been defined. Using an equine joint model, we investigated the effects of amikacin injected at three different doses (500, 125, and 31.25 mg) on the immune and cartilage responses in tibiotarsal joints. Synovial fluid (SF) was sampled at multiple time points over 24 h, the cell counts determined, and amikacin concentrations measured by liquid chromatography-mass spectrometry. Cytokine concentrations and collagen degradation products in SF were measured by ELISA and multiplex immunoassays. The mean amikacin concentrations in SF were greater than or equal to the minimum inhibitory concentration (MIC) (0.004 mg/ml) for most common equine joint pathogens at all time points tested to 24 h for all three amikacin doses evaluated. The inflammatory cytokines tumor necrosis factor-alpha (TNF-α) and interleukin-1 beta (IL-1β) increased significantly in SF in the highest amikacin dose group, despite the fact that increases in SF cell counts were not observed. Similarly, the biomarkers of cartilage type II collagen cleavage (C2C and C12C) were increased in SF following amikacin injection. Mechanistically, we further demonstrated using in vitro studies that chondrocytes and synoviocytes killed by exposure to amikacin underwent apoptotic cell death and were phagocytosed by macrophages in a non-inflammatory process resembling efferocytosis. Neutrophils and T cells were susceptible to amikacin cytotoxicity at clinically relevant doses, which may result in blunting of cellular inflammatory responses in SF and account for the lack of increase in total nucleated cell counts following amikacin injection. In summary, decisions on whether to inject cytotoxic antibiotics such as aminoglycosides intra-articularly and what doses to use should take into account the potential harm that antibiotics may cause and consider lower doses than those previously reported in equine practice.

The significance of plasma collagen degradation products as biomarkers for advanced hypertensive heart disease.

The significance of plasma collagen degradation products as biomarkers for advanced hypertensive heart disease.

A review on biomarkers in clinical osteoporosis - Significance of hydroxyproline

Osteoporosis is a chronic disorder in both men and women, where it decreases the bone quality and quantity, which may lead to a fall in bone mass density. It mostly occurs because of reduced peak bone mass or excessive formation or resorption during bone remodeling. Various degradation products of bone collagen are pyridinoline (Pyr), deoxypyridinoline, N-terminal telopeptides, and hydroxyproline (Hyp), which are excreted in the urine. Among that, Hyp is a crucial biomarker used to correlate osteoporosis condition. An overview of all the biomarkers and the signed note of Hyp with different analytical methods are provided in the current piece of work. To quantify amounts in biological samples, various methods have been developed for Hyp analysis, such as the colorimetric technique, gas chromatography, gas chromatography-mass spectrometry, and widely used high-performance liquid chromatography tests to detect the concentration of Hyp in different matrices. This review states the importance of biomarkers and Hyp's significance for detecting osteoporosis, clinical manifestation, and different analytical methods for Hyp. The study indicates that hydroxyproline is a unique biomarker used to detect osteoporosis. It is also showing the clinical manifestation of osteoporosis and discussed different analytical methods for hydroxyproline.

Association of Diabetic Kidney Disease Markers and Urinary Beta-crossLaps in Type 2 Diabetes

Introduction: Diabetic Kidney Disease (DKD) is a chronic complication in Type 2 diabetes. The Chronic Kidney Disease (CKD273) peptide classifier has been found to predict development of DKD even before microalbuminuria develops. Seventy four percent of peptides in the CKD273 classifier are Collagen degradation fragments. The Beta-CrossLaps (β-CTx) Enzyme Llinked Immunosorbent Assay (ELISA) assay detects the specific collagen degradation product, C terminal telopeptide of Type 1 collagen. In light of the Capillary Electrophoresis/Mass Spectrometry (CE-MS) findings, linking collagen degradation fragments excretion to early detection of DKD, the significance of urinary β-CTx levels as a DKD biomarker needs to be evaluated. Aim: To study the urinary excretion of β-CTx in type 2 diabetes patients and to evaluate its relation to Microalbuminuria status and estimated Glomerular Filtration Rate (eGFR) of the patients. Materials and Methods: This descriptive cross-sectional study was undertaken at a tertiary care hospital, with enrollment of 82 type 2 diabetes patients from the diabetes Out Patient Department (OPD) Participants were divided into groups based on their Urinary Albumin Creatinine Ratio (UACR) and eGFR levels. The study participants were tested for Urinary β-CTx level, UACR and eGFR. Mean or median was calculated for the parameters with normal and non-normal distribution, respectively. All statistical testing was performed on online calculators available at the site; https://www.socscistatistics.com/. Results: The median urinary β-CTx level observed was 100.6 ng/mmol of creatinine. Among the 82 participants, 15 participants had urinary β-CTx level 15pg/mL, the sensitivity of the kit. Among the remaining 67 participants, the minimum Urinary Beta-CrossLaps: Creatinine ratio observed was 2.6 ng/mmol and the maximum value observed was 2071 ng/mmol (i.e., 2.1 μg/mmol). The median urinary β-CTx level was highest (100.6 ng/mmol creatinine) in the patient group with eGFR in the normal range. The urinary β-CTx level was found to decline with decline in eGFR, with median urinary β-CTx 65.5 ng/mmol creatinine in the patient group with mildly decreased eGFR and 7.2 ng/mmol creatinine in the patient group with moderately decreased eGFR. Conclusion: The Urinary β-CTx concentration in type 2 diabetes patients is dispersed over a wide range. The Urinary β-CTx concentration correlates with the eGFR of the patient and is not influenced by age, gender or duration of diabetes. This parameter is a potential early DKD biomarker.

Elevated serum carboxy-terminal telopeptide of type I collagen predicts clinical outcome in patients with acute coronary syndrome who underwent percutaneous coronary intervention

Abstract Introduction Serum carboxy-terminal telopeptide of type I collagen (I-CTP) is a collagen degradation product of type I collagen in the extracellular matrix of the heart, blood vessels, and bone. The serum levels of I-CTP were reportedly a predictive marker for cardiac remodeling after acute myocardial infarction. However, it remains unclear whether I-CTP can predict poor clinical outcome in patient with acute coronary syndrome (ACS). Purpose The aim of this study was to investigate the association between serum levels of I-CTP and clinical outcome in patients with ACS. Methods Serum levels of I-CTP were measured in 200 patients with ACS who underwent percutaneous coronary intervention (PCI). All patients were prospectively followed during the median follow-up period of 1312 days with the end point of major adverse cardiovascular events (MACE). We divided the patients into tertiles according to serum I-CTP level: low I-CTP group (≤4.4 ng/ml, n=72), middle I-CTP group (4.4–6.4 ng/ml, n=65), and high I-CTP group (≥6.5 ng/ml, n=63). Results There were 44 MACE, including 24 all-cause death and 9 rehospitalization due to heart failure. I-CTP was significantly higher in patients with MACE than those without (4.90 [interquartile range (IQR): 3.80–6.38] ng/ml vs. 6.65 [IQR: 5.00–10.08] ng/ml, p&amp;lt;0.001). Kaplan-Meier analysis demonstrated that patients in the highest tertile of I-CTP had the greatest risk of MACE. In a univariate analysis, age, Albumin, estimated glomerular filtration rate (eGFR), low-density lipoprotein cholesterol (LDL-C), brain natriuretic peptide (BNP), high-sensitivity C-reactive protein (hsCRP) and I-CTP were significant predictors of MACE. A multivariate Cox proportional hazard analysis showed that the high I-CTP group had a higher risk for MACE (Hazard ratio [HR] 2.6, p=0.049) compared with the low I-CTP group after adjusting for confounding factors. Conclusions I-CTP was significantly associated with MACE, suggesting that I-CTP could be a reliable marker for clinical outcome in patients with ACS who underwent PCI. Figure 1 Funding Acknowledgement Type of funding source: None

Therapeutic efficacy of zoledronic acid combined with calcitriol in elderly patients receiving total hip arthroplasty or hemiarthroplasty for osteoporotic femoral neck fracture.

To validate the therapeutic efficacy of zoledronic acid (ZOL) in elderly patients with femoral neck fracture who received total hip arthroplasty (THA) or hemiarthroplasty (HA). Included in this study were 95 elderly patients with femoral neck fractures who received THA/HA between August 2015 and June 2018. They were randomized into a ZOL group and a control group. Patients in ZOL group received a yearly single dose of 5mg ZOL intravenous injection plus 0.5μg/day calcitriol and 1000mg/day calcium carbonate 2days before THA or HA. Patients in the control group were treated with the same dose of calcitriol and calcium carbonate only without ZOL. Bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry. Bone metabolism markers including the total extension of the peptide type I collagen amino end (P1NP) and beta collagen degradation product (β-CTX) were obtained by serum examination. The postoperative functional outcome was assessed using Harris Hip Score (HHS). During the follow-up period, BMD in the ZOL group was improved and significantly higher than that in the control group at 6 and 12months post-operation. Bone metabolism markers P1NP and β-CTX in ZOL group remained at a relatively low level as compared with that in the control group at 6months after treatment. No significant difference in the mean HHS and the excellent/good rate of joint function was observed during the follow-up period between the two groups. The occurrence of adverse events in the ZOL group was significantly higher than that in the control group. A single infusion of ZOL shows promise in improving BMD of the healthy side of the femoral neck, lumbar spine, and total hip and decreasing the level of bone markers, which may improve the clinical outcome of patients with osteoporotic femoral neck fractures receiving THA/HA.

A New Method of Gelatin Modified Collagen and Viscoelastic Study of Gelatin-Collagen Composite Hydrogel

Pure collagen materials are expensive with poor mechanical properties, which need modifications in most cases. As the degradation product of collagen, gelatin is cheap, degradable and biocompatible, but few literatures have reported the research about gelatin-collagen composite materials. This is because gelatin and collagen have different soluble temperatures—gelatin is soluble in hot water (≥30 °C) and swells in cold water. However, a low temperature (2–10 °C) is required to prepare and store collagen solution, and neutral collagen solution denatures quickly above the room temperature. In this study, gelatin was ground into powders and swelled in neutral bovine tendon pepsin-soluble collagen solution (BPSC) to form a homogeneous gelatin-collagen mixture, in light of the swelling characteristics of gelatin in cold water. The assembly properties and gel properties of this composite material were further studied. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) test results showed that the bovine tendon collagen had typical type-I collagen structural characterizations with two α chains of about 100 kDa and one β chain of about 200 kDa; while the SDS-PAGE pattern of gelatin displayed bands continuously distributed from 30 to 200 kDa. Amino acid composition analysis test indicated that the content of polar amino acids and the sum of acidic and base amino acids for gelatin were higher than that of BPSC. Studies on gel properties demonstrated that gelatin-collagen mixed solution had collagen-like assembly characteristics and assembly kinetics. The moduli of the assembled gel at 35 °C were equivalent to that of pure bovine tendon collagen system; moreover, the system moduli didn’t change with time with elastic moduli (G′) of about 40 Pa. However, at 25 °C, the moduli of gelatin-collagen composite hydrogel increased with the extension of time, its G′ increased about 18 times within 8 h, and the ratio of elastic modulus to viscous modulus (G″) increased 4.6 times, showing a significant aging effect of structural strength. Meanwhile, the mechanical strength of the composite hydrogel was also regulated by temperature—the gel was highly elastic (G′≈3,000 Pa, G′>>G″) at a low temperature (5 °C); as the temperature rose, the system moduli gradually decreased and the elastic gel transformed into waterlike fluid at 50 °C little by little. What’s more, gelatin-collagen composite hydrogel also had reversible sol-gel performances and self-healing capability similar to the gelatin hydrogel. This novel preparation method for preparing composite materials and the resultant composite hydrogel are expected to be used in the fields of natural food gels, injectable hydrogels, cell scaffolds, drug sustained-release materials and so on, and improve and promote the processing performances, price and large-scale production of collagen-based materials.

Prolidase deficiency in two dermatological patients in western Sicily

Prolidase deficiency is a rare disorder inherited through an autosomal recessive gene. The hallmark of the disorder are iminodipeptiduria, chronic skin ulcers, recurring infections, mental retardation and characteristic facial appearance, although prolidase deficiency can occur with no clinical manifestation. The primary biological function of the enzyme involves the metabolism of collagen degradation products and the recycling of proline for collagen resynthesis. We describe two patients with prolidase deficiency and review the different clinical manifestations suggesting the pathogenetic mechanism through few hypotheses.

Biomarkers in Painful Symptomatic Knee OA Demonstrate That MRI Assessed Joint Damage and Type II Collagen Degradation Products Are Linked to Disease Progression.

BackgroundOsteoarthritis (OA) is the most prevalent arthritis worldwide, but the evolution of pain in relation to joint damage and biochemical markers are not well understood. We evaluated the relation between clinical pain measures and evoked pain in relation to structural damage and biochemical biomarkers in knee OA.MethodsA cross-sectional study in people with knee OA and healthy controls was conducted. A total of 130 participants with advanced OA requiring total knee replacement (TKR) (n = 78), mild OA having standard care (n = 42) and non-OA controls (n = 6), with four drop-outs were assessed. Pain scoring was performed by the Western Ontario and McMaster Universities OA Index (WOMAC_P) and the Visual Analog Scale (VAS). Pain sensitization was assessed by pain pressure thresholds (PPTs). Knee magnetic resonance imaging (MRI) assessed joint damage using the MRI Knee OA Score (MOAKS). Overall MOAKS scores were created for bone marrow lesions (BMLs), cartilage degradation (CD), and effusion/Hoffa synovitis (tSyn). Type II collagen cleavage products (CTX-II) were determined by ELISA.ResultsThe advanced OA group had a mean age of 68.9 ± 7.7 years and the mild group 63.1 ± 9.6. The advanced OA group had higher levels of pain, with mean WOMAC_P of 58.8 ± 21.7 compared with the mild OA group of 40.6 ± 26.0. All OA subjects had pain sensitization by PPT compared with controls (p < 0.05). WOMAC_P correlated with the total number of regions with cartilage damage (nCD) (R = 0.225, p = 0.033) and total number of BMLs (nBML) (R = 0.195, p = 0.065) using body mass index (BMI), age, and Hospital Anxiety and Depression Scale (HADS) as covariates. Levels of CTX-II correlated with tSyn (R = 0.313, p = 0.03), nBML (R = 0.252, p = 0.019), number of osteophytes (R = 0.33, p = 0.002), and nCD (R = 0.218, p = 0.042), using BMI and age as covariates. A multivariate analysis indicated that BMI and HADS were the most significant predictors of pain scores (p < 0.05).ConclusionPeople with both mild and advanced OA show features of pain sensitization. We found that increasing MRI-detected joint damage was associated with higher levels of CTX-II, suggesting that increasing disease severity can be assessed by MRI and CTX-II biomarkers to evaluate OA disease progression.

Free urinary glycosylated hydroxylysine as an indicator of altered collagen degradation in the mucopolysaccharidoses.

Extracellular matrix (ECM) disruption is known to be an early pathological feature of the Mucopolysaccharidoses (MPS). Collagen is the main component of the ECM and its metabolism could act as a useful indicator of ECM disruption. We have measured the specific collagen breakdown products; urinary free hydroxylated (Lys-OH) and glycosylated hydroxylysines (Lys-O-Gal and Lys-O-GalGlc) in MPS patients using a tandem liquid chromatography tandem mass spectrometry assay. A pilot study cohort analysis indicated that concentrations of lysine and Lys-OH were raised significantly in MPS I (Hurler) disease patients. Lys-O-GalGlc was raised in MPS II and MPS VI patients and demonstrated a significant difference between MPS I Hurler and an MPS I Hurler-Scheie group. Further analysis determined an age association for glycosylated hydroxylysine in control samples similar to that observed for the glycosaminoglycans. Using defined age ranges and treatment naïve patient samples we confirmed an increase in glycosylated hydroxylysines in MPS I and in adult MPS IVA. We also looked at the ratio of Lys-O-Gal to Lys-O-GalGlc, an indicator of the source of collagen degradation, and noticed a significant change in the ratio for all pediatric MPS I, II, and IV patients, and a small significant increase in adult MPS IV. This indicated that the collagen degradation products were coming from a source other than bone such as cartilage or connective tissue. To see how specific the changes in glycosylated hydroxylysine were to MPS patients we also looked at levels in patients with other inherited metabolic disorders. MPS patients showed a trend towards increased glycosylated hydroxylysines and an elevated ratio compared to other metabolic disorders that included Battens disease, Fabry disease, Pyridoxine-dependent epilepsy (due to mutations in ALDH7A1), and Niemann Pick C disease.