Diameter Collagen Research Articles

Carbon Fiber-Mediated Electrospinning Scaffolds Can Conduct Electricity for Repairing Defective Tendon.

Partial or complete rupture of the tendon can damage the collagen structure, resulting in the disruption of the electrical signal pathway. It is a great challenge to reconstruct the original electrical signal pathway of the tendon and promote the regeneration and functional recovery of defective tendon. In this study, carbon fiber-mediated electrospinning scaffolds were fabricated by wrapping conductive, high-strength, loose single-bundle carbon fibers with nanofiber membranes. Due to the presence of nanofiber membranes, the maximum tensile force of the scaffolds was 2.4 times higher than that of carbon fibers, while providing excellent temporal and spatial prerequisites for tenocytes to adapt to electrical stimulation to accelerate proliferation and expression. The diameter of the carbon fiber monofilaments used in this study was 5.07 ± 1.20 μm, which matched the diameter of tendon collagen, allowing for quickly establishing the connection between the tendon tissue and the scaffold, and better promoting the recovery of the electrical signal pathway. In a rabbit Achilles tendon defect repair model, the carbon fiber-mediated electrospinning scaffold was almost filled with collagen fibers compared to a nonconductive polyethylene glycol terephthalate scaffold. Transcriptome sequencing revealed that fibromodulin and tenomodulin expression were upregulated, and their related proteoglycans and glycosaminoglycan binding proteins pathways were enhanced, which could regulate the TGF-β signaling pathway and optimize the extracellular matrix assembly, thus promoting tendon repair. Therefore, the scaffold in this study makes up for the shortage of conductive scaffolds for repairing tendon defects, revealing the potential impact of conductivity on the signaling pathway of tendon repair and providing a new approach for future clinical studies.

Microcurrent wave alleviates mouse intracranial arterial dolichoectasia development

Intracranial arterial dolichoectasia (IADE) is associated with the interaction of hypertension and inflammation, and microcurrent can be effective in hypertension. Therefore, this study aimed to investigate the therapeutic effect of microcurrent electrical stimulation in a mouse IADE model. This study randomly categorized 20 mice into five groups: group 1-C (healthy control), group 2-D (IADE model), group 3-M + D (microcurrent administration before nephrectomy and until brain surgery), group 4-D + M (microcurrent administration for 4 weeks following brain surgery), and group 5-M (microcurrent administration for 4 weeks). Cerebral artery diameter and thickness and cerebral arterial wall extracellular matrix components were assessed. Among the five groups, group 2-D showed significantly higher cerebral arterial wall diameter (117.79 ± 17.05 µm) and proportion of collagen (42.46 ± 14.12%) and significantly lower arterial wall thickness (9.31 ± 2.26 µm) and proportion of smooth muscle cell (SMC) and elastin in the cerebral arterial wall (SMC: 38.05 ± 10.32%, elastin: 11.11 ± 6.97%). Additionally, group 4-D + M exhibited a non-significantly lower diameter (100.28 ± 25.99 µm) and higher thickness (12.82 ± 5.17 µm). Group 5-M demonstrated no evidence of toxicity in the liver and brain. The pilot study revealed that microcurrent is effective in preventing IADE development, although these beneficial effects warrant further investigation.

A composite PET-matrix patch enhances tendon regeneration and tendon-to-bone integration for bridging repair of the chronic massive rotator cuff tears in a rabbit model.

In recent years, bridging repair has emerged as an effective approach for the treatment of massive rotator cuff tears (MRCTs). The objective of this study was to develop a composite patch that combines superior mechanical strength and biocompatibility and evaluate its potential for enhancing the outcomes of bridging repair for MRCTs. The composite patch, referred to as the PET-matrix patch (PM), was fabricated by immersing a plain-woven PET patch in decellularized matrix gel and utilizing the freeze-drying technique. The results demonstrated that the PM has reliable mechanical properties, with a maximum failure load of up to 480 N. The decellularized matrix sponge (DMS), present on the surface of the PM, displayed a loose and porous structure, with an average pore size of 62.51 μm and a porosity of 95.43%. In vitro experiments showed significant elongation of tenocytes on the DMS, with cells spanning across multiple pores and extending multiple protrusions as observed on SEM images. In contrast, tenocytes on the PET patch appeared smaller in size and lacked significant elongation. Additionally, the DMS facilitated the proliferation, migration and differentiation of tenocytes. In a rabbit model of chronic MRCTs, the PM group showed superior outcomes compared to the PET group at 4, 8 and 12 weeks after bridging repair. The PM group displayed significantly higher tendon maturing score, larger collagen diameter in the regenerated tendon and improved tendon-to-bone healing scores compared to the PET group (P < 0.05). Moreover, the maximum failure load of the tendon-bone complex in the PM group was significantly higher than that in the PET group (P < 0.05). In summary, the PM possesses reliable mechanical properties and excellent cytocompatibility, which can significantly improve the outcomes of bridging repair for chronic MRCTs in rabbits. Therefore, it holds great potential for clinical applications.

Mice Lacking PLAP-1/Asporin Show Alteration of Periodontal Ligament Structures and Acceleration of Bone Loss in Periodontitis.

Periodontal ligament-associated protein 1 (PLAP-1), also known as Asporin, is an extracellular matrix protein expressed in the periodontal ligament and plays a crucial role in periodontal tissue homeostasis. Our previous research demonstrated that PLAP-1 may inhibit TLR2/4-mediated inflammatory responses, thereby exerting a protective function against periodontitis. However, the precise roles of PLAP-1 in the periodontal ligament (PDL) and its relationship to periodontitis have not been fully explored. In this study, we employed PLAP-1 knockout mice to investigate its roles and contributions to PDL tissue and function in a ligature-induced periodontitis model. Mandibular bone samples were collected from 10-week-old male C57BL/6 (WT) and PLAP-1 knockout (KO) mice. These samples were analyzed through micro-computed tomography (μCT) scanning, hematoxylin and eosin (HE) staining, picrosirius red staining, and fluorescence immunostaining using antibodies targeting extracellular matrix proteins. Additionally, the structure of the PDL collagen fibrils was examined using transmission electron microscopy (TEM). We also conducted tooth extraction and ligature-induced periodontitis models using both wild-type and PLAP-1 KO mice. PLAP-1 KO mice did not exhibit any changes in alveolar bone resorption up to the age of 10 weeks, but they did display an enlarged PDL space, as confirmed by μCT and histological analyses. Fluorescence immunostaining revealed increased expression of extracellular matrix proteins, including Col3, BGN, and DCN, in the PDL tissues of PLAP-1 KO mice. TEM analysis demonstrated an increase in collagen diameter within the PDL of PLAP-1 KO mice. In line with these findings, the maximum stress required for tooth extraction was significantly lower in PLAP-1 KO mice in the tooth extraction model compared to WT mice (13.89 N ± 1.34 and 16.51 N ± 1.31, respectively). In the ligature-induced periodontitis model, PLAP-1 knockout resulted in highly severe alveolar bone resorption, with a higher number of collagen fiber bundle tears and significantly more osteoclasts in the periodontium. Our results demonstrate that mice lacking PLAP-1/Asporin show alteration of periodontal ligament structures and acceleration of bone loss in periodontitis. This underscores the significant role of PLAP-1 in maintaining collagen fibrils in the PDL and suggests the potential of PLAP-1 as a therapeutic target for periodontal diseases.

Role of Fibroblast Growth Factor Receptor 2 (FGFR2) in Corneal Stromal Thinning.

To determine the role of fibroblast growth factor receptor 2 (FGFR2)-mediated signaling in keratocytes during corneal development, a keratocyte-specific FGFR2-knockout (named FGFR2cKO) mouse model was generated, and its phenotypic characteristics were determined. A FGFR2cKO mouse model was generated by the following method: FGFR2 flox mice were crossed with the inducible keratocyte specific-Cre mice (Kera-rtTA/tet-O-Cre). Both male and female FGFR2cKO- and control mice (1 to 3-months-old) were analyzed for changes in corneal topography and pachymetry maps using the optical coherence tomography (OCT) method. The comparative TUNEL assay and immunohistochemical analyses were performed using corneas of FGFR2cKO and control mice to determine apoptotic cells, and expression of collagen-1 and fibronectin. Transmission electron microscopic analysis was conducted to determine collagen structures and their diameters in corneas of FGFR2cKO and control mice. OCT-analyses of corneas of FGFR2cKO mice (n = 24) showed localized central thinning and an increased corneal steepness compared to control mice (n = 23). FGFR2cKO mice further showed a decreased expression in collagen-1, decreased collagen diameters, acute corneal hydrops, an increased fibronectin expression, and an increased number of TUNEL-positive cells suggesting altered collagen structures and keratocytes' apoptosis in the corneas of FGFR2cKO mice compared to control mice. The FGFR2cKO mice showed several corneal phenotypes (as described above in the results) that are also exhibited by the human keratoconus corneas. The results suggested that the FGFR2cKO mouse model serves to elucidate not only the yet unknown role of FGFR2-mediated signaling in corneal physiology but also serves as a model to determine molecular mechanism of human keratoconus development.

Mechanical and matrix effects of short and long-duration exposure to beta-aminopropionitrile in elastase-induced model abdominal aortic aneurysm in mice

ObjectiveEvaluate the mechanical and matrix effects on abdominal aortic aneurysms (AAA) during the initial aortic dilation and after prolonged exposure to beta-aminopropionitrile (BAPN) in a topical elastase AAA model. MethodsAbdominal aortae of C57/BL6 mice were exposed to topical elastase with or without BAPN in the drinking water starting 4 days before elastase exposure. For the standard AAA model, animals were harvested at 2 weeks after active elastase (STD2) or heat-inactivated elastase (SHAM2). For the enhanced elastase model, BAPN treatment continued for either 4 days (ENH2b) or until harvest (ENH2) at 2 weeks; BAPN was continued until harvest at 8 weeks in one group (ENH8). Each group underwent assessment of aortic diameter, mechanical testing (tangent modulus and ultimate tensile strength [UTS]), and quantification of insoluble elastin and bulk collagen in both the elastase exposed aorta as well as the descending thoracic aorta. ResultsBAPN treatment did not increase aortic dilation compared with the standard model after 2 weeks (ENH2, 1.65 ± 0.23 mm; ENH2b, 1.49 ± 0.39 mm; STD2, 1.67 ± 0.29 mm; and SHAM2, 0.73 ± 0.10 mm), but did result in increased dilation after 8 weeks (4.3 ± 2.0 mm; P = .005). After 2 weeks, compared with the standard model, continuous therapy with BAPN did not have an effect on UTS (24.84 ± 7.62 N/cm2; 18.05 ± 4.95 N/cm2), tangent modulus (32.60 ± 9.83 N/cm2; 26.13 ± 9.10 N/cm2), elastin (7.41 ± 2.43%; 7.37 ± 4.00%), or collagen (4.25 ± 0.79%; 5.86 ± 1.19%) content. The brief treatment, EHN2b, resulted in increased aortic collagen content compared with STD2 (7.55 ± 2.48%; P = .006) and an increase in UTS compared with ENH2 (35.18 ± 18.60 N/cm2; P = .03). The ENH8 group had the lowest tangent modulus (3.71 ± 3.10 N/cm2; P = .005) compared with all aortas harvested at 2 weeks and a lower UTS (2.18 ± 2.18 N/cm2) compared with both the STD2 (24.84 ± 7.62 N/cm2; P = .008) and ENH2b (35.18 ± 18.60 N/cm2; P = .001) groups. No differences in the mechanical properties or matrix protein concentrations were associated with abdominal elastase exposure or BAPN treatment for the thoracic aorta. The tangent modulus was higher in the STD2 group (32.60 ± 9.83 N/cm2; P = .0456) vs the SHAM2 group (17.99 ± 5.76 N/cm2), and the UTS was lower in the ENH2 group (18.05 ± 4.95 N/cm2; P = .0292) compared with the ENH2b group (35.18 ± 18.60 N/cm2). The ENH8 group had the lowest tangent modulus (3.71 ± 3.10 N/cm2; P = .005) compared with all aortas harvested at 2 weeks and a lower UTS (2.18 ± 2.18 N/cm2) compared with both the STD2 (24.84 ± 7.62 N/cm2; P = .008) and ENH2b (35.18 ± 18.60 N/cm2; P = .001) groups. Abdominal aortic elastin in the STD2 group (7.41 ± 2.43%; P = .035) was lower compared with the SHAM2 group (15.29 ± 7.66%). Aortic collagen was lower in the STD2 group (4.25 ± 0.79%; P = .007) compared with the SHAM2 group (12.44 ± 6.02%) and higher for the ENH2b (7.55 ± 2.48%; P = .006) compared with the STD2 group. ConclusionsEnhancing an elastase AAA model with BAPN does not affect the initial (2-week) dilation phase substantially, either mechanically or by altering the matrix content. Late mechanical and matrix effects of prolonged BAPN treatment are limited to the elastase-exposed segment of the aorta. Clinical RelevanceThis paper explores the use of short- and long-term exposure to beta-aminopropionitrile to create an enhanced topical elastase abdominal aortic aneurysm model in mice. Readouts of aneurysm severity included loss of mechanical stability and vascular extracellular matrix composition reminiscent of what is seen in the course of human disease. Additionally, we show that the thoracic aorta, unlike the findings below the renal arteries, is not damaged in our animal model.

Differences in Collagen Fiber Diameter and Waviness between Healthy and Aneurysmal Abdominal Aortas.

Collagen plays a key role in the strength of aortic walls, so studying micro-structural changes during disease development is critical to better understand collagen reorganization. Second-harmonic generation microscopy is used to obtain images of human aortic collagen in both healthy and diseased states. Methods are being developed in order to efficiently determine the waviness, that is, tortuosity and amplitude, as well as the diameter, orientation, and dispersion of collagen fibers, and bundles in healthy and aneurysmal tissues. The results show layer-specific differences in the collagen of healthy tissues, which decrease in samples of aneurysmal aortic walls. In healthy tissues, the thick collagen bundles of the adventitia are characterized by greater waviness, both in the tortuosity and in the amplitude, compared to the relatively thin and straighter collagen fibers of the media. In contrast, most aneurysmal tissues tend to have a more uniform structure of the aortic wall with no significant difference in collagen diameter between the luminal and abluminal layers. An increase in collagen tortuosity compared to the healthy media is also observed in the aneurysmal luminal layer. The data set provided can help improve related material and multiscale models of aortic walls and aneurysm formation.

Comparative Effects of Exosomes and Ectosomes Isolated From Adipose-Derived Mesenchymal Stem Cells on Achilles Tendinopathy in a Rat Model

Background: Extracellular vesicles derived from mesenchymal stem cells (MSC-EVs) have gained momentum as a treatment for tendinopathy. Multiple studies have demonstrated significant differences in cargo composition between the 2 subtypes of MSC-EVs (ie, exosomes and ectosomes), which may result in different therapeutic effects. However, the effects of the 2 EV subtypes on tendinopathy have not yet been compared. Purpose: To compare the effects of adipose stem cell–derived exosomes (ASC-Exos) and ectosomes (ASC-Ectos) on Achilles tendinopathy. Study Design: Controlled laboratory study. Methods: Rats were administered collagenase injections to generate a model of Achilles tendinopathy. A week later, 36 rats were randomly assigned to 3 groups. In each group, Achilles tendons were injected with equal volumes of ASC-Exos, ASC-Ectos, or saline (12 legs/group). The healing outcomes were evaluated by magnetic resonance imaging, histology, immunohistochemistry, transmission electron microscopy, and biomechanical testing at 3 and 5 weeks after collagenase injection. Results: At 3 and 5 weeks, the ASC-Exo group had better histological scores (P = .0036 and P = .0276, respectively), a lower fibril density (P < .0001 and P = .0310, respectively), and a larger collagen diameter (P = .0052 and P < .0001, respectively) than the ASC-Ecto group. At 5 weeks, the expression of collagen type 1 and CD206 in the ASC-Exo group was significantly higher than that in the ASC-Ecto group (P = .0025 and P = .0010, respectively). Regarding biomechanical testing, the ASC-Exo group showed higher failure load (P = .0005), tensile stress (P < .0001), and elastic modulus (P < .0001) than the ASC-Ecto group. Conclusion: ASC-Exos had more beneficial effects on tendon repair than ASC-Ectos in a rat model of Achilles tendinopathy. Clinical Relevance: Administration of ASC-EVs may have the potential to treat Achilles tendinopathy, and delivery of ASC-Exos could provide additional benefits. It is necessary to compare the healing responses caused by different EV subtypes to further understand their effects on tendinopathy and to aid clinical decision making.

Upregulation of GLT25D1 in Hepatic Stellate Cells Promotes Liver Fibrosis via the TGF-β1/SMAD3 Pathway In Vivo and In vitro.

Collagen β(1-O) galactosyltransferase 25 domain 1 (GLT25D1) is associated with collagen production and glycosylation, and its knockout in mice results in embryonic death. However, its role in liver fibrosis remains elusive, particularly in hepatic stellate cells (HSCs), the primary collagen-producing cells associated with liver fibrogenesis. Herein, we aimed to elucidate the role of GLT25D1 in HSCs. Bile duct ligation (BDL)-induced mouse liver fibrosis models, primary mouse HSCs (mHSCs), and transforming growth factor beta 1 (TGF-β1)-stimulated LX-2 human hepatic stellate cells were used in in vivo and in vitro studies. Stable LX-2 cell lines with either GLT25D1 overexpression or knockdown were established using lentiviral transfection. RNA-seq was performed to investigate the genomic differences. HPLC-MS/MS were used to identify glycosylation sites. Scanning electronic microscopy (SEM) and second-harmonic generation/two-photon excited fluorescence (SHG/TPEF) were used to image collagen fibril morphology. GLT25D1 expression was upregulated in nonparenchymal cells in human cirrhotic liver tissues. Meanwhile, its knockdown attenuated collagen deposition in BDL-induced mouse liver fibrosis and inhibited mHSC activation. GLT25D1 was overexpressed in activated versus quiescence LX-2 cells and regulated in vitro LX-2 cell activation, including proliferation, contraction, and migration. GLT25D1 also significantly increased liver fibrogenic gene and protein expression. GLT25D1 upregulation promoted HSC activation and enhanced collagen expression through the TGF-β1/SMAD signaling pathway. Mass spectrometry showed that GLT25D1 regulated the glycosylation of collagen in HSCs, affecting the diameter of collagen fibers. Collectively, the upregulation of GLT25D1 in HSCs promoted the progression of liver fibrosis by affecting HSCs activation and collagen stability.

Effect Soursop (Annona muricata L.) Leaf Aqueous Extract (SLAE) on Remodeling of Ventricle Heart Tissue in Obesity Rats Model

A high-fat and high-fructose diet can lead to obesity which contributes to an increased risk of heart failure complications. Soursop leaves are known to have the potential of antioxidant, but research about the effects of soursop leaf on complications of heart failure has never been studied. This study evaluated whether soursop leaf aqueous extract can inhibit the total of cardiomyocyte necrosis, cardiomyocyte diameter, and density of cardiac collagen in obese rats. The heart organs of obese rats that have been paraffin, which randomly divided into normal group, obesity group, and soursop leaf aqueous extract (SLAE) group with dose I (100 mg/kgbw), II (200 mg/kgbw), and III (400 mg/kgbw) (n=6 rats). The rats were induced by high-fat high-fructose and SLI diets for 10 weeks. The number of necrosis and diameter cardiomyocytes were measured by using Hematoxylin Eosin staining, while the density of cardiac collagen was measured by Masson's Trichrome staining and then observed with a trinocular and dot side microscope at 200x and 400x magnification. Statistical analysis using One Way ANOVA and continued by LSD test. We found that group SLAE with doses I, II, and III significantly reduced the number of cardiomyocyte necrosis and cardiac collagen density compared to the obesity group. The SLI group with doses II and III significantly reduced the diameter of cardiomyocytes compared to the obesity group, while the SLAE group with the dose I was unable to reduce the diameter of cardiomyocytes. In conclusion, Soursop leaf aqueous extract can inhibit the increase of cardiac collagen density, number necrosis and diameter cardiomyocyte.

Stabilization by Nano Spray Dryer of Pioglitazone Polymeric Nanosystems: Development, In Vivo, Ex Vivo and Synchrotron Analysis.

Pioglitazone-loaded PLGA-PEG nanoparticles (NPs) were stabilized by the spray drying technique as an alternative to the treatment of ocular inflammatory disorders. Pioglitazone-NPs were developed and characterized physiochemically. Interaction studies, biopharmaceutical behavior, ex vivo corneal and scleral permeation, and in vivo bioavailability evaluations were conducted. Fibrillar diameter and interfibrillar corneal spacing of collagen was analyzed by synchrotron X-ray scattering techniques and stability studies at 4 °C and was carried out before and after the spray drying process. NPs showed physicochemical characteristics suitable for ocular administration. The release was sustained up to 46 h after drying; ex vivo corneal and scleral permeation profiles of pioglitazone-NPs before and after drying demonstrated higher retention and permeation through cornea than sclera. These results were correlated with an in vivo bioavailability study. Small-angle X-ray scattering (SAXS) analysis did not show a significant difference in the organization of the corneal collagen after the treatment with pioglitazone-NPs before and after the drying process, regarding the negative control. The stabilization process by Nano Spray Dryer B-90 was shown to be useful in preserving the activity of pioglitazone inside the NPs, maintaining their physicochemical characteristics, in vivo bioavailability, and non-damage to corneal collagen function after SAXS analysis was observed.

Measuring collagen fibril diameter with differential interference contrast microscopy

Collagen fibrils, linear arrangements of collagen monomers, 20–500 nm in diameter, comprising hundreds of molecules in their cross-section, are the fundamental structural unit in a variety of load-bearing tissues such as tendons, ligaments, skin, cornea, and bone. These fibrils often assemble into more complex structures, providing mechanical stability, strength, or toughness to the host tissue. Unfortunately, there is little information available on individual fibril dynamics, mechanics, growth, aggregation and remodeling because they are difficult to image using visible light as a probe. The principle quantity of interest is the fibril diameter, which is difficult to extract accurately, dynamically, in situ and non-destructively. An optical method, differential interference contrast (DIC) microscopy has been used to visualize dynamic structures that are as small as microtubules (25 nm diameter) and has been shown to be sensitive to the size of objects smaller than the wavelength of light. In this investigation, we take advantage of DIC microscopy’s ability to report dimensions of nanometer scale objects to generate a curve that relates collagen diameter to DIC edge intensity shift (DIC-EIS). We further calibrate the curve using electron microscopy and demonstrate a linear correlation between fibril diameter and the DIC-EIS. Using a non-oil immersion, 40x objective (NA 0.6), collagen fibril diameters between ~100 nm to ~ 300 nm could be obtained with ±11 and ±4 nm accuracy for dehydrated and hydrated fibrils, respectively. This simple, nondestructive, label free method should advance our ability to directly examine fibril dynamics under experimental conditions that are physiologically relevant.

Effectiveness of nuchal ligament autograft in the healing of an experimental superficial digital flexor tendon defect in equid.

This study aimed to investigate nuchal ligament (NL) autograft on experimental tendon defect healing in donkeys. Eight healthy donkeys were used. The left forelimb’s superficial digital flexor (SDF) tendon was assigned as treatment, and the right forelimb was allocated as the control group (without surgical intervention). A 3×1.5 cm segment of the funicular part of the NL was excised. A full thickness defect created in the treatment tendon and was grafted with the excised NL. The following parameters were evaluated in 120 days postoperatively: clinical, ultrasonography, radiography, histopathology, biomechanical properties, and scanning and electronic transmission microscopy. There were no significant changes observed in the neck angle so that it was confirmed this treatment regimen preserved the head and neck situation without any considerable neck swelling. Weight-bearing in gait and trot was similar between both forelimbs at the end of the study. Mild to moderate adhesion was detected in the dorsal surface of the SDF tendon. There was no significant difference in the echogenicity and fiber alignment, respectively, on days 90 and 120 after surgery. Treatment significantly amplified the collagen diameter and enhanced the collagen fibril diameter and density considerably compared to the NL. The transplanted tissue was mostly in the remodeling or maturation phase, on day 120 postoperatively. It seems that the NL is biocompatible, almost biodegradable, and effective in tendon healing without metaplasia or tissue rejection.

Tendon stem cell-derived exosomes regulate inflammation and promote the high-quality healing of injured tendon

BackgroundTendon stem cells (TSCs) have been reported to hold promises for tendon repair and regeneration. However, less is known about the effects of exosomes derived from TSCs. Therefore, we aimed to clarify the healing effects of TSC-derived exosomes (TSC-Exos) on tendon injury.MethodsThe Achilles tendons of Sprague-Dawley male rats were used for primary culture of TSCs and tenocytes, and exosomes were isolated from TSCs. The proliferation of tenocytes induced by TSC-Exos was analyzed using an EdU assay; cell migration was measured by cell scratch and transwell assays. We used western blot to analyze the role of the PI3K/AKT and MAPK/ERK1/2 signaling pathways. In vivo, Achilles tendon injury models were created in Sprague-Dawley rats. Rats (n = 54) were then randomly assigned to three groups: the TSC-Exos group, the GelMA group, and the control group. We used immunofluorescence to detect changes in the expression of inflammatory and apoptotic markers at 1 week after surgery. Histology and changes in expression of extracellular matrix (ECM)-related indices were assessed by hematoxylin-eosin (H&E) staining and immunohistochemistry at 2 and 8 weeks. The collagen fiber diameter of the healing tendon was analyzed at 8 weeks by transmission electron microscopy (TEM).ResultsTSC-Exos were taken up by tenocytes, which promoted the proliferation and migration of cells in a dose-dependent manner; this process may depend on the activation of the PI3K/AKT and MAPK/ERK1/2 signaling pathways. At 1 week after surgery, we found that inflammation and apoptosis were significantly suppressed by TSC-Exos. At 2 and 8 weeks, tendons treated with TSC-Exos showed more continuous and regular arrangement in contrast to disorganized tendons in the GelMA and control groups, and TSC-Exos may help regulate ECM balance and inhibited scar formation. Further, at 8 weeks, the TSC-Exos group had a larger diameter of collagen compared to the control group.ConclusionsOur data suggest that TSC-Exos could promote high-quality healing of injured tendon, which may be a promising therapeutic approach for tendon injury.

Glutamic acid concentration dependent collagen mineralization in aqueous solution

The aim of this research is to evaluate the effects of glutamic acid (Glu) on the process of collagen mineralization, the structure and property of mineralized composites. Collagen mineralization was initiated by introducing PBS to blended solutions containing 1 mg/mL collagen, 6 mmol/L calcium and Glu ranged from 0 to 550 mmol/L. The kinetic curves and quantitation analyses showed that Glu could delay the collagen mineralization, and reduce the crystalline size and the amount of hydroxyapatite. With the Glu concentration increased from 50 to 200 mmol/L, the collagen self-assembly was promoted, resulting in the improvement of hardness and thermal stability of mineralized composites. However, further increase in the Glu concentration to 400 mmol/L or above would significantly inhibit the self-assembly of collagen and reduce the hardness and thermal stability of mineralized composites. Scanning electron microscopy revealed that the diameter of collagen became thicker as the Glu concentration increased. Moreover, hydroxyapatite with spherical morphology was uniformly dispersed and well combined with collagen fibril at Glu concentration of 200 mmol/L. These results may provide a broader understanding of the potential mechanism of biomineralization and be critical in the design of biomimetic scaffolds for bone tissue engineering.

Efficacy of the Tent-Pole Technique in Horizontal Ridge Augmentation

Objective: To evaluate the efficacy of the technique for ridge augmentation prior to implant placement. Material and Methods: Six patients with nine sites presenting an initial ridge width of smaller than 4 mm were included in this study. The tent-pole technique (combination of osteosynthesis screws measuring 1.5 mm in diameter, ABBM, and resorbable collagen membrane) was applied at defect sites. After eight months of healing time, implants were placed in the treated sites. The primary outcomes (radiographic initial ridge width, re-entry ridge width, ridge width gain) were measured by cone-beam computed tomography in reference buccopalatal cross-sections and the secondary outcomes (clinical parameters) were recorded by a digital caliper at the first and second stages. Results: After grafting, the radiographic ridge width increased by 3.02 ± 1.11 mm (1.57-4.75 mm) at 2 mm below the crest and 3.32 ± 1.70 mm (0.23-5.66 mm) at 6 mm below the crest significantly (p< 0.05). For clinical results, the mean horizontal dimension gain was 3.21 ± 1.04 mm (1.83-4.57 mm), while the mean reduction in dimension was 0.38 ± 0.33 mm. These results were statistically significant (p<0.05). Uneventful healing and no infections or membrane exposure were recorded at all sites during the study process. Three of nine (33.33%) defect sites required additional bone grafting. Conclusion: The tent-pole technique is an effective method for increasing the horizontal ridge dimension, minimizing postoperative complications, and facilitating subsequent implant placement.

Dietary normalization from a fat, fructose and cholesterol-rich diet to chow limits the amount of myocardial collagen in a G\xf6ttingen Minipig model of obesity

BackgroundDietary interventions have been shown to attenuate some of the myocardial pathological alterations associated with obesity. This study evaluated the effect of dietary normalization from a fat/fructose/cholesterol-rich diet to chow on left ventricular (LV) myocardial fibrosis, fat infiltration and hypertrophy but also the specific influence of obesity, plasma lipids and glucose metabolism markers on heart morphology in a Göttingen Minipig model of obesity.MethodsForty castrated male Göttingen Minipigs were assigned to three groups fed either standard minipig chow (SD, n = 8) for 13 months, fat/fructose/cholesterol-rich diet (FFC, n = 16) for 13 months or fat/fructose/cholesterol-rich diet for 7 months and then changed to standard minipig chow for the remaining 6 months (FFC/SD, n = 16). Body weight, body fat percentage, plasma lipids and glucose metabolism markers were evaluated in all three groups after 6–7 months (prior to diet adjustment for FFC/SD) and again before termination. Further, biochemical quantification of myocardial collagen and triglyceride content, semi-quantitative histological evaluation of fibrosis and fat infiltration and quantitative histological analysis of collagen and cardiomyocyte diameter were performed and heart weight was obtained after termination. Group differences were evaluated using Kruskal-Wallis test and Fisher’s exact test for categorical variables. Pearson correlation analysis was performed to test for correlations between myocardial changes and selected explanatory variables. For non-parametric response variables, a Spearman correlation analysis was applied.ResultsMyocardial collagen content quantified biochemically was significantly lower in FFC/SD compared to FFC (P = 0.02). Furthermore, dietary normalization from a fat/fructose/cholesterol-rich diet to chow caused stagnation of body weight and body fat percentage, normalized intravenous glucose tolerance index (KG) and plasma lipid levels.ConclusionDietary normalization led to lower LV collagen content in obese Göttingen Minipigs. Despite gross obesity and significant deteriorations in glucose and lipid metabolism, only mild myocardial changes were found in this model of obesity and therefore further model optimization is warranted in order to induce more severe myocardial changes before dietary or pharmacological interventions.

Modulation of collagen population under honey assisted wound healing in diabetic rat model

Abstract Background Diabetic wound with impaired healing attributes owing to inappropriate synthesis and alignment of collagen fibrils with aberrant post translational modifications impose substantial economic burden. Honey: a recognized natural collagen pro-synthetic healing agent; stimulates rapid healing with its anti-inflammatory and anti-scarring approach. Present study investigates the effect of honey on collagen population in diabetic wound model. Methodology Full thickness wound was created on rat and divided into three groups: normal saline treated group (N), diabetic honey treated group (H), diabetic povidone iodine treated group (PI). Biopsies were collected from wound edges for collagen characterization. Main findings Histological attributes of collagen fibers in N and H group biopsy displayed proximity. Ultrastructure analysis using scanning electron microscope (SEM) revealed that the D-Spacing and collagen diameter under honey treatment were in close resemblance to normal skin collagen. FTIR spectra confirmed secondary structure of collagen with a greater peak height in PI group exhibiting excessive glycation than H group. Nano-indentation profiling showed decreased displacement in PI samples resulting from increased rigidity when compared to honey treated ones. Molecular expression of collagen I and III exhibited optimum ratio in H group similar to N group. Conclusions Based on these findings, it might be opined that honey predominantly helps in regulating synthesis, glycation and deposition followed by mechanical quality of collagen alike normal skin and its application also accelerated diabetic healing process.

The effect of gelatin hydrolysate from pygostyle of broiler produced by enzymatic hydrolysis process of Aspergillus niger to prophylaxis

Kuntana YP, Balia R, Murad S, Hidayat YA, Partasasmita R. 2017. The effect of gelatin hydrolysate from pygostyle of broiler produced by enzymatic hydrolysis process of Aspergillus niger to prophylaxis of osteoporosis white rat. Nusantara Bioscience 9: 361- 366. The pygostyle of the broiler is a cartilage tissue containing collagen, a mucoprotein being a plasma expander. The hydrolysis process of collagen by enzyme can produce gelatin hydrolysate differently compared to that from commercial gelatin because it has an amino acid composition with low average molecular weight. It becomes an advantage because it can be directly absorbed by the digestive tract. In addition, the enzymatic process can be done in short time with the low cost and also environment-friendly approach. This process can be assisted by Aspergillus niger, i.e., safe microorganisms (GRASS). A. niger has a protease enzyme that can digest collagen into gelatin hydrolysate. The enzyme is extracellularly removed so that its utilization does not require high input value expenditure. Gelatin hydrolysate can reduce the disease of osteoporosis. The objective of this study was to know the effect of gelatin hydrolysate produced from chicken bones in the favor of A. niger for increasing the benefit products. The specific targets are to find the amount of gelatin hydrolysate, precisely eliminated osteoporosis of rat (Rattus norvegicus). The experimental research was conducted in the laboratory with using complete randomized design followed by the descriptive analysis with 5 treatment and 5 replication. CCl4 dose 0,14 mL/g BW (P1 as a negative control), Potatoes dextrose Broth (PDB) dose 0,5 mL/g BW (P2 positive control), gelatin hydrolysate dose 0.25; 0.5; 0.75 mL/g BW (P3-P5). The observed parameters were including body weight and histological observation of musculoskeletal tissue in terms of measurement of fiber muscle, cartilage and collagen diameter. The test animal used was male white rat (R. norvegicus) Wistar strain aged 2-3 months with body weight 185-200 g. Data were analyzed using ANOVA and Duncan test. The results show that gelatin hydrolyzate as an environmentally friendly product serves as a prevention of immune diseases and degenerative diseases such as osteoporosis.

Studying Arterial Stiffness Using High-Frequency Ultrasound in Mice with Alzheimer Disease

Alzheimer disease (AD) is an irreversible, progressive brain disorder that causes slow loss of memory and thinking skills, normally leading to death in 3–9 y. The etiology of AD is not fully understood but is widely believed to be induced by the production and deposition of amyloid-β peptide in the brain. Recently, a correlation was discovered between amyloid-β deposition and atherosclerosis in the cerebral arteries of postmortem brains, indicating that amyloid-β promotes atherogenesis and that in turn atherosclerosis promotes brain amyloid-β accumulation. However, a direct measurement of arterial stiffness for AD is lacking. In the present study, the pulse wave velocity (PWV) of the carotid artery was measured non-invasively in young (3-mo-old) and middle-aged (9-mo-old) wild-type (WT) and modeled AD mice to obtain quantitative data of arterial stiffness by using a 35-MHz high-frequency dual-element transducer. Experimental results show that the PWVs were 1.6 ± 0.5 m/s for young and 2.4 ± 0.4 m/s for middle-aged WT mice and 1.7 ± 0.4 m/s for young and 3.2 ± 0.6 m/s for middle-aged AD mice. Middle-aged groups had higher PWVs (p < 0.0001), which were more pronounced in the AD mice (p < 0.001). The differences in PWVs were not caused by arterial lumen diameter, wall thickness or contents of elastin or collagen. These results imply that AD increases the stiffness of the carotid artery and introduce ultrasound as a potential tool for AD research and diagnosis.