Soluble Collagen Research Articles

A Comparative Study on Meat Quality Characteristics of Murrah Buffalo and Nellore Cattle Commercialized in Southeastern Brazil

Murrah buffalo and Nellore cattle meat commercialized in Southeastern Brazil were evaluated during aging. Ribeye steaks (Longissimus thoracis muscle) were analyzed during four wet aging times (0, 7, 14, and 21 days) stored at 2 ± 1 °C. The water holding capacity (WHC) decreased (p < 0.05) during aging (0.41 to 0.28), with buffalo meat having (p < 0.05) lower pH and a higher WHC than beef. Lower myofibrillar fragmentation index and shear force (WBsSF) values were observed (p < 0.05) in buffalo meat. Soluble collagen content increased (p < 0.05) during aging, with lower (p < 0.05) values in buffalo meat. Buffalo meat had (p < 0.05) higher metmyoglobin percentages, being darker (lower L* values) and with a higher red color intensity (higher C* values) than beef. There was a difference between buffalo meat and beef volatile compound profiles, with greater variation in the beef profile during aging, probably due to differences in lipid oxidation and proteolysis. It can be concluded that buffalo meat is very similar to that of Zebu cattle, with less variation during aging and greater tenderness than beef. Therefore, buffalo meat is a good alternative source for fresh marketing and processing of high-quality meat products.

Production of Partially Purified Collagenase from Bacillus licheniformis and It’s Use to Tenderize Aged Buffalo Meat

This study aimed to isolate and identify the Bacillus licheniformis bacteria that produces the enzyme collagenase. Due to the selective nature of this enzyme in breaking down and degrading collagen, it was used to tenderize aged buffalo meat. The bacterial isolates were screened using a selective medium and then grown on collagen-agar medium to identify enzyme-producing isolates. Thirteen enzyme-producing isolates were obtained based on the formation of a transparent corona around the colony. Biochemical assessments of the bacterial isolates were carried out using the ABIS online program. The isolates BL1–BL9, BL12 and BL13 were found to be related to B. licheniformis with a percentage match of 98%, while BL10 and BL11 were related to Bacillus pumilus and Bacillus subtilis with a percentage match of 95% and 95.3%, respectively. The enzyme activity of the isolates varied, with the highest activity reaching 200.71 units.ml-1 for isolate BL12. The enzyme of this isolate was chosen to complete the other tests. The optimal time required to obtain the highest enzymatic activity was found to be 20 h, which amounted to 210.25 units.ml-1, with a specific activity of 135.64 units.ml-1. The enzyme was partially purified using ammonium sulphate at a saturation rate of 20–70% as it showed an enzymatic activity and specificity of 355.81 units.ml-1 and a specific activity of 423.58 units.ml-1. Partially purified collagenase was used to tenderize aged buffalo meat. The results showed that treating aged buffalo meat with the enzyme solution for different immersion durations improved the pH, water-holding capacity (WHC) and cooking yield, with an increase in the concentration of hydroxyproline and a significant percentage of soluble collagen; the treated samples showed the best properties with a 60-min immersion duration.

Exploring the impact of various cooking techniques on the physicochemical and quality characteristics of camel meat product.

The objective of this study was to evaluate the effects of four different cooking techniques viz: boiling, grilling, microwave, and frying; on the physicochemical characteristics of camel meat. Protein composition and their degradation as well as biochemical and textural changes of camel meat as influenced by cooking methods were investigated. The highest cooking loss (52.61%) was reported in microwaved samples while grilled samples showed the lowest cooking loss (44.98%). The microwaved samples showed the highest levels of lipid oxidation as measured by thiobarbituric acid reactive substances, while boiled samples showed the lowest levels (4.5 mg/kg). Protein solubility, total collagen, and soluble collagen content were highest in boiled samples. Boiled camel meat had lower hardness values compared to the other treated samples. Consequently, boiling was the more suitable cooking technique for producing camel meat with a reduced hardness value and lower lipid oxidation level. The camel meat industry and camel meat consumer can benefit from this research by improving their commercial viability and making consumers aware about the effects of cooking procedures on the quality of camel meat. The results of this study will be of significance to researchers and readers who are working on the processing and quality of camel meat.

High-mobility group box-1 peptide ameliorates bronchopulmonary dysplasia by suppressing inflammation and fibrosis in a mouse model

BackgroundThis study aimed to examine the effect of the HMGB1 peptide on Bronchopulmonary dysplasia (BPD)-related lung injury in a mouse model. ResultsHMGB1 peptide ameliorates lung injury by suppressing the release of inflammatory cytokines and decreasing soluble collagen levels in the lungs. Single-cell RNA sequencing showed that the peptide suppressed the hyperoxia-induced inflammatory signature in macrophages and the fibrotic signature in fibroblasts. These changes in the transcriptome were confirmed using protein assays. ConclusionSystemic administration of HMGB1 peptide exerts anti-inflammatory and anti-fibrotic effects in a mouse model of BPD. This study provides a foundation for the development of new and effective therapies for BPD.

Royal jelly mediates fibrotic signaling, collagen cross-linking and cell proliferation in cardiac fibroblasts

Royal jelly (RJ) is a multifunctional bee product with a unique composition and wide-ranging biological properties, including antioxidant, anti-inflammatory and antiproliferative activities. Still, little is known about the possible myocardial protective properties of RJ. Considering that sonication could enhance RJ bioactivity, this study aimed to assess the effects of non-sonicated (NS) and sonicated (S) RJ on fibrotic signaling, cell proliferation, and collagen production in cardiac fibroblasts. S-RJ was produced by ultrasonication at 20 kHz. Ventricular fibroblasts isolated from neonatal rats were cultured and treated with different concentrations of NS-RJ or S-RJ (0, 50, 100, 150, 200, and 250 µg/well). S-RJ significantly depressed the expression levels of transglutaminase 2 (TG2) mRNA across all the concentrations tested and was inversely associated with the expression of this profibrotic marker. S-RJ and NS-RJ displayed distinct dose-dependent effects on mRNA expression of several other profibrotic, proliferation, and apoptotic markers. Unlike NS-RJ, S-RJ elicited strong negative dose-dependent relationships with the expression of profibrotic markers (TG2, COL1A1, COL3A1, FN1, CTGF, MMP-2, α-SMA, TGF-β1, CX43, periostin), as well as proliferation (CCND1) and apoptotic (BAX, BAX/BCL-2) markers, indicating that RJ dose-response effects were significantly modified by sonification. NS-RJ and S-RJ increased the content of soluble collagen, while decreasing collagen cross-linking. Collectively, these findings show that S-RJ has a greater range of action than NS-RJ for downregulating the expression of biomarkers associated with cardiac fibrosis. Reduced biomarker expression and collagen cross-linkages upon cardiac fibroblast treatment with specific concentrations of S-RJ or NS-RJ suggests putative roles and mechanisms by which RJ may confer some protection against cardiac fibrosis.

(158) In Vitro Efficacy of Intralesional Collagenase Clostridium Histolyticum for the Treatment of Calcified Peyronie’s Disease Plaques

Abstract Introduction Peyronie’s disease (PD), estimated to affect around 10% of the male population, is an acquired disorder of penile fibrotic tissue. The only FDA approved medical treatment for PD is Intralesional Collagenase Clostridium Histolyticum (CCh). CCh is a mixture of AUX1 and AUXII, two different forms of collagenase that cleave collage type I and type III, leading to plaque degradation. PD plaques, however, also contain fibrin and calcium which CCh is unable to digest. It is unknown if plaque calcification prevents collagenolysis with CCh. Objective To determine if calcification prevents collageolysis of PD plaques with CCh. Methods Calcified PD plaques were collected from five males undergoing excision and grafting for PD. Control tissue was collected from five males undergoing penile implant surgery for erectile dysfunction, with no known history of PD. Tissue samples were then divided into equal weights and incubated with CCh or PBS for 24 hours. The CCh or PBS was injected into the tissue (control or plaque) to mimic administration of CCH for treatment of PD. Following incubation, the fluid (CCh or PBS) was measured for soluble collagen amount and tissue was subjected to Picrosirius Red and H&E staining. Results There was a higher concentration of soluble collagen in the tissue (control and plaque) incubated in CCh (301ug, SD 21.1) compared to the issue incubated in PBS (32.5ug, SD 3.7) (p=0.02). Additionally, there was less type 1 and 3 collagen present in the CCh condition (mean 0.12, SD=0.08), when compared to the PBS condition (0.44, SD=0.17) (p=0.002). However, when comparing within the CCh condition to look at differences between tissue types (calcified vs. control) no differences were found. There was a similar amount of soluble collagen in the CCh control condition (316ug, SD 35) and the CCh calcified condition (286ug, SD 11) (p=0.06). Similarly, the PBS control condition (30ug, SD17) and the PBS calcified condition (35ug, SD 48) were found to have no significant differences in collagen concentration (p=0.4). Finally, there were no significant differences found in type I/III collagen staining between tissues in the CCh condition, with control (0.08, SD=0.02) and calcified (0.17, SD=0.09) demonstrating no significant difference (p=0.08). Tissues in the PBS condition, control (0.5, SD=0.23) and calcified (0.39, SD=0.39), similarly were not significant (p=0.2) (table 1). Conclusions CCh is capable of breaking down collagen in calcified plaques. Further research into CCh as a treatment option for PD calcified plaques is necessary. Disclosure Yes, this is sponsored by industry/sponsor: Endo Pharmaceuticals Clarification Industry funding only - investigator initiated and executed study Any of the authors act as a consultant, employee or shareholder of an industry for: Acerus Pharmaceuticals, Boston Scientific, Coloplast, Endo Pharmaceuticals, Empower Pharmacy, Nestle Health, Olympus, Hims, Inc.

(080) Exploring the Effects of Actinidin as a Novel Collagenase on Human Peyronie’s Disease Cells

Abstract Introduction Peyronie’s Disease (PD) is a connective tissue disorder of the penile tunica albuginea that affects approximately 10% of men. The plaques that hallmark PD contain excessive amounts of collagen and can result in penile curvature, painful erections, and/or erectile dysfunction. Intralesional Collagenase Clostridium histolyticum injection for PD are no longer available in Canada, making treatment options limited. Hayward kiwi fruit are rich in natural actinidin, a soluble enzyme that has the unique ability to hydrolyze various types of collagen and fibrinogen. Actinidin is considered a collagenase alternative; however, there have been no studies evaluating the application of actinidin on in vitro cellular PD models. Objective To determine the effectiveness of actinidin at reducing the collagen component of a 2D human PD cell model. Secondly, to determine the cytotoxicity of kiwifruit extract on human PD cells. Methods Human PD tissues obtained during penile prosthesis implantation were isolated for fibroblast cells and cultured using complete media. The cells were plated on a 96-well plate and treated with one of 4 treatment groups: media, saline, and low and high concentration actinidin treatment (0.5 mg/ml to 37.5 mg/ml). The actinidin treatments were prepared by dissolving freeze-dried kiwi powder in saline and then filtered; the low concentration was further diluted with saline. Cells were incubated at 37°C for 24 hours and then collagen was extracted and measured using a soluble collagen quantification assay kit and a fluorescent microplate reader. To measure cytotoxicity, cells were stained for the nucleus and actin filaments using DAPI and Phalloidin-conjugated FITC, respectively. A twelve-hour timelapse capturing images of FITC and DAPI fluorescein were carried through using a fluorescent confocal microscope. The total fluorescent area of each well was measured using ImageJ. Actin signals were normalized by DAPI signals, and all values were analyzed using Python and Prism10. Results The higher concentration of actinidin treatment significantly reduced cellular collagen of human PD fibroblasts compared to the media treatment (P < 0.05). The high concentration actinidin treatment had a significantly higher normalized actin signal than other treatment groups (P < 0.0001). This ratio allows us to observe the treated cells with compromised plasma membranes. The low concentration of actinidin did not have any significant difference from media and saline treatment groups. A significant increased level of normalized actin indicated that actinidin may have mechanisms to disturb the plasma membrane. Conclusions Our preliminary study suggests that the natural collagenase actinidin can break down PD cells by reducing the extracellular collagen content and compromising the cellular membrane. Further studies are underway to investigate treatment in 3D models and to determine the optimal actinidin concentration that can further reduce the collagen composition in human PD cell models. Actinidin has the potential to be a novel treatment option for patients suffering from PD. Disclosure Yes, this is sponsored by industry/sponsor: KiwiEnzyme.com Ltd Clarification No industry support in study design or execution

Kisspeptin regulates airway hyperresponsiveness and remodeling in a mouse model of asthma.

Asthma is a multifactorial disease of origin characterized by airway hyperresponsiveness (AHR) and airway remodeling. Several pieces of evidence from other pathologies suggest that Kisspeptins (Kp) regulate cell proliferation, migration, and invasion, mechanisms that are highly relevant to asthma. Our recent in vitro studies show Kp-10 (active peptide of Kp), via its receptor, KISS1R, inhibits human airway smooth muscle cell proliferation. Here, we hypothesize a crucial role for Kp-10 in regulating AHR and airway remodeling in vivo. Utilizing C57BL/6J mice, we assessed the effect of chronic intranasal Kp-10 exposure on mixed allergen (MA)-induced mouse model of asthma. MA-challenged mice showed significant deterioration of lung function compared to those exposed to vehicle (DPBS); Kp-10 treatment significantly improved the MA-altered lung functions. Mice treated with Kp-10 alone did not show any notable changes in lung functions. MA-exposed mice showed a significant reduction in KISS1R expression as compared to vehicle alone. MA-challenged mice showed significant alterations in immune cell infiltration in the airways and remodeling changes. Proinflammatory cytokines were significantly increased upon MA exposure, an effect abrogated by Kp-10 treatment. Furthermore, biochemical and histological studies showed Kp-10 exposure significantly reduced MA-induced smooth muscle mass and soluble collagen in the lung. Overall, our findings highlight the effect of chronic Kp-10 exposure in regulating MA-induced AHR and remodeling. © 2023 The Pathological Society of Great Britain and Ireland.

BRAF V600-Mutated Metastatic Melanoma and Targeted Therapy Resistance: An Update of the Current Knowledge.

Melanoma is the most common cause of death in skin cancer due to its high metastatic potential. While targeted therapies have improved the care of patients with metastatic melanoma harboring the BRAFV600E mutation, these treatments are associated with a high frequency of resistance. Resistance factors are related to cellular adaptation as well as to changes in the tumor microenvironment. At the cellular level, resistance involves mutations, overexpression, activation, or inhibition of effectors involved in cell signaling pathways such as MAPK, PI3K/AKT, MITF, and epigenetic factors (miRNAs). In addition, several components of the melanoma microenvironment, such as soluble factors, collagen, and stromal cells also play a crucial role in this resistance. In fact, extracellular matrix remodeling impacts the physical and chemical properties with changes in the stiffness and acidity, respectively of the microenvironment. The cellular and immune components of the stroma are also affected, including immune cells and CAF. The aim of this manuscript is to review the mechanisms responsible for resistance to targeted therapies in BRAFV600E-mutated metastatic melanoma.

Optimising aerosol jet printing of collagen inks for enhanced piezoelectricity and controlled surface potential

Collagen is a highly versatile protein used in tissue engineering constructs and as a model piezoelectric biomaterial. The piezoelectricity of collagen can be enhanced through the alignment of collagen domains and fibres, although most fabrication techniques used to form dense collagenous constructs do not allow for significant collagen alignment. The use of aerosol jet printing (AJP) mitigates the limitations of using soluble collagen inks for bioprinting or extrusion-based 3D printing, particularly if microfibrillar collagen suspensions are used as a cost-effective and scalable ink source. In this work, Type I and Type II microfibrillar collagen from different anatomical sources were successfully deposited using AJP with two different atomisation methods, namely pneumatic-AJP (p-AJP) and ultrasonic-AJP (u-AJP). The printing parameters were optimised for their piezoelectric amplitude and surface potential. Fourier transform infrared spectra of the films revealed that ultrasonic atomisation did not cause notable denaturation of collagen, although the process resulted in the fractionation and preferential deposition of the oligomeric and gelatinous components within the slurry. The printed collagen samples displayed a piezoelectric response that was four times higher than the values obtained from drop-cast collagen films, and their surface potential was found to be positively correlated to the roughness of the films which can be controlled through the mode of atomisation. These results indicate the ability to enhance and control the piezoelectricity and surface potential using p-AJP and u-AJP, which offers a promising physical modulation technique to tailor cell adhesion, proliferation or differentiation for collagen-based tissue engineering constructs.

Effect of Ultrasound Pre-Treatment on Extraction and Characterization of Collagen from Bactrian Camel Skin.

The objective of this study was to evaluate the effect of ultrasound pre-treatment on the characterization from Bactrian camel skin. It was possible to produce and characterize collagen extracted from Bactrian camel skin. The results showed that the yield of collagen was higher in ultrasound pre-treatment (UPSC) (41.99%) than the pepsin-soluble collagen extraction (PSC) (26.08%). All extracts were identified as type I collagens using sodium dodecyl sulfate polyacrylamide gel electrophoresis and retained their helical structure, as confirmed through Fourier transform infrared spectroscopy. The scanning electron microscopy analysis of UPSC revealed that some physical changes were caused by sonication. UPSC had smaller particle size than PSC. The viscosity of UPSC always plays a leading role in the range of 0-10 Hz. However, the contribution of elasticity to the solution system of PSC increased in the range of 1-10 Hz. Moreover, ultrasound-treated collagen had superior solubility property at pH 1-4 and at <3% (w/v) NaCl than non-ultrasound treated collagen. Therefore, the utilization of ultrasound for the extraction of pepsin soluble collagen is a good alternative technology to expand the application at industrial level.

Effect of Extraction Time on the Extractability and Physicochemical Properties of Pepsin—Soluble Collagen (PCS) from the Skin of Silver Catfish (Pangasius sp.)

The current study aimed to determine the effects of extraction time on the extractability and physicochemical properties of collagen from the skin of silver catfish (Pangasius sp.). Pepsin soluble collagen (PSC) was extracted for 24 and 48 h and analysed in terms of chemical composition, solubility, functional group, microstructure, and rheological properties. The yields of PSC at 24 h and 48 h extraction time were 23.64% and 26.43%, respectively. The chemical composition exhibited significant differences, with PSC extracted at 24 h showing better moisture, protein, fat, and ash content. Both collagen extractions indicated the highest solubility at pH 5. In addition, both collagen extractions exhibited Amide A, I, II, and III as fingerprint regions for collagen structure. The morphology of the extracted collagen appeared porous with a fibril structure. The dynamic viscoelastic measurements of complex viscosity (η*) and loss tangent (tan δ) decreased as temperature increased, and the viscosity increased exponentially as the frequency increased, whereas the loss tangent decreased. In conclusion, PSC extracted at 24 h showed similar extractability to that extracted at 48 h but with a better chemical composition and shorter extraction time. Therefore, 24 h is the best extraction time for PSC from silver catfish skin.

Preparation, Characterization and ex vivo Skin Permeability Evaluation of Type I Collagen-Loaded Liposomes.

In the present study, we prepared collagen liposomes with the addition of polyol, which is expected to not only increase the solubility of collagen but also improve skin penetration. Collagen liposomes were prepared by the film dispersion method, and their characteristics, integrity and biosafety were evaluated by Fourier transform infrared spectroscopy (FTIR), UV-VIS spectroscopy, polyacrylamide gel electrophoresis (SDS-PAGE), dynamic light scattering (DLS) and transmission electron microscope (TEM). The transdermal absorption of collagen and collagen liposomes were tested by an ex vivo horizontal Valia-Chien diffusion cell system. We first demonstrated that collagen extracted from bovine Achilles tendon was type I collagen. The results of DLS measurement and TEM observation showed that the collagen liposomes were spherical in shape with average diameter (75.34±0.93 nm) and maintained high stability at low temperature (4°C) for at least 42 days without toxicity. The encapsulation rate of collagen liposomes was 57.80 ± 0.51%, and SDS-PAGE analysis showed that collagen was intact in liposomes. Finally, permeability studies indicated that the collagen-loaded liposomes more easily penetrated the skin compared to collagen itself. This study proposed a new method to improve the bioavailability and permeability of bovine type I collagen, which improves the applicability of collagen in biomedicine, cosmeceuticals and pharmaceutical industries.

O/W/O multiple emulsions containing soluble collagen: in vitro and in vivo skin biophysical properties evaluation

The integrity of the stratum corneum is essential for skin hydration. In order to fulfill all of its functions, a balance of the water content in the stratum corneum is necessary, as well as maintaining the epidermis, an essential objective of cosmetic formulations. Cosmetic skin hydration can be achieved through the frequent application of cosmetic products that can act through basic mechanisms: occlusion, moisturization, and hydration or active moisturization. Therefore, when developing new cosmetic products, it is important to consider their positive effect on skin hydration and viscoelasticity, considering that the objective of a cosmetic product is to protect the skin and delay skin aging. Subsequently, multiple O/W/O emulsions were developed and skin hydration was evaluated in vitro and in vivo. The in vitro method for skin hydration employs gelatin whose composition mimics collagen and NMF of the skin, forming a surface (1 cm thick) through which internal water vapor can be released. The occlusion provided to this gelatin surface is evaluated, and the hydrating power of the applied products is determined. Due to the composition of the gelatin support cell and the evaporation of water inside it, it can simulate TEWL of the skin. The in vivo evaluation of hydration, pH, and sebum of the aqueous solution of proteins, simple O/W, and multiple O/W/O with collagen showed that O/W/O emulsions provided greater occlusive and oily content, maintaining the pH levels of the skin. Therefore, these O/W/O systems improve conditions for dry skin.

Gender, Dietary Copper and Carbohydrate Source Influence Cardiac Collagen and Lysyl Oxidase in Weanling Rats

The present investigation was conducted to determine the effects of consumption of diets containing fructose or cornstarch on cardiac collagen metabolism in weanling male and female rats fed copper-deficient or copper-adequate diets for 5 wk. Although both male and female rats that consumed the copper-deficient diet containing fructose were similarly copper deficient, only the males showed severe cardiac pathologies and two died prematurely due to heart-related abnormalities. These pathologies were accompanied by a significant reduction of cardiac lysyl oxidase activity and elevated soluble and total cardiac collagen concentrations compared with rats fed copper-adequate diets. These abnormalities were less severe in copper-deficient rats fed cornstarch. The data show that the activity of the copper-containing enzyme lysyl oxidase is affected by both dietary carbohydrate and gender. The pathologies of heart tissue could be the result of abnormal crosslinking of collagen induced by the combination of copper deficiency, fructose feeding and the sex of the rats.

Proteomics in bovine semitendinosus muscle to assess emerging strategies based on papain injection and ultrasounds on meat tenderization process

The impact of papain and/or ultrasound treatments on tenderization of semitendinosus muscle through a proteomic approach was studied. Sixteen bovine muscles were submitted to the following treatments: aging at 3 °C (Control), papain injection (PI), ultrasound (US), PI followed by US (PIUS) and US followed by PI (USPI). pH, myofibrillar fragmentation indices (MFI), soluble collagen, texture profile and changes of myofibrillar proteins were investigated after 2, 24, 48 and 96 h of storage. The highest MFI and soluble collagen content were found in PI, PIUS and USPI samples while control samples showed the lowest values. PI samples showed the lowest WBSF and hardness values until 48 h of storage while at 96 h meat from USPI treatment showed WBSF value comparable to PI treatment. The lowest values of cohesiveness, gumminess and chewiness were found in PI samples during all storage times.Proteomic analysis revealed a different quantity and expression of proteins among tenderization treatments. US treatment did not exhibit a significant ability to degrade muscle proteins, while, all treatments containing papain, showed a greater ability to hydrolyse and degrade myofibrillar proteins. PI promoted intense proteolysis leading to an early tenderization process; on the contrary, in PIUS and USPI treatments the sequence of treatments was relevant on meat tenderization. Particularly, USPI treatment, after 96 h, reached the same improvement in tenderness of enzymatic treatment but with slower hydrolysing rate; this could be determinant to preserve textural structure.

Functional Properties of Collagen Extracted From Catfish ( Silurus triostegus) Bone

Back ground: Collagen accounts for 25percent of all proteins found in vertebrates. The words &amp;quot;collagen&amp;quot;is created with Greek words &amp;quot;kolla&amp;quot; and &amp;quot;genos,&amp;quot; which respectively mean &amp;quot;glue&amp;quot; and &amp;quot;formation.&amp;quot; It hasa wide range of biological and pharmacological uses. Pig and cow skins provide the majority of thecollagen. Utilizing animal collagen has been restricted as a result of epidemics of certain animal diseasesincluding foot-and-mouth disease (FMD) and bovine spongiform encephalopathy (BSE) (FMD), whichpose a risk of transmission to humansObjective: The goal of this study is to extract collagen from catfish bone.Material and methods: Catfish (Silurus triostegus) bone was used to extract collagen that has beenpepsinized and acid-solubilized collagen (ASC) (PSC). The extracts&amp;#39; proximate composition (moisture,protein, fat, and ash) was measured, and dry weight was used to compute the yield of ASC and PSC. Themolecular weight of experimental collagen ranged from 97 to 200 KD, according to the SDS-PAGEpattern. In order to identify bone collagen, the Fourier Transform Infrared (FTIR) spectra analysis for theacquired ASC and PSC, as well as the standard collagen (type I, II), was used. Using a scanning electronmicroscope (SEM, the morphological structure of ASC and PSC was revealed), and the diffraction angles(2 θ) of ASC and PSC were determined using X-ray analysis. For quality control, the HPLC techniquewas used to determine the collagen.Rustles: The proximate composition of ASC and PSC bone of catfish. The ASC bone of catfish containedmoisture (75.22%), good amount of protein (16.65%) , fat (2.40%) and ash content (0.28 %), While thePSC bone had slightly lower moisture (68.50%) than ASC, it had higher protein (19.93%), fat (1.825%),and ash (0.58%) levels ,HPLC SEM ,FTIR ,Electrophoresis, X-ray.Conclusion: Fish bone was promising sources for preparing collagen, acid soluble collagen (ASC),pepsin soluble collagen (PSC) were successfully extracted by two methods. The collagen extracted frombone catfish, was of type I collagen, which are composed two α1, one α2 chains, and β chains as shownby SDS-PAGE patterns.

Baicalein alleviates fibrosis and inflammation in systemic sclerosis by regulating B-cell abnormalities

BackgroundSystemic sclerosis (SSc; also known as “scleroderma”) is an autoimmune disorder characterized by extensive fibrosis, vascular changes, and immunologic dysregulation. Baicalein (phenolic flavonoid derived from Scutellaria baicalensis Georgi) has been used to treat the pathological processes of various fibrotic and inflammatory diseases. In this study, we investigated the effect of baicalein on the major pathologic characteristics of SSc: fibrosis, B-cell abnormalities, and inflammation.MethodsThe effect of baicalein on collagen accumulation and expression of fibrogenic markers in human dermal fibroblasts were analyzed. SSc mice were produced by injecting bleomycin and treated with baicalein (25, 50, or 100 mg/kg). The antifibrotic features of baicalein and its mechanisms were investigated by histologic examination, hydroxyproline assay, enzyme-linked immunosorbent assay, western blotting and flow cytometry.ResultsBaicalein (5–120 μM) significantly inhibited the accumulation of the extracellular matrix and fibroblast activation in transforming growth factor (TGF)-β1- and platelet derived growth factor (PDGF)-induced human dermal fibroblasts, as evidenced by abrogated deposition of total collagen, decreased secretion of soluble collagen, reduced collagen contraction capability and downregulation of various fibrogenesis molecules. In a bleomycin-induced model of dermal fibrosis in mice, baicalein (25–100 mg/kg) restored dermal architecture, ameliorated inflammatory infiltrates, and attenuated dermal thickness and collagen accumulation in a dose-dependent manner. According to flow cytometry, baicalein reduced the proportion of B cells (B220+ lymphocytes) and increased the proportion of memory B cells (B220+CD27+ lymphocytes) in the spleens of bleomycin-induced mice. Baicalein treatment potently attenuated serum levels of cytokines (interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-17A, tumor necrosis factor-α), chemokines (monocyte chemoattractant protein-1, macrophage inflammatory protein-1 beta) and autoantibodies (anti-scleroderma 70 (Scl-70), anti-polymyositis-scleroderma (PM-Scl), anti-centromeres, anti-double stranded DNA (dsDNA). In addition, baicalein treatment can significantly inhibit the activation of TGF-β1 signaling in dermal fibroblasts and bleomycin-induce mice of SSc, evidenced by reducing the expression of TGF-β1 and IL-11, as well as inhibiting both small mother against decapentaplegic homolog 3 (SMAD3) and extracellular signal-related kinase (ERK) activation.ConclusionsThese findings suggest that baicalein has therapeutic potential against SSc, exerting modulating B-cell abnormalities, anti-inflammatory effects, and antifibrosis.

Biochemical and Microstructural Characteristics of Collagen Biopolymer from Unicornfish (Naso reticulatus Randall, 2001) Bone Prepared with Various Acid Types

Biopolymer-like collagen has great industrial potential in terms of its excellent properties, such as strong biocompatibility, high degradability, and low antigenicity. Collagen derived from fish by-products is preferable as it is safer (free from transmittable diseases) and acceptable to most religious beliefs. This study aimed to characterize the unicornfish (Naso reticulatus Randall, 2001) bone collagens prepared with different type of acids, i.e., acetic acid, lactic acid, and citric acid. A higher yield (Y) (p < 0.05) was obtained in the citric-acid-soluble collagen (CASC) (Y = 1.36%), followed by the lactic-acid-soluble collagen (LASC) (Y = 1.08%) and acetic-acid-soluble collagen (AASC) (Y = 0.40%). All extracted collagens were classified as type I due to the presence of 2-alpha chains (α1 and α2). Their prominent absorption spectra were located at the wavelengths of 229.83 nm to 231.17 nm. This is similar to wavelengths reported for other fish collagens. The X-ray diffraction (XRD) and infrared (IR) data demonstrated that the triple-helical structure of type I collagens was still preserved after the acid-extraction process. In terms of thermal stability, all samples had similar maximum transition temperatures (Tmax = 33.34-33.51 °C). A higher relative solubility (RS) of the unicornfish bone collagens was observed at low salt concentration (0-10 g/L) (RS > 80%) and at acidic condition (pH 1.0 to pH 3.0) (RS > 75%). The extracted collagen samples had an irregular and dense flake structure with random coiled filaments. Overall, bones of unicornfish may be used as a substitute source of collagen.

Unraveling the Relevance of Tissue-Specific Decellularized Extracellular Matrix Hydrogels for Vocal Fold Regenerative Biomaterials: A Comprehensive Proteomic and In Vitro Study.

Decellularized extracellular matrix (dECM) is a promising material for tissue engineering applications. Tissue-specific dECM is often seen as a favorable material that recapitulates a native-like microenvironment for cellular remodeling. However, the minute quantity of dECM derivable from small organs like the vocal fold (VF) hampers manufacturing scalability. Small intestinal submucosa (SIS), a commercial product with proven regenerative capacity, may be a viable option for VF applications. This study aims to compare dECM hydrogels derived from SIS or VF tissue with respect to protein content and functionality using mass spectrometry-based proteomics and in vitro studies. Proteomic analysis reveals that VF and SIS dECM share 75% of core matrisome proteins. Although VF dECM proteins have greater overlap with native VF, SIS dECM shows less cross-sample variability. Following decellularization, significant reductions of soluble collagen (61%), elastin (81%), and hyaluronan (44%) are noted in VF dECM. SIS dECM contains comparable elastin and hyaluronan but 67% greater soluble collagen than VF dECM. Cells deposit more neo-collagen on SIS than VF-dECM hydrogels, whereas neo-elastin (~50 μg/scaffold) and neo-hyaluronan (~ 6 μg/scaffold) are comparable between the two hydrogels. Overall, SIS dECM possesses reasonably similar proteomic profile and regenerative capacity to VF dECM. SIS dECM is considered a promising alternative for dECM-derived biomaterials for VF regeneration.