Two Novel Angiotensin-Converting Enzyme (ACE) Inhibitory and ACE2 Upregulating Peptides from the Hydrolysate of Pumpkin (Cucurbita moschata) Seed Meal.

Pumpkin (Cucurbita moschata) seed meal (PSM), the major byproduct of pumpkin seed oil industry, was used to prepare angiotensin-converting enzyme (ACE) inhibitory and angiotensin-converting enzyme 2 (ACE2) upregulating peptides. These peptides were isolated and purified from the PSM hydrolysate prepared using Neutrase 5.0 BG by ultrafiltration, Sephadex G-15 column chromatography, and reversed-phase high-performance liquid chromatography. Two peptides with significant ACE inhibition activity were identified as SNHANQLDFHP and PVQVLASAYR with IC50 values of 172.07 and 90.69 µM, respectively. The C-terminal tripeptides of the two peptides contained Pro, Phe, and Tyr, respectively, and PVQVLASAYR also had Val in its N-terminal tripeptide, which was a favorable structure for ACE inhibition. Molecular docking results declared that the two peptides could interact with ACE through hydrogen bonds and hydrophobic interactions. Furthermore, the two peptides performed protective function on EA.hy926 cells by decreasing the secretion of endothelin-1, increasing the release of nitric oxide, and regulating the ACE2 activity. In vitro simulated gastrointestinal digestion showed the two peptides exhibited good stability against gastrointestinal enzyme digestion. In conclusion, PSM is a promising material for preparing antihypertensive peptides.

Cluster-determinant 36 (CD36) mediates intestinal absorption of dietary astaxanthin and affects its secretion.

The functional activity of dietary astaxanthin is closely related to its absorption, and the absorption of dietary carotenoids mainly mediated by transmembrane transport protein (TTP) has become the mainstream research direction in recent years. However, the main TTP mediating astaxanthin absorption and its potential mechanisms are still unclear. Hence, based on the preliminary screening results, this study aims to elucidate the role of cluster-determinant 36 (CD36) mediating astaxanthin absorption from the perspective of expression levels through in vitro cell model, in situ single-pass intestinal perfusion model and in vivo mice model. The results showed that astaxanthin uptake was significantly increased by 45.13% in CD36 overexpressing cells and decreased by 20.92% in the case of sulfo-N-succinimidyl oleate (SSO) inhibition. A similar trend also appeared in the duodenum and jejunum by in situ model. Moreover, astaxanthin uptake in the small intestine of CD36 knockout mice was significantly reduced by 88.22%. Furthermore, the inhibition or knockout of CD36 suppressed the expression of other transporters (SR-BI and NPC1L1). Interestingly, CD36 was also involved in the downstream secretion pathway, which is manifested by interfering with the expression of related proteins (ERK1/2, MTP, ApoB48, and ApoAI). Therefore, these results indicate the important role of CD36 in astaxanthin transmembrane transport for the first time, providing vital exploration way for the absorption of dietary fat-soluble substances.

Elucidating the molecular mechanism of water migration in myosin gels of Nemipterus virgatus during low pressure coupled with heat treatment.

The relationship between myosin denaturation, aggregation and water migration in Nemipterus virgatus myosin gels with different treatment processes under optimal low pressure coupled with heat treatment was investigated to clarify the molecular mechanism of water migration. With the different treatment processes, the proportion of bound water of the myosin gels increased significantly (P < 0.05). Denaturation of myosin S1 sub-fragments and α-helical unfolding during different treatment processes led to an increase in ß-sheets content. These promote increased exposure of Try residues and hydrophobic groups of myosin, formation of clathrate hydrates, and reduced mobility of bound water. Furthermore, hydrophobic interactions and disulfide bonds caused the head-head and head-hinge to coalesce into a 3D honeycomb network with greater fractal dimension, less lacunarity, smaller water hole diameter and more water holes. This increased the capillary pressure experienced by the bound water, causing immobile water to migrate towards the bound water. The present study may be necessary to improve the mechanism of water migration in protein gel systems and to promote the industrial application of high pressure processing technology in surimi-based foods.

Review of recent advances on health benefits, microbial transformations, and authenticity identification of Citri reticulatae Pericarpium bioactive compounds.

The extensive health-promoting effects of Citri Reticulatae Pericarpium (CRP) have attracted researchers' interest. The difference in storage time, varieties and origin of CRP are closely related to the content of bioactive compounds they contain. The consitituent transformation mediated by environmental microorganisms (bacteria and fungi) and the production of new bioactive components during the storage process may be the main reason for 'the older, the better' of CRP. In addition, the gap in price between different varieties can be as large as 8 times, while the difference due to age can even reach 20 times, making the 'marketing young-CRP as old-CRP and counterfeiting origin' flood the entire market, seriously harming consumers' interests. However, so far, the research on CRP is relatively decentralized. In particular, a summary of the microbial transformation and authenticity identification of CRP has not been reported. Therefore, this review systematically summarized the recent advances on the main bioactive compounds, the major biological activities, the microbial transformation process, the structure, and content changes of the active substances during the transformation process, and authenticity identification of CRP. Furthermore, challenges and perspectives concerning the future research on CRP were proposed.

Gastroprotective Effects of the Aqueous Extract of Finger Citron Pickled Products against Ethanol-Induced Gastric Damage: In Vitro and In Vivo Studies.

Finger citron pickled products (FCPP), as folk remedies, are famous in southern China for protecting gastric mucosa. However, the gastric mucosa protection of FCPP has not been reported yet, and its effective mechanism is unclear. In this study, the protective mechanism of FCPP aqueous extract on gastric mucosa was investigated in vitro and in vivo for the first time, using human gastric mucosa epithelial cells (GES-1) and acute alcoholic gastric ulcer rat model respectively. Furthermore, we also investigated the main substances in the aqueous extract that exert gastroprotective activity using a GES-1 scratch test and basic chemical composition analysis. FCPP aqueous extract was found to play a protective and reparative role in GES-1 by promoting the secretion of trefoil factor thyroid transcription factor 2 (TFF2) and inhibiting the secretion of tumor necrosis factor-α (TNF-α) in cells damaged by alcohol. The ulcer index of gastric tissue induced by alcohol was significantly decreased (p < 0.01) after pretreatment with FCPP aqueous extract, indicating that FCPP aqueous extract had a good protective effect on the stomach mucosa. Moreover, FCPP aqueous extract could increase superoxide dismutase (SOD) activity and inhibit malondialdehyde (MDA) content, exhibiting good antioxidant capacity. Aqueous extract of FCPP could also effectively inhibit the increase of cytokines TNF-α, interleukin-1ß (IL-1ß) and interleukin-6 (IL-6) in serum of rats, and promote the increase of anti-inflammatory cytokines interleukin-10 (IL-10) to some extent. Furthermore, FCPP aqueous extract could inhibit the expression of nuclear factor kappa-B (NF-κB/P65) protein, caspase-1 protein and IL-1ß protein in the gastric tissue of rats, while promoting the expression of IκBα protein, indicating that the gastric mucosa protection effects of FCPP aqueous extract were mainly dependent on the NF-κB/caspase-1/IL-1ß axis. The polysaccharides in FCPP aqueous extract might be the main components that exerted gastroprotective activity, as demonstrated by GES-1 cell scratch assay. This study confirmed that FCPP aqueous extract presented promising potential in protecting gastric mucosa and avoiding gastric ulcers, which could provide an experimental basis for further utilizing the medicinal value and developing new products of FCPP.

Structural characterization and immunomodulatory activity of a polysaccharide from finger citron extracted by continuous phase-transition extraction.

FCP-2-1, a water-soluble polysaccharide rich in galacturonic acid was isolated by continuous phase-transition extraction and purified with DEAE-52 cellulose and Sephadex G-100 column chromatography from finger citron with essential oil and flavonoids removed. The structural characterization and immunomodulatory activity of FCP-2-1 were further investigated in this work. FCP-2-1 with a Mw and Mn of 1.503 × 104 g/mol and 1.125 × 104 g/mol, respectively, was predominantly composed of galacturonic acid, galactose, and arabinose in a molar ratio of 0.685: 0.032: 0.283. The main linkage types of FCP-2-1 were proved to be →5)-α-L-Araf-(1→ and →4)-α-D-GalpA-(1→ based on methylation and NMR analysis. Moreover, FCP-2-1 was demonstrated to have significant immunomodulatory effects on macrophages in vitro by improving the cell viability, and enhancing phagocytic activity and secretion of NO and cytokines (IL-1ß, IL-6, IL-10 and TNF-α), indicating that FCP-2-1 could be used as a natural agent in immunoregulation functional foods.

Effect of sodium bicarbonate on textural properties and acceptability of gel from unwashed Asian sea bass mince.

Asian sea bass mince gels having different adjusted moisture/water content (80 and 85%; w/w) were prepared with addition of sodium bicarbonate (SB) at various concentrations (0, 0.05 and 0.1%; w/w). Fish mince gels of 80% water content added with 0.05 and 0.1% SB (G80-0.05 and G80-0.1, respectively) had the highest increase (135-139%) in breaking force (BrF) than the respective control gel (G80) (P < 0.05). For gel with 85% water content, a lower increase (17-28%) in BrF was found with the addition of SB as compared to their corresponding control (G85). Whiteness of all samples was continuously decreased with increasing amount of SB, however the water holding capacity was increased drastically with augmenting levels of SB, regardless of the water content (P < 0.05). A loss in the elasticity of gel was attained with the addition of SB as indicated by decreasing storage modulus. A finer and more compact network was detected in a gel containing SB, irrespective of water content. Based on sensory scores, gel having 85% water content added with 0.05 and 0.1% SB had similar acceptability to the control gel (G80) containing 80% water content (commercial level). Therefore, SB at the appropriate level could improve the gelling properties with higher water holding ability of the mince gel with high acceptability.

The potential mechanisms of bergamot-derived dietary fiber alleviating high-fat diet-induced hyperlipidemia and obesity in rats.

This study is aimed to investigate the health-associated benefits of bergamot-dietary fibers (DFs) with a special emphasis on weight loss and lipid-lowering effects, as well as the potential mechanisms involved. The feeding experiment of Sprague-Dawley (SD) rats for 6 weeks showed that DFs had dose-dependent regulatory effects against metabolic syndrome and they controlled obesity by slowing down the rate of weight growth, and reduced body mass index (BMI) and Lee's index without affecting appetite. Furthermore, DFs inhibited increment in TG, TC, LDL-C levels and AI index caused by a high-fat diet, and improved the pathological abnormality of the liver. Western blot results showed that DFs significantly up-regulated the protein expression levels of LXRα and CYP7A1, and down-regulated the levels of SREBP-1c, FAS, ACC and SREBP-2 in the liver. QRT-PCR results showed that DFs up-regulated PGC-1α, PRDM16, UCP-1, and PPARγ in brown adipose tissue. These results suggest that DFs played an effective role in reducing weight and lipids levels by promoting the decomposition and transport of lipids in liver, increasing the energy consumption of brown adipose tissue. DFs intervention reduced the difference in the intestinal microflora between rats fed with a normal diet and those fed with a high-fat diet. Soluble dietary fiber (SDF) and total dietary fiber (TDF) showed better weight loss and hypolipidemic potential compared to insoluble dietary fiber (IDF) at the same dose. In conclusion, bergamot-derived DFs demonstrated the potential to lower blood cholesterol and body weight and could be used to develop novel functional foods for the prevention or treatment of obesity and hyperlipidemia.

Insight into the mechanism of optimal low-level pressure coupled with heat treatment to improve the gel properties of Nemipterus virgatus surimi combined with water migration.

The effect of optimal low-level pressure coupled with heat treatment (OPH treatment) on the gel properties and water migration of Nemipterus virgatus surimi was studied and compared with optimal high-pressure processing treatment (OP treatment) and traditional two-stage heat treatment (H treatment). Furthermore, the mechanism of OPH treatment in improving the gel properties were explored based on myosin. OPH treatment was found to be more conducive in improving the gel strength and water-holding capacity (WHC) of surimi gel than H or OP treatments. Moreover, OPH treatment induced an increase in the proportion of myosin ß-sheets and exposed more intramolecular Tyr residues as compared to the other two treatments, which promoted myosin to form large protein clusters through disulfide bonds and hydrophobic interactions, and a honeycomb three-dimensional network structure with larger fractal dimension, lower porosity, smaller water hole diameter, and a greater number of water holes, was obtained. These helped the OPH-induced surimi gel lock in more unfrozen bound water and immobile water, and ultimately rendered better gel properties.

Omicron variant (B.1.1.529) of SARS-CoV-2: Mutation, infectivity, transmission, and vaccine resistance.

The appearance of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variant Omicron (B.1.1.529) has caused panic responses around the world because of its high transmission rate and number of mutations. This review summarizes the highly mutated regions, the essential infectivity, transmission, vaccine breakthrough and antibody resistance of the Omicron variant of SARS-CoV-2. The Omicron is highly transmissible and is spreading faster than any previous variant, but may cause less severe symptoms than previous variants. The Omicron is able to escape the immune system's defenses and coronavirus disease 2019 vaccines are less effective against the Omicron variant. Early careful preventive steps including vaccination will always be key for the suppression of the Omicron variant.

Insight into the Effect of Ice Addition on the Gel Properties of Nemipterus virgatus Surimi Gel Combined with Water Migration.

The effect of the amount of ice added (20-60%) on the gel properties and water migration of Nemipterus virgatus surimi gel obtained with two-stage heat treatment was studied. The gel strength and water-holding capability (WHC) of the surimi gel with 30% ice added were significantly higher than those of other treatment groups (p < 0.05). The addition of 30% ice was conducive to the increase of protein ß-sheet proportion during heat treatment, exposing more reactive sulfhydryl groups. These promoted the combination of protein-protein through disulfide bonds and hydrophobic-hydrophobic interactions, forming an ordered three-dimensional gel network structure. Meanwhile, the increase in hydrogen bonds promoted the protein-water interaction. Low-field nuclear magnetic resonance analysis showed that more bound water was locked in the gel system, reducing the migration of immobile water to free water and finally showing better gel properties. When the amount of ice added was insufficient (20%), the gel structure lacked the support of immobile water, resulting in deterioration of gel strength. However, excessive addition of ice (>30%) was not conducive to the combination of protein-protein and protein-water, forming a large and rough gel structure, resulting in the migration of immobile water to free water and ultimately exhibited weak gel properties.

An "off-on-off" fluorescence chemosensor for the sensitive detection of Cu2+ in aqueous solution based on multiple fluorescence emission mechanisms.

A new organosiloxane precursor ((E)-3-hydroxy-4-((2-(2-hydroxy-4-(3-(3-(triethoxysilyl)propyl)ureido)benzoyl)hydrazono)methyl)phenyl(3-(triethoxysilyl)propyl)carbamate, hereinafter referred to as AHBH-Si) and tetraethylorthosilicate (TEOS) were mixed as the mixed Si source, and bridged periodic mesoporous organic silica (AHBH-PMOs) nanoparticles were obtained through the co-condensation reaction. AHBH-PMO nanoparticles possess mechanisms of "Aggregation Induced Emission" (AIE) and "Intramolecular Charge Transfer" (ICT), which originate from the molecular structure of AHBH having "C[double bond, length as m-dash]N" bond, ortho hydroxyl groups, etc.. Therefore, the optical properties of AHBH are excellent with respect to the solvent effect and enhanced fluorescence. For hybrid materials, the silica framework provides a rigid environment that restricts the rotation of AHBH, thereby turning on the fluorescence of AHBH due to the regulation by the AIE effect. In particular, AHBH-PMOs are no longer restricted by organic solvents and could really achieve the response to Cu2+ with high sensitivity and selectivity in aqueous solutions of a wide pH range. In addition, the detection limit is as low as 3.26 × 10-9 M. Methods such as Fourier transform infrared spectroscopy, proton nuclear magnetic resonance spectroscopy, and high-resolution mass spectrometry have shown the coordination interaction between AHBH and Cu2+. The Gaussian 09 software of density functional theory to calculate the reducing changes of energy gaps among AHBH and AHBH-Si before and after the addition of Cu2+ showed that coordination interaction exists in the system. These results indicate that AHBH-PMO hybrid materials have potential applications in the field of environmental monitoring.

Variations of volatile flavour compounds in finger citron (Citrus medica L. var. sarcodactylis) pickling process revealed by E-nose, HS-SPME-GC-MS and HS-GC-IMS.

The pickled products of finger citron are famous in southern China for their unique taste and flavor. Although pickling process involves complex treatments including salting, desalting, sugaring, cooking and drying, extended shelf-life up to ten years after pickling can be achieved. In this study, the variations of volatile flavour components in the pickling process of finger citron were investigated by electronic nose (E-nose), headspace solid phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) and headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS). HS-SPME-GC-MS identified 85 substances, and HS-GC-IMS identified 81 substances, including terpenoids (21), aromatic hydrocarbons (11), alcohols (11), aldehydes (10), esters (7), phenols (6), acids (5), ethers (2), ketones (2), and other species (10). Linalool, limonene, (E)-3,7-dimethyl-1,3,6-octatriene, myrcene, 3-carene, ß-pinene, α-pinene, terpinolene, 1-methyl-4-(1-methylethyl)-1,4-cyclohexadiene, α-terpinene, (S)-ß-bisabolene, 1-isopropyl-2-methylbenzene and 1-methyl-4-(1-methylethenyl)-benzene were the stable substances at relatively high contents in finger citron at different pickling process. Salting and drying steps in the pickling process exerted greatest influence on the volatile components of finger citron. Salting promoted the generation of aldehydes, esters and acids, but led to the disappearance of alcohols, while drying promoted the generation of alcohols, phenols, aldehydes and acids at the expense of reduction in terpenoids. Our study revealed that the characteristic volatile compounds of finger citron pickled products was mainly formed by the biological reactions in the salting stage and thermal chemical transformations in the drying stage. This study also validated the suitability of E-nose combined with HS-SPME-GC-MS and HS-GC-IMS in tracking the changes of volatile components in finger citron during the pickling process.

The gut microbiota alteration and the key bacteria in Astragalus polysaccharides (APS)-improved osteoporosis.

Osteoporosis is a worldwide epidemic, and certain functional foods can alleviate osteoporosis with great efficiency. Here, in a dexamethasone-induced osteoporosis rat model, it was proved that APS could restore the bone mineral density (BMD) and repair the impairment of bone microarchitecture, two major features of osteoporosis. In APS-treated rats, acid phosphatase 5 (ACP5) and pro-inflammatory cytokines (TNF-α and IL-2) were significantly decreased. This suggested that APS might improve osteoporosis by inhibiting osteoclastogenesis and preventing inflammation. Further analysis on the bacterial community revealed that the structure of gut microbiota was dramatically changed by APS, and 13 bacteria (such as c_Bacteroidia, p_Bacteroidetes, and g_Allpprevotella) could serve as biomarkers for APS-improved osteoporosis. Furthermore, five genera (uncultured_bacterium_f_Ruminococcaceae, Alloprevotella, Ruminococcaceae_UCG-014, Blautia and Lactobacillus) were inferred as the key bacteria in APS-improved osteoporosis. In conclusion, APS-modified gut microbiota and the potential key bacteria to alleviate osteoporosis, as well as its relationship with improved osteoporosis, were investigated in our present study. Our results will help to understand how APS improves osteoporosis by regulating gut microbiota and contribute to the development and application of functional foods to alleviate refractory osteoporosis by regulating targeted intestinal bacteria.

Identification of Flavoanoids From Finger Citron and Evaluation on Their Antioxidative and Antiaging Activities.

Finger citron (Citrus medica L. var. sarcodactylis Swingle) is a traditional Chinese herb and considered as a healthy food. Flavonoids are the major bioactive substances in Finger citron. In this study, the major flavonoids of finger citron (FFC) were purified with AB-8 macroporous resins, and then three of them were identified as diosmetin-6-8-di-C-glucoside, hesperidin and diosmetin-6-C-glucoside, and other two were preliminarily inferred as limocitrol 3-alpha-l-arabinopyranosyl-(1->3)-galactoside and scutellarein 4'-methyl ether 7-glucoside by high-performance liquid chromatography and ultraperformance liquid chromatography to quadrupole time-of-flight mass spectrometry. Further, their antioxidation and antiaging activities were determined in vitro and in vivo. In vitro, chemical assays revealed that the purified FFC had strong antioxidative activity as demonstrated by its strong DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS [2,2-azinobis (3-ethyl-benzothiazoline-6-sulphonic acid) diammonium salt] radical scavenging activities and ORAC (oxygen radical absorbance capacity). In vivo, the purified FFC significantly increased the mean and maximum lifespan of Caenorhabditis elegans by 31.26 and 26.59%, respectively, and showed no side effects on their physiological functions. Under normal and oxidative stress conditions, purified FFC reduced the accumulation of reactive oxygen species (ROS) and malondialdehyde, while increased superoxide dismutase (SOD) and catalase (CAT) enzyme activities in C. elegans. Together, we successfully identified three major substances in purified FFC of finger citron and determined the excellent antiaging activity of FFC, which is attributed to its strong antioxidative activity and effect on homeostasis of ROS.

Optimization in continuous phase-transition extraction of crude flavonoids from finger citron fruit and evaluation on their antiaging activities.

The development of antiaging functional products is a hot topic in the field of functional foods. However, the efficient extraction of functional ingredients is the limiting step for the functional food industry. Continuous phase-transition extraction (CPE) is a new extraction technique that combines the advantages of Soxhlet extraction and supercritical extraction, which may have a distinct advantage over traditional methods in the extraction of flavonoids. In our study, the Box-Behnken design combined with response surface methodology was used to optimize CPE of crude flavonoids from finger citron fruit. The antiaging activities of finger citron crude flavonoids (FCCF) were evaluated by Caenorhabditis elegans (C. elegans) model. The optimal extraction conditions for CPE were as follows: ethanol concentration 85%, temperature 90°C, time 120 min, and pressure 0.2 MPa. Compared with the heat reflux extraction, the extraction rate and content of FCCF extracted by CPE increased by 24.28% and 33.22% (p < .05), respectively. FCCF extended the lifespan of C. elegans by 14.94% without causing adverse effects on their reproduction and locomotion ability. A further analysis suggested that FCCF prolonged the lifespan of nematodes under normal and oxidative stress by increasing the activity of major enzymes in endogenous antioxidant defense system and reducing the accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA). The results confirmed the effectiveness of CPE in extracting crude flavonoids from finger citron fruit, and the extracted FCCF exhibited strong antiaging activities.

Preliminary characterization of the structure and immunostimulatory and anti-aging properties of the polysaccharide fraction of Haematococcus pluvialis.

In this study, a polysaccharide fraction (HPP-c3-s1) was obtained from Haematococcus pluvialis using DEAE-52 anion exchange and Sephacryl S400 chromatographies. The structure of HPP-c3-s1 was partially characterized and its biological activity was investigated. HPP-c3-s1 is a homogeneous polysaccharide with a molecular weight of 23 413 kDa as determined by high-performance gel permeation chromatography and polyacrylamide gel electrophoresis. Periodate oxidation and Smith degradation analysis combined with GC-MS determined that HPP-c3-s1 contains 1 → 2, 1 → 3, 1 → 4, and probably a few 1 → 6 glycosyl linkages. Fourier transform infrared spectroscopy and nuclear magnetic resonance analyses indicate HPP-c3-s1 is a pyranose containing an amino and O-acetyl group. The sugar chains in HPP-c3-s1 contain both α and ß glycosidic configurations, where the ß configuration is the primary form. It was observed that the HPP-c3-s1 nanostructure had linear and branched forms at a concentration of 5 µg mL-1 and created different sized aggregates at a higher concentration of 50 µg mL-1. Furthermore, HPP-c3-s1 had significant immunostimulatory effects on splenocytes and B lymphocytes. In assays assessing anti-aging effects, HPP-c3-s1 extended the mean survival of Caenorhabditis elegans without adversely affecting reproduction. In addition, treatment with HPP-c3-s1 resulted in delays in age-related physiologic parameters, including body movement, rates of head swing and body bending, and accumulation of intestinal lipofuscinosis in C. elegans. Overall, these results suggest HPP-c3-s1 has remarkable immunomodulatory and anti-aging properties that may serve as the basis for development of functional foods and dietary supplements.

[Research Progress of Collagen-based Three-dimensional Porous Scaffolds Used in Skin Tissue Engineering].

Collagen is a kind of natural biomedical material and collagen based three-dimensional porous scaffolds have been widely used in skin tissue engineering. However, these scaffolds do not meet the requirements for artificial skin substitutes in terms of their poor mechanical properties, short supply, and rejection in the bodies. All of these factors limit their further application in skin tissue engineering. A variety of methods have been chosen to meliorate the situation, such as cross linking and blending other substance for improving mechanical properties. The highly biomimetic scaffolds either in structure or in function can be prepared through culturing cells and loading growth factors. To avoid the drawbacks of unsafety attributing to animals, investigators have fixed their eyes on the recombinant collagen. This paper reviews the the progress of research and application of collagen-based 3-dimensional porous scaffolds in skin tissue engineering.

Comparison of two proanthocyanidin cross-linked recombinant human collagen-peptide (RHC) - chitosan scaffolds.

Cross-linking plays an important role in tissue engineering, which involves the alternative of cross-linker and the way of components interaction. We compared two proanthocyanidin (PA) cross-linked recombinant human collagen-peptide - chitosan scaffolds: immerse cross-linking (I-CLS) and premix cross-linking (P-CLS). Both of the scaffolds presented homogeneous pore structure with mean pore size of 110-115 µm. The swelling ratio was decreased to 29.6 in I-CLS, but increased to 37.1 in P-CLS while porosity of the two scaffolds was reduced about 8% comparing to 94.3% before cross-linking. The cross-linked scaffolds exhibited enhanced resistance to enzyme degradation and improved compressive modulus (I-CLS > P-CLS). The scaffolds transformed from elastic region to plastic region until the strain reached 60%, and the stress was 40.5, 133.2 and 84.1 kPa of uncross-linking scaffold, I-CLS and P-CLS individually. Thermal stability indicated molecular bonding between PA and the scaffold components, simultaneously, Fourier transform infrared spectroscopy mainly presented hydrogen bonding between the protein amide carbonyl and the phenolic hydroxyl with a particular transform due to pyrrolidine rings of proline in P-CLS. Both of the I-CLS and P-CLS could promote human umbilical vein endothelial cells attachment and proliferation. The characterization suggested in situ biodegradable application of P-CLS, while a potential long-term utilization of I-CLS in tissue engineering.

Pore architecture and cell viability on freeze dried 3D recombinant human collagen-peptide (RHC)-chitosan scaffolds.

Pore architecture of 3D scaffolds used in tissue engineering plays a critical role in the maintenance of cell survival, proliferation and further promotion of tissue regeneration. We investigated the pore size and structure, porosity, swelling as well as cell viability of a series of recombinant human collagen-peptide-chitosan (RHCC) scaffolds fabricated by lyophilization. In this paper, freezing regime containing a final temperature of freezing (Tf) and cooling rates was applied to obtain scaffolds with pore size ranging from 100µm to 120µm. Other protocols of RHC/chitosan suspension concentration and ratio modification were studied to produce more homogenous and appropriate structural scaffolds. The mean pore size decreased along with the decline of Tf at a slow cooling rate of 0.7°C/min; a more rapid cooling rate under 5°C/min resulted to a smaller pore size and more homogenous microstructure. High concentration could reduce pore size and lead to thick well of scaffold, while improved the ratio of RHC, lamellar and fiber structure coexisted with cellular pores. Human umbilical vein endothelial cells (HUVECs) were seeded on these manufactured scaffolds, the cell viability represented a negative correlation to the pore size. This study provides an alternative method to fabricate 3D RHC-chitosan scaffolds with appropriate pores for potential tissue engineering.