Combined Use of Autologous Sustained-Release Scaffold of Adipokines and Acellular Adipose Matrix to Construct Vascularized Adipose Tissue.

BACKGROUND: Adipose tissue engineering plays a key role in the reconstruction of soft-tissue defects. The acellular adipose matrix (AAM) is a promising biomaterial for the construction of engineered adipose tissue. However, AAM lacks sufficient adipoinduction potency because of the abundant loss of matrix-bound adipokines during decellularization. METHODS: An adipose-derived extracellular matrix collagen scaffold, "adipose collagen fragment" (ACF), was prepared using a novel mechanical method that provides sustained release of adipokines. Here, the authors used label-free proteomics methods to detect the protein components in AAM and ACF. In vivo, ACF was incorporated into AAM or acellular dermal matrix and implanted into nude mice to evaluate adipogenesis. Neoadipocytes, neovessels, and corresponding gene expression were evaluated. The effects of ACF on adipogenic differentiation of human adipose-derived stem cells and tube formation by human umbilical vein endothelial cells were tested in vitro. RESULTS: Proteomics analysis showed that ACF contains diverse adipogenic and angiogenic proteins. ACF can release diverse adipokines and induce highly vascularized, mature adipose tissue in AAM, and even in nonadipogenic acellular dermal matrix. Higher expression of adipogenic markers peroxisome proliferator-activated receptor gamma and CCAAT/enhancer-binding protein alpha and greater numbers of tubule structures were observed in ACF-treated groups in vitro. CONCLUSION: The combination of ACF and AAM could serve as a novel and promising strategy to construct mature, vascularized adipose tissue for soft-tissue reconstruction. CLINICAL RELEVANCE STATEMENT: The combined use of AAM and ACF has been proven to induce a highly vascularized, mature, engineered adipose tissue in the nude mouse model, which may serve as a promising strategy for soft-tissue reconstruction.

Corrigendum to 'Methoxy polyethylene glycol modification promotes adipogenesis by inducing the production of regulatory T cells in xenogeneic acellular adipose matrix' [Materials Today Bio 12 (2021) 100161].

[This corrects the article DOI: 10.1016/j.mtbio.2021.100161.].

Adipose Collagen Fragment: A Novel Adipose-Derived Extracellular Matrix Concentrate for Skin Filling.

BACKGROUND: Skin filler is an option for treating skin aging and wrinkles; however, currently used fillers are limited by poor biocompatibility, rapid degradation, and possible hypersensitivity reactions. Autologous adipose tissue-derived products have been recognized as promising options for skin rejuvenation. OBJECTIVES: This study aimed to develop a novel adipose-derived product for skin filling. METHODS: Adipose collagen fragment (ACF) was prepared through pulverization, filtration, and centrifugation. The macrography, structure, types of collagen, and cell viability of ACF were evaluated by immunostaining, western blotting, and cell culture assays. ACF, nanofat, and phosphate-buffered saline (9 spots/side, 0.01 mL/spot) were intradermally injected in the dorsal skin of 36 female BALB/c nude mice; the skin filling capacity and the collagen remodeling process were then investigated. Twenty-one female patients with fine rhytides in the infraorbital areas were enrolled and received clinical applications of ACF treatment. Therapeutic effects and patients' satisfaction scores were recorded. RESULTS: The mean [standard deviation] yield of ACF from 50 mL of Coleman fat was 4.91 [0.25] mL. ACF contained nonviable cells and high levels of collagen I, collagen IV, and laminin. Fibroblasts and procollagen significantly increased in ACF and ACF-treated dermis (P < 0.05). Overall, 85.7% of patients were satisfied with the therapy results, and no infections, injection site nodules, or other unwanted side effects were observed. CONCLUSIONS: ACF significantly improved dermal thickness and collagen synthesis and may serve as a potential autologous skin filler.

Macrophage-mediated extracellular matrix remodeling after fat grafting in nude mice.

Clinical unpredictability and variability following fat grafting remain non-negligible problems due to the unknown mechanism of grafted fat retention. The role of the extracellular matrix (ECM), which renders cells with structural and biochemical support, has been ignored. This study aimed to clarify the ECM remodeling process, related cellular events, and the spatiotemporal relationship between ECM remodeling and adipocyte survival and adipogenesis after fat grafting. Labeled Coleman fat by the matrix-tracing technique was grafted in nude mice. The ECM remodeling process and cellular events were assessed in vivo. The related cytokines were evaluated by qRT-PCR. An in vitro cell migration assay was performed to verify the chemotactic effect of M2-like macrophages on fibroblasts. The results demonstrated that in the periphery, most of the adipocytes of the graft survived or regenerated, and the graft-derived ECM was gradually replaced by the newly-formed ECM. In the central parts, most adipocytes in the grafts died shortly after, and a small part of the graft-derived and newly-formed ECM was expressed with irregular morphology. Adipose ECM remodeling is associated with increased infiltration of macrophages and fibroblasts, as well as up-regulated expression of cytokines in the adipose tissue. To sum up, our results describe the various preservation mode of fat grafts after transplantation and underscore the importance of macrophage-mediated ECM remodeling in graft preservation after fat grafting. The appreciation and manipulation of underlying mechanisms that are operant in this setting stand to explore new therapeutic approaches and improve clinical outcomes of fat grafting.

Inflammation-mediated matrix remodeling of extracellular matrix-mimicking biomaterials in tissue engineering and regenerative medicine.

Extracellular matrix (ECM)-mimicking biomaterials are considered effective tissue-engineered scaffolds for regenerative medicine because of their biocompatibility, biodegradability, and bioactivity. ECM-mimicking biomaterials preserve natural microstructures and matrix-related bioactive components and undergo continuous matrix remodeling upon transplantation. The interaction between host immune cells and transplanted ECM-mimicking biomaterials has attracted considerable attention in recent years. Transplantation of biomaterials may initiate injuries and early pro-inflammation reactions characterized by infiltration of neutrophils and M1 macrophages. Pro-inflammation reactions may lead to degradation of the transplanted biomaterial and drive the matrix into a fetal-like state. ECM degradation leads to the release of matrix-related bioactive components that act as signals for cell migration, proliferation, and differentiation. In late stages, pro-inflammatory cells fade away, and anti-inflammatory cells emerge, which involves macrophage polarization to the M2 phenotype and leukocyte activation to T helper 2 (Th2) cells. These anti-inflammatory cells interact with each other to facilitate matrix deposition and tissue reconstruction. Deposited ECM molecules serve as vital components of the mature tissue and influence tissue homeostasis. However, dysregulation of matrix remodeling results in several pathological conditions, such as aggressive inflammation, difficult healing, and non-functional fibrosis. In this review, we summarize the characteristics of inflammatory responses in matrix remodeling after transplantation of ECM-mimicking biomaterials. Additionally, we discuss the intrinsic linkages between matrix remodeling and tissue regeneration. STATEMENT OF SIGNIFICANCE: Extracellular matrix (ECM)-mimicking biomaterials are effectively used as scaffolds in tissue engineering and regenerative medicine. However, dysregulation of matrix remodeling can cause various pathological conditions. Here, the review describes the characteristics of inflammatory responses in matrix remodeling after transplantation of ECM-mimicking biomaterials. Additionally, we discuss the intrinsic linkages between matrix remodeling and tissue regeneration. We believe that understanding host immune responses to matrix remodeling of transplanted biomaterials is important for directing effective tissue regeneration of ECM-mimicking biomaterials. Considering the close relationship between immune response and matrix remodeling results, we highlight the need for studies of the effects of clinical characteristics on matrix remodeling of transplanted biomaterials.

Long-acting microneedle patch loaded with adipose collagen fragment for preventing the skin photoaging in mice.

Skin photoaging is one of the most serious public health problems in the 21st century that may lead to thin, saggy, and structurally weakened skin. Adipokine therapy toward skin photoaging is always associated with poor permeability, biologic stability and the short in vivo release duration. Our laboratory previously extracted an extracellular matrix component of adipose tissue by purely physical methods, namely "adipose collagen fragment (ACF)", which holds promise for preventing skin photoaging. However, the injection treatment of ACF requires repeated preparation processes and injection procedures, which may be time-consuming and painful. Therefore, we describe the fabrication and assessment of a detachable ACF-microneedle (ACF-MN) patch that creates minimally invasive dermal microtrauma upon application. And we evaluated the morphology characterization, mechanical properties and puncture performance in vitro. The delivery efficiency of ACF from the patches was estimated in vitro and vivo. Then, the therapeutic efficacy was identified through applying ACF-MN patches into the dermis of UVA-induced photoaging mice and the related detection of skin photoaging was estimated. Our results demonstrated that ACF-MN exhibited well skin puncture performance and could release ACF component slowly. Meanwhile, this microneedle device loaded with ACF exhibited the treatment efficiency on skin photoaging in a mouse model. Therefore, implantation of the microtrauma-mediated, long-acting ACF-MN system can be utilized as a potential candidate for preventing skin photoaging in the clinic.

Impact of wheat bran micronization on dough properties and bread quality: Part II - Quality, antioxidant and nutritional properties of bread.

To investigate the impact of superfine grinding of wheat bran on bread quality, antioxidant and nutritional properties, bran with different particle sizes (coarse, D50 of 362.3 µm; medium, 60.4 µm; superfine, 11.3 µm) were produced and fortified to white bread at three levels (10, 20 and 30%). At 20% fortification, compared to coarse bran, superfine bran increased the hardness and reduced the brightness of bread crumb by 56.3 and 3.30%, respectively, while it decreased bread's cell size by 10.7% and insignificantly impacted on bread's specific volume and porosity. Superfine bran retarded bread staling by 8.3% than coarse bran. It resulted in significantly better sensory attributes of bread in taste, texture and general palatability, and the fortified bread was overall acceptable (score > 6). Moreover, faster release of antioxidants (285-353% higher k), slower release of glucose (10.8% lower k), 3.76% less rapidly digestible starch, 5.65% more slowly digestible starch and 13.2% more resistant starch were found in the superfine group than the coarse one. Results demonstrated the potential of 20% fortification of superfine bran in developing fibre-enriched bread with satisfactory quality, increased antioxidant property and improved glycaemic modulation.

Light-regulated nitric oxide release from hydrogel-forming microneedles integrated with graphene oxide for biofilm-infected-wound healing.

Nitric oxide (NO) is an antimicrobial agent that possesses tissue-regenerating ability. However, it also has a short half-life and storage difficulties as disadvantages to its application. To overcome these limitations, a new type of hydrogel-forming microneedle (HFMN) is proposed that can be fabricated by integrating polyvinyl alcohol (PVA) hydrogels (a highly biocompatible drug carrier) with S-nitrosoglutathione (GSNO, a NO releasing agent), and graphene oxide (GO) at freezing temperatures (GO-GNSO-HFMNs). Results show that GSNO-GO-HFMNs release NO gradually with increasing temperature and, more importantly, can be warmed up by mild infrared irradiation to accelerate subcutaneous release of NO from the heat-sensitive GSNO. Biofilm-infected wounds often present obstacles to drug delivery, whereas the microneedle (MN) structure disrupts the biofilm and directly releases NO into the wound. This inhibits bacterial growth and increases tissue regeneration while shortening the healing time of biofilm-infected wounds. Therefore, this type of patch can be regarded as a novel, heat-sensitive, light-regulated, NO-releasing MN patch.

Ethnicity impact on oral processing behaviour and glycemic response to noodles: Chinese (Asian) vs. New Zealander (Caucasian).

There is an increasing awareness of the link between food breakdown during chewing and its nutrient release and absorption in the gastrointestinal tract. However, how oral processing behaviour varies among different ethnic groups, and how such difference further impacts on bolus characteristics and consequently glycemic response (GR) are not well understood. In this study, we recruited a group of Asian (Chinese) subjects in China (n = 32) and a group of Caucasian subjects in New Zealand (n = 30), both aged between 18 and 30 years, and compared their blood glucose level (BGL) over 120 min following consumption of a glucose drink and cooked white noodles. We also assessed their chewing behaviour, unstimulated saliva flow rate, and ready-to-swallow bolus characteristics to determine whether these measures explain the ethnic differences in postprandial glycaemia. Compared to New Zealand subjects, the Chinese subjects showed 35% slower saliva flow rate but around 2 times higher salivary α-amylase activity in the unstimulated state. During consumption of noodles, Chinese subjects on average took a larger mouthful size, chewed each mouthful for longer and swallowed a larger number of particles with a smaller particle size area. Total GR measured by area under the curve (IAUC) was higher among the Chinese subjects. They also experienced higher BGL at 15 min, as well as higher peak BGL. There were strong correlations observed between oral processing and GR parameters. Results of this study confirmed the significance of oral processing in determining food digestion, and will provide new insights on the role of ethnicity in influencing people's physiological response to food.

Adipose Collagen Fragment: A Novel Adipose-Derived Extracellular Matrix Concentrate for Skin Filling.

BACKGROUND: Skin filler is an option for treating skin aging and wrinkles; however, currently used fillers are limited by poor biocompatibility, rapid degradation, and possible hypersensitivity reactions. Autologous adipose tissue-derived products have been recognized as promising options for skin rejuvenation. OBJECTIVES: This study aimed to develop a novel adipose-derived product for skin filling. METHODS: Adipose collagen fragment (ACF) was prepared through pulverization, filtration, and centrifugation. The macrography, structure, types of collagen, and cell viability of ACF were evaluated by immunostaining, western blotting, and cell culture assays. ACF, nanofat, and phosphate-buffered saline (9 spots/side, 0.01 mL/spot) were intradermally injected in the dorsal skin of 36 female BALB/c nude mice; the skin filling capacity and the collagen remodeling process were then investigated. Twenty-one female patients with fine rhytides in the infraorbital areas were enrolled and received clinical applications of ACF treatment. Therapeutic effects and patients' satisfaction scores were recorded. RESULTS: The mean [standard deviation] yield of ACF from 50 mL of Coleman fat was 4.91 [0.25] mL. ACF contained nonviable cells and high levels of collagen I, collagen IV, and laminin. Fibroblasts and procollagen significantly increased in ACF and ACF-treated dermis (P < 0.05). Overall, 85.7% of patients were satisfied with the therapy results, and no infections, injection site nodules, or other unwanted side effects were observed. CONCLUSIONS: ACF significantly improved dermal thickness and collagen synthesis and may serve as a potential autologous skin filler.

Methoxy polyethylene glycol modification promotes adipogenesis by inducing the production of regulatory T cells in xenogeneic acellular adipose matrix.

Acellular adipose matrix (AAM) has emerged as an important biomaterial for adipose tissue regeneration. Current decellularization methods damage the bioactive components of the extracellular matrix (ECM), and the residual immunogenic antigens may induce adverse immune responses. Here, we adopted a modified decellularization method which can protect more bioactive components with less immune reaction by methoxy polyethylene glycol (mPEG). Then, we determined the adipogenic mechanisms of mPEG-modified AAM after xenogeneic transplantation. AAM transplantation caused significantly lesser adipogenesis in the wild-type group than in the immune-deficient group. The mPEG-modified AAM showed significantly lower immunogenicity and higher adipogenesis than the AAM alone after xenogeneic transplantation. Furthermore, mPEG modification increased regulatory T (Treg) cell numbers in the AAM grafts, which in turn enhanced the M2/M1 macrophage ratio by secreting IL-10, IL-13, and TGF-ß1. These findings suggest that mPEG modification effectively reduces the immunogenicity of xenogeneic AAM and promotes adipogenesis in the AAM grafts. Hence, mPEG-modified AAM can serve as an ideal biomaterial for xenogeneic adipose tissue engineering.

Bread breakdown pathways during mastication: impact of wheat bran fortification.

The aim of this study was to investigate the impact of wheat bran fortification on the mastication process of bread. White wheat bread (WB) and bran-fortified wheat bread (BB) were consumed by eighteen panellists. The bolus was collected at four different mastication stages and characterized by properties of hydration, particle size, and texture. The results showed that there was no difference between the two bread samples in terms of swallowable bolus moisture. BB with a harder and denser texture produced more small particles and had a slightly shorter chewing time than WB during mastication. Moreover, bolus heterogeneity (D75/D25) indicated a distinct difference among mastication stages and revealed different disintegration pathways between the two samples: BB bolus exhibited a monotonous particle size reduction during mastication with reducing D50 and D75/D25; whereas, WB displayed a combination pattern of disintegration and agglomeration featuring relatively steady D50 and fluctuating D75/D25. It was concluded that bran fortification changed the bread breakdown pathways in terms of bread disintegration and bolus formation during the mastication process. This information offers new guidelines for fortifying innovative materials to manufacture foods specifically targeted for health.

An Adipose-Derived Injectable Sustained-Release Collagen Scaffold of Adipokines Prepared Through a Fast Mechanical Processing Technique for Preventing Skin Photoaging in Mice.

Ultraviolet A (UVA) radiation is the major contributor to skin photoaging, associated with increased collagen degradation and reactive oxygen species (ROS) expression. Adipokines have been proven as promising therapeutic agents for skin photoaging. However, adipokine therapy is generally limited by the short in vivo release duration and biological instability. Therefore, developing a treatment that provides a sustained release of adipokines and enhanced therapeutic effects is desirable. In this study, we developed a novel mechanical processing technique to extract adipose tissue-derived ECM components, named the "adipose collagen fragment" (ACF). The physical characterization, injectability, collagen components, residual DNA/RNA and adipokine release pattern of ACF were identified in vitro. L929 cells were treated with ACF or phosphate-buffered saline for 24 h after UVA irradiation in vitro. The expression of senescence-associated xß-galactosidase (SA-ß-gal), ROS and antioxidase were investigated. Then, we evaluated its therapeutic efficacy by injecting ACF and phosphate-buffered saline, as a control, into the dermis of photoaging nude mice and harvesting skin samples at weeks 1, 2, and 4 after treatment for assessment. The content of adipokines released from ACF was identified in vivo. The collagen synthesis and collagen degradation in ACF implants were evaluated by immune staining. Dermal thickness, fibroblast expression, collagen synthesis, ROS level, antioxidase expression, capillary density, and apoptotic cell number were evaluated by histological assessment, immune staining, and polymerase chain reaction in the skin samples. We demonstrated that ACF is the concentrated adipose extracellular matrix collagen fragment without viable cells and can be injected through fine needles. The lower expression of SA-ß-gal, ROS and higher expression of antioxidase were observed in the ACF-treated group. ACF undergoes collagen degradation and promotes neocollagen synthesis in ACF implants. Meanwhile, ACF serves as a sustained-release system of adipokines and exhibits a significantly higher therapeutic effect on mouse skin photoaging by enhancing angiogenesis, antioxidant abilities, antiapoptotic activities, and collagen synthesis through sustainedly releasing adipokines. To sum up, ACF is an adipokines-enriched, sustained-release extracellular matrix collagen scaffold that can prevent UVA-induced skin photoaging in mice. ACF may serve as a novel autologous skin filler for skin rejuvenation applications in the clinic.

Impact of wheat bran micronization on dough properties and bread quality: Part I - Bran functionality and dough properties.

This study aimed to investigate the effect of wheat bran micronization on its functionality including physicochemical and antioxidant properties, and dough properties. Coarse bran (D50 = 362.3 ± 20.5 µm) was superfine ground to medium (D50 = 60.4 ± 10.1 µm) and superfine (D50 = 11.3 ± 2.6 µm) bran, accompanied with increasing specific surface area and breakdown of aleurone layers. Bran micronization increased its soluble dietary fibre content, ferulic acid liberation, and antioxidant properties including total polyphenol content, ABTS•+ and DPPH• scavenging activities, while decreased its water retention capacity and insoluble dietary fibre content. Moreover, bran micronization impacted dough rheological properties. The dough with superfine bran had higher water absorption and gelatinization temperature, peak viscosity, final viscosity and setback value, lower stability time, resistance to extension, and extensibility than the dough with coarse bran. This dough furthermore exhibited more solid-like properties characterized by decreased loss moduli and frequency dependence (n').

Whole-wheat flour particle size influences dough properties, bread structure and in vitro starch digestibility.

Whole-wheat flour (WWF) is increasingly popular because of the health benefits of whole grains. This study investigated the effect of WWF particle size on dough properties, bread quality and in vitro starch digestibility. WWF was made from intact whole grain directly. Three WWF particle sizes were examined, including coarse, medium and fine with a mean size of 1315, 450 and 199 µm, respectively. The dough made from WWF of a larger particle size exhibited lower extensibility and stability, and subsequently the bread had a more compact structure (i.e. lower open porosity and thicker cell thickness), smaller specific volume and harder texture, which were regarded as poor quality attributes. On the other hand, the bread made from the fine WWF exhibited a higher amount of released glucose than those made from the coarse and medium WWFs. Moreover, the particle size of bread bolus showed no significant effect on in vitro starch digestion. The whole study demonstrated that the particle size of WWF plays a critical role in determining both bread quality and digestibility.

In vitro digestion of bread: How is it influenced by the bolus characteristics?

This study aimed to understand the impact of in vitro oral processing methods on bolus formation and the kinetics of starch hydrolysis of refined white bread during in vitro gastrointestinal digestion. Four in vitro oral processing methods (i.e., cut, cut-and-pestle, blend, and grind) were performed at two levels of disintegration (less and more intensive) and compared with human mastication. Boluses prepared using the in vitro methods had a larger particle size (20-69 mm2 vs. 14 mm2 ), a higher moisture content (64-68% vs. 47%), a softer texture (1.3-2.3 N vs. 6.3 N) and a less adhesive surface (0.3-1.0 vs. 1.6 N•s) as compared to the in vivo masticated ones. Moreover, in vitro prepared blouses were digested more rapidly than in vivo masticated ones during the stimulated intestinal digestion from 150 min onward, with a higher hydrolysis rate (0.011-0.012 mg/mL • min vs. 0.010 mg/mL • min) and a higher equilibrium concentration of reducing sugar (5.5-6.3 mg/mL vs. 4.9 mg/mL). Among all the in vitro methods, the blending and grinding methods produced boluses that most closely resemble the in vivo masticated ones in terms of their physical characteristics. The blending method also produced boluses having the highest amount of reducing sugar released (6.32 mg/mL). The amount of reducing sugar present in the PBS buffer outside the dialysis tube might be controlled by the diffusion efficiency at the beginning of the digestion (≤120 min) and then be largely influenced by the particle size of the bolus in the latter stage of the digestion. PRACTICAL APPLICATION: Studying the in vitro starch amyloysis is valuable for predicting the postprandial glycemic potential of starchy food. This work provides novel insights on the role of in vitro oral processing in the prediction of the glycemic potential of carbohydrate-rich staple food. Blending method is recommended because of its ability to produce boluses with similar physical characteristics as the in vivo masticated boluses. But the excessive structural breakdown occurred during blending also resulted in a higher enzymatic accessibility and a higher rate of starch digestion. Further study is needed to propose a new in vitro method that stimulates multiple actions occurred during mastication (cutting, grinding, and shearing), in order to match both physical properties and digestion profiles. Moreover, the amount of artificial saliva added should be adjusted according to the specific type of food.