Fibrosis in burns: an overview of mechanisms and therapies.

Scar development remains a common occurrence and a major healthcare challenge affecting the lives of millions of patients annually. Severe injuries to the skin, such as burns can lead to pathological wound healing patterns, often characterized by dermal fibrosis or excessive scarring, and chronic inflammation. The two most common forms of fibrotic diseases following burn trauma are hypertrophic scars (HSCs) and keloids, which severely impact the patient's quality of life. Although the cellular and molecular mechanisms are similar, HSC and keloids have several distinct differences. In this review, we discuss the different forms of fibrosis that occur postburn injury, emphasizing how the extent of burn influences scar development. Moreover, we highlight how a systemic response induced by a burn injury drives wound fibrosis, including both the role of the inflammatory response, as well as the fate of fibroblast during skin healing. Finally, we list potential therapeutics aimed at alleviating pathological scar formation. An understanding of the mechanisms of postburn fibrosis will allow us to effectively move studies from bench to bedside.

New Insights into the Effects of Microbial Muramidase Addition in the Diets of Broiler Chickens.

The study aimed to explore how broiler chickens' blood biochemistry, breast muscles' fatty acid profile, growth, intestinal morphology, and immune status would be influenced by adding microbial muramidase (MUR) to the diet. Four hundred 3-day-old male broiler chickens were allocated to a completely randomized design consisting of four nutritional treatments (n = 100 per treatment, 10 chicks/replicate), each containing MUR at levels of 0 (control group), 200, 400, and 600 mg Kg-1 diet, with enzyme activity 0, 12,000, 24,000, and 36,000 LSU(F)/kg diet, respectively. The 35-day experiment was completed. The findings showed that adding MUR to broiler meals in amounts of 200, 400, or 600 mg/kg had no impact on growth performance (p > 0.05) during the periods of 4-10, 11-23, and 24-35 days of age. MUR supplementation quadratically impacted the feed conversion ratio of broiler chicks at 11 and 23 days of age (p = 0.02). MUR addition to the diet significantly and level-dependently enhanced the percentage of n-3 and n-6 polyunsaturated fatty acids (PUFA) in breast muscles (p ≤ 0.01), with no alterations to the sensory characteristics of the breast muscles. Dietary MUR increased most of the morphometric dimensions of the small intestine, with the best results recorded at the 200 and 400 mg Kg-1 levels. MUR supplementation at 200, 400, and 600 mg kg-1 linearly lowered the total cholesterol, triglycerides, and low-density lipoprotein cholesterol level (p < 0.01). Still, it significantly increased the high-density lipoprotein cholesterol and very-low-density lipoprotein cholesterol contents compared with the unsupplemented group. Compared to controls, there was a substantial rise in the blood concentration of total protein, albumin, globulin, IL10, complement 3, and lysozyme activity as MUR levels increased (p < 0.01). Moreover, MUR addition significantly increased the immunoexpression of lymphocyte subpopulation biomarkers. We could conclude that MUR can be added to broiler chicken diets up to 600 mg kg -1 to improve broiler chickens' fatty acid profile in breast muscles, immunity, and blood biochemistry. MUR addition had no positive influence on the bird's growth.

Effect of 1,3-Beta Glucans Dietary Addition on the Growth, Intestinal Histology, Blood Biochemical Parameters, Immune Response, and Immune Expression of CD3 and CD20 in Broiler Chickens.

This experiment evaluated the impact of the dietary addition of 1,3-ß-glucans (GLU) on broiler chickens' growth, intestinal histology, blood biochemical parameters, and immunity. Two hundred three-day-old male broilers (Ross 308) (97.93 ± 0.19 g/chick) were randomly assigned into four treatments with five replicates, each containing ten birds, in a complete randomized design. The four treatments were formulated with 0, 50, 100, and 150 mg 1,3-ß-glucans kg−1 in broiler chicken diets. During the study, no significant impacts (p > 0.05) were observed in weight gain and feed conversion ratio (FCR) between treatment groups. Based on the results of total body weight gain and FCR, the optimal level of 1,3-ß-glucan is 120 mg Kg−1. The intestinal histomorphology was improved by GLU supplementation, as indicated by increased villi height and villi height to crypt depth ratio (p < 0.01). All levels of supplemental ß-1,3 glucan decreased the serum total cholesterol (TC), triglyceride levels, and low-density lipoprotein cholesterol (LDL-C) (p < 0.05). The serum levels of growth hormones (GH), triiodothyronine (T3), and thyroxine (T4) were increased in GLU-supplemented groups (p < 0.05). The serum immune indices (lysozyme activity, interleukin 10 (IL10), complement 3 (C3), and total protein levels) were increased in the GLU-supplemented groups (p < 0.05). Dietary GLU up-regulated the immunoexpression of CD3 (T-cell marker) and CD20 (B-cell marker) in the spleen of birds (p < 0.01). It can be concluded that 1,3-ß-glucan can be added to broiler chicken diets for improving the development and integrity of the intestine and enhancing the bird's immune status. The optimal level for 1,3-ß-glucan dietary supplementation was 120 mg Kg−1. Dietary 1,3-ß-glucan has a hypolipidemic effect and improves the hormonal profile of birds without affecting their growth rate.

Evaluation of Spray-Dried Bovine Hemoglobin Powder as a Dietary Animal Protein Source in Nile Tilapia, Oreochromis niloticus.

The present study evaluated the potential effects of dietary inclusion of spray-dried bovine hemoglobin powder (SDBH) on the growth, gene expression of peptide and amino acid transporters, insulin growth factor-1 (IGF-1) and myostatin, digestive enzymes activity, intestinal histomorphology and immune status, immune-related gene expression, and economic efficiency in Nile tilapia, Oreochromis niloticus. Two hundred twenty-five fingerlings (32.38 ± 0.05 g/fish) were distributed into five treatments with five dietary inclusion levels of SDBH: 0, 2.5, 5, 7.5, and 10% for a ten-week feeding period. Dietary inclusion of SDBH linearly increased the final body weight (FBW), total weight gain (TWG), specific growth rate (SGR), and protein efficiency ratio (PER). Additionally, a linear decrease in feed conversion ratio (FCR) and daily feed intake relative to the daily BW was reported in the highest inclusion levels (7.5 and 10%). Dietary inclusion of SDBH was associated with a significant increase in the intestinal villous height (VH), villous width (VW), villous height: crypt depth ratio (VH: CD), and muscle coat thickness (MCT), with the highest values reported in SDBH7.5 group. Increased serum growth hormone levels and decreased serum leptin hormone levels were also reported by increasing the SDBH level. The serum glucose level was decreased in the SDBH7.5 and SDBH10 groups. The digestive enzymes' activity (amylase and protease) was increased by increasing the SDBH inclusion level. An up-regulation in the expression of peptide and amino acid transporters, IGF-1, and down-regulation of myostatin was reported in the SDBH2.5 to SDBH7.5 groups. Spleen sections showed more lymphoid elements, especially in the SDBH2.5 and SDBH7.5 groups. The SDBH inclusion increased the serum lysozyme activity, nitric oxide (NO), and complement 3 (C3) levels, with the highest values recorded in the SDBH5 group. The phagocytic % and the phagocytic index were increased by increasing the SDBH inclusion %. The expressions of immune-related genes (transforming growth factor-beta (TGF-ß), Toll-like receptor 2 (TLR2), and interleukin 10 (IL10)) were up-regulated by SDBH inclusion with the highest expression in the SDBH5 group. Economically, the feed costs and feed costs/kg gain were linearly decreased in the SDBH7.5 and SDBH10 diets. In conclusion, spray-dried bovine hemoglobin powder could be used as a protein source for up to 10% of the diets of Nile tilapia for better growth and immune status of fish.

New Perylene Diimide Ink for Interlayer Formation in Air-Processed Conventional Organic Photovoltaic Devices.

Roll-to-roll coating of conventional organic photovoltaic architectures in air necessitates low work function, electron-harvesting interlayers as the top interface, termed cathode interlayers. Traditional materials based on metal oxides are often not compatible with coating in air and/or green solvents, require thermal annealing, and are limited in feasibility due to interactions with underlying layers. Alternatively, perylene diimide materials offer easily tunable redox properties, are amenable to air coating in green solvents, and are considered champion organic-based cathode interlayers. However, underlying mechanisms of the extraction of photogenerated electrons are less well understood. Herein, we demonstrate the utilization of two N-annulated perylene diimide materials, namely, PDIN-H and CN-PDIN-H, in air-processed conventional organic photovoltaic devices, using the now standard PM6:Y6 photoactive layer. The processing ink formulation using cesium carbonate as a processing agent to solubilize the perylene diimides in suitable green solvents (1-propanol and ethyl acetate) for uniform film formation using spin or slot-die coating on top of the photoactive layer is critical. Cesium carbonate remains in the film, creating hybrid organic/metal salt cathode interlayers. Best organic photovoltaic devices have power conversion efficiencies of 13.2% with a spin-coated interlayer and 13.1% with a slot-die-coated interlayer, superior to control devices using the classic conjugated polyelectrolyte PFN-Br as an interlayer (ca. 12.8%). The cathode interlayers were found to be semi-insulating in nature, and the device performance improvements were attributed to beneficial interfacial effects and electron tunneling through sufficiently thin layers. The efficiencies beyond 13% achieved in air-processed organic photovoltaic devices utilizing slot-die-coated cathode interlayers are among the highest reported so far, opening new opportunities for the fabrication of large-area solar cell modules.

Long-Term Feeding with Curcumin Affects the Growth, Antioxidant Capacity, Immune Status, Tissue Histoarchitecture, Immune Expression of Proinflammatory Cytokines, and Apoptosis Indicators in Nile Tilapia, Oreochromis niloticus.

The impact of dietary curcumin (CUR) on the growth, antioxidant activity, histomorphology of certain organs, proinflammatory cytokine production, and immune status of Oreochromis niloticus was evaluated. The fingerlings (n = 225, 41.60 ± 0.09 g/fish) were randomly allotted into five experimental groups in triplicate. Fish were fed basal diets complemented with 0, 200, 400, 600, or 800 mg curcumin/kg diet (CUR0, CUR200, CUR400, CUR600, and CUR800, respectively) for 10 weeks. An increase in fish growth was reported in the CUR200 and CUR400 groups. The feed conversion ratio was enhanced by 15% in the CUR400 group. Fish body protein content was increased in the CUR600 group (p ≤ 0.01). Body fat was decreased, and ash content was increased by CUR supplementation in a level-related way (p < 0.05). The villus height was increased in the CUR400 and CUR600 groups. The villus width was increased by CUR supplementation, with the best result found in the CUR600 group. The liver of CUR-fed fish displayed comparatively normal hepatocytes. TNF-α and caspase-3 were significantly upregulated by dietary CUR in a level-related way. The serum catalase activity and GSH level were increased in CUR200 and CUR400 groups. Curcumin supplementation boosted the serum SOD activity and reduced the MDA level. IL10 and IgM levels were increased in the CUR200 and CUR400 groups. Lysozyme activity was increased in the CUR200−400 groups. Serum complement 3 level was increased in the CUR400 group. The percentage survival of O. niloticus challenged with Aeromonas hydrophila was highest in the CUR200-CUR600 groups (100%) and decreased in the CUR800 group (80%). This study concluded that CUR could be added to Nile tilapia diets up to 400 mg·kg−1 to achieve better growth, antioxidant capacity, immune response, and intestinal histology. Long feeding periods on high levels of CUR (600 and 800 mg·kg−1) stimulate inflammatory reactions in fish tissues.

Development of Tetrameric N-Annulated Perylene Diimides Using "Click" Chemistry.

Herein, we report the design, synthesis, and characterization of two novel N-annulated perylene diimide (NPDI) tetramer arrays that were developed using copper catalyzed alkyne-azide cycloaddition. Despite the optoelectronic properties of both tetramers being nearly identical, the two tetramers exhibited very different molecular geometries. The twisted spirobifluorene NPDI tetramer (sbfNPDI4 ) was found to have an extended and flexible geometry, while the planar pyrene NPDI tetramer (pyrNPDI4 ) exhibited a highly congested and conformationally locked geometry. Organic photovoltaic devices were constructed to demonstrate the use of both new compounds as electron acceptor materials, where slightly higher power conversion efficiencies were achieved with pyrNPDI4 than sbfNPDI4 . This study highlights the viability of using "click" chemistry as a facile synthetic strategy towards the development of new multicomponent perylene diimide materials for organic electronic applications.

Mechanotransductive Mechanisms of Biomimetic Hydrogel Cues Modulating Meckel's Cartilage Degeneration.

Meckel's cartilage, a cartilage rod present in the mandible during developmental stages, shows a unique developmental fate: while the anterior and posterior portions undergo ossification, the middle part degenerates. Previously, it was shown that a stiff environment promoted cartilage degeneration in the middle region, while a soft environment enhanced the mineralization in the anterior region of Meckel's cartilage. This study aims to elucidate the spatio-temporal changes in the mechanosensing properties of Meckel's cartilage during its early developmental stages and clarify the mechanotransduction-related mechanisms involved in its degeneration. The results show that the expression of Hippo pathway effector yes-associated protein (YAP) is only detectable in the Meckel's cartilage onward embryonic day (E)14.5, indicating that mechanosensing is dependent on the tissue developmental stage. Consistently, microenvironmental stiffness-induced cartilage degeneration can only be induced in cartilages onward E14.5, but not in those at earlier developmental stages. Expressions of integrin-ß1 and cartilage matrix-degrading enzymes, matrix metalloproteinase 1 (MMP-1) and MMP-13, are significantly enhanced in the degeneration area. Moreover, verteporfin (YAP inhibitor) and integrin-ß1 antibody block the substrate stiffness-induced degeneration by suppressing the expressions of MMP-1 and MMP-13. These data provide new insights into the interplay between biochemical and mechanical cues determining the fate of Meckel's cartilage.

Tin Oxide Electron Transport Layers for Air-/Solution-Processed Conventional Organic Solar Cells.

Commercialization of organic solar cells (OSC) is imminent. Interlayers between the photoactive film and the electrodes are critical for high device efficiency and stability. Here, the applicability of SnO2 nanoparticles (SnO2 NPs) as the electron transport layer (ETL) in conventional OSCs is evaluated. A commercial SnO2 NPs solution in butanol is mixed with ethanol (EtOH) as a processing co-solvent to improve film formation for spin and slot-die coating deposition procedures. When processed with 200% v/v EtOH, the SnO2 NPs film presents uniform film quality and low photoactive layer degradation. The optimized SnO2 NPs ink is coated, in air, on top of two polymer:fullerene-based systems and a nonfullerene system, to form an efficient ETL film. In every case, addition of SnO2 NPs film significantly enhances photovoltaic performance, from 3.4 and 3.7% without the ETL to 6.0 and 5.7% when coated on top of PBDB-T:PC61BM and PPDT2FBT:PC61BM, respectively, and from 3.7 to 7.1% when applied on top of the PTQ10:IDIC system. Flexible, all slot-die-coated devices, in air, are also fabricated and tested, demonstrating the versatility of the SnO2 NPs ink for efficient ETL formation on top of organic photoactive layers, processed under ambient condition, ideal for practical large-scale production of OSCs.

Chronic exposure to FGF2 converts iPSCs into cancer stem cells with an enhanced integrin/focal adhesion/PI3K/AKT axis.

We previously demonstrated the conversion of normal stem cells, including induced pluripotent stem cells (iPSCs), into cancer stem cells (CSCs) without genetic manipulation. Herein, we designed a meta-analysis to assess gene expression profiles in different breast cancer cell lines focusing on the secretory factors responsible for conversion. As a result, fibroblast growth factor 2 (FGF2) was found to be the best candidate in T47D and BT549 cells, of which conditioned medium was previously successful in inducing CSCs. When treated with 3.1 µg/ml FGF2, mouse iPSCs not only maintained survival without LIF for three weeks but also acquired growth ability independent of FGF2. The resultant cells exhibited expression of stemness and cancer stem cell markers, sphere-forming ability, differentiation, and tumorigenicity with malignancy. The primary cultures of the tumor confirmed the signatures of CSCs with two different phenotypes with or without GFP expression under control of the Nanog promoter. Bioinformatic analysis of gene expression profiles suggested constitutive autocrine activation of the FGF receptor, integrins, focal adhesions, and PI3K/AKT pathways. FGF2 could potently initiate cancer as a component of the inflammatory microenvironment.

Hybrid Tetrameric Perylene Diimide Assemblies.

Organic photovoltaics have found utility as indoor light recycling devices providing an opportunity for the sustainable powering of IoT sensors and related smart electronics. In the report, two organic π-conjugated molecules consisting of four perylene diimide (PDI) chromophores each are presented and used as non-fullerene acceptors in indoor photovoltaic devices. The new materials consist of a dimeric N-annulated PDI core with single PDIs grafted onto the pyrrolic N-atom positions of the core. Compounds PDI4 e and PDI4 i are PDI tetramers and differ with PDI4 e having the terminal N-annulated PDI with pyrrolic N-atom distal to the core and PDI4 i having the terminal N-annulated PDI with pyrrolic N-atom proximal to the core. The structural and optoelectronic properties were investigated using NMR spectroscopy, optical absorption and emission spectroscopy, and cyclic voltammetry. The compounds exhibit typical optical signatures for PDIs but notable is that the addition of grafted PDI molecules prevents significant aggregation of the dimeric PDI core, as compared to a reference dimer. Use as non-fullerene acceptors in ternary bulk-heterojunction blends with the polymer FBT and fullerene PC61 BM lead to increased open-circuit voltages and power conversion efficiencies upwards of 13.7 % at 2000 lux light intensity.

Slot-Die-Coated Ternary Organic Photovoltaics for Indoor Light Recycling.

Efficient organic photovoltaics (OPVs) based on slot-die-coated (SD) ternary blends were developed for low-intensity indoor light harvesting. For active layers processed in air and from eco-friendly solvents, our device performances (under 1 sun and low light intensity) are the highest reported values for fluoro-dithiophenyl-benzothiadiazole donor polymer-based OPVs. The N-annulated perylene diimide dimer acceptor was incorporated into a blend of donor polymer (FBT) and fullerene acceptor (PC61BM) to give ternary bulk heterojunction blends. SD ternary-based devices under 1 sun illumination showed enhanced power conversion efficiency (PCE) from 6.8 to 7.7%. We observed enhancement in the short-circuit current density and open-circuit voltage of the devices. Under low light intensity light-emitting device illumination (ca. 2000 lux), the ternary-based devices achieved a PCE of 14.0% and a maximum power density of 79 µW/cm2 compared to a PCE of 12.0% and a maximum power density of 68 µW/cm2 for binary-based devices. Under the same illumination conditions, the spin-coated (SC) devices showed a PCE of 15.5% and a maximum power density of 88 µW/cm2. Collectively, these results demonstrate the exceptional promise of a SD ternary blend system for indoor light harvesting and the need to optimize active layers based on industry-relevant coating approaches toward mini modules.

Fibronectin-induced ductal formation in salivary gland self-organization model.

BACKGROUND: Recent advances in tissue regeneration approaches including 3D organoids, were based on various 3D organogenesis models. However, 3D models are generally technique-sensitive and time-consuming. Thus, we utilized an existing model of submandibular salivary gland (SMG) to modify a simple and highly reproducible in vitro 3D culture model of primary SMG cells self-organization into a well-developed cell spheroid inside Matrigel substrate. We used this model to observe the collective multicellular behavior during spheroid formation. Further, we applied various quantitative approaches including real-time live imaging and immune histochemical image analysis to dissect the cellular dynamics during tissue patterning. RESULTS: On a time-scale of hours, we observed marked size and shape transformations in the developed 3D spheroid which resulted in a spatially-controlled growth differential from the canter to the periphery of the formed aggregates. Moreover, we investigated the effect of fibronectin (FN) on SMG cells self-organization using our simplified culture model. Interestingly, we discovered a novel role of FN in inducing duct-like elongation during initial stages of SMG bud formation. CONCLUSION: This in vitro model provides an excellent tool for analyzing the intercellular dynamics during early SMG tissue development as well as revealing a novel role of FN in SMG ductal expansion.

Fabrication of single gel with different mechanical stiffness using three-dimensional mold.

The hydrogel is considering as functional substrates for three-dimensional (3D) environment mimicking the native tissue in vitro. To get the cell or tissue culture result in different stiffness, researchers used separate gel at different times. Sometimes these results are manipulated by surrounding environment. To overcome this, we prepared a single hydrogel with different young modulus using a 3 D printed mold and cell culture and tissue culture was performed to check the functional capacity. In this proposed device we successfully produced a multiproperties agarose hydrogel on a single platform. We designed different mold pattern to confirm that this gel formation technique can be used for any types of design and many different concentrated hydrogels can be combined together. The cell and tissue culture results showed that even in same planer surface, each gel solely maintains their own physical properties and control the cell and tissue adherence and proliferation behavior. The protocol is fairly simple and reproducible. The design helps to produce consistent gel thickness, shape, and size. This 3 D mold has provided a new way tostudy the fundamental cellular responses to engineered microenvironments that may have a high implementation in both biological and healthcare-related applications. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 107A: 6-11, 2019.

MSCs feeder layers induce SMG self-organization and branching morphogenesis.

Dysfunction of salivary glands leads to several oral health problems, including dental caries, mastication and swallowing dysfunctions and multiple oral infections. Conventional treatments for such condition fell short of providing satisfying therapeutic results. Recent advances in organ regeneration therapy which utilize tissue stem cells to fabricate bioengineered 3D organ buds, have introduced a promising therapeutic tool for full functional organ regeneration. However, finding a sustainable and easily accessible cell source for such approaches is still challenging, especially in case of severely atrophied tissues such as irradiated salivary glands. In response to this, we hypothesized that bone marrow derived mesenchymal stem cells (MSCs) could be used as feeder cells to induce salivary epithelial tissues/cells branching. Indeed, in 2D cultures, MSCs supported branching of embryonic submandibular salivary gland (SMG) epithelium. Interestingly, this enhancing effect was dependent on the initial number of MSC feeder cells. In addition, MSCs supported the self-assembly of SMG epithelial progenitor cells into well-patterned and branched 3D salivary organoids. Therefore, these findings propose MSCs as a valuable candidate cell source for induced SMG epithelial branching, which can potentially be applied in future methods for SMG regeneration approaches.

MCSF orchestrates branching morphogenesis in developing submandibular gland tissue.

The importance of macrophages in tissue development and regeneration has been strongly emphasized. However, the specific roles of macrophage colony-stimulating factor (MCSF), the key regulator of macrophage differentiation, in glandular tissue development have been unexplored. Here, we disclose new macrophage-independent roles of MCSF in tissue development. We initially found that MCSF is markedly upregulated at embryonic day (E)13.5, at a stage preceding the colonization of macrophages (at E15.5), in mouse submandibular gland (SMG) tissue. Surprisingly, MCSF-induced branching morphogenesis was based on a direct effect on epithelial cells, as well as indirectly, by modulating the expression of major growth factors of SMG growth, FGF7 and FGF10, via the phosphoinositide 3-kinase (PI3K) pathway. Additionally, given the importance of neurons in SMG organogenesis, we found that MCSF-induced SMG growth was associated with regulation of neurturin expression and neuronal network development during early SMG development in an in vitro organogenesis model as well as in vivo These results indicate that MCSF plays pleiotropic roles and is an important regulator of early SMG morphogenesis.

Synergistic Effects of Morphological Control and Complementary Absorption in Efficient All-Small-Molecule Ternary-Blend Solar Cells.

In this study, we combined two small-molecule donors-a diketopyrrolopyrrole-based small molecule (SMD) and a benzodithiophene-based molecule (BDT6T)-with [6,6]-phenyl-C61-butyric acid methyl ester (PC61BM) to form ternary blend solar cells. The power conversion efficiency of the binary SMD:PC61BM bulk heterojunction solar cell improved from 4.57 to 6.28% after adding an appropriate amount BDT6T as a guest. We attribute this 37% improvement in device performance to the complementary absorption behavior of BDT6T and SMD, as evidenced by the increase in the short circuit current. After addition of BDT6T to form the ternary blend, the crystallinity and morphology of the active layer were enhanced. For example, the features observed in the ternary active layers were finer than those in the binary blends. This means that BDT6T as a third component in the ternary blend has effective role on both the absorption and the morphology. In addition, adding BDT6T to form the ternary blend also led to an increase in the open-circuit voltage. Our findings suggest that the preparation of such simple all-small-molecule ternary blends can be an effective means of improving the efficiency of photovoltaic devices.

Peptide-modified Substrate for Modulating Gland Tissue Growth and Morphology In Vitro.

In vitro fabricated biological tissue would be a valuable tool to screen newly synthesized drugs or understand the tissue development process. Several studies have attempted to fabricate biological tissue in vitro. However, controlling the growth and morphology of the fabricated tissue remains a challenge. Therefore, new techniques are required to modulate tissue growth. RGD (arginine-glycine-aspartic acid), which is an integrin-binding domain of fibronectin, has been found to enhance cell adhesion and survival; it has been used to modify substrates for in vitro cell culture studies or used as tissue engineering scaffolds. In addition, this study shows novel functions of the RGD peptide, which enhances tissue growth and modulates tissue morphology in vitro. When an isolated submandibular gland (SMG) was cultured on an RGD-modified alginate hydrogel sheet, SMG growth including bud expansion and cleft formation was dramatically enhanced. Furthermore, we prepared small RGD-modified alginate beads and placed them on the growing SMG tissue. These RGD-modified beads successfully induced cleft formation at the bead position, guiding the desired SMG morphology. Thus, this RGD-modified material might be a promising tool to modulate tissue growth and morphology in vitro for biological tissue fabrication.