Sustained Release of Insulin-Like Growth Factor-1 from Bombyx mori L. Silk Fibroin Delivery for Diabetic Wound Therapy

The goals of this study are to develop a high purity patented silk fibroin (SF) film and test its suitability to be used as a slow-release delivery for insulin-like growth factor-1 (IGF-1). The release rate of the SF film delivering IGF-1 followed zero-order kinetics as determined via the Ritger and Peppas equation. The release rate constant was identified as 0.11, 0.23, and 0.09% h−1 at 37 °C for SF films loaded with 0.65, 6.5, and 65 pmol IGF-1, respectively. More importantly, the IGF-1 activity was preserved for more than 30 days when complexed with the SF film. We show that the IGF-1-loaded SF films significantly accelerated wound healing in vitro (BALB/3T3) and in vivo (diabetic mice), compared with wounds treated with free IGF-1 and an IGF-1-loaded hydrocolloid dressing. This was evidenced by a six-fold increase in the granulation tissue area in the IGF-1-loaded SF film treatment group compared to that of the PBS control group. Western blotting analysis also demonstrated that IGF-1 receptor (IGF1R) phosphorylation in diabetic wounds increased more significantly in the IGF-1-loaded SF films group than in other experimental groups. Our results suggest that IGF-1 sustained release from SF films promotes wound healing through continuously activating the IGF1R pathway, leading to the enhancement of both wound re-epithelialization and granulation tissue formation in diabetic mice. Collectively, these data indicate that SF films have considerable potential to be used as a wound dressing material for long-term IGF-1 delivery for diabetic wound therapy.

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An Insulin-like Growth Factor-1 Conjugated Bombyx mori Silk Fibroin Film for Diabetic Wound Healing: Fabrication, Physicochemical Property Characterization, and Dosage Optimization In Vitro and In Vivo

Pharmaceutics ◽

10.3390/pharmaceutics13091459 ◽

2021 ◽

Vol 13 (9) ◽

pp. 1459

Author(s):

Meng-Jin Lin ◽

Mei-Chun Lu ◽

Yun-Chen Chan ◽

Yu-Fen Huang ◽

Hwan-You Chang

Keyword(s):

Wound Healing ◽

Growth Factor ◽

Silk Fibroin ◽

Epithelial Tissue ◽

Dependent Manner ◽

Insulin Like Growth Factor ◽

Diabetic Wound ◽

Tissue Area ◽

Diabetic Wound Healing ◽

Low Dosage

This study aimed to develop a silk fibroin (SF)-film for the treatment of chronic diabetic wounds. Silk fibroin was purified through a newly developed heating degumming (HD) process and casted on a hydrophobic surface to form SF-films. The process allowed the fabricated film to achieve a 42% increase in transparency and a 32% higher proliferation rate for BALB/3T3 fibroblasts compared to that obtained by conventional alkaline degumming treatment. Fourier transform infrared analysis demonstrated that secondary structure was retained in both HD- and alkaline degumming-derived SF preparations, although the crystallinity of beta-sheet in SF-film after the HD processing was slightly increased. This study also investigated whether conjugating insulin-like growth factor-1 (IGF-1) would promote diabetic wound healing and what the optimal dosage is. Using BALB/3T3 cells grown in hyperglycemic medium as a model, it was demonstrated that the optimal IGF-1 dosage to promote the cell growth was approximately 0.65 pmol. Further analysis of wound healing in a diabetic mouse model indicated that SF-film loaded with 3.25 pmol of IGF-1 showed significantly superior wound closure, a 13% increase at the 13th day after treatment relative to treatment with 65 pmol of free IGF-1. Improvement in diabetic wound healing was exerted synergistically by SF-film and IGF-1, as reflected by parameters including levels of re-epithelialization, epithelial tissue area, and angiogenesis. Finally, IGF-1 increased the epithelial tissue area and micro-vessel formation in a dose-dependent manner in a low dosage range (3.25 pmol) when loaded to SF-films. Together, these results strongly suggest that SF-film produced using HD and loaded with a low dosage of IGF-1 is a promising dressing for diabetic wound therapy.

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Role of Nerve Growth Factor in Cutaneous Wound Healing: Accelerating Effects in Normal and Healing-impaired Diabetic Mice

Journal of Experimental Medicine ◽

10.1084/jem.187.3.297 ◽

1998 ◽

Vol 187 (3) ◽

pp. 297-306 ◽

Cited By ~ 148

Author(s):

Hiroshi Matsuda ◽

Hiromi Koyama ◽

Hiroaki Sato ◽

Junko Sawada ◽

Atsuko Itakura ◽

...

Keyword(s):

Wound Healing ◽

Nerve Growth Factor ◽

Growth Factor ◽

Granulation Tissue ◽

Messenger Rna ◽

Biological Activities ◽

Serum Levels ◽

Blood Stream ◽

Diabetic Mice ◽

Nerve Growth

Four full-thickness skin wounds made in normal mice led to the significant increase in levels of nerve growth factor (NGF) in sera and in wounded skin tissues. Since sialoadenectomy before the wounds inhibited the rise in serum levels of NGF, the NGF may be released from the salivary gland into the blood stream after the wounds. In contrast, the fact that messenger RNA and protein of NGF were detected in newly formed epithelial cells at the edge of the wound and fibroblasts consistent with the granulation tissue produced in the wound space, suggests that NGF was also produced at the wounded skin site. Topical application of NGF into the wounds accelerated the rate of wound healing in normal mice and in healing-impaired diabetic KK/Ta mice. This clinical effect of NGF was evaluated by histological examination; the increases in the degree of reepithelialization, the thickness of the granulation tissue, and the density of extracellular matrix were observed. NGF also increased the breaking strength of healing linear wounds in normal and diabetic mice. These findings suggested that NGF immediately and constitutively released in response to cutaneous injury may contribute to wound healing through broader biological activities, and NGF improved the diabetic impaired response of wound healing.

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Application of Sustained-release Carrier of NanoAg-Epidermal Growth Factor in Promoting Refractory Diabetic Wound Healing

Letters in Drug Design & Discovery ◽

10.2174/1570180812666150630183147 ◽

2015 ◽

Vol 13 (2) ◽

pp. 135-140

Author(s):

Ning-ning Tang ◽

Feng Hu ◽

Jin-yan Liu ◽

Can Liu ◽

Chao-qi Yin ◽

...

Keyword(s):

Wound Healing ◽

Growth Factor ◽

Epidermal Growth Factor ◽

Sustained Release ◽

Diabetic Wound ◽

Diabetic Wound Healing ◽

Epidermal Growth

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Adipose-Derived Stromal Cell-Sheets Sandwiched, Book-Shaped Acellular Dermal Matrix Capable of Sustained Release of Basic Fibroblast Growth Factor Promote Diabetic Wound Healing

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.646967 ◽

2021 ◽

Vol 9 ◽

Author(s):

Xin Shi ◽

Liyuan Jiang ◽

Xin Zhao ◽

Bei Chen ◽

Wei Shi ◽

...

Keyword(s):

Stem Cells ◽

Wound Healing ◽

Growth Factor ◽

Sustained Release ◽

Fibroblast Growth Factor ◽

Stromal Cell ◽

Fibroblast Growth ◽

Diabetic Wound ◽

Dermal Matrix ◽

Diabetic Wound Healing

The management of diabetic wounds is a therapeutic challenge in clinical settings. Current tissue engineering strategies for diabetic wound healing are insufficient, owing to the lack of an appropriate scaffold that can load a large number of stem cells and induce the interaction of stem cells to form granulation tissue. Herein we fabricated a book-shaped decellularized dermal matrix (BDDM), which shows a high resemblance to native dermal tissue in terms of its histology, microstructure, and ingredients, is non-cytotoxic and low-immunogenic, and allows adipose-derived stromal cell (ASC) attachment and proliferation. Then, a collagen-binding domain (CBD) capable of binding collagen was fused into basic fibroblast growth factor (bFGF) to synthetize a recombinant growth factor (termed as CBD–bFGF). After that, CBD–bFGF was tethered onto the collagen fibers of BDDM to improve its endothelial inducibility. Finally, a functional scaffold (CBD–bFGF/BDDM) was fabricated. In vitro and in vivo experiments demonstrated that CBD–bFGF/BDDM can release tethered bFGF with a sustained release profile, steadily inducing the interaction of stem cells down to endothelial differentiation. ASCs were cultured to form a cell sheet and then sandwiched by CBD–bFGF/BDDM, thus enlarging the number of stem cells loaded into the scaffold. Using a rat model, the ASC sheets sandwiched with CBD–bFGF/BDDM (ASCs/CBD–bFGF/BDDM) were capable of enhancing the formation of granulation tissue, promoting angiogenesis, and facilitating collagen deposition and remodeling. Therefore, the findings of this study demonstrate that ASCs/CBD–bFGF/BDDM could be applicable for diabetic wound healing.

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Promotion of wound healing by insulin-like growth factor-1-loaded Bombyx mori L. silk fibroin films in diabetic mice

10.21203/rs.3.rs-400183/v1 ◽

2021 ◽

Author(s):

Meng-Jin Lin ◽

Mei-Chun Lu ◽

Yun-Chen Chan ◽

Hwan-You Chang

Keyword(s):

Wound Healing ◽

Growth Factor ◽

Bombyx Mori ◽

Silk Fibroin ◽

Hydrophobic Surface ◽

Diabetic Mouse ◽

Alkaline Treatment ◽

Epithelial Tissue ◽

Insulin Like Growth Factor ◽

3T3 Fibroblasts

Abstract This study aimed to engineer an advanced wound-dressing combining Bombyx mori L. silk fibroin (SF) with insulin-like growth factor-1 (IGF-1). Silk fibroin was purified through a newly-developed high-temperature degumming process and cast on a hydrophobic surface to form SF-films. The process allowed the fabrication of a film exhibiting a 42% increase in transparency and a 32% higher proliferation rate of BALB/3T3 fibroblasts compared to that obtained by conventional alkaline treatment. This study demonstrated that the optimal concentration of IGF-1 to promote BALB/3T3 cell growth in hyperglycemic medium was approximately 130 nM. Further analysis of wound healing in a diabetic mouse model indicated that SF-film loaded with 3.25 pmol IGF-1 showed significantly superior wound closure (13% increase) at 13 days after treatment compared to treatment with 65 pmol free IGF-1. We clearly observed improvement in diabetic wound healing exerted synergistically by SF-film and IGF-1, as reflected by parameters including degree of re-epithelialization, epithelial tissue area, and angiogenesis. These results strongly suggest the great potential of IGF-1-loaded SF-film as a dressing for the treatment of diabetic wounds.

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Development of stabilized dual growth factor-loaded hyaluronate collagen dressing matrix

Journal of Tissue Engineering ◽

10.1177/2041731421999750 ◽

2021 ◽

Vol 12 ◽

pp. 204173142199975

Author(s):

Jihyun Kim ◽

Kyoung-Mi Lee ◽

Seung Hwan Han ◽

Eun Ae Ko ◽

Dong Suk Yoon ◽

...

Keyword(s):

Wound Healing ◽

Growth Factor ◽

Type I ◽

Diabetic Mice ◽

Patients With Diabetes ◽

Diabetic Wound Healing ◽

Epidermal Growth ◽

Wound Healing Effect ◽

Growth Factor Production

Patients with diabetes experience impaired growth factor production such as epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF), and they are reportedly involved in wound healing processes. Here, we report dual growth factor-loaded hyaluronate collagen dressing (Dual-HCD) matrix, using different ratios of the concentration of stabilized growth factors—stabilized-EGF (S-EGF) and stabilized-bFGF (S-bFGF). At first, the optimal concentration ratio of S-EGF to S-bFGF in the Dual-HCD matrix is determined to be 1:2 in type I diabetic mice. This Dual-HCD matrix does not cause cytotoxicity and can be used in vivo. The wound-healing effect of this matrix is confirmed in type II diabetic mice. Dual HCD enhances angiogenesis which promotes wound healing and thus, it shows a significantly greater synergistic effect than the HCD matrix loaded with a single growth factor. Overall, we conclude that the Dual-HCD matrix represents an effective therapeutic agent for impaired diabetic wound healing.

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Polyvinyl Alcohol/Chitosan Composite Hydrogels with Sustained Release of Traditional Tibetan Medicine for Promoting Chronic Diabetic Wound Healing

Biomaterials Science ◽

10.1039/d1bm00346a ◽

2021 ◽

Author(s):

Xingyu Chen ◽

Zuxin Wang ◽

Shan Gao ◽

Wanlin Zhang ◽

Hanwen Gong ◽

...

Keyword(s):

Wound Healing ◽

Polyvinyl Alcohol ◽

Sustained Release ◽

Skin Diseases ◽

Tibetan Medicine ◽

Diabetic Wound ◽

Composite Hydrogels ◽

Diabetic Wound Healing ◽

Traditional Tibetan Medicine ◽

Chitosan Composite

The Tibetan eighteen flavor dangshen pills (TEP) are composed of 18 traditional Tibetan medicines, which are commonly used in the treatment of skin diseases in the Tibetan medicine system. They...

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