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1.
Einstein (Sao Paulo) ; 18: eAO4876, 2020.
Artigo em Inglês, Português | MEDLINE | ID: mdl-31576909

RESUMO

OBJECTIVE: To investigate the effects of sericin extracted from silkworm Bombyx mori cocoon on morphophysiological parameters in mice with obesity induced by high-fat diet. METHODS: Male C57Bl6 mice aged 9 weeks were allocated to one of two groups - Control and Obese, and fed a standard or high-fat diet for 10 weeks, respectively. Mice were then further subdivided into four groups with seven mice each, as follows: Control, Control-Sericin, Obese, and Obese-Sericin. The standard or high fat diet was given for 4 more weeks; sericin (1,000mg/kg body weight) was given orally to mice in the Control-Sericin and Obese-Sericin Groups during this period. Weight gain, food intake, fecal weight, fecal lipid content, gut motility and glucose tolerance were monitored. At the end of experimental period, plasma was collected for biochemical analysis. Samples of white adipose tissue, liver and jejunum were collected and processed for light microscopy analysis; liver fragments were used for lipid content determination. RESULTS: Obese mice experienced significantly greater weight gain and fat accumulation and had higher total cholesterol and glucose levels compared to controls. Retroperitoneal and periepididymal adipocyte hypertrophy, development of hepatic steatosis, increased cholesterol and triglyceride levels and morphometric changes in the jejunal wall were observed. CONCLUSION: Physiological changes induced by obesity were not fully reverted by sericin; however, sericin treatment restored jejunal morphometry and increased lipid excretion in feces in obese mice, suggesting potential anti-obesity effects.


Assuntos
Fármacos Antiobesidade/uso terapêutico , Dieta Hiperlipídica , Obesidade/tratamento farmacológico , Sericinas/uso terapêutico , Tecido Adiposo/patologia , Animais , Fármacos Antiobesidade/farmacologia , Peso Corporal/efeitos dos fármacos , Colesterol/análise , Dieta Hiperlipídica/efeitos adversos , Ingestão de Alimentos/efeitos dos fármacos , Fígado Gorduroso/patologia , Trânsito Gastrointestinal/efeitos dos fármacos , Teste de Tolerância a Glucose , Fígado/metabolismo , Masculino , Camundongos Endogâmicos C57BL , Camundongos Obesos , Obesidade/etiologia , Obesidade/fisiopatologia , Reprodutibilidade dos Testes , Sericinas/farmacologia , Fatores de Tempo , Resultado do Tratamento , Triglicerídeos/análise , Ganho de Peso/efeitos dos fármacos
2.
Photochem Photobiol Sci ; 18(10): 2497-2508, 2019 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-31432056

RESUMO

Ultraviolet radiation (UVR)-induced redox imbalance in melanocytes triggers the activation of tyrosinase that results in melanogenesis and its related skin disorders. Supplementation of biological reductants or anti-tyrosinase compounds inhibits such melanogenesis. Silk sericin (SS), a globular protein, is known to possess antioxidant and anti-tyrosinase activities along with other biological attributes. However, its inhibitory activity against UVR-induced melanogenesis has yet to be explored. In the current study, we have scientifically explored the inhibitory activity of SS against UVR-induced melanogenesis. Anti-tyrosinase activity of SS was assessed using mushroom tyrosinase, showing that Antheraea assamensis sericin (AAS) and Philosamia ricini sericin (PRS) inhibited 50% of its activity. Inhibitory activity of SS against UVR-induced melanogenesis was assessed by measuring the cellular melanin content, intracellular tyrosinase activity, and reactive oxygen species (ROS) levels in mouse melanoma. SS pretreatment significantly reduced cellular melanin and ROS production in UV irradiated melanocytes compared with SS untreated cells. AAS treatment before UVA or UVB irradiation significantly inhibited tyrosinase activity. Rheological studies showed that the skin care formulation prepared by the addition of AAS to the basic formulation minimally affected its flow properties. Altogether, our results validate that AAS efficiently inhibited UVR-induced melanogenesis and it could be used as a potential antioxidant molecule in skin care cosmeceutics.


Assuntos
Cosmecêuticos/química , Melaninas/metabolismo , Sericinas/química , Raios Ultravioleta , Animais , Antioxidantes/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos da radiação , Cosmecêuticos/metabolismo , Cosmecêuticos/farmacologia , Melanócitos/citologia , Melanócitos/metabolismo , Melanócitos/efeitos da radiação , Camundongos , Monofenol Mono-Oxigenase/antagonistas & inibidores , Monofenol Mono-Oxigenase/metabolismo , Morus/química , Morus/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Reologia/efeitos dos fármacos , Sericinas/metabolismo , Sericinas/farmacologia
3.
BMC Complement Altern Med ; 19(1): 168, 2019 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-31291937

RESUMO

BACKGROUND: Several immunological pathways, particularly skin inflammation via various pro-inflammatory cytokines have been reported to be involved in the pathogenesis and clinical manifestations of psoriasis. The aim of the study was to investigate the potential role of naringin from Citrus maxima (Burm.) Merr and sericin from Bombyx mori combination in the treatment of psoriasis. Inhibitory effects on the expression of mRNA and the production of pro-inflammatory cytokines (TNF-α, IL-6, IL-23, and IL-12p40) were investigated. METHODS: Human peripheral blood mononuclear cells (hPBMCs) were isolated from 10 healthy subjects and 10 patients with psoriasis. The hPBMCs from each group were exposed to naringin or sericin alone, and the combination of naringin and sericin. The expression levels of mRNA and the production of all cytokines were determined using quantitative RT-PCR and ELISA, respectively. RESULTS: Naringin/sericin combination significantly decreased the expression of mRNA and the production of all pro-inflammatory cytokines in hPBMCs from patients with psoriasis. The potency of inhibitory activity was markedly higher than naringin or sericin alone. CONCLUSION: The activity of naringin/sericin combination on down-regulation of these pro-inflammatory cytokines suggested its potential clinical use in psoriasis as well as other inflammation-associated diseases. The combination might be used as a complementary therapy with conventional treatment in psoriasis to improve clinical efficacy and tolerability.


Assuntos
Citrus/química , Flavanonas/farmacologia , Leucócitos Mononucleares/efeitos dos fármacos , Extratos Vegetais/farmacologia , Psoríase/tratamento farmacológico , Sericinas/farmacologia , Adulto , Anti-Inflamatórios/farmacologia , Citocinas/biossíntese , Citocinas/genética , Quimioterapia Combinada , Feminino , Flavanonas/uso terapêutico , Humanos , Técnicas In Vitro , Mediadores da Inflamação/metabolismo , Masculino , Pessoa de Meia-Idade , Extratos Vegetais/uso terapêutico , Sericinas/uso terapêutico , Adulto Jovem
4.
Mater Sci Eng C Mater Biol Appl ; 102: 502-510, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31147021

RESUMO

As a first step towards the production of functional cell sheets applicable for the regeneration of gut muscle layer, microstructured bacterial cellulose (mBC) was assessed for its ability to support the growth of enteric nervous system (ENS) and gut smooth muscle cells (SMCs). To improve the cellular response, mBC was modified with silk sericin (SS) which has renowned abilities in supporting tissue regeneration. While SS did not impair the line structures imparted to BC by PDMS templates, similarly to the patterns, it affected its physical properties, ultimately leading to variations in the behavior of cells cultured onto these substrates. Enabled by the stripes on mBC, both SMCs and ENS cells were aligned in vitro, presenting the in vivo-like morphology essential for peristalsis and gut function. Interestingly, cell growth and differentiation remarkably enhanced upon SS addition to the samples, indicating the promise of the mBC-SS constructs as biomaterial not only for gut engineering, but also for tissues where cellular alignment is required for function, namely the heart, blood vessels, and similars.


Assuntos
Celulose/química , Trato Gastrointestinal/patologia , Sericinas/farmacologia , Engenharia Tecidual/métodos , Tecidos Suporte/química , Cicatrização , Animais , Materiais Biocompatíveis/química , Cristalização , Sistema Nervoso Entérico/efeitos dos fármacos , Feminino , Trato Gastrointestinal/inervação , Gluconacetobacter/química , Humanos , Masculino , Camundongos Endogâmicos BALB C , Miócitos de Músculo Liso/efeitos dos fármacos , Espectroscopia de Infravermelho com Transformada de Fourier , Cicatrização/efeitos dos fármacos
5.
Int J Biol Macromol ; 137: 545-553, 2019 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-31220499

RESUMO

The damage to the skin is most prominent and evident as it is our first line of defense and unremittingly under attack by biological and environmental factors. The restoration of the skin is dependent on the extent of the injury. To explore the prospects of new biomimetic material, bi-layered skin construct is fabricated in vitro with nonmulberry silk protein sericin and chitosan hydrogels using human dermal fibroblasts and keratinocytes. The in vitro analysis of the hydrogels showed enhanced adhesion, proliferation, and migration of skin cells with coordinated interaction amongst themselves leading to the synthesis of collagen IV and matrix metalloproteinase (MMP2 and MMP9). The in vivo evaluation indicates the regeneration of skin with densely packed collagen and formation of matured blood vessels in the animals implanted with sericin containing hydrogels. Moreover, the local and systemic immune response determined in vivo exhibits the biosafety of sericin based hydrogels. In addition, the cross-sectional analysis of the implanted hydrogels displays infiltration of the skin tissue cells into the hydrogels marking their biocompatibility and non-toxicity. The cumulative analysis of the in vitro and in vivo observations demonstrates that the sericin based hydrogels are non-inflammatory, supports the regeneration and repair of the skin tissue.


Assuntos
Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Hidrogéis/química , Regeneração/efeitos dos fármacos , Sericinas/química , Sericinas/farmacologia , Pele/efeitos dos fármacos , Animais , Bombyx , Matriz Extracelular/efeitos dos fármacos , Matriz Extracelular/metabolismo , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Queratinócitos/citologia , Queratinócitos/efeitos dos fármacos , Masculino , Teste de Materiais , Ratos , Ratos Wistar , Pele/citologia
6.
Carbohydr Polym ; 212: 403-411, 2019 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-30832874

RESUMO

Developing biomaterials based on the natural biomacromolecule silk sericin from Bombyx mori cocoon is of great interest for biomedical application. Dialdehyde carboxymethyl cellulose (DCMC) is derived from periodate oxidation of carboxy- methyl cellulose. Here, we developed a novel strategy of cross-linking of sericin with DCMC via the Schiff's base reaction. Fourier transform infrared spectroscopy and scanning electron microscopy indicated the formation of Schiff's base via the blending of sericin and DCMC. The mechanical properties tests suggested the covalent cross-linking effectively enhanced the tensile strength of sericin. The swelling test and water contact angle indicated the DCMC/SS film had excellent hydrophilicity, swellability. Additionally, we demonstrated the DCMC/SS film had excellent blood compatibility, cytocompatibility and promoting cell proliferation activity by the hemolysis ratio analysis, cell adhesion, cells viability and proliferation assays. The prepared DCMC/SS film has shown great promise in biomedical applications such as wound dressing, artificial skin and tissue engineering.


Assuntos
Carboximetilcelulose Sódica/síntese química , Celulose/análogos & derivados , Reagentes para Ligações Cruzadas/síntese química , Sericinas/síntese química , Seda/síntese química , Animais , Bandagens , Bombyx , Carboximetilcelulose Sódica/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Sobrevivência Celular/fisiologia , Celulose/síntese química , Celulose/farmacologia , Reagentes para Ligações Cruzadas/farmacologia , Portadores de Fármacos/síntese química , Portadores de Fármacos/farmacologia , Camundongos , Células NIH 3T3 , Sericinas/farmacologia , Seda/farmacologia , Engenharia Tecidual/tendências
7.
J Insect Sci ; 19(1)2019 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-30690536

RESUMO

The silk sericin hydrolysate (SSH) from the waste of silk processing as a substitute of fetal bovine serum (FBS) was used for the culture of Chinese hamster ovary (CHO) cells and Henrietta Lacks (Hela) strain of human cervical cancer cells. The survival ratio of these cells cultured in SSH media were similar to or higher than those in FBS media. Especially after the serum was replaced by low concentration of SSH at 15.0 µg/ml for 5 d, the proliferation of both cells was also similar to or higher than that of FBS group; the percentages of CHO and Hela cells in S-phase were 28.9 and 28.0%, respectively. The former is nearly two times that of FBS group, the latter is also higher than the control group. Reverse transcription-polymerase chain reaction (RT-PCR) revealed that among the differentially expressed genes, the relative expression of CXCL12 gene of CHO cells in SSH group increased, was three times that of serum group, and the relative expression of LCN2 gene of Hela cells increased 2.8 times, indicating that these related genes were activated to promote cell growth and proliferation. These results fully illustrated the hydrolysated sericin has a potential use as serum substitutes in cell culture.


Assuntos
Técnicas de Cultura de Células/métodos , Meios de Cultura Livres de Soro/farmacologia , Sericinas/farmacologia , Animais , Células CHO , Bovinos , Proliferação de Células/efeitos dos fármacos , Cricetulus , Expressão Gênica , Células HeLa , Humanos , Soro , Seda/química
8.
Food Chem Toxicol ; 123: 275-287, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30391273

RESUMO

Pro-oxidative stress induced by dietary polyphenols elevates reactive oxygen species (ROS) production in cancer cells, which subsequently leads to oxidative stress-mediated apoptosis. Sericin, a principal component of silk is associated with a mixture of polyphenols and flavonoids, possesses various biomedical attributes including anticancer activity. In the present study, we have evaluated the pro-oxidative effect of Bombyx mori sericin (BMS), Antheraea assamensis sericin (AAS), and Philosamia ricini sericin (PRS) against different cancer cells. Cytotoxicity of silk sericin (SS) evaluated using A431, SAS, and MCF-7 cells showed ≥50% reduction in their viability at 4 mg/mL. Intracellular ROS levels, cell cycle arrest, and apoptosis assessed using flow cytometry corroborated that SS treatment elevated the intracellular ROS levels, caused cell cycle arrest at the sub-G1 phase and resulted in apoptotic cell death. SS treated A431 and SAS cells showed upregulation of p53 and dysregulation of Bax and Bcl-2 gene expression. Whereas, AAS treated MCF-7 cells showed upregulation of Bax and downregulation of Bcl-2 gene expression. AAS treated MCF-7 and SAS cells showed downregulation of Bcl-2 protein expression in comparison to their control cells. Thus, the present study demonstrates that the pro-oxidative effect induced by SS suppresses the cancer growth indicating its potential anticancer activity.


Assuntos
Apoptose/efeitos dos fármacos , Proteínas de Insetos/farmacologia , Neoplasias/fisiopatologia , Estresse Oxidativo/efeitos dos fármacos , Sericinas/farmacologia , Seda/química , Animais , Bombyx , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Humanos , Células MCF-7 , Mariposas , Neoplasias/genética , Neoplasias/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
9.
Biol Res ; 51(1): 54, 2018 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-30497518

RESUMO

BACKGROUND: Hyperpigmentation disorders such as post-inflammatory hyperpigmentation are major concerns not only in light-skinned people but also in Asian populations with darker skin. The anti-tyrosinase and immunomodulatory effects of sericin have been known for decades. However, the therapeutic effects of sericin on hyperpigmentation disorders have not been well documented. METHODS: In this study, we used an in vitro model to study the anti-tyrosinase, tolerogenic, and anti-melanogenic effects of sericin on Staphylococcus aureus peptidoglycan (PEG)-stimulated melanocytes, dendritic cells (DCs), and artificial skin (MelanoDerm™). Enzyme-linked immunosorbent assay, conventional and immunolabeled electron microscopy, and histopathological studies were performed. RESULTS: The results revealed that urea-extracted sericin has strong anti-tyrosinase properties as shown by a reduction of tyrosinase activity in melanin pigments both 48 h and 10 days after allergic induction with PEG. Anti-inflammatory cytokines including interleukin (IL)-4, IL-10, and transforming growth factor-ß were upregulated upon sericin treatment (10, 20, and 50 µg/mL), whereas production of allergic chemokines, CCL8 and CCL18, by DCs was diminished 48 h after allergic induction with PEG. Moreover, sericin lowered the expression of micropthalmia-associated transcription factor (MITF), a marker of melanogenesis regulation, in melanocytes and keratinocytes, which contributed to the reduction of melanin size and the magnitude of melanin deposition. However, sericin had no effect on melanin transport between melanocytes and keratinocytes, as demonstrated by a high retention of cytoskeletal components. CONCLUSION: In summary, sericin suppresses melanogenesis by inhibition of tyrosinase activity, reduction of inflammation and allergy, and modulation of MITF function.


Assuntos
Hiperpigmentação/tratamento farmacológico , Queratinócitos/efeitos dos fármacos , Melanócitos/efeitos dos fármacos , Monofenol Mono-Oxigenase/antagonistas & inibidores , Sericinas/farmacologia , Células Cultivadas , Humanos , Hipersensibilidade , Inflamação , Queratinócitos/ultraestrutura , Melanócitos/ultraestrutura , Fator de Transcrição Associado à Microftalmia , Microscopia Eletrônica , Transdução de Sinais/efeitos dos fármacos , Fatores de Transcrição/efeitos dos fármacos
10.
Sci Rep ; 8(1): 14943, 2018 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-30297713

RESUMO

We recently demonstrated that in addition to its protective effect on pancreatic and adrenal biosynthesis, antioxidant properties of sericin decrease blood cholesterol levels and improve the liver mitochondrial architecture. However, little is known about the detailed mechanisms underlying these effects. Using proteomics and electron microscopy, we identified mitochondrial proteins that play important roles in the preservation of the mitochondrial ultrastructure and cholesterol-lowering properties of sericin. Our results showed that sericin maintains the mitochondrial architecture during conditions of high blood cholesterol by regulating apoptotic (NADH-ubiquinone oxidoreductase 75 kDa subunit) and autophagic (mitochondrial elongation factor Tu and prohibitin-2) proteins as well as energy maintenance proteins [haloacid dehalogenase-like hydrolase domain-containing protein 3, succinate dehydrogenase (ubiquinone) flavoprotein subunit, ATP synthase-α subunit precursor, enoyl-CoA hydratase domain-containing protein 3 and electron transfer flavoprotein subunit-α]. Sericin also exerts anti-oxidative properties via aconitate hydratase and Chain A, crystal structure of rat carnitine palmitoyltrasferase 2 proteins. Together, these activities may reduce hepatocytic triglyceride deposition, thereby decreasing steatosis, as demonstrated by the modulatory effects on ornithine aminotransferase, mitochondrial aspartate aminotransferase, acyl-CoA synthase, hydroxyacyl-CoA dehydrogenase and D-beta-hydroxybutyrate dehydrogenase. Sericin activity further balanced nitrogenous waste detoxification, characterised by carbamoyl-phosphate synthase (ammonia), aldehyde dehydrogenase and uricase, or folate biosynthesis via sarcosine dehydrogenase and dimethyl glycine dehydrogenase. These results suggest that sericin maintains the hepatic mitochondrial architecture through apoptotic, autophagic, energy maintenance and anti-oxidative mitochondrial proteins for alleviating hepatic steatosis and promoting liver function under conditions of hypercholesterolaemia.


Assuntos
Anticolesterolemiantes/farmacologia , Antioxidantes/farmacologia , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Bombyx/química , Mitocôndrias Hepáticas/efeitos dos fármacos , Sericinas/farmacologia , Animais , Anticolesterolemiantes/química , Antioxidantes/química , Metabolismo Energético/efeitos dos fármacos , Fígado Gorduroso/tratamento farmacológico , Fígado Gorduroso/metabolismo , Fígado Gorduroso/patologia , Feminino , Mitocôndrias Hepáticas/metabolismo , Mitocôndrias Hepáticas/patologia , Proteômica , Ratos , Ratos Wistar , Sericinas/química , Triglicerídeos/metabolismo
11.
Biomater Sci ; 6(11): 2859-2870, 2018 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-30259043

RESUMO

Full-thickness skin injury affects millions of people worldwide each year. It often leads to scar formation and loss of skin appendages even after clinical treatment. The majority of wound dressings currently used cannot achieve scarless skin regeneration with complete recovery of appendages such as hair follicles and sebaceous glands. Functional regeneration of these skin appendages is a great challenge. However, we achieved this goal by the successful development and utilization of a photo-crosslinkable sericin hydrogel (SMH) as a new type of wound dressing for repairing full-thickness skin injury. SMH implanted in a mouse full-thickness skin injury model promoted scarless wound healing with effective regeneration of hair follicles and sebaceous glands. By employing techniques of molecular biology, biochemistry, and in vivo cell tracing, we revealed the underlying repair mechanisms: SMH inhibited inflammation, stimulated angiogenesis during healing process, prevented scar tissue formation via regulating the expressions of TGF-ß1 and TGF-ß3, and recruited mesenchymal stem cells to injury sites for regeneration of skin appendages. Collectively, in this study, we developed a sericin-based hydrogel as a wound dressing for full-thickness skin injury repair, uncovered the functional roles of sericin hydrogels in promoting scarless skin regeneration along with effective recovery of skin appendages, and thus unveiled sericin's potential for skin wound healing.


Assuntos
Derme/efeitos dos fármacos , Folículo Piloso/efeitos dos fármacos , Hidrogéis/química , Regeneração/efeitos dos fármacos , Glândulas Sebáceas/efeitos dos fármacos , Sericinas/farmacologia , Cicatrização/efeitos dos fármacos , Animais , Materiais Biocompatíveis/química , Materiais Biocompatíveis/farmacologia , Folículo Piloso/citologia , Folículo Piloso/metabolismo , Folículo Piloso/fisiologia , Camundongos , Células NIH 3T3 , Glândulas Sebáceas/fisiologia , Sericinas/química , Células-Tronco/efeitos dos fármacos , Células-Tronco/metabolismo , Fator de Crescimento Transformador beta1/metabolismo , Fator de Crescimento Transformador beta3/metabolismo , Regulação para Cima/efeitos dos fármacos
12.
Carbohydr Polym ; 200: 572-582, 2018 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-30177201

RESUMO

Chronic wounds suffer impaired healing that results in psychological effect and morbity in addition to economic burden on the patient. These wounds are characterized by prolonged inflammatory phase, poor collagen production, hampered angiogenesis, upregulation in reactive oxygen species (ROS) and matrix metalloproteases (MMP). In the current work, three different concentrations (03:01, 05:01 and 10:01) of flexible agar-sericin-glycerol hydrogel films with sericin as minor component were fabricated to improve collagen production and capture free radical species to expediate chronic wound healing. The films exhibited improved collagen production with a p-value ≤ 0.001 for all the three concentrations. With regards to free radical scavenging, the 03:01 and 05:01 films exhibited improved performance with a p-value ≤ 0.01, while, 10:01 film showed improvement with a p-value ≤ 0.05. This was confirmed by Sirius red staining and 2,2-diphenyl-1-picryl hydrazyl (DPPH) assay. In addition, these films showed an adequate tensile strength of around 35 MPa with elongation at break higher than 25%. The as cast films exhibited satisfactory swelling behavior of around 320% in simulated wound fluid (SWF).


Assuntos
Ágar/química , Bandagens , Portadores de Fármacos/química , Hidrogéis/química , Fenômenos Mecânicos , Sericinas/química , Cicatrização/efeitos dos fármacos , Células 3T3 , Animais , Bandagens/microbiologia , Proliferação de Células/efeitos dos fármacos , Colágeno/metabolismo , Portadores de Fármacos/farmacologia , Liberação Controlada de Fármacos , Humanos , Hidrogéis/farmacologia , Cinética , Camundongos , Sericinas/farmacologia , Vapor , Resistência à Tração
13.
World J Gastroenterol ; 24(30): 3398-3413, 2018 Aug 14.
Artigo em Inglês | MEDLINE | ID: mdl-30122879

RESUMO

AIM: To develop a novel hepatocyte serum-free medium based on sericin, and to explore the effect of sericin on the hepatocyte transcriptome. METHODS: A controlled trial comparing novel serum-free medium and other media: C3A cells were cultured in our novel serum-free medium, HepatoZYME, complete medium (DMEM/F12 with 100 mL/L FBS), and DMEM/F12, and then cell attachment, proliferation, and function as well as the biocompatibility of the media were assessed. A comparative study of serum-free media with or without 2 mg/mL sericin: the effect of sericin on C3A growth was assessed by cell viability and proliferation, the effect of sericin on C3A cell cycle distribution was determined by flow cytometry, and the effect of sericin on the C3A transcriptome was assessed by gene-chip array and RT-qPCR. RESULTS: More C3A cells attached to the plate containing our serum-free medium than to those containing HepatoZYME and DMEM/F12 at 24 h post-seeding. Both the viability and proliferation rate of C3A cells in sericin-based serum-free medium were superior to those of cells in HepatoZYME and DMEM/F12 (P < 0.001). The content of albumin and urea in our serum-free medium was significantly higher than that in HepatoZYME and DMEM/F12 throughout the whole culture period (P < 0.001) and was similar to that in complete medium at day 3, 4, and 5. In part 2, cell viability and proliferation were greater in the presence of 2 mg/mL sericin (P < 0.001), as was the proportion of cells in S phase (16.21% ± 0.98% vs 12.61% ± 0.90%, P < 0.01). Gene-chip array analysis indicated that the expression of CCR6, EGFR, and FOS were up-regulated by 2 mg/mL sericin, and RT-qPCR revealed that the expression of CCR6, EGFR, FOS, AKT1, JNK1, NFkB1, MMP-9, MEK2, ERK1/2 and MYC was up-regulated by 2 mg/mL sericin (P < 0.05). CONCLUSION: We developed a novel hepatocyte serum-free medium. Sericin probably enhances cell attachment through the CCR6-Akt-JNK-NF-κB pathway and promotes cell proliferation through CCR6-mediated activation of the ERK1/2-MAPK pathway.


Assuntos
Técnicas de Cultura de Células/métodos , Hepatócitos/efeitos dos fármacos , Sericinas/farmacologia , Transcriptoma/efeitos dos fármacos , Adesão Celular/efeitos dos fármacos , Divisão Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Meios de Cultura Livres de Soro/química , Meios de Cultura Livres de Soro/farmacologia , Perfilação da Expressão Gênica/métodos , Hepatócitos/metabolismo , Humanos , Análise de Sequência com Séries de Oligonucleotídeos/métodos
14.
Cryobiology ; 84: 33-39, 2018 10.
Artigo em Inglês | MEDLINE | ID: mdl-30098997

RESUMO

Cryopreservation results in substantial deterioration of heat shock protein 70 (HSP70) and ultra-structural changes in sperm organelles, resulting in a marked reduction in post-thaw semen quality. The present study was aimed to explicate the effect of sericin supplementation on expression profile of HSP70, redox status and post-thaw semen quality in Barbari goat. Five Barbari bucks were used to collect thirty semen ejaculates by using artificial vagina and each ejaculate was divided into three aliquots to which sericin was supplemented at 0% (Control), 0.25% (T1) and 0.50% (T2). Further, extended semen samples were equilibrated followed by their cryopreservation. Post-thaw semen characteristics, redox status of seminal plasma, enzyme leakage and HSP70 gene/protein expression in spermatozoa were assessed in all the groups. Per cent progressive motile spermatozoa, spermatozoa having intact plasma membrane (HOST + ve) and intact acrosomes in post-thaw spermatozoa were significantly (p < 0.01) higher in T1 and T2 as compared to control. A significant (p < 0.01) reduction in abnormal spermatozoa was found in T1 as compared to T2. Sericin supplementation significantly (p < 0.05) improved the antioxidative status (SOD, GST, CAT), reduced lipid peroxidation (MDA) and also prevented enzyme (ALT, LDH) leakage as compared to control samples. qRT-PCR results revealed that HSP70 mRNA expression was significantly (p < 0.01) upregulated in T1 and T2 group as compared to control. The positive effect of sericin on expression of HSP70 was further confirmed by immunoblotting followed by densitometry revealing higher expression in T1 and T2 compared to control. Inclusion of 0.25% w/v sericin in semen extender ameliorated the post-thaw semen quality by improving antioxidative status and minimizing the leakage of intracellular enzymes. Sericin supplementation had a beneficial effect on HSP70/HSP70 mRNA expression either by induction or by protection of HSP70/HSP70 mRNA as evident from the gene expression and immunoblotting studies.


Assuntos
Criopreservação/métodos , Crioprotetores/farmacologia , Proteínas de Choque Térmico HSP70/biossíntese , Preservação do Sêmen/métodos , Sericinas/farmacologia , Animais , Antioxidantes/farmacologia , Cabras , Masculino , Sêmen , Análise do Sêmen , Motilidade Espermática , Espermatozoides/metabolismo
15.
Acta Biomater ; 79: 239-252, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30149211

RESUMO

Sericin, as the major component of Bombyx mori silk, is a useful biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Here, we report the fabrication of a human acidic fibroblast growth factor (FGF1)-functionalized sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its sericin layer. Sericin, together with FGF1, were simultaneously extracted from the silk fiber and then exposed to cold-induced hydrogel formation without additional crosslinking. The fabricated FGF1 sericin hydrogels demonstrated injectability, useful mechanical properties and a porous microstructure, which contributed to cell adhesion and survival. In addition, FGF1 achieved long-term storage in the sericin hydrogels over a wide range of temperatures. Further, the sericin-FGF1 demonstrated sustained release to promote cell proliferation and wound healing. Furthermore, cellular inflammatory responses showed that the FGF1 sericin hydrogels exhibited biocompatibility and no immunogenicity. This study revealed the successful exploration of FGF1-functionalized sericin hydrogels as a new protein-based biomaterial to expand applications of FGF1 and sericin in tissue and medical engineering. Further, we demonstrated a strategy for the predesign of exogenous protein-functionalized sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale. STATEMENT OF SIGNIFICANCE: Sericin from the Bombyx mori silk, is regarded as a desirable biomaterial for tissue engineering due to its hydrophilicity, biocompatibility and biodegradability. Genetically engineering the sericin with functional exogenous proteins would enhance its biofunctions and further expand its application in tissue engineering. In this study, we demonstrated a method to fabricate a human acidic fibroblast growth factor (FGF1)-functionalized sericin hydrogel using a transgenic silkworm spun silk with FGF1 incorporated in its sericin layer. The fabricated FGF1 sericin hydrogels demonstrated injectability, porous microstructure, biocompatibility and no immunogenicity which contributed to cell adhesion and survival. Remarkably, FGF1 could achieve a long-term stability in the sericin hydrogels over a wide range of temperatures and sustained release to promote cell proliferation and wound healing. This study revealed the successful exploration of FGF1-functionalized sericin hydrogels as a new protein-based biomaterial in tissue and medical engineering application, and provided a strategy for the predesign of exogenous protein-functionalized sericin hydrogels through genetically modifying silk fibers as sources for their cost effective production at a large scale.


Assuntos
Bombyx/genética , Proliferação de Células/efeitos dos fármacos , Fator 1 de Crescimento de Fibroblastos/farmacologia , Engenharia Genética/métodos , Hidrogéis/farmacologia , Sericinas/farmacologia , Animais , Animais Geneticamente Modificados , Adesão Celular/efeitos dos fármacos , Preparações de Ação Retardada/farmacologia , Fator 1 de Crescimento de Fibroblastos/química , Inflamação/patologia , Camundongos , Células NIH 3T3 , Estrutura Secundária de Proteína , Células RAW 264.7 , Espectroscopia de Infravermelho com Transformada de Fourier
16.
Acta Biomater ; 78: 137-150, 2018 09 15.
Artigo em Inglês | MEDLINE | ID: mdl-30059800

RESUMO

Recently, the progress in biomaterials for biomedical applications brings the focus of the research community toward nanomaterials. The nanofibrous matrices offer certain advantages (structural similarity to extracellular matrix, high surface area-to-volume ratio, increased elasticity, biostability, and strength) compared to other prevalent type of materials. This affirms their superiority and flexibility to be used in regenerative medicine. We have fabricated nonmulberry (Antheraea mylitta) silk protein sericin-based nanofibrous matrices (fiber thickness; 80-400 nm) with improved mechanical strength and desired stability (>4 weeks) as required for tissue reconstruction. These matrices support the adhesion, proliferation, and cellular interconnection of human keratinocytes. These are minimally hemolytic, nonimmunogenic, and capable of wound healing in vivo. Antibiotic (cephalexin hydrate [CH])-loaded nanofibrous matrices accelerate the full-thickness wound repair with minimal inflammation and without any signs of infection. The histological analysis authenticates skin restoration with re-epithelialization, generation of associated skin appendages, and synthesis of dense collagen fibrils. In addition, analysis of inflammatory genes and immunohistochemical assays have proved their biocompatibility and wound healing potential. Angiogenesis is also prevalent in the animal tissue treated with nanofibrous matrices. The results of in vitro and in vivo experimentations indicate a clear prospect of the fabricated sericin-based nanofibrous matrices to be used for skin regeneration. STATEMENT OF SIGNIFICANCE: Nonmulberry silk protein sericin-based nanofibrous matrix is a useful biomaterial for wound healing, collagen production, and skin tissue repair. It has been used in different formulations including hydrogels and nanofibrous membranes with chitosan (CS) and polyvinyl alcohol (PVA). No experiments have been carried out to evaluate sericin-based nanofibrous membranes for skin tissue engineering application. The present study shows that the nanofibrous matrices fabricated by electrospinning nonmulberry silk protein sericin with CS and PVA mimic the architectural environment of the extracellular matrix fibrils. These matrices are minimally hemolytic, are nonimmunogenic, and support better growth of human keratinocytes in vitro and wound healing in vivo with re-epithelialization of the skin tissue and angiogenesis. This work indicates that these nonmulberry sericin-based nanofibrous matrices with CS may be used as an ideal physical environment and biological cues for the promotion of skin tissue reconstruction and repair.


Assuntos
Inflamação/patologia , Nanofibras/química , Sericinas/farmacologia , Seda/farmacologia , Cicatrização/efeitos dos fármacos , Quitosana/química , Humanos , Queratinócitos/metabolismo , Testes de Sensibilidade Microbiana , Álcool de Polivinil/química , Tecidos Suporte/química
17.
Int J Biol Macromol ; 118(Pt A): 792-799, 2018 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-29959012

RESUMO

Intervertebral disk degeneration is an oxidative and inflammatory pathological condition that induces viability and functionality reduction of Nucleus Pulposus cells (NPs). Cellular therapies were previously proposed to repair and substitute the herniated disk but low proliferative index and pathological conditions of NPs dramatically reduced the efficacy of this approach. To overcome these problems we proposed, for the first time, a therapeutic system based on the association of silk sericin microparticles and platelet-derived products. Silk sericin (SS) is a bioactive protein with marked antioxidant properties, while platelet lysate (PL) and platelet poor plasma (PPP) represent a source of growth factors able to support cell viability and to promote tissue regeneration. We demonstrated that the mixture PL + PPP promoted NPs proliferation with a significant reduction of cellular doubling time. SS microparticles, alone or in combination with PPP, presented the higher ROS-scavenging activity while, SS microparticles and PL resulted as the best association able to protect NPs against oxidative stress induce by hydroxide peroxide. Based on these results, the authors are confident that, with the ever increasing need of efficacious tools for regenerative medicine purposes, SS microparticles and PL + PPP association could represent an effective approach for the development of low impact and non-invasive therapies.


Assuntos
Plaquetas/química , Proliferação de Células/efeitos dos fármacos , Disco Intervertebral/fisiologia , Regeneração/efeitos dos fármacos , Sericinas , Humanos , Disco Intervertebral/patologia , Sericinas/química , Sericinas/farmacologia
18.
Int J Biol Macromol ; 119: 37-47, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30048722

RESUMO

Silk sericin (SS), a globular and water-soluble protein, showed to in vitro sustain the proliferation of different mammalian cell lines, and to in vitro protect them from the oxidative stress because of its free-radical-scavenging activity. Also, platelet-derived products, such as platelet gel, platelet rich plasma or platelet lysate (PL) are used in cell cultures as a serum substitute due to their great content of growth factors. In the clinical practice, both SS and PL were employed for various reparative treatments and showed to sustain the wound repair process regulating fibroblast recruitment, migration, proliferation and differentiation. For these reasons, the combination of PL and SS could lead to effective and innovative products, either for the ex vivo cell expansion, or for the production of active wound healing advanced medications.


Assuntos
Plaquetas/metabolismo , Sericinas/química , Sericinas/farmacologia , Seda/química , Cicatrização , Animais , Proliferação de Células , Humanos , Plasma Rico em Plaquetas/metabolismo , Regeneração
19.
Mol Reprod Dev ; 85(8-9): 696-708, 2018 08.
Artigo em Inglês | MEDLINE | ID: mdl-29947084

RESUMO

We investigated the effects of sericin on the developmental competence of bovine embryos exposed to heat stress (HS). Putative zygotes were cultured with sericin and subjected to HS (40.5°C for 6 hr) on Day 2 or 7 followed by continuous culture at 38.5°C until Day 8. Day 2 HS significantly decreased blastocyst development on Day 8 as well as mitochondrial activity, and significantly increased the amount of intracellular reactive oxygen species and terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling (TUNEL)-positive cells, whereas Day 7 HS only significantly decreased mitochondrial activity and increased the number of TUNEL-positive cells in Day 8 blastocysts. These detrimental effects were neutralized by sericin supplementation. Next, to investigate the potential production of blastocysts with high viability in terms of thermotolerance, embryos were cultured with sericin until Day 7, and then exposed to HS in the sericin-free medium. TUNEL-positive cell numbers were significantly lower in blastocysts produced by sericin culture than in control blastocysts. Transcript abundance for HSPA1A and BAX was significantly decreased but IFNT2 levels were increased in blastocysts produced by sericin culture. In conclusion, these findings demonstrate the anti-oxidative and anti-apoptotic activities of sericin, and the potential use of sericin to produce embryos with high viability in vitro.


Assuntos
Antioxidantes/metabolismo , Antioxidantes/farmacologia , Técnicas de Cultura Embrionária/métodos , Desenvolvimento Embrionário/efeitos dos fármacos , Resposta ao Choque Térmico/efeitos dos fármacos , Sericinas/metabolismo , Sericinas/farmacologia , Animais , Apoptose/efeitos dos fármacos , Blastocisto/efeitos dos fármacos , Blastocisto/metabolismo , Bovinos , Embrião de Mamíferos/metabolismo , Fertilização In Vitro , Transtornos de Estresse por Calor/prevenção & controle , Temperatura Alta/efeitos adversos , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Termotolerância/efeitos dos fármacos , Fatores de Tempo , Zigoto/metabolismo
20.
Drug Deliv ; 25(1): 1103-1116, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29742945

RESUMO

Drug resistance is the major challenge facing cancer chemotherapy and nanoscale delivery systems based on natural materials, such as sericin, are a promising means of overcoming drug resistance. Yet, no attempt of introducing synthetic poly(γ-benzyl-L-glutamate) (PBLG) onto sericin polypeptide to fabricate a facile biocompatible and biodegradable micelle has been tried. Here, we prepared a polypeptide-based amphiphilic polymer containing hydrophilic sericin polypeptide backbone and PBLG side chains via ring-opening polymerization (ROP) strategy. The introduction of PBLG side chains remarkably enhances the stability of sericin micelles in water. Meanwhile, the micelles exhibited a high loading capacity and pH-responsive release ability for antitumor drug doxorubicin (DOX), called sericin-PBLG-DOX. Owing to the excellent cell membrane penetration of sericin-PBLG, the cellular uptake of DOX when loaded into micelles was improved. Subsequently, sericin-PBLG-DOX was transferred into perinuclear lysosomes, where the release rate of DOX was accelerated. Compared to the same dose of DOX, sericin-PBLG-DOX could induce a more efficient anti-tumor effect both in vitro and in vivo, and these micelles have promise for future clinical applications in overcoming cancer drug resistance with good biosafety, enhanced cellular uptake, pH-triggered drug release, efficient anti-tumor effects, and minimized systemic toxicity.


Assuntos
Antineoplásicos/farmacologia , Doxorrubicina/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Nanopartículas/química , Neoplasias/tratamento farmacológico , Sericinas/farmacologia , Animais , Bombyx , Linhagem Celular Tumoral , Portadores de Fármacos/química , Sistemas de Liberação de Medicamentos/métodos , Liberação Controlada de Fármacos , Células Hep G2 , Humanos , Concentração de Íons de Hidrogênio , Interações Hidrofóbicas e Hidrofílicas , Células MCF-7 , Masculino , Micelas , Peptídeos/química , Ácido Poliglutâmico/análogos & derivados , Ácido Poliglutâmico/química , Polímeros/química , Ratos , Ratos Sprague-Dawley
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