Metformin loaded injectable silk fibroin microsphere for the treatment of spinal cord injury.

The repair of spinal cord injury is a great challenge in clinical. Improving the microenvironment of the injured site is the key strategy for accelerating axon regeneration and synaptic formation. Herein, a kind of silk fibroin microspheres functionalized by metformin through dopamine was developed using water-in-oil emulsification-diffusion method and surface modification technique, and the effect on cortical neuron was evaluated. The results showed that the microspheres showed a uniform size distribution with the diameter of around 60 µm and a concave structure. Moreover, the microspheres possessed good injectability and stability. In addition, the metformin could be successfully immobilized in the silk fibroin microspheres. The cell culture results displayed that the growth and morphology of cortical neurons on the microspheres with metformin concentration of 5 mg/mL and 10 mg/mL were obviously better than that on other samples. Notably, the spread area of single cortical cell on silk fibroin microspheres was increased with the ascending metformin concentration. Therefore, the results indicated that the metformin loaded silk fibroin microsphere could obviously improve the growth and spreading behavior of cortical neuron. The study may provide an important experimental basis for the development of drug loaded injectable biomaterials scaffolds for the treatment of spinal cord injury and have great potential for spinal cord regeneration.

Supplementation with a Natural Source of Amino Acids, Sil-Q1 (Silk Peptide), Enhances Natural Killer Cell Activity: A Redesigned Clinical Trial with a Reduced Supplementation Dose and Minimized Seasonal Effects in a Larger Population.

The aim of this study was to re-validate the changes in natural killer (NK) cell cytotoxicity and cytokines related to T cells after Sil-Q1 (SQ; silk peptide) supplementation in a larger pool of Korean adults with minimized daily dose of SQ and controlling seasonal influence compared to the previous study. A total of 130 subjects were randomly assigned (1:1) to consume either 7.5 g of SQ or placebo for 8 weeks. NK cell cytotoxicity and cytokines were measured at T0 (baseline) and T8 (follow-up). Comparing the NK cell cytotoxicity values at T0 and T8 within each group, the cytotoxicity at all effector cell (E) to target cell (T) ratios of 10:1, 5:1, 2.5:1, and 1.25:1 was significantly increased in the SQ group at T8. Additionally, significant differences in the changed value (Δ, subtract baseline values from follow-up values) comparison between the groups at E:T = 10:1, 5:1, and 2.5:1 were found. As a secondary endpoint, the interleukin (IL)-12 level in the SQ group was significantly increased for 8 weeks, and Δ IL-12 in the SQ group was greater than in the placebo group. In conclusion, the present study showed considerable practical implications of SQ supplementation. Thus, SQ is an effective and safe functional food supplement for enhancing immune function.

Nanosilk Increases the Strength of Diabetic Skin and Delivers CNP-miR146a to Improve Wound Healing.

Diabetes mellitus is a metabolic disorder associated with properties and an increased risk of chronic wounds due to sustained pro-inflammatory response. We have previously of radical scavenging cerium oxide nanoparticles (CNP) conjugated to the anti-inflammatory microRNA (miR)-146a, termed CNP-miR146a, improves diabetic wound healing by synergistically lowering oxidative stress and inflammation, and we sought to evaluate this treatment in a topical application. Silk fibroin is a biocompatible polymer that can be fabricated into nanostructures, termed nanosilk. Nanosilk is characterized by a high strength-to-density ratio and an ability to exhibit strain hardening. We therefore hypothesized that nanosilk would strengthen the biomechanical properties of diabetic skin and that nanosilk solution could effectively deliver CNP-miR146a to improve diabetic wound healing. The ability of nanosilk to deliver CNP-miR146a to murine diabetic wounds and improve healing was assessed by the rate of wound closure and inflammatory gene expression, as well as histologic analysis. The effect of nanosilk on the properties of human diabetic skin was evaluated by testing the biomechanical properties following topical application of a 7% nanosilk solution. Diabetic murine wounds treated with topical nanosilk and CNP-miR146a healed by day 14.5 compared to day 16.8 in controls (p = 0.0321). Wounds treated with CNP-miR146a had higher collagen levels than controls (p = 0.0126) with higher pro-fibrotic gene expression of TGFß-1 (p = 0.0092), Col3α1 (p = 0.0369), and Col1α2 (p = 0.0454). Treatment with CNP-miR146a lowered pro-inflammatory gene expression of IL-6 (p = 0.0488) and IL-8 (p = 0.0009). Treatment of human diabetic skin with 7% nanosilk solution resulted in significant improvement in maximum load and modulus (p < 0.05). Nanosilk solution is able to strengthen the biomechanical properties of diabetic skin and can successfully deliver CNP-miR146a to improve diabetic wound healing through inhibition of pro-inflammatory gene signaling and promotion of pro-fibrotic processes.

Assembly of FN-silk with laminin-521 to integrate hPSCs into a three-dimensional culture for neural differentiation.

Three-dimensional (3D) neural tissue cultures recapitulate the basic concepts during development and disease better than what can be obtained using conventional two-dimensional cultures. Here, we use a recombinant spider silk protein functionalized with a cell binding motif from fibronectin (FN-silk) in combination with a human recombinant laminin 521 (LN-521) to create a fully defined stem cell niche in 3D. A novel method to assemble silk blended with LN-521 together with human pluripotent stem cells (hPSC) is used to create centimeter-sized foams, which upon cultivation develop into 3D cell constructs supported by a microfibrillar network. After initial cell expansion, neural differentiation was induced to form a homogenous layer of continuous neuroectodermal tissue that allows further differentiation into neuronal subtypes. The silk-supported 3D cell constructs could then be detached from the bottom of the well and cultured as floating entities, where cells appeared in distinctive radial organization resembling early neural tube. This shows that the neural progenitors retain their cellular self-organization ability in the FN-silk/LN-521-supported 3D culture. Calcium imaging demonstrated spontaneous activity, which is important for the formation of neuronal networks. Together, the results show that hPSCs integrated into FN-silk/LN-521 foam develop into neural progenitors and that these stay viable during long-term differentiations. FN-silk/LN-521 also supports morphogenesis mimicking the human brain development and can serve as base for engineering of hPSC-derived neural tissue.

Injectable hydrogel systems with multiple biophysical and biochemical cues for bone regeneration.

Bone regeneration is a complex process in which angiogenesis and osteogenesis are crucial. Introducing multiple angiogenic and osteogenic cues simultaneously into a single system and tuning these cues to optimize the niche remains a challenge for bone tissue engineering. Herein, based on our injectable biomimetic hydrogels composed of silk nanofibers (SNF) and hydroxyapatite nanoparticles (HA), deferoxamine (DFO) and bone morphogenetic protein-2 (BMP-2) were loaded on SNF and HA to introduce more angiogenic and osteogenic cues. The angiogenesis and osteogenesis capacity of injectable hydrogels could be regulated by tuning the delivery of DFO and BMP-2 independently, resulting in vascularization and bone regeneration in cranial defects. The angiogenesis and osteogenesis outcomes accelerated the regeneration of vascularized bones toward similar composition and structure to natural bones. Therefore, the multiple biophysical and chemical cues provided by the nanofibrous structures, organic-inorganic compositions, and chemical and biochemical angiogenic and osteogenic inducing cues suggest the potential for clinical applicability of these hydrogels in bone tissue engineering.

Cervical Augmentation with an Injectable Silk-Based Gel: Biocompatibility in a Rat Model of Pregnancy.

The aim of this study was to study the biocompatibility of an injectable silk gel in the cervix in a rat model of pregnancy. The rationale is to study an injectable gel as an alternate treatment for cervical insufficiency. We further aimed to perform cervical injections via a vaginal route to mimic the clinical procedure of a cervical cerclage. We performed an in vivo study in pregnant female Sprague Dawley rats. Cervical procedures were performed using a customized speculum under general anesthesia. Injections were performed on gestational day 16. The responses to silk gel injections were compared to polyethylene terephthalate suture and saline controls on gestational day 19 and postpartum. The inflammatory response was evaluated by histology, PCR for inflammatory gene expression, and ELISA for protein levels of proinflammatory mediators. Silk gel injections were performed on 13 animals. All animals tolerated the procedure. Silk gel occupied 5% of the stroma after injection. Injected silk gel caused neither preterm birth nor prolonged pregnancy and had no effect on the kits. When comparing inflammatory responses, expression of inflammatory genes and proinflammatory proteins in the silk gel group was intermediate between saline (lowest) and cerclage suture (highest). Injectable silk gel was more inflammatory compared to saline injections but less inflammatory compared to the suture material used for cervical cerclage. This study is an important step toward development of an alternative treatment for cervical insufficiency.

Natural Nanofiber Shuttles for Transporting Hydrophobic Cargo into Aqueous Solutions.

Hydrophobic biomolecules realize their functions in vivo in aqueous environments, often through a delicate balance of amphiphilicity and chaperones. Introducing exogenous hydrophobic biomolecules into in vivo aqueous systems is a challenge in drug delivery and regenerative medicine, where labile linkers, carriers, and fusions or chimeric molecules are often designed to facilitate such aqueous interfaces. Here, we utilize naturally derived silk nanofiber shuttles with the capacity to transport hydrophobic cargos directly into aqueous solutions. These nanofibers disperse in organic solvents and in aqueous solutions because of their inherent amphiphilicity, with enriched hydrophobicity and strategically interspersed negatively charged groups. Hydrophobic molecules loaded on these shuttles in organic solvent-water systems separated from the solvent after centrifugation. These concentrated hydrophobic molecule-loaded nanofibers could then be dispersed into aqueous solution directly without modification. These shuttle systems were effective for different hydrophobic molecules such as drugs, vitamins, and dyes. Improved biological stability and functions of hydrophobic cargos after loading on these nanofibers suggest potential applications in drug delivery, cosmetology, medical diagnosis, and related health fields, with a relatively facile process.

Acid Hydrolyzed Silk Peptide Consumption Improves Anti-Diabetic Symptoms by Potentiating Insulin Secretion and Preventing Gut Microbiome Dysbiosis in Non-Obese Type 2 Diabetic Animals.

: Silk fibroin hydrolysates have been reported to reduce hyperglycemia, but the mechanism has not been determined in Asian type 2 diabetes (T2DM). We hypothesized that the consumption of acid hydrolyzed silk peptides (SPs) alleviates hyperglycemia by improving insulin sensitivity and subsequently normalizing glucose-stimulated insulin secretion in T2DM. We investigated this hypothesis in a partial pancreatectomized (Px) rat model. Px rats was assigned randomly to the following six groups and fed assigned diet for 8 weeks: the Px-CON (0.5 g/kg/day dextrin), the SP-L (0.05 g/kg/day), the SP-M (0.1 g/kg/day), the SP-H (0.5 g/kg/day), the positive-CON (30 mg/kg/day metformin), or the normal-CON (sham-operated rats; 0.5 g/kg/day dextrin). SPs contained high levels of glycine, alanine, and serine. We found SPs dose-dependently increased food efficiency and body weight gain in Px rats. Animals in the Px-control group rats exhibited lower glucose metabolism, as evidenced by impaired glucose-stimulated insulin secretion coupled with impaired insulin sensitivity, and reduced bone mineral density (BMD) and lean body mass (LBM), compared to normal-CON. SPs and metformin similarly partially protected against Px-induced BMD loss in the lumbar spine and femur. Px-induced decreases in LBM were dose-dependently prevented by SPs, and muscle forces in the SP-M and SP-H groups were maintained at the normal-CON level. Glucose tolerance was dose-dependently improved by SPs as determined by oral glucose tolerance and oral maltose tolerance tests, and glucose tolerances were similar in the SP-H and positive-CON groups. Insulin tolerance, an index of insulin sensitivity, was dose-dependently enhanced by SPs, and the SP-H group exhibited better insulin tolerance than the positive-CON group as determined by intraperitoneal insulin sensitivity testing. Insulin secretory capacity assessed using a hyperglycemic clamp improved in the following order: Px-control

Potential Photoprotective Effect of Dietary Corn Silk Extract on Ultraviolet B-Induced Skin Damage.

Ultraviolet B (UVB) irradiation causes adverse effects on the skin. Corn silk contains flavonoids and other bioactive compounds and antioxidants, which may prevent skin photoaging through antioxidant and anti-inflammatory effects. We aimed to investigate the potential photoprotective effects of dietary corn silk on UVB-induced skin damage in mice and the mechanisms behind these effects on human skin cells. Oral administration of corn silk water extract (CS) (2 or 4 g/kg/day) for 19 weeks decreased epidermal thickness, wrinkle formation, and positive staining for PCNA, Ki67, and 8-OHdG, and increased collagen staining in UVB-irradiated SKH-1 hairless mice compared with controls. The pro-inflammatory NF-κB target genes (IL-1ß, iNOS, and COX-2) and MMP-9 expressions were lower in the CS groups, and TGF-ß/Smad signaling increased. Low skin lipid peroxidation and blood DNA oxidation levels and high blood glutathione were detected. Antioxidant transcription factor Nrf2-related catalase and SOD1 proteins and glutaredoxin mRNA levels increased. The results of CS extract treatment and UVB irradiation in HaCaT cells showed the same results in Nrf2 and NF-κB target genes. An LC-MS/MS analysis showed that the CS extract contained potential antioxidants, which might have contributed to its anti-photoaging effects in tissues and cells. CS extract may reduce UVB-induced skin damage through antioxidant and anti-inflammatory mechanisms.

Injectable Silk Protein Microparticle-based Fillers: A Novel Material for Potential Use in Glottic Insufficiency.

OBJECTIVES AND HYPOTHESIS: A novel, silk protein-based injectable filler was engineered with the intention of vocal fold augmentation as its eventual intended use. This injectable filler leverages the unique properties of silk protein's superior biocompatibility, mechanical tunability, and slow in vivo degradation to one day better serve the needs of otolaryngologists. This paper intends to demonstrate the mechanical properties of the proposed novel injectable and to evaluate its longevity in animal models. MATERIALS AND METHODS: Experimental. The mechanical properties of silk bulking agents were determined to characterize deformation resistance and recovery compared with commercially available calcium hydroxylapatite through rheologic testing. Fresh porcine vocal fold tissue was used for injectable placement to simulate the mechanical outcomes of native tissue after bulking procedures. In vivo subcutaneous rodent implantation examined immune response, particle migration, and volume retention. RESULTS: Porous, elastomeric silk microparticles demonstrate high recovery (>90% original volume) from compressive strain and mimic the native storage modulus of soft tissues (1-3 kPa). Injectable silk causes only a slight increase in porcine vocal fold stiffness immediately after injection (20%), preserving the native mechanics of bulked tissue. In the subcutaneous rat model, silk demonstrated biocompatibility and slow degradation, thus enabling host cell integration and tissue deposition. CONCLUSIONS: The presented novel silk injectable material demonstrates favorable qualities for a vocal fold injection augmentation material. An in vivo long-term canine study is planned.

A novel silk-based vocal fold augmentation material: 6-month evaluation in a canine model.

OBJECTIVES: Ideal long-term vocal fold augmentation materials should be biocompatible, easily administered, allow tissue integration for long-term effect, and remain at the site of injection. A novel silk protein particle suspended in hyaluronic acid (Silk-HA) has been developed specifically for vocal fold augmentation to address this unmet need. This article presents the 6-month, preclinical findings of a canine vocal fold injection trial for Silk-HA. METHODS: Twelve beagle dogs were injected transorally in the lateral/deep aspect of their right thyroarytenoid muscles with 0.3 cc of Silk-HA or calcium hydroxylapatite in carboxymethyl cellulose (CaHA-CMC). The Silk-HA particle injectable was delivered via a custom catheter, whereas CaHA-CMC was delivered through a commercially available malleable needle. The six dogs from each material group were sacrificed 6 months from the injection date for the evaluation of implant longevity, immune response, and material migration. RESULTS: Silk-HA provides immediate medialization of the right vocal fold, lasting for a minimum of 6 months in a canine model. Silk-HA and CaHA-CMC both demonstrate similar inflammatory responses. The Silk-HA was shown to remain without migration at the site of injection in all six canine subjects, whereas CaHA-CMC demonstrated migration in four of the six canines. In two canines implanted with CaHA-CMC, material was discovered to migrate to the retropharyngeal lymph nodes. CONCLUSION: In a canine subject model, the Silk-HA material compares favorably in terms of longevity and immune response to CaHA-CMC. The lack of migration of the Silk-HA material demonstrates a promising potential for vocal fold injection in the clinic. LEVEL OF EVIDENCE: NA Laryngoscope, 129:1856-1862, 2019.

Immunomodulatory injectable silk hydrogels maintaining functional islets and promoting anti-inflammatory M2 macrophage polarization.

Islet transplantation is considered the most promising treatment for type 1 diabetes. However, the clinical success is limited by islet dysfunction in long-term culture. In this study, we have utilized the rapid self-gelation and injectability offered by blending of mulberry silk (Bombyx mori) with non-mulberry (Antheraea assama) silk, resulting in a biomimetic hydrogel. Unlike the previously reported silk gelation techniques, the differences in amino acid sequences of the two silk varieties result in accelerated gelation without requiring any external stimulus. Gelation study and rheological assessment depicts tuneable gelation as a function of protein concentration and blending ratio with minimum gelation time. In vitro biological results reveal that the blended hydrogels provide an ideal 3D matrix for primary rat islets. Also, A. assama fibroin with inherent Arg-Gly-Asp (RGD) shows significant influence on islet viability, insulin secretion and endothelial cell maintenance. Furthermore, utility of these hydrogels demonstrate sustained release of Interleukin-4 (IL-4) and Dexamethasone with effective M2 macrophage polarization while preserving islet physiology. The immuno-informed hydrogel demonstrates local modulation of inflammatory responses in vivo. Altogether, the results exhibit promising attributes of injectable silk hydrogel and the utility of non-mulberry silk fibroin as an alternative biomaterial for islet encapsulation.

The potential of Antheraea pernyi silk for spinal cord repair.

One of the most challenging applications for tissue regeneration is spinal cord damage. There is no cure for this, partly because cavities and scar tissue formed after injury present formidable barriers that must be crossed by axons to restore function. Natural silks are considered increasingly for medical applications because they are biocompatible, biodegradable and in selected cases promote tissue growth. Filaments from wild Antheraea pernyi silkworms can support axon regeneration in peripheral nerve injury. Here we presented evidence that degummed A. pernyi filaments (DAPF) support excellent outgrowth of CNS neurons in vitro by cell attachment to the high density of arginine-glycine-aspartic acid tripeptide present in DAPF. Importantly, DAPF showed stiffness properties that are well suited to spinal cord repair by supporting cell growth mechano-biology. Furthermore, we demonstrated that DAPF induced no activation of microglia, the CNS resident immune cells, either in vitro when exposed to DAPF or in vivo when DAPF were implanted in the cord. In vitro DAPF degraded gradually with a corresponding decrease in tensile properties. We conclude that A. pernyi silk meets the major biochemical and biomaterial criteria for spinal repair, and may have potential as a key component in combinatorial strategies for spinal repair.

Prevention of preterm birth: Novel interventions for the cervix.

Preterm birth is the leading cause of neonatal mortality and morbidity worldwide. Spontaneous preterm birth is a complex, multifactorial condition in which cervical dysfunction plays an important role in some women. Current treatment options for cervical dysfunction include cerclage and supplemental progesterone. In addition, cervical pessary is being studied in research protocols. However, cerclage, supplemental progesterone and cervical pessary have well known limitations and there is a strong need for alternate treatment options. In this review, we discuss two novel interventions to treat cervical dysfunction: (1) injectable, silk protein-based biomaterials for cervical tissue augmentation (injectable cerclage) and (2) a patient-specific pessary. Three-dimensional computer simulation of the cervix is performed to provide a biomechanical rationale for the interventions. Further development of these novel interventions could lead to new treatment options for women with cervical dysfunction.

Drug delivery systems improving chemical and physical properties of anticancer drugs currently investigated for treatment of solid tumors.

Cancer is the second leading cause of death worldwide. Conventional cancer treatment like chemotherapy do not fulfil the expectations of both patients and physicians and there is a pressing need for a new kind of therapies that will increase drug delivery to the tumor mass. Standard chemotherapy does not show either specific tumor-targeting, or selective mode of action for cancer cells. Moreover, tumor microenvironments additionally disturb drug perfusion and diffusion. Currently approved anticancer drugs have many limitations and therefore special delivery systems improving their chemical and physical properties are beneficial. In the present review paper we discuss various drug delivery systems for solid tumors that are actually at various stages of pre-clinical tests or approved for therapy.

Curcumin-functionalized silk biomaterials for anti-aging utility.

Curcumin is a natural antioxidant that is isolated from turmeric (Curcuma longa) and exhibits strong free radical scavenging activity, thus functional for anti-aging. However, poor stability and low solubility of curcumin in aqueous conditions limit its biomedical applications. Previous studies have shown that the anti-oxidation activity of curcumin embedded in silk fibroin films could be well preserved, resulting in the promoted adipogenesis from human mesenchymal stem cells (hMSCs) cultured on the surface of the films. In the present study, curcumin was encapsulated in both silk fibroin films (silk/cur films) and nanoparticles (silk/cur NPs), and their anti-aging effects were compared with free curcumin in solution, with an aim to elucidate the mechanism of anti-aging of silk-associated curcumin and to better serve biomedical applications in the future. The morphology and structure of silk/cur film and silk/cur NP were characterized using SEM, FTIR and DSC, indicating characteristic stable beta-sheet structure formation in the materials. Strong binding of curcumin molecules to the beta-sheet domains of silk fibroin resulted in the slow release of curcumin with well-preserved activity from the materials. For cell aging studies, rat bone marrow mesenchymal stem cells (rBMSCs) were cultured in the presence of free curcumin (FC), silk/cur film and silk/cur NP, and cell proliferation and markers of aging (P53, P16, HSP70 gene expression and ß-Galactosidase activity) were examined. The results indicated that cell aging was retarded in all FC, silk/cur NP and silk/cur film samples, with the silk-associated curcumin superior to the FC.

Silk fibroin scaffold as a potential choice for female pelvic reconstruction: A study on the biocompatibility in abdominal wall, pelvic, and vagina.

The aim of this study was to compare the tissue reactions to silk fibroin scaffolds in the abdominal wall, vagina, and pelvic vesico-uterine of rats. Silk fibroin scaffolds were implanted subcutaneously in the abdominal, pelvic vesico-uterine space, and under the vaginal mucosa of 16 rats. The animals were euthanized at 2, 4, 8, and 12 weeks postoperatively. Hematoxylin and eosin staining was performed to evaluate cellular infiltration, the percentage of macrophages and granulocytes inside and around the scaffolds. The amounts of M1/M2 macrophages at the interface of scaffolds and host tissue were evaluated through an immunofluorescence assay. The degree of acute inflammation was similar among the three groups, and lasted no more than 4 weeks. A faster ingrowth of fibroblasts was found in the abdominally implanted silk fibroin scaffolds, while vaginal implanted scaffolds committed a slower tissue ingrowth rate and much more macrophages infiltration than the pelvic and abdominal group. However, a significantly higher amount of M2 cells were seen in the three groups. In general, silk fibroin has nice biocompatibility in the abdominal, vagina, and pelvic tissue, eliciting healthy tissue formation, and might be a potential choice for female pelvic reconstruction. Microsc. Res. Tech. 80:291-297, 2017. © 2016 Wiley Periodicals, Inc.

Silk fibroin hydrogel as physical barrier for prevention of post hernia adhesion.

BACKGROUND: Adhesion formation remains a major complication following hernia repair surgery. Physical barriers though effective for adhesion prevention in clinical settings are associated with major disadvantages, therefore, needs further investigation. This study evaluates silk fibroin hydrogel as a physical barrier on polypropylene mesh for the prevention of adhesion following ventral hernia repair. STUDY DESIGN: Peritoneal explants were cultured on silk fibroin scaffold to evaluate its support for mesothelial cell growth. Full thickness uniform sized defects were created on the ventral abdominal wall of rabbits, and the defects were covered either with silk hydrogel coated polypropylene mesh or with plain polypropylene mesh as a control. The animals were killed after 1 month, and the adhesion formation was graded; healing response of peritoneum was evaluated by immunohistochemistry with calretinin, collagen staining of peritoneal sections, and expression of PCNA, collagen-I, TNFα, IL6 by real time PCR; and its adverse effect if any was determined. RESULTS: Silk fibroin scaffold showed excellent support for peritoneal cell growth in vitro and the cells expressed calretinin. A remarkable prevention of adhesion formation was observed in the animals implanted with silk hydrogel coated mesh compared to the control group; in these animals peritoneal healing was complete and predominantly by mesothelial cells with minimum fibrotic changes. Expression of inflammatory cytokines decreased compared to control animals, histology of abdominal organs, haematological and blood biochemical parameters remained normal. CONCLUSION: Therefore, silk hydrogel coating of polypropylene mesh can improve peritoneal healing, minimize adhesion formation, is safe and can augment the outcome of hernia surgery.

Implantable chemotherapy-loaded silk protein materials for neuroblastoma treatment.

Neuroblastoma is the most common extracranial childhood solid tumor. Treatment of high risk tumors require intense multicycle chemotherapies, resulting in short- and long-term toxicities. Here, we present treatment of an orthotopic neuroblastoma mouse model, with silk fibroin materials loaded with vincristine, doxorubicin or the combination as a intratumoral, sustained release system. The materials, loaded with vincristine with or without doxorubicin, significantly decreased neuroblastoma tumor growth compared to materials loaded without drug or doxorubicin only as well as intravenous (IV) drug treatment. The intratumoral drug concentration was significantly higher with intratumoral delivery versus IV. Furthermore, intratumor delivery decreased the maximum plasma concentration compared to IV delivery, reducing systemic exposure and possibly reduing long-term side effects of chemotherapy exposure. Histopathologically, tumors with remission periods >25 days before recurrence transformed from a "small-round-blue cell" (SBRC) to predominantly "large cell" neuroblastoma (LCN) histopathology, a more aggressive tumor subtype with unfavorable clinical outcomes. These results show that intratumoral chemotherapy delivery may be a treatment strategy for pediatric neuroblastoma, potentially translatable to other focal tumors types. Furthermore, this treatment modality allows for a clinically relevant mouse model of tumor transformation that may be used for studying the phenotypical tumor recurrence and developing more effective treatment strategies for recurrent tumors.

The Efficacy of Cyclic Injection of Bone Morphogenetic Protein-2 in Large-Scale Calvarial Bone Defects.

Bone morphogenetic protein-2 (BMP-2) appears to be one of the most potent growth factors thus far studied. However, recent publications on the clinical application of BMP-2 revealed that its correct control is the paramount issue in clinical practice. For improving BMP-2 delivery, the cyclic administration might be an alternative. Accordingly, the authors cyclically injected BMP-2 in a cyclic injection model of large cranial defects to maintain the proper dosage during the bone healing process. A 10-mm diameter calvarial bone defect was produced using a round drill in 8-week-old Sprague-Dawley rats. Silk-hydroxyapatite scaffolds soaked in the appropriate concentration of BMP-2 were implanted into the defect. The animals were split into 4 single-injection groups and 3 multiple-injection groups; the latter groups received weekly subcutaneous injections of BMP-2 solution (1, 5, and 10 µg/mL) for 4 weeks, whereas the former groups received a single injection of BMP-2 at these concentrations. Each rat underwent computed tomography at 8 weeks. In terms of total volumes of the new bone, the 5 µg/mL multiple-injection BMP-2 group had significantly greater increases in bone volume than the single-injection groups. In terms of bone thickness, the multiple-injection groups had better outcomes than the single-injection groups. Thus, the cyclic injection protocol restored the original thickness without overgrowth. Cyclic injection of BMP-2 permits more accurate dosage control than single injection and improves thickness and dense bone regeneration. Therefore, it may represent a promising approach for future clinical trials. Further investigation using a greater number of animals is required.