Recombinant Epidermal Growth Factor Research Articles

Preparation of chitosan‐embedded recombinant human epidermal growth factor nanoparticles as accelerating compounds for skin remodelling in chronic lesions

AbstractThe chronic lesion has become a major biological difficulty. Using nanoparticles as drug delivery systems is remarkable nowadays. The unique properties of chitosan in combination with epidermal growth factor (EGF) can accelerate the wound‐healing process.In this study, Chitosan‐EGF (CS‐EGF) nanoparticles were manipulated and characterized by Transmission Electron Microscopy (TEM) microscopy, dynamic light scattering (DLS) method, and Fourier Transform Infrared Spectroscopy (FTIR) radiation. The antibacterial effect was estimated by minimum inhibitory concentration (MIC)/minimum bactericidal concentration (MBC) methods and the proliferation assay was measured on the HFF‐1 (human fibroblast cell line). Then, migration assay was accomplished and the gene expression analysis for transforming growth factor‐beta (TGF‐β), vascular endothelial growth factor (VEGF), and platelets‐derived growth factor (PDGF) was manipulated by the real‐time‐PCR method. The obtained results were considered statistically significant with P < 0.05.Obtained results illustrated no toxic effect on the HFF‐1 cell line treated with CS‐EGF. In cellular proliferation and migration assays, CS‐EGF nanoparticles demonstrated 2‐folds higher than the control. In the duration of 72 h of the experiment, and concentration of 10 µM, 90 ±10% of cells were migrated and the whole scratch was covered by fibroblasts. The real‐time‐PCR analysis also showed 7.5, 4.5‐, and 7‐fold upregulation of all TGF‐β, VEGF, and PDGF genes in comparison with the control group.This study confirmed that using chitosan nanoparticles as a carrier for EGF can accelerate cellular remodelling and annihilate bacterial infection in the process of treatment. As CS‐EGF nanoparticles in the acceleration of the skin remodelling process showed promising results, subsequent studies might be useful.

Final results from a phase II trial of CIMAvax-EGF and nivolumab as second-line (2L) therapy after platinum-based chemotherapy in advanced non-small cell lung cancer (NSCLC).

9135 Background: CIMAvax-EGF (C-E) is a recombinant anti-human epidermal growth factor (EGF) depleting immunotherapy which has previously shown increased survival as maintenance after platinum-based chemotherapy in patients (pts) with advanced NSCLC. The primary objective of this single-arm phase II trial was to evaluate the 12-month overall survival (OS) in pts receiving C-E in combination with Nivolumab(N) as 2L therapy for advanced NSCLC. Methods: Pts with previously treated, immunotherapy-naive advanced NSCLC received 2.4 mg C-E IM every 2 weeks(w) for 4 doses (loading phase) in combination with N 240mg IV every 2 w, then continued monthly maintenance C-E combined with N 240mg IV every 2 w. Enrollment to this arm was terminated before estimated sample size was met due to poor accrual as immunotherapy became incorporated into 1st line therapy. We present OS and progression-free survival (PFS) data [determined using a Kaplan-Meier test with 90% confidence intervals (CI)] of pts who were able to complete the loading phase per protocol (PP). Results: 21 out of 23 enrolled pts were included in the PP analysis. Among the 21 pts, 17 (81.0%) had non-squamous(nsq) histology, 12 (57.1%) were KRAS wildtype (9.5% unknown status), 13 (61.9%) had PD-L1 tumor proportion score 0%. 43% pts (n=3) with known KRAS mutation (n=7) had co-mutated STK11. Disease control rate was 47.6% (n=10) defined as pts who had stable disease or partial response as best response per RECIST v1.1. The 21 PP pts had a 29% 3-year(yr) OS rate (90% CI 14, 45; intention-to-treat [ITT] population in 23 pts with 26% 3-yr OS rate, 90% CI 13, 42). Median(m) OS for PP pts was 11.9 months, 90% CI 8.0 – 23.9 months (ITT mOS 10.4 months, 90% CI 6.8-13.6). Pts with squamous histology had a better 3-yr OS rate compared to those with nsq histology [50% (90% CI 10, 81) vs 24% (90% CI 9, 41), respectively]. Pts with PD-L1 expression ≥1% had higher 3-yr OS [38% (90% CI 12, 63)] and 3-year PFS [38% (90% CI 12, 63)] compared to pts with no PD-L1 expression (3-yr OS 23% [90% CI 8, 44] and 3-yr PFS 8% [90% CI 1, 25]). mOS, 1-yr and 3-yr OS for EGFR/ALK/KRAS wildtype pts was higher [31.7 months (90% CI 5.9, NR), 67% (90% CI 4, 84), 50% (90% CI 24,71), respectively) compared to KRAS mutated NSCLC [10.1 months (90% CI 6.5, 12.1), 29% (90% CI 6, 56), 0% (90% CI 1, 41), respectively]. Conclusions: NSCLC pts who were able to complete the PP combination of C-E plus N at the minimum had numerically better OS compared to historical study cohorts with N as 2L monotherapy. Among pts who completed PP treatment, pts with KRAS wildtype NSCLC had the longest mOS observed. C-E is currently being investigated in combination with pembrolizumab as maintenance therapy after completing 1L chemoimmunotherapy for NSCLC with PD-L1 < 50% and as 1L therapy in combination with pembrolizumab for EGFR/ALK wildtype NSCLC and PD-L1 ≥ 50%. Clinical trial information: NCT02955290 .

Phase 2 trial of epidermal growth factor (EGF) vaccine CIMAvax in combination with pembrolizumab in first line and maintenance setting in advanced non–small cell lung cancer patients.

TPS2677 Background: CIMAvax-EGF is a novel growth-factor depleting immunotherapy consisting of human recombinant EGF conjugated to recombinant P64k derived from Neisseria meningitidis, that elicits an anti-EGF antibody response, resulting in reduction of circulating EGF levels. A randomized phase 3 study of CIMAvax-EGF administered as switch maintenance therapy after first-line platinum-based chemotherapy in patients with advanced NSCLC demonstrated overall survival (OS) advantage in comparison to best supportive care alone, particularly in patients with high baseline serum EGF levels. A phase 1 trial combining CIMAvax-EGF with nivolumab, an anti-PD1 immune checkpoint inhibitor, showed that the combination is safe and appears to induce higher rate of good anti-EGF antibody response within the first 8 weeks of therapy compared to historical comparison group receiving CIMAvax-EGF alone. CIMAvax-EGF is currently being investigated in combination with pembrolizumab in a multi-arm phase II trial as 1L therapy in combination with pembrolizumab for patients with EGFR/ALK wildtype NSCLC and PD-L1 > 50% (Cohort C) and as maintenance therapy after completing 1L chemoimmunotherapy for patients with PD-L1 < 50% (Cohorts D and E). Accrual to this trial is ongoing (NCT02955290). Methods: This is an open-label, non-randomized, multi-site, phase 2 basket trial. CIMAvax-EGF (2.4 mg IM every 2 weeks x 4 doses, then every 4 weeks thereafter) is being tested as first-line therapy in combination with pembrolizumab (200 mg IV every 4 weeks) in patients with advanced NSCLC with PD-L1 ≥ 50% (Cohort C), using a Simon two stage design (n up to 41). A safety lead-in consisting of the first 6 NSCLC patients was completed. There were no G3 or higher adverse events attributed to the combination within the first 4 weeks of protocol treatment. Patients with advanced squamous NSCLC (Cohort D, n=50) and non-squamous NSCLC (without EGFR/ALK/ROS-1/KRAS mutations; Cohort E, n=45) with PD-L1 expression <50% and no evidence of progression of disease by RECIST 1.1 criteria after at least 4 cycles of induction platinum-based doublet with pembrolizumab, prior to initiation of standard of care maintenance therapy will be enrolled to receive protocol treatment with pembrolizumab in combination with CIMAvax-EGF as maintenance therapy. The primary endpoint is to evaluate objective response rate (Cohort C), and 12-month PFS in Cohorts D and E. Secondary endpoints include 12-month overall survival in all cohorts. Exploratory objectives include characterizing tissue-based immune and EGFR signaling profile as well as serum EGF levels, anti-EGF antibody levels and other blood-based biomarkers in relation to clinical outcomes. Clinical trial information: NCT02955290 .

Multifunctional Double-Layer and Dual Drug-Loaded Microneedle Patch Promotes Diabetic Wound Healing.

Chronic nonhealing diabetic wounds are a serious complication of diabetes, with a high morbidity rate that can cause disability or death. The long period of inflammation and dysfunctional angiogenesis are the main reasons for wound-healing difficulty in diabetes. In this study, a multifunctional double-layer microneedle (DMN) is constructed to control infection and promote angiogenesis, meeting the multiple demands of the healing process of a diabetic wound. The double-layer microneedle is consisted in a hyaluronic acid substrate and a mixture of carboxymethyl chitosan and gelatin as the tip. The antibacterial drug tetracycline hydrochloride (TH) is loaded into the substrate of the microneedle to achieve rapid sterilization and promote resistance to external bacterial infections. The microneedle tip loaded with recombinant human epidermal growth factor (rh-EGF) is inserted into the skin, in response to gelatinase produced by resident microbe and disassociate to achieve the enzymatic response release. The double-layer drug-loaded microneedles (DMN@TH/rh-EGF) have antibacterial and antioxidant effects, and promote cell migration and angiogenesis in vitro. In an in vivo diabetic wound model, using rats, the DMN@TH/rh-EGF patch is able to inhibit inflammation, promote angiogenesis, collagen deposition, and tissue regeneration during the wound healing process, promoting its healing.

Crystalline silica-exposed human lung epithelial cells presented enhanced anchorage-independent growth with upregulated expression of BRD4 and EZH2 in autocrine and paracrine manners.

Crystalline silica-induced inflammation possibly facilitates carcinogenesis. Here, we investigated its effect on lung epithelium damage. We prepared conditioned media of immortalized human bronchial epithelial cell lines (hereinafter bronchial cell lines) NL20, BEAS-2B, and 16HBE14o- pre-exposed to crystalline silica (autocrine crystalline silica conditioned medium), a phorbol myristate acetate-differentiated THP-1 macrophage line, and VA13 fibroblast line pre-exposed to crystalline silica (paracrine crystalline silica conditioned medium). As cigarette smoking imposes a combined effect on crystalline silica-induced carcinogenesis, a conditioned medium was also prepared using the tobacco carcinogen benzo[a]pyrene diol epoxide. Crystalline silica-exposed and growth-suppressed bronchial cell lines exhibited enhanced anchorage-independent growth in autocrine crystalline silica and benzo[a]pyrene diol epoxide conditioned medium compared with that in unexposed control conditioned medium. Crystalline silica-exposed nonadherent bronchial cell lines in autocrine crystalline silica and benzo[a]pyrene diol epoxide conditioned medium showed increased expression of cyclin A2, cdc2, and c-Myc, and of epigenetic regulators and enhancers, BRD4 and EZH2. Paracrine crystalline silica and benzo[a]pyrene diol epoxide conditioned medium also accelerated the growth of crystalline silica-exposed nonadherent bronchial cell lines. Culture supernatants of nonadherent NL20 and BEAS-2B in crystalline silica and benzo[a]pyrene diol epoxide conditioned medium had higher EGF concentrations, whereas those of nonadherent 16HBE14o- had higher TNF-α levels. Recombinant human EGF and TNF-α promoted anchorage-independent growth in all lines. Treatment with EGF and TNF-α neutralizing antibodies inhibited cell growth in crystalline silica conditioned medium. Recombinant human TNF-α induced BRD4 and EZH2 expression in nonadherent 16HBE14o-. The expression of γH2AX occasionally increased despite PARP1 upregulation in crystalline silica-exposed nonadherent lines with crystalline silica and benzo[a]pyrene diol epoxide conditioned medium. Collectively, crystalline silica- and benzo[a]pyrene diol epoxide-induced inflammatory microenvironments comprising upregulated EGF or TNF-α expression may promote crystalline silica-damaged nonadherent bronchial cell proliferation and oncogenic protein expression despite occasional γH2AX upregulation. Thus, carcinogenesis may be cooperatively aggravated by crystalline silica-induced inflammation and genotoxicity.

Layer-by-Layer Microneedle-Mediated rhEGF Transdermal Delivery for Enhanced Wound Epidermal Regeneration and Angiogenesis

Appropriate treatments for acute traumas tend to avoid hemorrhages, vascular damage, and infections. However, in the homeostasis-imbalanced wound microenvironment, currently developed therapies could not precisely and controllably deliver biomacromolecular drugs, which are confronted with challenges due to large molecular weight, poor biomembrane permeability, low dosage, rapid degradation, and bioactivity loss. To conquer this, we construct a simple and effective layer-by-layer (LBL) self-assembly transdermal delivery patch, bearing microneedles (MN) coated with recombinant human epidermal growth factor (LBL MN-rhEGF) for a sustained release to wound bed driven by typical electrostatic force. Pyramidal LBL MN-rhEGF patches hold so enough mechanical strength to penetrate the stratum corneum, and generated microchannels allow rhEGF direct delivery in situ. The administrable delivery of biomacromolecular rhEGF through hierarchically coated MN arrays follows the diffusion mechanism of Fick's second law. Numerous efforts further have illustrated that finger-pressing LBL MN-rhEGF patches could not only promote cell proliferation of normal human dermal fibroblasts (NHDF) and human umbilical vein endothelial cells (HUVEC) in vitro but also take significant effects (regenerative epidermis: ∼144 μm; pro-angiogenesis: higher CD31 expression) in accelerating wound healing of mechanically injured rats, compared to the traditional dressing, which relies on passive diffusion. Our proof-of-concept features novel LBL biomacromolecular drug-delivery systems and self-administrated precision medicine modes at the point of care.

Biopotentials of Collagen Scaffold Impregnated with Plant-Cell-Derived Epidermal Growth Factor in Defective Bone Healing.

The combination of scaffolds with recombinant human epidermal growth factor (rhEGF) protein can enhance defective bone healing via synergistic activation to stimulate cellular growth, differentiation, and survival. We examined the biopotentials of an rhEGF-loaded absorbable collagen scaffold (ACS) using a mouse model of calvarial defects, in which the rhEGF was produced from a plant cell suspension culture system because of several systemic advantages. Here, we showed a successful and large-scale production of plant-cell-derived rhEGF protein (p-rhEGF) by introducing an expression vector that cloned with its cDNA under the control of rice α-amylase 3D promoter into rice calli (Oryza sativa L. cv. Dongjin). Implantation with p-rhEGF (5 μg)-loaded ACSs into critical-sized calvarial defects enhanced new bone formation and the expression of osteoblast-specific markers in the defected regions greater than implantation with ACSs alone did. The potency of p-rhEGF-induced bone healing was comparable with that of Escherichia coli-derived rhEGF protein. The exogenous addition of p-rhEGF increased the proliferation of human periodontal ligament cells and augmented the induction of interleukin 8, bone morphogenetic protein 2, and vascular endothelial growth factor in the cells. Collectively, this study demonstrates the successful and convenient production of p-rhEGF, as well as its potency to enhance ACS-mediated bone regeneration by activating cellular responses that are required for wound healing.

Abstract 174: Organoids developed from synovial sarcoma patient-derived xenografts

Abstract Background: Soft tissue sarcoma (STS) is a heterogeneous family of rare malignancies. Despite this heterogeneity, systemic treatment options for patients with advanced STS are rather uniform and have very limited efficacy. There is an unmet medical need to develop more active, subtype-specific treatment options. Three-dimensional (3D) cell culture models (organoids) have been successfully used for drug testing in carcinoma research. However, the development of STS organoids is lagging behind. We attempted to identify the best growth conditions for organoids derived from synovial sarcoma (SySa), which is one of the more common translocation-related subtypes of STS. Methods: Human synovial sarcoma tissue was implanted subcutaneously into immunodeficient mice and grown to create a patient-derived xenograft (PDX). Fresh tumor tissue from the PDX model was collected and enzymatically and mechanically dissociated into small cell clusters. The cells were seeded into a 3D scaffold with advanced Dulbecco's modified Eagle’s medium (DMEM)/F12 and different growth factors were added. In order to obtain optimal results, different scaffolds and growth factors were compared. The growth of the 3D cultures was evaluated by bright-field microscopy and a cell viability assay using PrestoBlue. The organoids were characterized by histopathological stainings and the presence of the SSX-SS18 translocation was molecularly confirmed. Finally, we attempted to create organoids from PDX-derived material stored in liquid nitrogen. Results: A scaffold consisting of 50% collagen type 1, 30% Matrigel, 20% DMEM 10x and sodium hydroxide was superior to Matrigel alone. The addition of human recombinant epidermal growth factor (EGF, 10 ng/ml), human insulin-like growth factor 1 (IGF-1, 20 ng/ml), human recombinant fibroblast growth factor 2 (FGF-2, 10 ng/ml), and N-acetylcysteine (NAC, 500 mmol) to advanced DMEM/F12 stimulated the growth of SySa organoids the best. The Rho-kinase inhibitor Y-27632 was added to prevent apoptosis during early passages. So far, attempts to create organoids from frozen material have been unsuccessful. Conclusions: At present, the success rate of establishing organoids from synovial sarcoma cells is limited if the culture conditions are not adapted to their needs. We identified that growth factors such as EGF, FGF-2, IGF-1, and NAC are beneficial for growth. The availability of fresh samples plays an important role in the development of these 3D models. Despite these limitations, organoids can be an interesting tool for studying this rare disease and aid in the development of new drugs. Citation Format: Lore De Cock, Agnieszka Wozniak, Kimberly Verbeeck, Patrick Schöffski. Organoids developed from synovial sarcoma patient-derived xenografts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 174.

RECOMBINANT HUMAN EPIDERMAL GROWTH FACTOR IN TREATING DIABETIC FOOT ULCERS-A RANDOMISED CONTROLLED TRIAL

Background: Diabetic foot ulcers are among the most common complications in patients with poor glycemic control. Diabetic foot ulcers are the result of poor glycemic control, underlying neuropathy, peripheral vascular disease, or poor foot care. Diabetic foot ulcers are a debilitating complication of diabetes mellitus that leads to increased overall morbidity in patients. When the recombinant human epidermal growth factor (rhEGF) is used as the topical application it stimulates epithelial proliferation, differentiation, and migration. As a result of this, there is an acceleration of epithelial regeneration and wound healing of diabetic ulcers. The Aim and Objective: aim is to study the efcacy of recombinant human epidermal growth factor by comparing the time taken for healing, cost-effectiveness, patient compliance, and duration of hospital stay after topical application of epidermal growth factor over a control group treated with conventional dressings of saline and hydrogen peroxide, which acts as the control group. This study is a randomized controlled trial conducted in the Methods: Department of General Surgery, Kanyakumari Government Medical College, from September 2022 to February 2023. A total of 60 patients were divided into two groups: 30 patients in the interventional group treated with rhEGF and 30 patients in the control group receiving conventional dressings with saline and hydrogen peroxide were monitored, and the wound healing time, patient compliance, cost effectiveness, and duration of hospital stay were studied. A total of Results: 60 patients of which 38 (63.3%) were males and 22(36.6%) were female patients. The wound healing was better in the group treated with rhEGF and the duration of hospital stay was signicantly shorter. Patient compliance was found to be similar in both groups & conventional wound dressing was found to be cost effective as rhEGF topical application is a costly affair. This study suggests that recombinant human Conclusion: epidermal growth factor helps in the acceleration of epithelial regeneration and wound healing in diabetic ulcers. Patients who were treated with recombinant human epidermal growth factor had a signicant reduction in wound healing time and decreased duration of hospital stay.

A Comparative Study on Therapeutic Efficacy of Autologous Platelet-rich Plasma, Autologous Platelet-rich Fibrin Matrix, Recombinant Human Epidermal Growth Factor, and Collagen Particles in Nonhealing Leg Ulcers.

Nonhealing leg ulcers are challenging to manage and cause significant patient morbidity. To promote healing, newer techniques focus on delivering/enhancing dermal matrix components. The aim of this study was to compare the therapeutic efficacy of autologous platelet-rich plasma (PRP), autologous platelet-rich fibrin matrix (PRFM), recombinant human epidermal growth factor (rhEGF), and collagen particles in treating nonhealing leg ulcers. Open, randomized prospective study was conducted in a single tertiary center over 2 years where after fulfilling the criteria, randomization was done into four groups. Group A: Autologous PRP (double spin, manual method, weekly); Group B: Autologous PRFM (weekly); Group C: rhEGF (daily application); and Group D: Collagen particles (weekly) along with cleansing, debris removal, and wound dressing. Treatment endpoints were complete healing/6 months of treatment, whichever was earlier. Follow-up was done two weekly by clinical assessment, photographs, and measurement of the ulcer area. Epi info 7 software was used for statistical analysis. A total of 48 patients completed the study, 12 in each group, with mean age: 42.37 ± 4.56 years and male-to-female ratio 2.6:1. Underlying etiology was varicosities (43.75%), traumatic (25%), diabetes (22.91%), and leprosy (8.34%). At baseline, all groups were comparable in terms of patient and ulcer characteristics. Complete healing was seen in 79.17% at the end of 12 weeks: 91.67% of patients from Groups A and B each, and 66.67% from Groups C and D each. The mean time to complete healing was 6.9 ± 2.5 weeks, the least in Group B (4.73 ± 2.3 weeks). Differences between excellent (≥75%) ulcer healing across all groups were statistically significant at the end of 8 weeks where Group B showed maximum improvement. No major adverse events were seen. PRFM resulted in relatively faster ulcer healing compared with other modalities.

Intralesional epidermal growth factor therapy in recalcitrant diabetic foot ulcers

Diabetic foot ulcers (DFUs) cause high morbidity and mortality despite best treatment. Thus, new products are urgently needed to treat DFUs. Intralesional epidermal growth factor (EGF) (Heberprot-p) is considered to be an adjuvant therapy to standard of care (SOC) in DFUs. In the present study, the effect of Heberprot-p treatment on wound healing is compared to standard treatment. The data of patients with DFUs were retrospectively analysed. The patients who had had DFUs of at least four weeks' duration and who had been treated in the wound clinic between January 2014 and 2017 were included in the study. The patients were divided into study and control groups. The study group consisted of patients in whom intralesional recombinant human EGF, Heberprot-p 75μg, was applied; the control group consisted of the remaining patients in whom EGF was not applied. The efficacy of Heberprot-p treatment in Wagner 2 and 3 DFUs were retrospectively investigated. The study group (n=29 patients) who received Heberprot-p treatment was found to have shorter treatment times and higher rates of wound healing than the control group (n=22 patients). Although the amputation rate in the study group was less than the control group, the difference was not statistically significant. Heberprot-p therapy is a promising treatment in DFUs, which can be routinely used as an adjunct to standard care.

The effect of human recombinant epidermal growth factor on capsule contraction in an irradiated rat model

The effect of human recombinant epidermal growth factor on capsule contraction in an irradiated rat model

Effect of combined use of sodium hyaluronate and rhEGF on xerophthalmia in the elderly after cataract surgery

Purpose: To determine the effect of combined use of sodium hyaluronate and recombinant human epidermal growth factor (rhEGF) on xerophthalmia in elderly patients after cataract surgery, as well as levels of serum inflammatory factors.
 Methods: A total of 84 elderly patients diagnosed with xerophthalmia after phacoemulsification were retrospectively studied. The patients comprised two groups: control group consisting of 40 patients given basic therapy using anti-inflammatory drugs and sodium hyaluronate eye drops (1 drop 3 times a day), and combination group (n = 44) who additionally received rhEGF. The expression levels of interleukin-6 (IL-6), high-sensitivity C-reactive protein (hs-CRP), and tumor necrosis factor (TNF-α) in tears were determined using enzyme-linked immunosorbent assay (ELISA). After 4 weeks of treatment, clinical efficacy was assessed.
 Results: There was significantly higher clinical efficacy in the combination group than in the control group (p < 0.05). The values of tear break-up time (BUT) and Schirmer test (Sit) in both groups increased significantly with time, while fluorescein corneal staining (FL), symptom score, and expression levels of IL-6, hs-CRP and TNF-α significantly decreased (p < 0.05). At 2 weeks and 4 weeks of therapy, the combination group had higher values of BUT and Sit than the control group (p < 0.01). However, FL value, symptom score, and levels of IL-6, hs-CRP and TNF-α were significantly lower in the combination group than in the control group (p < 0.01).
 Conclusion: Sodium hyaluronate, in combination with rhEGF, improves treatment efficacy in elderly xerophthalmia patients after operation, reduces inflammatory reactions, and enhances disease alleviation. However, a multi-center prospective study will be required to validate the findings of this study.

Development, Validation and Application of a Bridging ELISA for Detection of Antibodies against GQ1001 in Cynomolgus Monkey Serum.

Immunogenicity is a major issue associated with the PK, efficacy, and safety evaluation of therapeutic protein products during pre-clinical and clinical studies. A multi-tiered approach consisting of screening, confirmatory, and titration assays has been widely adopted for anti-drug antibody testing. GQ1001, a recombinant humanized anti-human epidermal growth factor receptor 2 monoclonal antibody covalently linked to a cytotoxin of DM1, possesses a novel format of antibody-drug conjugates. In this study, we reported the development, validation, and application of an acid-dissociation bridging enzyme-linked immunosorbent assay for the detection of antibodies against GQ1001 in cynomolgus monkey serum. The sensitivity of the screening assay was 126.141 ng/mL in undiluted serum. The screening assay and confirmatory assay were neither affected by the naïve monkey serum nor by 2% and 5% (v/v) erythrocyte hemolysates. Moreover, the assay was not subject to interference by 2500 ng/mL of human IgG1 in the samples. Drug interference at low positive control (150 ng/mL) and high positive control (8000 ng/mL) of anti-GQ1001 antibodies was not observed when GQ1001 concentrations were below 3.125 μg/mL and 100 μg/mL, respectively. Furthermore, no hook effect was observed for the positive antibodies in the concentration range of 8 to 64 μg/mL. The validated assay was, thereafter, successfully applied to a single-dose toxicity study of GQ1001. Anti-drug antibody positive rates among dosing animals and testing samples were reported, and no significant impact was found on toxicokinetic outcomes.

Efficient expression of fusion human epidermal growth factor in tobacco chloroplasts

BackgroundChloroplast transformation is a robust technology for the expression of recombinant proteins. Various types of pharmaceutical proteins including growth factors have been reported in chloroplasts via chloroplast transformation approach at high expression levels. However, high expression of epidermal growth factor (EGF) in chloroplasts with the technology is still unavailable.ResultsThe present work explored the high-level expression of recombinant EGF, a protein widely applied in many clinical therapies, in tobacco chloroplasts. In this work, homoplastic transgenic plants expressing fusion protein GFP-EGF, which was composed of GFP and EGF via a linker, were generated. The expression of GFP-EGF was confirmed by the combination of green fluorescent observation and Western blotting. The achieved accumulation of the recombinant fusion GFP-EGF was 10.21 ± 0.27% of total soluble proteins (1.57 ± 0.05 g kg− 1 of fresh leaf). The chloroplast-derived GFP-EGF was capable of increasing the cell viability of the NSLC cell line A549 and enhancing the phosphorylation level of the EGF receptor in the A549 cells.ConclusionThe expression of recombinant EGF in tobacco chloroplasts via chloroplast transformation method was achieved at considerable accumulation level. The attempt gives a good example for the application of chloroplast transformation technology in recombinant pharmaceutical protein production.

Retracted: Effects of Recombinant Human Epidermal Growth Factor Eye Drops Combined with Phacoemulsification on Short‐ and Long‐Term Visual Acuity Recovery and Related Dry Eye Complications in Patients with Senile Cataract

Retracted: Effects of Recombinant Human Epidermal Growth Factor Eye Drops Combined with Phacoemulsification on Short‐ and Long‐Term Visual Acuity Recovery and Related Dry Eye Complications in Patients with Senile Cataract

A biocompatible polyethylene glycol/alginate composite hydrogel with significant reactive oxygen species consumption for promoting wound healing.

In areas of wound inflammation, excessive reactive oxygen species (ROS) may worsen the infection and lead to tissue damage, resulting in a vicious circle. Therefore, many hydrogels with sensitive ROS consumption capabilities and antibacterial properties have been widely developed and applied. These hydrogels usually achieve their ROS consumption capacity by introducing reactive active groups: however, these materials normally require complicated preparation procedures and have high potential toxicity. Inspired by these limitations, an integrated polyethylene glycol/alginate-based hydrogel (itg-PEGDA@SA) has been developed via a simple two-step process, where the inner PEGDA hydrogel (hdg-PEGDA) acts as a ROS scavenger and the outer sodium alginate hydrogel (SA) can be degraded to act as a recombinant human epidermal growth factor (rhEGF)-loaded platform to enhance the functionality of this composite hydrogel. Altogether, the itg-PEGDA@SA hydrogel showed significant ROS consumption and biocompatibility in vitro, and when applied for wound healing, the formation of uniform and ordered collagen fibers (stained using aniline blue) can be achieved. This hydrogel showed favorable attributes in ROS scavenging, and it can be a promising material for use in wound dressings and biomaterial fields.

The use of epidermal growth factor in dermatological practice.

Epidermal growth factor (EGF) is a growth factor that plays a pivotal role in wound healing and maintaining tissue homeostasis by regulating cell survival, proliferation, migration, and differentiation. Exogenous administration of bioidentical human recombinant epidermal growth factor (rhEGF) has been known to promote skin wound healing, although rhEGF is increasingly being used in drug delivery systems and nanotechnology. However, despite considerable attention being focused on the potential clinical applications of rhEGF in several dermatological conditions beyond wound healing, the number of studies still remains relatively low. Herein, we conducted a literature search of PubMed/Medline and Google Scholar databases to retrieve published literature related to rhEGF and summarised the effects of rhEGF in the treatment of various wound types, radiotherapy or chemotherapy-related skin reactions, atopic dermatitis, skin aging, and post-inflammatory hyperpigmentation.

Влияние интравитреального введения эпидермального фактора роста на морфофункциональное состояние глаза кролика

Purpose. To study the morphofunctional state of the rabbit eye after intravitreal injection of epidermal growth factor. Material and methods. In an experiment on rabbits, intravitreal course administration of Eberprot-P containing recombinant epidermal growth factor (EGF) at a dose of 100 ng was carried out. 7 and 30 days after each injection, biomicroscopy and ophthalmoscopy of the eye, optical coherence tomography (OCT) of the fundus in vivo were performed. Morphological studies were performed according to the generally accepted method with staining of histological preparations with hematoxylin and eosin. The cell density of the ganglionic layer, the inner and outer nuclear layers of the retina was assessed. SPSS software for Windows was used for statistical analysis. Results. Biomicroscopic and ophthalmoscopic studies of the intraocular structures of the experimental eyes did not reveal signs of edema or intraocular inflammation, toxic or irritant effects of intravitreal injection. There were no pathological changes in the form of cataracts, Tyndall's phenomenon in the anterior chamber moisture, vitreous opacities, retinal infiltrates. According to OCT data, the retinal thickness remained unchanged: 158 ± 5µm versus the control 158 ± 3µm (p = 1.0), no retinal wrinkling effects were detected. The morphological picture of the intraocular tissues of the eye after intravitreal injection was characterized by the absence of any pathological changes in the architectonics of the layers and cellular structures of the retina. Histomorphometric analysis showed no difference in the cell density of the ganglion layer, inner and outer nuclear layers of the retina of the experimental and control eyes of animals. Conclusion. Intravitreal application of EGF in rabbits does not cause intraocular specific inflammation or any destructive effects in the retina, which indicates the potential for its clinical use in the treatment of dystrophic retinal diseases. Keywords: epidermal growth factor, retina, intravitreal injection, morphology, histomorphometry

The effect of recombinant human epidermal growth factor on radiation dermatitis in rectal and anal cancer patients: a self-controlled study

BackgroundOur previous study reported that recombinant human epidermal growth factor (rhEGF)-triggered EGFR internalization promoted radioresistance. Here, we aimed to evaluate the effect of rhEGF on the skin protection of rectal and anal cancer patients receiving radiotherapy.MethodsOne hundred and ninety-three rectal and anal cancer patients who received radiotherapy were prospectively enrolled from January 2019 to December 2020. To perform self-controlled study, the left and right pelvic skin area (separated by midline) were randomly assigned to the rhEGF and control side. The association between radiation dermatitis and factors including rhEGF, the dose of radiotherapy and tumor distance from anal edge were analyzed.ResultsAmong 193 enrolled patients, 41 patients (21.2%) did not develop radiation dermatitis, and 152 patients (78.8%) suffered radiation dermatitis on at least one side of pelvic skin at the end of radiotherapy. For the effect on radiation dermatitis grade, rhEGF had improved effect on 6 (4.0%) patients, detrimental effect on 2 (1.3%) patients, and no effect on 144 (94.7%) patients. Whereas for the effect on radiation dermatitis area, rhEGF showed improved effect on the radiation dermatitis area of 46 (30.2%) patients, detrimental effect on 15 (9.9%) patients, and no effect on 91 (59.9%) patients. The radiation dermatitis area of rhEGF side was significantly smaller than that of control side (P = 0.0007).ConclusionsrhEGF is a skin protective reagent for rectal and anal cancer patients receiving radiotherapy.Trial registrationChinese Clinical Trial Registry identifier: ChiCTR1900020842; Date of registration: 20/01/2019.