Nanofiber Dressings Topically Delivering Molecularly Engineered Human Cathelicidin Peptides for the Treatment of Biofilms in Chronic Wounds.

Biofilms of multidrug-resistant bacteria in chronic wounds pose a great challenge in wound care. Herein, we report the topical delivery of molecularly engineered antimicrobial peptides using electrospun nanofiber dressings as a carrier for the treatment of biofilms of multidrug-resistant bacteria in diabetic wounds. Molecularly engineered human cathelicidin peptide 17BIPHE2 was successfully encapsulated in the core of pluronic F127/17BIPHE2-PCL core-shell nanofibers. The in vitro release profiles of 17BIPHE2 showed an in initial burst followed by a sustained release over 4 weeks. The peptide nanofiber formulations effectively killed methicillin-resistant Staphylococcus aureus (MRSA) USA300. Similarly, the 17BIPHE2 peptide containing nanofibers could also effectively kill other bacteria including Klebsiella pneumoniae (104 to 106 CFU) and Acinetobacter baumannii (104 to 107 CFU) clinical strains in vitro without showing evident cytotoxicity to skin cells and monocytes. Importantly, 17BIPHE2-containing nanofiber dressings without debridement caused five-magnitude decreases of the MRSA USA300 CFU in a biofilm-containing chronic wound model based on type II diabetic mice. In combination with debridement, 17BIPHE2-containing nanofiber dressings could completely eliminate the biofilms, providing one possible solution to chronic wound treatment. Taken together, the biodegradable nanofiber-based wound dressings developed in this study can be utilized to effectively deliver molecularly engineered peptides to treat biofilm-containing chronic wounds.

Aesthetic Reconstruction Based on Facial Subunit Principle for Basal Cell Carcinoma of the Face: A Retrospective Analysis.

Background and objective Basal cell carcinoma (BCC) is the most common malignancy of the skin. Reconstruction of post-excisional defects in BCC should follow the subunit principle for better outcomes. The location of BCC of the face is determined based on facial units; however, very few studies have described the involvement of multiple units and multiple subunits in BCC. In this study, we aimed to provide valuable insights into the management of BCC involving various facial units and subunits, thereby contributing to improved patient care and outcomes. Materials and methods We conducted a retrospective study at the Plastic Surgery Department of the SCB Medical College in Cuttack, Odisha, from January 2020 to January 2022, after obtaining ethical approval from the SCB Medical College IRB (no: 1155). We examined 35 patients with BCC of the face. The inclusion criteria were as follows: patients with early-stage and primary tumors that were mobile, not attached to underlying bone or cartilage, and amenable to surgical resection. Conversely, patients with late-stage, neglected, and recurrent tumors, fixed tumors, or those infiltrating the underlying bone or cartilage were excluded from the study. Data collection involved retrieving pertinent information from medical records, including parameters such as age, sex, tumor site, type of flap utilized, follow-up, and any complications observed. The tumor sites were further divided into six separate groups based on facial aesthetic units: the forehead, the nose, the area around the eyes, the cheek, the mouth, and the area around the ear, each with its own subunits. Results A total of 35 patients were included in this study, comprising 15 males (42.85%) and 20 females (57.15%), with a male-to-female ratio of 1:1.33. The ages of the patients ranged from 42 to 68 years. Among the facial units, the nose was the most commonly involved (in seven cases), while the lip was the least commonly affected (in one case). In 24 cases, a single unit was involved, while 11 cases involved multiple units. Furthermore, single subunits were affected in 18 cases, double subunits in 10 cases, three subunits in five cases, four subunits in one case, and five subunits in another case. Notably, no cases exhibited flap necrosis, wound dehiscence, wound hematoma, or seroma, indicating excellent surgical outcomes. All flaps remained viable, and all patients were followed up for a minimum of one year, with no reported recurrence during the follow-up period ranging from 6 to 18 months, reaffirming the effectiveness of the treatment approach. Conclusions For small, superficial lesions, full-thickness skin grafts (FTSG) are a suitable treatment option. However, when dealing with larger lesions that encompass multiple subunits, the preferred approach involves reconstructing with locoregional flaps. It is essential to plan the procedure carefully, taking into account the goal of positioning the final scar along the junction of facial subunits. This strategic plan aims to achieve superior aesthetic outcomes.

Peripheral Odontogenic Myxoma of Zygoma and Orbital Region - A Unique Case Report with Review of Literature.

Rationale: Peripheral odontogenic myxoma (POM) is a rare mesenchymal tumour and it is the first case report of POM involving orbital and zygoma region. Patient Concerns: A 16-year-old male presented with a painless, slow-growing swelling over his left infratemporal region. Diagnosis: The histopathological examination of the tumour was diagnosed as POM. Treatment: The patient was treated by surgical removal of tumour under general anaesthesia. Outcomes: The patient has been under follow-up for the past 2.5 years and there has been no recurrence. Take-away Lessons: POM is a rare mesenchymal tumour. To our knowledge, this is only the second report of a POM of the infratemporal region and the first report of a myxoma, which extends into the zygomatic region and lateral wall of the orbit.

Bactericidal and biocatalytic temperature responsive microgel based self-cleaning membranes for water purification.

The present study focuses on creating an antimicrobial and biocatalytic smart gating membrane by synthesizing unique core-shell microgels. The core-shell microgels are synthesized by grafting short chains of poly(ethylenimine) (PEI) onto a poly((N-isopropyl acrylamide)-co-glycidyl methacrylate)) (P(NIPAm-co-GMA)) core. Subsequently, the produced microgels are utilized as a substrate for synthesizing and stabilizing silver nanoparticles (Ag NPs) through an in-situ approach. These Ag NPs immobilized microgels are then suction filtered over a polyethylene terephthalate (PET) track-etched support to create cross-linked composite microgel membranes (CMMs). After structural and permeation characterization of the prepared CMMs, the laccase enzyme is then covalently grafted to the surface of the membrane and tested for its effectiveness in degrading Reactive red-120 dye. The laccase immobilized biocatalytic CMMs show effective degradation of the Reactive red-120 by 71%, 48%, and 34% at pH 3, 4, and 5, respectively. Furthermore, the immobilized laccase enzyme showed better activity and stability in terms of thermal, pH, and storage compared to the free laccase, leading to increased reusability. The unique combination of Ag NPs and laccase on a thermoresponsive microgel support resulted in a responsive self-cleaning membrane with excellent antimicrobial and dye degradation capabilities for environmentally friendly separation technology.

Drug-Induced Oromandibular Dystonia Presenting as Chronic Temporomandibular Joint Dislocation: A Rare Case Report.

Approximately 15%-30% of patients receiving neuroleptic medication for a longer duration develop drug-induced dystonia. There are many variations of oromandibular dystonia (OMD), but the most common one is involuntary jaw-opening dystonia. A rare case of chronic mandibular dislocation under long-term neuroleptic therapy is reported with clinical features, diagnosis, and various treatment modalities. Chronic dislocation leads to changes in associated soft tissue and muscles. Therefore, besides alteration of bony articular surfaces (eminectomy), soft tissue remodeling is required to achieve the perfect balance for temporomandibular joint (TMJ) working and occlusion. Drug-induced orofacial dystonia presenting as chronic TMJ dislocation is rare. Therefore, in long-standing chronic dislocation cases during treatment, biomechanics of TMJ, its complex neurological system, and the physiology of the masticatory system should be considered to customize the treatment plan.

Metastasis in the mandible involving gingiva: An intriguing case with a perplexing pathology.

Oral metastasis, although rare, tends to involve jawbones, particularly the posterior region of the mandible, and involvement of oral soft tissues, even when less likely, is most often seen on the gingiva and tongue. Clinically, the soft-tissue masses tend to mimic pyogenic granuloma, peripheral giant cell granuloma or an epulis and thus are difficult to diagnose and identify. The jaw bone is preferred by prostate carcinoma as a metastatic target. Prostate malignancy, which is more common in Western countries than in India, may be adenocarcinomas or carcinomas. Oftentimes, metastatic lesions develop in the alveolar region and are a cause for tooth mobility, yet, they tend to be detected only after extraction of the affected tooth. In such cases, the symptomatic presentation therefore, is vague and indicative of tooth mobility secondary to periodontal pathology unless, a detailed history and follow-up is done. We report a case of a male patient who presented to our department with a proliferative, painful, swelling postextraction of the left first molar region, and the lesion was seen at the extraction site as well as in the mandibular anterior tooth region. The swelling was associated with palpable lymph nodes. Orthopantomogram showed an irregular, radiolucent lesion extending from the lower left central incisor to the left first molar region in the mandibular alveolus. Incisional biopsy tissue came with provisional diagnosis of osteomyelitis or squamous cell carcinoma as the patient was a habitual bidi smoker for more than 20 years. Histologically, it was an undifferentiated tumor with tumor cells seen in deep connective tissue with a lack of lineage differentiation. An undifferentiated malignant tumor represents either a metastasis of unknown origin or a primary neoplasia without obvious cell line of differentiation. Immunohistochemistry (IHC) of undifferentiated tumors helps to categorize them into small round blue cell tumors or large cell tumors. The oral pathologist was perplexed as there was no mention of any other malignancy in the patient's history, which, however, was noted by the surgeons few days later. Hence, initially, a hematopoietic malignancy was suspected which was ruled out by IHC, and later, staining with cytokeratin 7 (CK7), CK-high molecular weight and P63 confirmed prostate metastases as all three were negative.

Osteosarcoma of jaws: Challenges in diagnosis.

Osteosarcoma (OS) accounts for about 20% of all sarcomas with gnathic involvement seen in about 6%-10% of all OSs. The clinical presentation of OSs in the jaws is different from that of long bones as swelling is the most common complaint in patients with jaw OS followed by pain. The histopathologic variables seen are more favorable in OSs of jaws. Low-grade tumors are Stage I, high-grade tumors are Stage II and metastatic tumors (regardless of grade) are Stage III. A 17-year-old male patient reported with a complaint of the presence of an intra-oral growth gradually increasing in size in the right buccal mucosa region soft tissue enveloping the occlusal aspect of the right mandibular second molar. Extraorally swelling was present on the right side of the face for 4 months. Radiographically, there was a radiolucency from the distal aspect of right Mandibular second molar extending into the ramus region of the mandible with ill-defined borders. Hemi-mandibulectomy was done with the removal of the right mandible from the premolar region to condyle and coronoid processes. Microscopic evaluation of the sections after hematoxylin and eosin staining revealed interlacing fascicles of spindle-shaped cells arranged in a biphasic pattern and some areas of attempted bone formation evident in deeper sections. Tumor was an osteoblastic variety consisting of tumor osteoid surrounded by bizarrely arranged fibroblast-like cells. It showed positive staining with α-smooth muscle actin and Vimentin, suggesting a malignant tumor of mesenchymal cells with high myofibroblastic activity. Our case had small-cell histology; therefore, differential diagnosis was important.

Design and surface immobilization of short anti-biofilm peptides.

Short antimicrobial peptides are essential to keep us healthy and their lasting potency can inspire the design of new types of antibiotics. This study reports the design of a family of eight-residue tryptophan-rich peptides (TetraF2W) obtained by converting the four phenylalanines in temporin-SHf to tryptophans. The temporin-SHf template was identified from the antimicrobial peptide database (http://aps.unmc.edu/AP). Remarkably, the double arginine variant (TetraF2W-RR) was more effective in killing methicillin-resistant Staphylococcus aureus (MRSA) USA300, but less cytotoxic to human skin HaCat and kidney HEK293 cells, than the lysine-containing dibasic combinations (KR, RK and KK). Killing kinetics and fluorescence spectroscopy suggest membrane targeting of TetraF2W-RR, making it more difficult for bacteria to develop resistance. Because established biofilms on medical devices are difficult to remove, we chose to covalently immobilize TetraF2W-RR onto the polyethylene terephthalate (PET) surface to prevent biofilm formation. The successful surface coating of the peptide is supported by FT-IR and XPS spectroscopies, chemical quantification, and antibacterial assays. This peptide-coated surface indeed prevented S. aureus biofilm formation with no cytotoxicity to human cells. In conclusion, TetraF2W-RR is a short Trp-rich peptide with demonstrated antimicrobial and anti-biofilm potency against MRSA in both the free and immobilized forms. Because these short peptides can be synthesized cost effectively, they may be developed into new antimicrobial agents or used as surface coating compounds. STATEMENT OF SIGNIFICANCE: It is stunning that the total deaths due to methicillin-resistant Staphylococcus aureus (MRSA) infection are comparable to AIDS/HIV-1, making it urgent to explore new possibilities. This study deals with this problem by two strategies. First, we have designed a family of novel antimicrobial peptides with merely eight amino acids, making it cost effective for chemical synthesis. These peptides are potent against MRSA USA300. Our study uncovers that the high potency of the tryptophan-rich short peptide is coupled with arginines, whereas these Trp- and Arg-rich peptides are less toxic to select human cells than the lysine-containing analogs. Such a combination generates a more selective peptide. As a second strategy, we also demonstrate successful covalent immobilization of this short peptide to the polyethylene terephthalate (PET) surface by first using a chitosan linker, which is easy to obtain. Because biofilms on medical devices are difficult to remove by traditional antibiotics, we also show that the peptide coated surface can prevent biofilm formation. Although rarely demonstrated, we provide evidence that both the free and immobilized peptides target bacterial membranes, rendering it difficult for bacteria to develop resistance. Collectively, the significance of our study is the design of novel antimicrobial peptides provides a useful template for developing novel antimicrobials against MRSA. In addition, orientation-specific immobilization of the same short peptide can prevent biofilm formation on the PET surface, which is widely used in making prosthetic heart valves cuffs and other bio devices.

Antimicrobial functionalization of silicone surfaces with engineered short peptides having broad spectrum antimicrobial and salt-resistant properties.

Catheter-associated urinary tract infections (CAUTIs) are often preceded by pathogen colonization on catheter surfaces and are a major health threat facing hospitals worldwide. Antimicrobial peptides (AMPs) are a class of new antibiotics that hold promise in curbing CAUTIs caused by antibiotic-resistant pathogens. This study aims to systematically evaluate the feasibility of immobilizing two newly engineered arginine/lysine/tryptophan-rich AMPs with broad antimicrobial spectra and salt-tolerant properties on silicone surfaces to address CAUTIs. The peptides were successfully immobilized on polydimethylsiloxane and urinary catheter surfaces via an allyl glycidyl ether (AGE) polymer brush interlayer, as confirmed by X-ray photoelectron spectroscopy and water contact angle analyses. The peptide-coated silicone surfaces exhibited excellent microbial killing activity towards bacteria and fungi in urine and in phosphate-buffered saline. Although both the soluble and immobilized peptides demonstrated membrane disruption capabilities, the latter showed a slower rate of kill, presumably due to reduced diffusivity and flexibility resulting from conjugation to the polymer brush. The synergistic effects of the AGE polymer brush and AMPs prevented biofilm formation by repelling cell adhesion. The peptide-coated surface showed no toxicity towards smooth muscle cells. The findings of this study clearly indicate the potential for the development of AMP-based coating platforms to prevent CAUTIs.

Immobilization studies of an engineered arginine-tryptophan-rich peptide on a silicone surface with antimicrobial and antibiofilm activity.

With the rapid rise of antibiotic-resistant-device-associated infections, there has been increasing demand for an antimicrobial biomedical surface. Synthetic antimicrobial peptides that have excellent bactericidal potency and negligible cytotoxicity are promising targets for immobilization on these target surfaces. An engineered arginine-tryptophan-rich peptide (CWR11) was developed, which displayed potent antimicrobial activity against a broad spectrum of microbes via membrane disruption, and possessed excellent salt resistance properties. A tethering platform was subsequently developed to tether CWR11 onto a model polymethylsiloxane (PDMS) surface using a simple and robust strategy. Surface characterization assays such as attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR), X-ray photoelectron spectroscopy (XPS), and energy-dispersive X-ray spectroscopy (EDX) confirmed the successful grafting of CWR11 onto the chemically treated PDMS surface. The immobilized peptide concentration was 0.8 ± 0.2 µg/cm(2) as quantitated by sulfosuccinimidyl-4-o-(4,4-dimethoxytrityl) butyrate (sulfo-SDTB) assay. Antimicrobial assay and cytotoxic investigation confirmed that the peptide-immobilized surface has good bactericidal and antibiofilm properties, and is also noncytotoxic to mammalian cells. Tryptophan-arginine-rich antimicrobial peptides have the potential for antimicrobial protection of biomedical surfaces and may have important clinical applications in patients.