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Extracellular vesicles (EVs) are considered essential mediators of regenerative roles of autologous platelet- and extracellular vesicle-rich plasma (PVRP) and platelet- and extracellular vesicle-rich gel (PVRG). PVRP and PVRG are novel blood-derived products gaining attraction in regenerative medicine. However, despite their reported good efficacy, their preparation protocols are too time-consuming. Moreover, patient-tailored preparation protocols are desired to optimize platelet and EV count in PVRP and PVRG. This article presents the clinical implementation of one-step, patient-tailored erythrocyte sedimentation rate (ESR)-based, PVRP and PVRG preparation protocols through the presentation of three cases: (1) large chronic tympanic membrane (TM) perforation, (2) osteoradionecrosis of the lateral skull base, and (3) cerebrospinal fluid (CSF) leak in the sphenoid sinus. These were treated with PVRP and PVRG, prepared according to our preclinically constructed mathematical sedimentation model of cells and EVs based on the patient's ESR. (1) TM healed completely after the treatment with 3.6 mL of PVRP and PVRG (high platelet and EV protocol). The speech discrimination score and air conduction pure tone average improved from 75% to 95% and from 65 to 25 dB, respectively. (2) The osteoradionecrotic surface area decreased from 46 to 18 cm

Platelets are a natural source of signaling molecules, growth factors, cytokines and extracellular vesicles that modify the pericellular microenvironment in favor of tissue healing and regeneration. Autologous platelet-rich products, such as platelet-rich plasma are used in many fields of clinical medicine to benefit from these effects. Platelet-rich products are ideally used in more than a single application, but repetitive harvesting of concentrated platelets is time-consuming and impractical for patients and clinicians. In order to maximize the utility of priorly made products and create a supply of patients’ own preparations, investigations of appropriate storage of platelet products have emerged. Recent explorations of hypothermic preservation, such as cryopreservation or freeze-drying have shown positive results, providing an efficient possibility of platelet-rich product storage retaining their growth factor, cytokine and chemokine activity. Keywords: Platelet-rich plasma, Cryopreservation, Freeze-drying, Platelet storage, Cyto-kines, Growth factors, Healing

The diagnostic and therapeutic potential of extracellular vesicles (EVs) has been recognised in many fields of medicine for several years. More recently, it has become a topic of increasing interest in otorhinolaryngology, head and neck surgery (ORL-HNS). With this narrative review, we have aspired to determine different aspects of those nanometrically sized theranostic particles, which seem to have promising potential as biomarkers in some of the most common diseases of the ORL-HNS by being available via less invasive diagnostic methods. At the same time, a better understanding of their activity provides us with new possibilities for developing specific target treatments. So far, most research has been oriented towards the role of EVs in the progression of head and neck cancer, notably head and neck squamous cell cancer. Nonetheless, some of this research has focused on chronic diseases of the ears, nose and paranasal sinuses. However, most research is still in the preclinical or experimental phase. It therefore requires a further and more profound understanding of EV content and behaviour to utilise their nanotheranostic capacities to their fullest potential.

In order to prepare optimal platelet and extracellular vesicle (EV)-rich plasma for the treatment of chronic temporal bone inflammation, we studied effects of centrifugation parameters on redistribution of blood constituents in blood samples of 23 patients and 20 volunteers with no record of disease. Concentrations of blood cells and EVs were measured by flow cytometry. Sample content was inspected by scanning electron microscopy. A mathematical model was constructed to interpret the experimental results. The observed enrichment of plasma in platelets and EVs after a single spin of blood depended on the erythrocyte sedimentation rate, thereby indicating the presence of a flow of plasma that carried platelets and EVs in the direction opposite to settling of erythrocytes. Prolonged handling time correlated with the decrease of concentration of platelets and larger EVs in platelet and EV-rich plasma (PVRP), R = -0.538, p = 0.003, indicating cell fragmentation during the processing of samples. In further centrifugation of the obtained plasma, platelet and EV enrichment depended on the average distance of the sample from the centrifuge rotor axis. Based on the agreement of the model predictions with observations, we propose the centrifugation protocol optimal for platelet and EV enrichment and recovery in an individual sample, adjusted to the dimensions of the centrifuge rotor, volume of blood and erythrocyte sedimentation rate.[Figure: see text].

Background The aim of the study was to identify the value of extensive resection and reconstruction with flaps in the treatment of locoregionally advanced lateral skull-base cancer. Patients and methods The retrospective case review of patients with lateral skull-base cancer treated surgically with curative intent between 2011 and 2019 at a tertiary otorhinolaryngology referral centre was made. Results Twelve patients with locoregionally advanced cancer were analysed. Lateral temporal bone resection was performed in nine (75.0%), partial parotidectomy in six (50.0%), total parotidectomy in one (8.3%), ipsilateral selective neck dissection in eight (66.7%) and ipsilateral modified radical neck dissection in one patient (8.3%). The defect was reconstructed with anterolateral thigh free flap, radial forearm free flap or pectoralis major myocutaneous flap in two patients (17.0%) each. Mean overall survival was 3.1 years (SD = 2.5) and cancer-free survival rate 100%. At the data collection cut-off, 83% of analysed patients and 100% of patients with flap reconstruction were alive. Conclusions Favourable local control in lateral skull-base cancer, which mainly involves temporal bone is achieved with an extensive locoregional resection followed by free or regional flap reconstruction. Universal cancer registry should be considered in centres treating this rare disease to alleviate analysis and multicentric research.

Platelet- and extracellular vesicle-rich plasma (PVRP) and platelet- and extracellular vesicle-rich gel (PVRG) are blood-derived products gaining attraction in regenerative medicine. However, despite their reported good efficacy, their preparation protocols are too time-consum-ing. Moreover, patient-tailored preparation protocols are desired to optimise platelet and extra-cellular vesicle (EV) count in PVRP and PVRG. This scientific contribution presents a clinical implementation of mathematical model for calculation of the desired total blood volume for PVRG preparation. PVRG was prepared according to previously derived mathematical model of blood cell and EV sedimentation during centrifugation based on the patient's erythrocyte sedi-mentation rate (ESR) (Božič et al., 2021). We estimated the volume of blood required for the treat-ment of the surface area of the individual patient, prepared the PVRG accordingly and applied it to the patient’s wound. After six applications of 13 mL to 65 mL of PVRG, the osteoradi-onecrotic surface area decreased from 46 cm2 to 18 cm2, and infection was eradicated. The math-ematical modelling of total blood volume needed to prepare PVRG proved useful to prepare the therapeutic amount of PVRG and also optimized time for the PVRG preparation protocol. Keywords: Extracellular vesicles; Platelet-rich plasma; Regenerative medicine; Osteitis; Osteora-dionecrosis; Osteomyelitis; Temporal bone

The facial nerve and its reconstructive surgical procedures are complex and challenging. The main function of facial nerve is namely motor innervation of facial muscles and its dysfunction presents as facial paralysis. Depending on the extent of facial nerve injury (neurapraxia, axonotmesis, neurotmesis) and consequently a physiological phenomenon of Wallerian degeneration, mechanism, location of the injury, time course of the paralysis and medical condition we decide about the type of the reconstructive surgery. Generally, possible surgical interventions to improve facial nerve functioning are mainly nerve decompression, neurorrhaphy/end-to-end anastomosis, interposition (cable) grafts and nerve rerouting. Moreover, most commonly nerves undergoing facial reconstruction are great auricular and sural nerves. In addition, nerve rehabilitation can be improved by using platelet-rich plasma (PRP/PVRP), applied directly to nerve. There are many roles of PVRP, described in the literature such as neuroprotective, neurogenic, neuroinflammatory, angiogenic role and improving hemostasis. Also, its neoplastic and proliferative effects were not reported. Considering all these features implementing PVRP in the facial nerve regenerative treatment has strong potential in the future. Keywords: Facial nerve; Reeconstructive surgery; Platelet and extracellular vesicle rich plasma; Nerve regeneration

To provide physicians and patients with the tools needed to evaluate patients' problems and health-related quality of life by cross-culturally adapting and validating the Chronic Otitis Media Questionnaire 12 (COMQ-12), the Dizziness Handicap Inventory (DHI), the Neuropsychological Vertigo Inventory (NVI) and the Tinnitus Handicap Inventory (THI).COMQ-12, DHI, NVI and THI were translated into the Slovenian language and completed by patients treated at our department for chronic otitis media, vertigo or tinnitus. The control group for each questionnaire consisted of healthy volunteers. Internal consistency, test-retest reliability, discriminant validity, diagnostic accuracy and cut-off value were determined for each questionnaire.Test-retest reliability was excellent for DHI (ICC A=0.946) and NVI (p=0.315, ICC A=0.975), good to excellent for COMQ-12 (p=0.680, ICC A=0.858) and satisfactory for THI (p=0.120). Discriminant validity was confirmed for each questionnaire (p0.05) using the Mann-Whitney U test (COMQ-12, DHI, THI) or the Welch t-test (NVI). COMQ-12 had acceptable (α=0.796) and DHI (α=0.910), NVI (α=0.950) and THI (α=0.924) perfect internal consistency. COMQ-12 and DHI had excellent, NVI acceptable and THI perfect diagnostic accuracy (AUC=0.987, AUC=0.999, AUC=0.781 and AUC=1.000 respectively). Cut-off values determined by Youden's index were 7, 7, 9 and 56 for COMQ-12, THI, DHI and NVI, respectively.Slovenian COMQ-12, DHI, NVI and THI are a valid and accurate tool for the diagnosis and measurement of health-related quality of life in patients with chronic otitis media, vertigo and tinnitus. They could aid general practitioners, occupational health specialists, neurologists and otorhinolaryngologists.Orodja za oceno težav bolnikov s kroničnimi vnetji srednjega ušesa, težav z ravnotežjem in tinitusom ter vpliv slednjih na kakovost življenja v slovenščini ne obstajajo. S projektom smo jih želeli zagotoviti zdravnikom in bolnikom s temi težavami. Vrzel bi zapolnili z medkulturno prilagoditvijo in potrditvijo vprašalnika o kroničnem vnetju srednjega ušesa (COMQ-12), vrtoglavici (DHI), nevropsiholoških vplivih vrtoglavice (NVI) in o obremenjenosti zaradi tinitusa (THI).Vprašalniki so bili prevedeni iz angleščine v slovenščino skladno s priporočili. COMQ-12 so izpolnili bolniki s kroničnim vnetjem srednjega ušesa, DHI in NVI z vrtoglavico in THI s tinitusom. Kontrolno skupino za vsak vprašalnik so sestavljali zdravi prostovoljci. Vsakemu vprašalniku je bila določena stopnja notranje skladnosti, zanesljivost pri ponovnem testiranju, diskriminantna validnost, diagnostična natančnost in mejna vrednost.Zanesljivost pri ponovnem testiranju je bila odlična za DHI (ICCA = 0,946) in NVI (p = 0,315, ICCA = 0,975), dobra do odlična za COMQ-12 (p = 0,680, ICCA = 0,858) in zadovoljiva za THI (p = 0,120). Diskriminantna validnost je bila potrjena (p0,05) za vsak vprašalnik bodisi z uporabo testa Mann-Whitney U (COMQ-12, DHI, THI) bodisi z Welchovim t-testom (NVI). COMQ-12 je imel sprejemljivo (α = 796), DHI (α = 0,910), NVI (α = 0,950) in THI (α = 0,924) pa popolno notranjo skladnost. COMQ-12 in DHI sta imela odlično (0,987 in 0,999), NVI sprejemljivo (0,781) in THI popolno (1,000) diagnostično natančnost glede na vrednost površine pod krivuljo ROC. Mejne vrednosti so bile določene objektivno z Youdonovim indeksom (J) in so znašale 7 za COMQ-12 (J = 0,90) in THI (J = 1,00), 9 za DHI (J = 0,95) in 56 za NVI (J = 0,43).Slovenske različice vprašalnikov COMQ-12, DHI, NVI in THI so medkulturno prilagojene, potrjene in uporabne kot pomembna merila za oceno z zdravjem povezane kakovosti življenja. Hkrati so tudi natančen diagnostični pripomoček pri bolnikih s kroničnim vnetjem srednjega ušesa, vrtoglavico ali tinitusom, ki bo v pomoč zdravnikom specialistom družinske medicine, specialistom medicine dela, prometa in športa, nevrologom ter otorinolaringologom.

This study aims to present characteristics of pediatric patients with peripheral vertigo and dizziness and their clinical workup results in the tertiary otorhinolaryngology center. We wanted to investigate whether the detailed history could replace the extensive vestibular testing and whether the clinical presentation could guide the first contact physician to appropriately directed specialist referral.Retrospective case review of consecutive pediatric vertigo and dizziness patients referred to the tertiary otorhinolaryngology center from 2015 to 2020. The data about the signs and symptoms of vertigo and dizziness and the results of audiological and vestibular tests were collected.Of 257 children aged 10.9±4.3 years (R: 1-17 years), 32 (12.5%) had peripheral, and 49 (19%) had central vertigo and dizziness. Acute vestibulopathy was diagnosed in 22/257 (8.5%) children, sudden sensorineural hearing loss and benign paroxysmal positional vertigo in 5/257 (2%) children each. 60% of children with peripheral vertigo and dizziness had emesis, and 55.6% had nausea. 8% of children had spontaneous horizontal-rotatory nystagmus that followed Alexander's law. Goodman and Kruskal's л for determining whether the type of nystagmus could predict the type of vertigo and dizziness (central or peripheral) was 0.481 (p = 0.001). 12/26 (60%) of children with peripheral vertigo and dizziness had emesis compared to 14 (30.04%) children without emesis, a difference in proportions of 0.296 (p = 0.024, chi-square test of homogeneity). Binomial logistic regression to ascertain the effects of duration, nausea and emesis on the likelihood of the presence of peripheral vertigo was statistically significant (χ2(3) = 10.626, p = 0.014).Unlike adults, peripheral vestibular causes of vertigo and dizziness in children may be rare but have the same typical signs and symptoms. The detailed history and careful clinical examination are crucial in differentiating between peripheral and central causes. This guides the first contact physician for further referral to appropriate specialists included in a multidisciplinary workup. Namely, nausea, emesis, horizontal nystagmus and a longer duration of symptoms in a child with vertigo and dizziness indicate a peripheral etiology. Therefore, a referral to an otorhinolaryngologist is reasonable.

Lice zavzema največji del obraza in je pogosto mesto nastanka pridobljenih mehkotkivnih sprememb. Najpogostejši vzroki so poškodbe in kožni malignomi. Slednje je potrebno izrezati z varnostnim robom, kar ima za posledico velike tkivne vrzeli. Za kritje take vrzeli na licu primarni šiv rane ni mogoč, zato moramo razmisliti o načinu rekonstrukcije z VY, cervikofacialnim ali submentalnim režnjem. Cervikofacialni reženj smo uporabili za kritje vrzeli vzdolž celotne nazolabialne gube v 1. estetski podenoti lica, VY reženj za kritje velike vrzeli superomedialnega dela te podenote, submentalni pa za kritje velike vrzeli v 2. estetski podenoti. Prispevek temeljito opisuje njihove značilnosti, tehnike priprave in klinične primere. Za boljše razumevanje opiše tudi estetske podenote, kožne linije in del anatomije obraza.

Background and Objectives: Numerous authors have reported that the commonest type of vertigo in children is migraine-associated vertigo (vestibular migraine and benign paroxysmal vertigo of childhood—BPV). We aimed to provide the possible etiological background of vertigo and dizziness in Slovenian children. Materials and Methods: A retrospective case series of pediatric vertigo and dizziness children referred to the tertiary pediatric otorhinolaryngology center from 2015 to 2020. Children received a complete audiological and vestibular workup and were referred to pediatric specialists depending on the clinical presentation. Results: Of 257 children (42% male, 58% female) aged 1–17 years (M = 10.9, SD = 4.3 years) in 19.1% vertigo and dizziness were classified as central, in 12.4% as a peripheral vestibular, in 10.9% as a hemodynamic, in 5.8% as a psychological and none as visual by pediatric neurologists, otorhinolaryngologists, cardiologists, psychologists or ophthalmologists, respectively. 40.8% (20) children with central vertigo had BPV (7.8% of all children) and 8.2% (4) migrainous vertigo. In 43.6% (112 children), the etiology remained unclassified. Conclusions: After a thorough multidisciplinary workup, the etiology of vertigo and dizziness was unraveled in the majority of children referred to our tertiary otorhinolaryngology center. The most common cause was central, however, in a considerable number, the etiology remained unclassified. The latter could be attributed to the self-limiting nature of vertigo spells. Hence, a child presenting with dizziness and vertigo requires a multidisciplinary approach, in which referral to a neurologist is, in most cases, essential.

Extracellular vesicles (EVs), also known as microvesicles, exosomes, nanovesicles, microparticles etc. are a heterogeneous group of nanometrically sized cell-derived membranous structures which can originate from any cell, including plant cell or bacteria. They were found in many body fluids including platelet-rich plasma (PRP). PRP is a blood-derived product with immune, hemostatic and regenerative effects. PRP is expected to contain important concentrations of EVs which could be important contributors to PRPs effects. For that reason, we call the preparation from PRP a platelet- and extracellular vesicle-rich plasma (PVRP). PVRP has been used in various medical fields including orthopedics, traumatology, wound care surgery, plastic, reconstructive and esthetic surgery, hand surgery, maxillofacial and oral surgery, gynecology, urology, ophthalmology, cardiothoracic surgery, neurosurgery and otorhinolaryngology. Otorhinolaryngology is a field in which PVRP has a great potential in the treatment of diseases of the face, neck, nose, paranasal sinuses, anterior skull base, lips, oral cavity, pharynx, esophagus, salivary glands, ear and lateral skull base. To present this potential and also hitherto gathered results in the field of otorhinolaryngology, this chapter describes the role of platelets and EVs in diagnostics and treatment with an emphasis on otorhinolaryngology. Also, we discuss future perspectives of EVs and PVRP in treatment of moist radical cavities after cholesteatoma surgery.

Within the micro and nano world, tiny membrane-enclosed bits of material are more or less free to move and act as communication tools within cells, between cells, between different tissues and between organisms in global environment. Based on the mechanism of membrane budding and vesiculation that includes all types of cells, in this review, we attempted to present a review on SARS-CoV-2 virus actions in compartments of different scales (cells and their surroundings, tissues, organisms and society). Interactions of the virus with cells on a molecular level, with neural system, endothelium, hematopoietic system, gastrointestinal system and genitourinary system. Transmission route between organisms and between mother and fetus are considered. Also, transmission of virus through contact with materials and with environment, the suggested measures to prevent contamination with the virus and to support the organism against the disease are given.

Background Extracellular vesicles (EVs) are submicron cellular fragments that mediate intercellular communication. EVs have in the last decade attracted major interest as biomarkers or platforms for biomarkers of health and disease. To better understand the reasons why despite great expectations and considerable effort, EV-based methods have not yet been introduced into clinical practice, we present a systematic analysis of published results of clinical studies. Materials and methods Clinical studies on populations of body fluid samples, published from 2010 to including 2015, applying centrifugation of fluid human samples with centrifuge accelerations up to about 25 000 g and flow cytometry for detection of EVs were analysed with respect to statistical significance (p), statistical power (P), clinical significance (CS), defined as the difference between the means divided by the sum of standard deviations, and size of the populations (Nmin), defined as the number of samples in the smaller group. Results Final analysis included 65 publications with 716 comparisons reporting 308 (43%) statistically significant differences (P 0·8 and 88 (12%) had clinical importance CS > 1·96. None of comparison with CS > 1·96 included populations in which the smaller group consisted of 50 or more samples. Conclusions To fulfil claimed expectations for EV-based methods as promising diagnostic tools, more evidence on EV-based mechanisms of diseases should be gathered. Also, the methods of EV harvesting and assessment should be improved to yield better repeatability and thus allow clinical studies with larger number of samples.