Pig Skin Research Articles

In vitro study on the transmission of multidrug-resistant bacteria from textiles to pig skin.

The survival of microorganisms on textiles and specifically on healthcare professionals' (HCP) attire has been demonstrated in several studies. The ability of microorganisms to adhere and remain on textiles for up to hours or days raises questions as to their possible role in transmission from textile to skin via HCP to patients. To evaluate the presence, survival and transmission of different multidrug-resistant bacteria (MDRB) from HCP attire onto skin. Three MDRB [methicillin-resistant Staphylococcus aureus (MRSA); vancomycin-resistant Enterococcus faecium (VRE); carbapenem-resistant Klebsiella pneumoniae, CRKP)] were inoculated on textiles from scrubs (60% cotton-40% polyester) and white coat (100% cotton) at concentrations of 108 colony-forming units (CFU), 105 CFU, and 103 CFU per mL. The inoculation of swatches was divided in time intervals of 1 min, 5 min, 15 min, 30 min, 1 h, 2 h, 3 h, 4 h, 5 h, and 6 h. At the end of each period, textiles were imprinted onto pig skins and each skin square was inverted onto three different selective chromogenic media. Growth from the pig skin squares was recorded for the 3 MDRB at the three above concentrations, for the whole length of the 6-h experiment. MRSA was recovered from pig skins at all concentrations for the whole duration of the 6-h study. VRE was recovered from the concentration of 108 CFU/mL for 6 h and from 105 CFU/mL for up to 3 h, while showing no growth at 103 CFU/mL. CRKP was recovered from 108 CFU/mL for 6 h, up to 30 min from 105 CFU/mL and for 1 min from the concentration of 103 CFU/mL. Evidence from the current study shows that MRSA can persist on textiles and transmit to skin for 6 h even at low concentrations. The fact that all MDRB can be sustained and transferred to skin even at lower concentrations, supports that textiles are implicated as vectors of bacterial spread.

Carbon dots-based fluorescent films to act as a potential antioxidant agent and pH ratiometric sensor for skin applications

The wound healing process is accompanied by changes in pH values. Monitoring this physicochemical parameter can indicate the effectiveness of the applied treatment and act as early identification of wound infection. This study focuses on the development of a fluorescent film-based polyvinyl alcohol (PVA) and carbon dots (CDs) derived from lemon bagasse (CD-L) and ortho-phenylenediamine (CD-oPD) named to act as antioxidants and potential ratiometric fluorescent pH sensors, in wound applications. The I460/I550 intensity ratio, as a function of pH value for the dual-system CDs prepared from the mixture of CD-L and CD-oPD, named CD-L/oPD, was investigated. The fit corresponded to a sigmoidal function in the pH range of 5–10, with a relationship having a r2 = 0.992. The variation in the values of the I460/I550 ratio allows for the visualization of the color change from yellowish-green to green with increasing pH. Through a simulated ex vivo pig skin model, it was possible to note that the films prepared from mixed of the CD-L and CD-oPD carbon dots incorporated in a matrix PVA named CD-L/oPD-F was more efficient at visually discriminating color in relation to changes in pH than the films prepared from both individual CD-L (CD-L-F) and CD-oPD (CD-oPD-F) carbon dots. CD-L and CD-oPD demonstrated antioxidant capacity against reactive oxygen species (ROS). The IC50 values for CD-L and CD-oPD were 56.7 and 39.5 μg mL−1 in the DPPH● inhibition assay, and 25.1 and 63.4 μg mL−1 in the HOCl scavenging one, respectively. MTT viability assays using human non-tumoral skin fibroblast (HFF-1) cell showed a cell survival rate of over 80% for both CDs up to a concentration of 1,000 μg mL−1. Finally, the developed films can act in a bifunctional way, by monitoring healing through pH changes and by acting as an antioxidant agent in the treatment of wounds.

Comparison of the agar contact method and the wet-dry double swabbing method for determining the total viable bacterial count on pig carcass surfaces

A practical and representative sampling method for microbiological examination of the slaughter process is useful for identifying abattoir-specific risk factors within the pig slaughter line. The aim of this study was to examine the suitability of an agar contact method (ACM), where the agar was homogenized before the microbiological processing, in comparison with the wet-dry double swabbing method (WDSM) for quantitative determination of total viable counts (TVC) on pig skin surfaces. In our experimental trial, pig skin pieces were artificially contaminated at 2 levels (3 log and 7 log cfu/ml) with a suspension of bacteria species commonly found on pig skin and cultivated in vitro. Within our field trial, pig carcasses were investigated at pre-chilling in an abattoir under standard processing conditions. For both sampling methods, TVC was determined, and statistical equivalence tests were calculated. Linear regression models showed the similarity of the sampling methods, with coefficient of determination (R2) > 90% and slope parameters of nearly 1 for both trials separately. Statistically significant equivalence between the 2 sampling methods was proven in both trials (with p < 0.0001 within an equivalence range of ± 0.5 log cfu/ml, respectively). The field trial revealed TVC on carcass surfaces sometimes at or below the lower detection limit for the ACM, while TVC from all carcasses were able to be determined by WDSM. Overall, low contamination levels were less reliably detectable by ACM than by WDSM. The ACM can be seen as an additional and suitable sampling procedure for pig skin and can contribute to the identification of abattoir specific risk factors for investigations of the hygienic status at process stages along the pig slaughter line.

Chitosan-based multifunctional hydrogel with bio-adhesion and antioxidant properties for efficient wound hemostasis

Benefiting from the biocompatibility, adhesiveness, and natural extracellular matrix-mimicking ability, hydrogels have received increasing research in recent years. In this study, a hydrogel system composed of dopamine, quaternized ammoniated chitosan (QCS), and polyvinylpyrrolidone was reported to exhibit fast hemostatic properties in Sprague-Dawley rat tail amputation and liver bleeding models. The results showed that this hydrogel had good hemostatic properties. The designed hydrogel showed high swelling ratios in H2O, PBS, and 0.9 % NaCl solution, indicating its capability to absorb tissue residual exudate and form a stable hydrogel. Compared with the control group, the blood loss in Sprague-Dawley rat tail amputation and liver bleeding were reduced by nearly 78 % and 76 %, respectively. Interestingly, dopamine endowed the hydrogel with antioxidant properties, thus holding a great application promise in inflammatory wounds. Furthermore, the designed hydrogel demonstrated good and reversible adhesion properties (12.23 ± 0.22 kPa–24.31 ± 0.55 kPa), ensuring its firm attachment to bleeding wounds of pig skin in wet environments. This research points out a novel path for designing chitosan-based hydrogels for biomedical applications.

Detection of fragrances on the skin and study of their interaction using infrared and Raman spectroscopy

Nowadays, fragrances belong to the widely used cosmetics. Their composition is designed in a way that it evolves and changes over time. In this work, the effect of fragrances on the skin was studied – the interactions between pig skin samples and fragrances and the possibility of their detection and mutual differentiation. Non-invasive techniques of vibrational spectroscopy were used to obtain the data, namely FT-IR spectroscopy with attenuated total reflection accessory and Raman microspectroscopy. Vibrational spectra were measured within 8 h with different time intervals and after 22 h from the application of fragrance for FT-IR and Raman measurements, respectively. The obtained spectra were pre‑processed and subsequently evaluated by multivariate statistical methods. The study showed that skin treated by fragrances is well distinguishable from untreated skin, even after 22 h. In addition, it is possible to differentiate individual fragrances from each other; therefore, the use of spectroscopical techniques could be a potential tool for forensic analysis of fragrances.

Administration of Moringa (Moringa oleifera L) Leaf Ethanol Extract Cream Inhibited the Increase of Tyrosinase Enzyme Levels and Amount of Melanin in Male Guinea Pigs (Cavia porcellus) Skin Exposed to Ultraviolet-B

Introduction: The purpose of this study was to prove the effect of administering Moringa leaf ethanol cream extract in inhibiting the increase in tyrosinase level and the amount of melanin in the skin of male guinea pigs (Cavia porcellus) exposed to ultraviolet-B (UVB). Methods: This research was an experimental study using a randomized post-test only control group design method. The research subjects were 30 male guinea pigs divided randomly into five groups: group without cream, group with basic cream, group with 5%, 10% and 20% Moringa leaf ethanol cream extract. The five groups were exposed to UVB lamps for 15 days, and skin samples were taken 48 hours after the last exposure to UVB to examine the amount of melanin using Masson Fontana staining and tyrosinase enzyme using ELISA method. Results: The best reduction in tyrosinase enzyme was in the group given 20% of Moringa leaf extract cream (10.63 ± 6.53 ng/mL) with a p value of 0.004 (p&lt;0.05). Based on the amount of melanin, the highest mean amount of melanin was found in the group without cream (37.40 ± 16.77) and the lowest was in the treatment group given 20% Moringa leaf cream extract (10.63 ± 6.53) with a p value of 0.008 (p&lt;0.05). Conclusion: Administration of Moringa leaf (Moringa oleifera L.) ethanol cream extract inhibits the increase in tyrosinase enzyme level and the amount of melanin in the skin of male guinea pigs (Cavia porcellus) exposed to UVB with an appropriate concentration for safe use of the cream is between 5% and 10%. Keywords: Moringa leaf (Moringa oleifera L.); tyrosinase enzyme; melanin; guinea pig (Cavia porcellus); ultraviolet-B

Glycopolymer-Based Antiswelling, Conductive, and Underwater Adhesive Hydrogels for Flexible Strain Sensor Application.

With the fast development of soft electronics, underwater adhesion has become a highly desired feature for various sensing uses. Currently, most adhesive hydrogels are based on catechol-based structures, such as polydopamine, pyrogallol, and tannic acid, with very limited structural variety. Herein, a new type of glycopolymer-based underwater adhesive hydrogel has been prepared straightforwardly by random copolymerization of acrylic acid, acetyl-protected/unprotected glucose, and methacrylic anhydride in dimethyl sulfoxide (DMSO). By employing a DMSO-water solvent exchange strategy, the underwater adhesion was skillfully induced by the synergetic effects of hydrophobic aggregation and hydrogen bonding, leading to excellent adhesion behaviors on various surfaces, including pig skins, glasses, plastics, and metals, even after 5 days of storage in water. In addition, the underwater adhesive hydrogels with simple and low-cost protected/unprotected carbohydrate compositions showed good mechanical and rheological properties, together with cytocompatibility and antiswelling behavior in water, all of which are beneficial for underwater adhesions. In application as a flexible strain sensor, the adhesive hydrogel exhibited stable and reliable sensing ability for monitoring human motion in real time, suggesting great potential for intelligent equipment design.

Catechol chitosan coated dual-loaded liposomes based on oxidation and saccharification mechanisms for enhancing skin anti-aging effects

Skin aging has become a major urgent problem to be solved. Evidence reveals that oxidation and glycosylation are two dominant inducements of aging. Resveratrol (RES) with outstanding anti-oxidant effect and carnosine (CAR) with superb anti-glycation property were selected as two model drugs to evaluate the feasibility of their synergistic anti-aging effect. RES and CAR at the most desired mass ratio, supplying the most superior synergistic anti-aging effects were further encapsulated in liposomes (LP), which were separately coated with chitosan (CS) and catechol chitosan (Cat-CS) to increase the transdermal penetration. Their anti-aging efficacy was explored in human skin fibroblast (HSF) and human immortalized keratinocytes (HaCaT) cells, as well as the back skin of guinea pigs. Herein, RES and CAR at the mass ratio of 2:1 exhibited the most ideal synergistic anti-aging effect. The constructed liposomes have been shown to possess excellent fundamental properties and sustained-release properties. The aging-related indicator levels in the two cells and guinea pigs were obviously improved for the RES + CAR@Cat-CS-LP group. Additionally, skin appearance, tissue morphology, and collagen content were visibly improved, indicating its perfect anti-aging effect. In conclusion, RES + CAR@Cat-CS-LP is expected to be exploited as a potential anti-aging drug delivery system.

Exploring Alkyl Ester Salts of L-Amino Acid Derivatives of Ibuprofen: Physicochemical Characterization and Transdermal Potential.

This research presents novel ibuprofen derivatives in the form of alkyl ester salts of L-amino acids with potential analgesic, anti-inflammatory, and antipyretic properties for potential use in transdermal therapeutic systems. New derivatives of (RS)-2-[4-(2-methylpropyl)phenyl]propionic acid were synthesized using hydrochlorides of alkyl esters (ethyl, propyl, isopropyl, butyl, sec-butyl, tert-butyl, and pentyl) of L-glutamine. These were further transformed into alkyl esters of L-amino acid ibuprofenates through neutralization and protonation reactions. Characterization involved spectroscopic methods, including nuclear magnetic resonance and Fourier-transform infrared spectroscopy. Various physicochemical properties were investigated, such as UV-Vis spectroscopy, polarimetric analysis, thermogravimetric analysis, differential scanning calorimetry, X-ray diffraction, water solubility, octanol/water partition coefficient, and permeability through pig skin using Franz diffusion cells. The research confirmed the ionic structure of the obtained hydrochlorides of alkyl esters of L-amino acids and ibuprofenates of alkyl esters of L-glutamic acid. It revealed significant correlations between ester chain length and thermal stability, crystallinity, phase transition temperatures, lipophilicity, water solubility, skin permeability, and skin accumulation of these compounds. Compared to the parent ibuprofen, the synthesized derivatives exhibited higher water solubility, lower lipophilicity, and enhanced skin permeability. This study introduces promising ibuprofen derivatives with improved physicochemical properties, highlighting their potential for transdermal therapeutic applications. The findings shed light on the structure-activity relationships of these derivatives, offering insights into their enhanced solubility and skin permeation, which could lead to more effective topical treatments for pain and inflammation.

CHANGES IN THE AMINO ACID COMPOSITION OF GELATIN AFTER TREATMENT OF BOVINE COLLAGEN WITH ENZYME PREPARATION

Recently, more attention has been paid to the study of the amino acid composition of gelatins, which is associated with the quality of the corresponding gels as intermediates for human and animal nutrition. In a brief review, a modification of the general method of acid extraction of collagens for the preparation of gelatins using enzymes (such as papain, actinidin, and others) is considered and the corresponding changes in the amino acid composition of gelatins are discussed. It is clear that there are changes in the content of glycine in gelatins from any collagens, but in all cases the content of glycine is about a third of the content of all amino acids (as in the original collagens). It is important that the content of imino acids (the sum of proline and hydroxyproline, which largely determines the properties of gels) in gelatins from any collagens with the use of all the studied enzymes is much higher than without them. In addition, the content of imino acids in gelatin from the bovine skin of cows with the use of any enzymes is significantly higher than in gelatins from the skin of pigs and fish. This holds true for other key “proteinogenic” amino acids as well. The reverse trend is observed only for a few amino acids: serine, threonine, tyrosine, phenylalanine, the content of which is low in gelatins from any collagens.

A self-assembled hydrogel dressing as multi-target therapeutics to promote wound healing

Wound healing is one of the major challenges in biomedicine owing to the absence of ideal wound dressings which can integrate easy absorption, satisfied injectability, rapid self-healing property, and multiple therapeutic effects into one system. Here, we explore a self-assembled supramolecular hydrogel composed of herbal cinnamaldehyde (CA) and folic acid (FA). This enables the Schiff base and the Ionic crosslinking as a dynamic double cross-linked network. This nano-scale hydrogel (C-FA-gel) maintains good adhesion and is apt to get peeled off from tissues without any residuals. Further experiments show a percutaneous absorption within 12 h after the hydrogel dressing perfectly fills the irregular wound by injection on pig skin. For muti-target therapeutics, the drug sustained-release of C-FA-gel exhibits broad-spectrum antibacterial activities without evident cytotoxicity. Besides, the topical dressing on rat skin promotes wound healing via multifunctional effects including inflammatory reduction, facilitation of the angiogenesis improvement, and regeneration promotion of skin appendages. Moreover, the C-FA-gel induces anti-inflammatory macrophages from pro-inflammatory macrophages. Therefore, we successfully present a self-assembled hydrogel using the ‘one system’ strategy. This study offers a green and efficient design for the potential to be applied to complicated wound treatment.

Infrared Raman spectroscopy enables noninvasive biochemical assessment of skin tissue and the thermal stability

Raman-active modes of human skin and pork belly have been studied systematically by a near-infrared Raman spectrometer with an exciting laser of 1064nm. The main components and quantitative determination of pork belly are extracted by fitting the Raman spectra with the normalized Raman spectra of biochemical reagents such as collagen, elastin, triolein, fibronectin, fibrin, and hyaluronic acid. It demonstrates that the main components and quantity are various at different locations of pork belly, while the main components of human skin are similar to those of pig skin. In a further step, the evolution of the heating time-dependent Raman modes of isolated pig skin has been investigated for the mechanism of burnt skin. One can find that the spatial structure and main components of skin have an excellent thermal stability in the temperature range from -120 to 200∘C, which is confirmed by the temperature dependent Raman spectra of isolated pig skin, microporous acellular dermal matrix (MADM) as well as their corresponding biochemical reagents (collagen, elastin, triolein, etc.). These results help understand the mechanism of the living skin burnt by fire or hot water, and supplies an alternative technology for surgeons to diagnose the depth of a burn injury in time.

Characterization of spatial lipidomic signatures in tick-bitten guinea pig skin as a model for host-vector-pathogen interaction profiling.

Here, we demonstrate the adaptability of spatial "omics" methods to identify interphylum processes regulated at the vector-host interface of ticks during a mammalian blood meal. This approach enables a better understanding of complex bipartite or tripartite molecular interactions between hosts, arthropod vectors and transmitted pathogens, and contributes toward the development of spatially aware therapeutic target discovery and description.

In vitro and in vivo immune assessments of genetically-engineered pig skin grafts in New World (squirrel) monkeys.

Half a million patients in the USA alone require treatment for burns annually. Following an extensive burn, it may not be possible to provide sufficient autografts in a single setting. Genetic manipulations (GM) of pigs offer the possibility of reducing primate humoral and cellular rejection of pig skin xenografts and thus extending graft survival. We compared the survival of skin grafts from pigs with 9-GM with that of autografts and allografts in squirrel monkeys. Monitoring for rejection was by (1) macroscopic examination, (2) histopathological examination of skin biopsies, and (3) measurement of anti-monkey and anti-pig IgM and IgG antibodies. Autografts (n = 5) survived throughout the 28 days of follow-up without histopathological features of rejection. Median survival of allografts (n = 6) was 14 days and of pig xenografts (n = 12) 21 days. Allotransplantation was associated with an increase in anti-monkey IgM, but the anticipated subsequent rise in IgG had not yet occurred at the time of euthanasia. Pig grafts were associated with increases in anti-pig IgM and IgG. In all cases, histopathologic features of rejection were similar. 9-GM pig skin xenografts survive at least as long as monkey skin allografts (and trended to survive longer), suggesting that they are a realistic clinical option for the temporary treatment of burns. Although monkeys with pig skin grafts developed anti-pig IgM and IgG antibodies, these did not cross-react with monkey antigens, indicating that a primary 9-GM pig skin graft would not be detrimental to a subsequent monkey skin allograft.

Development of Essential Oil-Loaded Polymeric Nanocapsules as Skin Delivery Systems: Biophysical Parameters and Dermatokinetics Ex Vivo Evaluation.

Essential oils (EOs) are natural antioxidant alternatives that reduce skin damage. However, EOs are highly volatile; therefore, their nanoencapsulation represents a feasible alternative to increase their stability and favor their residence time on the skin to guarantee their effect. In this study, EOs of Rosmarinus officinalis and Lavandula dentata were nanoencapsulated and evaluated as skin delivery systems with potential antioxidant activity. The EOs were characterized and incorporated into polymeric nanocapsules (NC-EOs) using nanoprecipitation. The antioxidant activity was evaluated using the ferric thiocyanate method. The ex vivo effects on pig skin were evaluated based on biophysical parameters using bioengineering techniques. An ex vivo dermatokinetic evaluation on pig skin was performed using modified Franz cells and the tape-stripping technique. The results showed that the EOs had good antioxidant activity (>65%), which was maintained after nanoencapsulation and purification. The nanoencapsulation of the EOs favored its deposition in the stratum corneum compared to free EOs; the highest deposition rate was obtained for 1,8-cineole, a major component of L. dentata, at 1 h contact time, compared to R. officinalis with a major deposition of the camphor component. In conclusion, NC-EOs can be used as an alternative antioxidant for skin care.

Simultaneous Extraction of Oil and Protein from Silkworm (Bombyx mori L.) Pupae (Lueng Parroj var.) and Their In Vitro Skin Moisturization.

Oil and protein from silkworm (Bombyx mori var. Leung Pairoj) pupae, by-product from sericulture, were extracted and evaluated for their potential uses as skin biomoisturizer. The silkworm pupae (SWP) oil and protein were simultaneously extracted by using three-phase partitioning (TPP) method and determined for their physicochemical properties including fatty acid and amino acid content, respectively. The highest yields of oil and protein at 8.24 ± 0.21% and 8.41 ± 0.26% w/w, respectively were obtained from 18 h extraction. Fatty acid analysis of SWP oil was rich in linolenic acid (37.81 ± 0.34%), oleic acid (28.97 ± 0.13%), palmitic acid (21.27 ± 0.05%), stearic acid (6.60 ± 0.09%) and linoleic acid (4.73 ± 0.21%). The clear yellow SWP oil possessed saponification value of 191.51 mg/g, iodine value of 119.37 g I2/g and peroxide value of 2.00 mg equivalent O2/kg. The SWP protein composed of 17 amino acids which aspartic acid, glutamic acid, glycine and serine were the major residues. SDS-PAGE analysis revealed that the SWP protein consisted of distinct protein at around 51, 70, 175 and over 175 kDa. Cytotoxicity of the SWP oil and protein was evaluated by using MTT assay and they showed low cytotoxicity toward keratinocyte cell (HaCat cell line). The SWP oil provided moisturizing effect on pig skin comparable to olive oil, while 1% and 2% of SWP protein showed higher moisturizing efficacy than 3% hydrolyzed collagen. The study indicated that the SWP oil and protein could be potential biomoisturizers for cosmetic products.

Agar Contact Method as a Valuable Tool to Identify Slaughter Hygiene Deficiencies along the Slaughter Process by Longitudinally Sampling Pig Skin Surfaces.

Examinations of total viable counts (TVCs) and Salmonella spp. on the skin of individual pigs during the slaughter process are useful to identify abattoir-specific risk factors for (cross-)contamination. At seven process stages (lairage to before chilling), pigs were bacteriologically investigated by repeatedly sampling the same animals using the agar contact method. The mean TVC of all pigs increased significantly at the first three tested process stages (mean count, after delivery: 5.70 log cfu/cm2, after showering: 6.27 log cfu/cm2, after stunning: 6.48 log cfu/cm2). Significant mean TVC reductions occurred after scalding/dehairing (mean count: 3.71 log cfu/cm2), after singeing/flaming (2.70 log cfu/cm2), and after evisceration (2.44 log cfu/cm2) compared with the respective preceding process stages. At the end of the slaughter line and before chilling, the mean TVC was 2.33 log cfu/cm2, showing that the slaughter process reduced contamination significantly. The slaughter process effectively reduced even very high levels of incoming TVCs, since at the individual animal level, at the end of the slaughter process, there was no difference in the TVCs of animals with initially high and initially low TVCs. Additionally, 12 Salmonella spp. isolates were recovered from 12 different pigs, but only until the stage after scalding/dehairing. Overall, the agar contact method used is valuable for detecting hygiene deficiencies at slaughter, and is animal-equitable, practical, and suitable for use on live animals.

In vitro, ex vivo, and in vivo appraisal of clobetasol propionate microparticles embedded topical bigel for psoriasis management

The current work aims to develop a bigel-containing clobetasol propionate-loaded PLGA microparticles and further assess its in vitro, ex vivo, and in vivo behavior for psoriasis management. PLGA microparticles (2%w/w) containing clobetasol propionate were developed by solvent evaporation technique. For the developed particles, size, drug loading, entrapment efficiency, and surface morphology were determined. Further, the particles were suspended into bigel developed from oleogel and hydrogel (40:60) and assessed for stability, in vitro drug release, ex vivo skin permeation, retention, and in vivo study in BALB c mice for antipsoriatic potential. PLGA microparticles developed by solvent evaporation were found to have a mean particle size of 19.45 ± 1.74 μm. Drug loading and entrapment efficiency for clobetasol were found to be 43.5 ± 4.94% and 94.5 ± 3.68%, respectively. For the developed bigel, sustained in vitro drug release was observed till 7 h. Ex vivo skin permeation with the developed formulation was found to be higher than the marketed formulation and drug suspended in bigel. A retention study performed on pig skin suggested the topical skin retention of the drug. Further, the psoriatic model was developed in BALB c mice with IMQ cream, and it was observed that the developed formulation helped considerably reduce the irritation score and PASI scoring in mice as compared to the standard treatment. The same is corroborated with the help of histopathological studies performed by Hematoxylin and Eosin (H&E) staining in the same animals. The study findings suggest that clobetasol microparticle-loaded bigel formulation appeared to be a promising therapy option for psoriasis.

228.7: In vitro and in vivo immune assessments of TKO pig skin grafts in New World (squirrel) monkeys

228.7: In vitro and in vivo immune assessments of TKO pig skin grafts in New World (squirrel) monkeys

Feasibility and Clinical Implementation of Electron FLASH Radiation Therapy in the Yorkshire Swine Model

Preclinical studies have shown FLASH radiation therapy (RT) increases the therapeutic index through reduction in normal tissue toxicity but with retained tumor control compared to conventional dose rate (CONV) RT. Dosimetry in FLASH beams is challenging and complex as beam monitoring and proper dosimetry analysis remain uncertain and under investigation. Despite these limitations, clinical translation of FLASH RT has already begun. For translation of FLASH RT from the preclinical stage, it is critical that robust clinical workflows and dosimetry methods be confidently established to ensure patient safety. Here, we present the clinical workflow for the Yorkshire pig, an animal that resembles the body dimension, weight, and biology of a human patient, with the goal to establish standard operating procedures to ensure a safe and robust clinical translation in our upcoming phase I study in cutaneous tumors. The study determines feasibility and safety while finding incidence of dose-limiting toxicities and maximum tolerable dose for future Phase II trials. All procedures were approved by the institutional animal care and use committee. 6 pigs (40-50 kg) were placed under general anesthesia and underwent CT imaging for radiation therapy simulation purposes. The skin was first shaven, and targets on the dorsolateral flanks were marked with tattoos and BBs for CT visualization. Vacloc immobilization was used to allow for reproducible setup on the treatment couch. A treatment planning model was established for treatment planning and dose evaluation purposes. CONV and FLASH single and fractionated dose regimens were prescribed to the 90% isodose line in a 9 MeV beam. Skin collimation and bolus minimized beam penumbra and increased skin dose. Treatment time and pulse repetition frequency were constant between all FLASH fields. Prescription levels were varied via dose per pulse. Calibration and verification of these settings were performed utilizing a multi-dosimeter method for verification in solid water. Output of the beam was verified on the day of the treatment using beam current transformers. This same multi-dosimeter method was used as in-vivo dosimetry on treatment day and compared to the dose verification ensure full dose was received. Variation between the three dosimeter methods was found to be within 5% among all pigs within the study. The maximum percent difference between dose verification and dose delivery was 6%. Consideration must be taken in dosimeter readout error due to the surface of the pig skin. FLASH and CONV toxicity results are currently under evaluation and will be published upon completion of the study. Establishing guidelines and protocols for electron FLASH clinical translation is important to instill confidence in patient safety with this new technique. This study has further optimized and developed dosimetry tools and setup to be used in future clinical trials.