Tracking new insights into antifungal and anti-mycotoxigenic properties of a biofilm forming Pediococcus pentosaceus strain isolated from grain silage.

The present study offers detailed insights into the antifungal and anti-mycotoxigenic potential of a biofilm forming lactic acid bacterium (Pediococcus pentosaceus) against one atoxigenic (Aspergillus flavus) and two toxigenic (Aspergillus nomius and Fusarium verticillioides) fungal strains. The antifungal effect of P. pentosaceus LBM18 strain was initially investigated through comparative analysis of fungi physiology by macroscopic visual evaluations and scanning electron microscopy examinations. The effects over fungal growth rate and asexual sporulation were additionally accessed. Furthermore, analytical evaluations of mycotoxin production were carried out by HPLC-MS/MS to provide insights on the bacterial anti-mycotoxigenic activity over fungal production of the aflatoxins B1, B2, G1 and G2 as well as fumonisins B1 and B2. Finally, reverse transcription quantitative real-time PCR (RT-qPCR) analysis was employed at the most effective bacterial inoculant concentration to evaluate, at the molecular level, the down-regulation of genes aflR, aflQ and aflD, related to the biosynthesis of aflatoxins by the strain of Aspergillus nomius. The effects over mycotoxin contamination were thought to be result of a combination of several biotic and abiotic factors, such as interaction between living beings and physical-chemical aspects of the environment, respectively. Several possible mechanisms of action were addressed along with potentially deleterious effects ascribing from P. pentosaceus misuse as biopesticide, emphasizing the importance of evaluating lactic acid bacteria safety in new applications, concentrations, and exposure scenarios.

Evaluating the potential of Pediococcus pentosaceus as a biocontrol agent against tenuazonic acid-producing Alternaria alternata on livestock feeds.

AIMS: This study aims to demonstrate the potential of the lactic acid bacteria (LAB) Pediococcus pentosaceus LBM18 against the mycotoxin-producing Alternaria alternata TEF-1A and highlight its application as an effective grain silage inoculant to control mycotoxin contamination. METHODS AND RESULTS: The antifungal properties of Ped. pentosaceus lyophilized (PPL) were assessed by evaluating its effect on A. alternata TEF-1A grown in a corn silage-based medium, which included morphological changes by Scanning Electron Microscopy (SEM) observations, growth rate, conidia production assays, and inhibition of Tenuazonic acid (TeA) production by high-performance liquid chromatography (HPLC-MS/MS) analyses. Furthermore, TeA biosynthesis was monitored for changes at the molecular level by PKS gene expression. The growth and sporulation processes of A. alternata TEF-1A were affected by Ped. pentosaceus LBM18 in a concentration-dependent manner. Moreover, a significant inhibition of TeA production (74.3%) and the transcription level of the PKS gene (42.9%) was observed. CONCLUSIONS: Ped. pentosaceus is one of the promising LAB to be applied as an inoculant for corn silage preservation, aiming to inhibit mycotoxigenic fungi growth and their mycotoxin production. SIGNIFICANCE AND IMPACT OF THE STUDY: Ped. pentosaceus could be used as an inoculant to reduce fungal and mycotoxins contamination in grain silage production.

Structural and Ultrastructural Characteristics of the Spix's Yellow-Toothed Cavy (Galea spixii, Wagler, 1831) Tongue.

The tongue is a fundamental organ in feeding, vocalization, and grooming. It is characterized by evolutionary adaptations reflected by diet, habitat, and function. Rodents are a very diverse mammalian order and the tongue's morphology varies in size, form, and presence of papillae. This work aimed to describe the morphological and ultrastructural aspects of the tongue of Spix's yellow-toothed cavy (Galea spixii, Wagler, 1831). Tongues of Spix's yellow-toothed cavies were analyzed with light microscopy, scanning, and transmission electron microscopy. The results showed that the tongue was divided into apex, body, and root. There were different types of papillae, such as vallate, foliate, laterally placed fungiform, fungiform, filiform, and robust filiform. The epithelium was organized into layers, including keratinized, granulous, spinous, and basal, below were lamina propria, and musculature, which evolved mucous and serous gland clusters. The tongue of Spix's yellow-toothed cavy was structurally and ultrastructurally similar to other rodents and had papillae with similar morphologies to other Caviidae species. However, the presence of robust filiform papillary lines and laterally placed fungiform papillae showed the main differences from other species. This was the first description of the tongue of Spix's yellow-toothed cavy.

Structural and Ultrastructural Changes in the Tongue of mdx Mice.

The mdx mouse is an experimental model of Duchenne muscular dystrophy, a genetic disorder characterized by progressive muscular degeneration which affects the oral cavity musculature, and promotes difficulty in swallowing. This study aimed to describe morphological, structural, and ultrastructural changes in the tongue mucosa and musculature of mdx mice. Forty six-month-old mice were divided into two groups: Control C57bl/10 (n = 20) and mdx C57bl/10mdx (n = 20). The tongue was dissected and analyzed with light microscopy, scanning electron microscopy, and transmission electron microscopy techniques. Our results showed conical and triangular filiform, fungiform, foliate, and vallate papillae, and their connective tissue cores. The epithelium layers identified were corneum, granulosum, spinosum, and basale. The mdx group had a thicker epithelium. Lamina propria was composed of reddish and greenish collagen. In mdx, collagen was present in the musculature of the tongue's body and in the muscular tissue between mucous and serous glands of the caudal region. Musculature was also characterized by a shorter length of sarcoplasmic invaginations, myocytolysis in mitochondrial groupings, and inflammatory focus. In conclusion, the tongue of 6-month-old mdx mice had morphology, structure, and ultrastructure revealed, showing higher wear of filiform papillae indirect reflex from the muscular degeneration process.

Morphological Changes in the Myotendinous Junction of mdx Mice.

The myotendinous junction (MTJ) is the interface between muscle and tendon, and it is the main area of force transmission of the locomotor apparatus. Dystrophic processes promote pathological injury which affects the skeletal muscle and can influence the morphology of the MTJ. This study aimed to investigate the adaptations in MTJ morphology of mdx mice in the tibialis anterior muscle. Male mice (n = 24) were divided into Control­C57bl/10 and mdx­C57bl/10mdx (Duchenne muscular dystrophy experimental model). In the mdx group, centralized nuclei with a large area and greater deposition of type III collagen (fibrosis) were observed. Also, shorter sarcomeres and sarcoplasmatic projections of MTJ were observed. We concluded that the adaptations in mdx mice demonstrated extensive impairment in the MTJ region with reduced ultrastructures.

Aquatic Training after Joint Immobilization in Rats Promotes Adaptations in Myotendinous Junctions.

The myotendinous junction (MTJ) is the muscle-tendon interface and constitutes an integrated mechanical unit to force transmission. Joint immobilization promotes muscle atrophy via disuse, while physical exercise can be used as an adaptative stimulus. In this study, we aimed to investigate the components of the MTJ and their adaptations and the associated elements triggered with aquatic training after joint immobilization. Forty-four male Wistar rats were divided into sedentary (SD), aquatic training (AT), immobilization (IM), and immobilization/aquatic training (IMAT) groups. The samples were processed to measure fiber area, nuclear fractal dimension, MTJ nuclear density, identification of telocytes, sarcomeres, and MTJ perimeter length. In the AT group, the maintenance of ultrastructure and elements in the MTJ region were observed; the IM group presented muscle atrophy effects with reduced MTJ perimeter; the IMAT group demonstrated that aquatic training after joint immobilization promotes benefits in the muscle fiber area and fractal dimension, in the MTJ region shows longer sarcomeres and MTJ perimeter. We identified the presence of telocytes in the MTJ region in all experimental groups. We concluded that aquatic training is an effective rehabilitation method after joint immobilization due to reduced muscle atrophy and regeneration effects on MTJ in rats.

Structural and Ultrastructural Characteristics of the Red-Rumped Agouti (Dasyprocta leporina-Linnaeus, 1758) Palatine Epithelium.

The palate is a fundamental region in food swallowing and presents different adaptations in species. This research aimed to describe structural and ultrastructural characteristics of the palatine epithelium and the connective tissue cores (CTCs) of ten red-rumped agoutis (Dasyprocta leporina­Linnaeus, 1758) using macroscopic, light microscopy, scanning electron microscopy, and transmission electron microscopy. We found nine palatine ridges in the diastema and hard palate, and a smooth surface in the soft palate. Stratified squamous keratinized epithelium with projections of lamina propria and soft palate had gland clusters. Epithelial removal revealed CTCs with a conical shape with high density in the hard palate and the sides of the soft palate. Near the CTCs were nerve fibers in the hard palate, and the soft palate had muscular tissue below the gland clusters. The structural and ultrastructural characteristics enable stability of the hard palate and fixation to the soft palate sides, while the soft palate center has greater mobility thus assisting in food swallowing. We concluded that structural characteristics are similar to other mammals, although the morphology of agouti's palate differs in the amount and disposition of palatine ridges, and the conical CTC's morphology.

Myotendinous Junction Components of Different Skeletal Muscles Present Morphological Changes in Obese Rats.

Obesity is characterized by excess adipose tissue and chronic inflammation and promotes extensive changes that can compromise skeletal muscles' structural and functional integrity. Obesity can seriously impact the force transmission region between the muscle and the tendon, the myotendinous junction (MTJ). The present study aimed to investigate the plasticity of muscle fibers and MTJ regions in high-fat diet-induced obesity in rat tibialis anterior (TA) and soleus (SO) muscles. Wistar rats were divided into control and obese groups (induced by a high-fat diet). The samples of TA and SO muscles were prepared for histochemical and ultrastructural analysis (sarcomeres and MTJ projection). In the muscle fiber, similar adaptations were observed between the muscles of the smaller fiber (types I and IIa) in the obesity results. The MTJ region demonstrated different adaptations between the analyzed muscles. The TA­MTJ region has shorter ultrastructures, while in the SO­MTJ region, the ultrastructures were larger. We conclude that obesity induced by a high-fat diet promotes similar adaptation in the muscle fibers; however, in the MTJ region, the sarcoplasmatic projections and adjacent sarcomere demonstrate different adaptations according to distinct muscle phenotypes.

Ecomorphological, space, and mineral relations of dermal denticles in angular angel shark (Squatina guggenheim).

Shark skin is predominantly specialized for swimming and protection, with the dermal denticle being the main structure associated with these abilities. The dermal denticle is a mineral structure with a unique morphology for each species, which allows its use as a taxonomic tool. Few studies have investigated the microscopy aspects of skin and dermal denticles, considering the high diversity of sharks. Here, we investigated the three-dimensional morphoquantitative aspects and mineral composition of dermal denticles in different regions of the angular angel shark, Squatina guggenheim, using scanning electron microscopy and dispersive energy system. With the microscopy, we were able to observe that the dermal denticle morphology changes according to the area it is located. It was possible to describe the dermal denticles individually, from root to the crown, highlighting all of their individualities. Through the dispersive energy system, we showed the proportions of each mineral found in the denticle, by area, demonstrating the composition and the particularities of crown, body, and root, where whitlockite was described for the first time in elasmobranchs. In this way, the present study presented the specificities of the dermal denticles of S. guggenheim, as well sought to understand the different structure functions for the animal, thus assisting future research in animal morphology.

Structural and ultrastructural characterization of the palatine epithelium of the Guinea pig: A new record of telocytes in the oral cavity.

The morphology of the oral cavity of mammals relates to diet, habitat, and function. The palate is an important region with adaptations for oral somatosensation and mechanical loads due to the pressure of the tongue with food. The research aimed to describe the structural and ultrastructural characteristics of the epithelium and the connective tissue cores of the guinea pig palate using macroscopic, light microscopy, scanning electron microscopy, and transmission electron microscopy analysis. The hard palate had conical and filiform papillae, and the soft palate had open salivary ducts. After the removal of the epithelium, the connective tissue cores revealed thin filaments and laminar projections in the hard palate, and opening ducts were evidenced in the soft palate. The palatine epithelium was keratinized and organized by layers, lamellated corpuscles were found in lamina propria of the hard palate. In contrast, the soft palate had glands clusters associated with nerve fibers, and in both regions were identified telocytes. We concluded that the hard palate presented conical and filiform papillae that differ from other mammals. Besides, it is a new description of the connective tissue cores morphology and the first record of the telocytes in this anatomical region for mammals.

Myotendinous junction adaptations to ladder-based resistance training: identification of a new telocyte niche.

The present study shows chronic adjustments in the myotendinous junction (MTJ) in response to different ladder-based resistance training (LRT) protocols. Thirty adult male Wistar rats were divided into groups: sedentary (S), calisthenics (LRT without additional load [C]), and resistance-trained (LRT with extra weight [R]). We demonstrated longer lengths of sarcoplasmatic invaginations in the trained groups; however, evaginations were seen mainly in group R. We showed a greater thickness of sarcoplasmatic invaginations in groups C and R, in addition to greater evaginations in R. We also observed thinner basal lamina in trained groups. The support collagen layer (SCL) adjacent to the MTJ and the diameters of the transverse fibrils were larger in R. We also discovered a niche of telocytes in the MTJ with electron micrographs of the plantar muscle and with immunostaining with CD34+ in the gastrocnemius muscle near the blood vessels and pericytes. We concluded that the continuous adjustments in the MTJ ultrastructure were the result of tissue plasticity induced by LRT, which is causally related to muscle hypertrophy and, consequently, to the remodeling of the contact interface. Also, we reveal the existence of a collagen layer adjacent to MTJ and discover a new micro anatomic location of telocytes.

A new insight of Platelet-Rich Fibrin clots morphology and their elemental composition.

This study analyzed the architecture of Platelet-Rich Fibrin (PRF) clots and assessed their elemental composition in order to provide new insight into this biomaterial. Five surplus PRF clots (2,700 RPM, 12 min.) donated by patients (63.6 ±â€¯12.3 years old) were prepared for use in dental clinical procedures. The internal three-dimensional morphology of the red zones and the thirds of the yellow zones of the clots were analyzed by Variable Pressure Scanning Electron Microscope (VPSEM) after sample preparation by two methods: 1. Fixation (2.5% gluataraldehyde); and 2. Fixation with subsequent partial removal of extracellular elements (8 N, HCl). Semi-quantitative elemental analysis was performed by energy-dispersive X-ray spectrometry (EDX). VPSEM analysis showed erythrocytes in both the red zone and the yellow zone, which consisted mainly of fibrin. Removal of extracellular elements enriched the morphology of both zones; the organization of the fibrin was observed to differ in the thirds of the yellow zone, with increasing density and organization to distal. The elements that compose organic substances (C-Carbon, N-Nitrogen, O-Oxygen, Na-Sodium and P-Phosphorus) and halogens (Cl-Chloride and S-Sulfur) were detected; the highest concentrations were of C, followed by O (p < 0.05), in the proximal region of the fibrin. The results of the present study suggest organization of fibrin in the PRF clot, and also reveal the distribution of the elements present in the different regions of the clot. Improved understanding of these characteristics may favor the use of this biomaterial by increasing its efficiency and functionality.

Antibacterial and antifungal activity of crude and freeze-dried bacteriocin-like inhibitory substance produced by Pediococcus pentosaceus.

Pediococcus pentosaceus LBM 18 has shown potential as producer of an antibacterial and antifungal bacteriocin-like inhibitory substance (BLIS). BLIS inhibited the growth of spoilage bacteria belonging to Lactobacillus, Enterococcus and Listeria genera with higher activity than Nisaplin used as control. It gave rise to inhibition halos with diameters from 9.70 to 20.00 mm, with Lactobacillus sakei being the most sensitive strain (13.50-20.00 mm). It also effectively suppressed the growth of fungi isolated from corn grain silage for up to 25 days and impaired morphology of colonies by likely affecting fungal membranes. These results point out that P. pentosaceus BLIS may be used as a new promising alternative to conventional antibacterial and antifungal substances, with potential applications in agriculture and food industry as a natural bio-controlling agent. Moreover, cytotoxicity and cell death induction tests demonstrated cytotoxicity and toxicity of BLIS to human colon adenocarcinoma Caco-2cells but not to peripheral blood mononuclear cells, with suggests possible applications of BLIS also in medical-pharmaceutical applications.

Repercussions on sarcomeres of the myotendinous junction and the myofibrillar type adaptations in response to different trainings on vertical ladder.

The myofibrillary types establish to the skeletal muscle functional and adaptive properties that influence the sarcomeric arrangement during muscle contraction and may have repercussions on an important related force transmission region of the locomotor apparatus, the myotendinous junction (MTJ). This study aimed to describe changes in myofibrillary type and sarcomeric lengths in the belly muscle and MTJ of the soleus and plantaris muscles associated with training protocols in vertical ladder. Thirty adults male Wistar rats were divided into three groups (n = 10): Control (CTR), No-load Training (NLT), and Load Training (LT). Morphoquantitative analysis of different fibers types and sarcomere lengths were performed in distinct regions of plantaris and soleus muscles. In the plantaris muscle with both trainings, there was an increase in the cross-sectional area (CSA) in Type I and II fibers (p < .0001) while sarcomeric lengths revealed greater lengths in the proximal and distal sarcomeres of NLT, although in the LT we found greater lengths in the belly and MTJ sarcomeres. The soleus muscle showed an increase in CSA muscle fiber only in the NLT (p < .0001) and revealed alterations in belly and MTJ sarcomere lengths with training. We concluded that plantaris muscle has an adaptive effect directly associated with training load, with hypertrophy in both trainings and sarcomere length inverse from belly and MTJ, in LT associated with increased force generation and transmission at the MTJ, although soleus muscle has a lower adaptive response to training stimuli with variation in the belly and distal sarcomere of the MTJ.

Morphological and ultrastructural characteristics of the tongue of wild boar.

The present study aimed to describe the structural and ultrastructural morphological characteristics of the lingual epithelium and the connective tissue cores (CTCs) of wild boar (Sus scrofa). The tongues were processed for light microscopy, scanning electron microscopy, and transmission electron microscopy. In this study, we revealed the filiform, fungiform, foliate, and vallate papillae. The filiform papilla is elongated with a conical shape and its CTC has a conical shape; the fungiform papilla is rounded with a dome-shape and its CTC is flower bud; the foliate papilla is formed by four pairs of epithelial folds and irregular grooves, and its CTC is thin with adjacent conjunctive projections, and taste buds and serous glands in the epithelial layer have been evidenced; and the vallate papilla is oval surrounded by a groove with increases of epithelium surface, and the CTC is formed by numerous connective projections lined. Also noted were serous gland and taste buds on the medial wall of the vallate papilla. The epithelium has the keratinized, granular, spinous, basal, and lamina propria layers. In conclusion, we found new descriptions and shapes of the CTCs of the lingual papillae. In addition, we demonstrated the epithelium structural characteristics, the nuclear distribution between the epithelial layers, and the ultrastructural aspects of the dorsal epithelium of the tongue.

Beneficial effects of benzodiazepine on masticatory muscle dysfunction induced by chronic stress and occlusal instability in an experimental animal study.

Psychological stress and occlusal alteration are important etiologic factors for temporomandibular/masticatory muscular disorders. In particular, the exact physiologic mechanism underlying the relation by occlusal alteration and temporomandibular disorders remains unclear. Our purpose was to test the hypothesis that benzodiazepine therapy is able to prevent metabolic and vascular changes in the medial pterygoid muscle of rats under chronic stress after 14 days of unilateral exodontia. Adult Wistar rats were submitted to unpredictable chronic mild stress (10 days) and/or unilateral exodontia and their plasma and medial pterygoid muscles were removed for analysis. A pre-treatment with diazepam was used to verify its effect on stress. The parameters evaluated included anxiety behavior, plasma levels of corticosterone, metabolic activity by succinate dehydrogenase, capillary density by laminin staining and ultrastructural findings by transmission electron microscopy. Occlusal instability induced anxiety-like behavior on elevated plus-maze test and diazepam administration blocked the appearance of this behavior. Unilateral exodontia promoted in the contralateral muscle an increase of oxidative fibers and capillaries and modification of sarcoplasmic reticulum. Chronic stress caused increased glycolytic metabolism, reduced capillary density and morphological changes in mitochondria on both sides. Association of both factors induced a glycolytic pattern in muscle and hemodynamic changes. Pharmacological manipulation with diazepam inhibited the changes in the medial pterygoid muscle after stress. Our results reveal a preventive benzodiazepine treatment for stress and occlusal instability conditions affecting masticatory muscle disorders. In addition, provide insights into the mechanisms by which chronic stress and exodontia might be involved in the pathophysiology of masticatory muscular dysfunctions.

Protective Malaria Vaccine in Mice Based on the Plasmodium vivax Circumsporozoite Protein Fused with the Mumps Nucleocapsid Protein.

Plasmodium vivax is the most common species of human malaria parasite found outside Africa, with high endemicity in Asia, Central and South America, and Oceania. Although Plasmodium falciparum causes the majority of deaths, P. vivax can lead to severe malaria and result in significant morbidity and mortality. The development of a protective vaccine will be a major step toward malaria elimination. Recently, a formulation containing the three allelic variants of the P. vivax circumsporozoite protein (PvCSP-All epitopes) showed partial protection in mice after a challenge with the hybrid Plasmodium berghei (Pb) sporozoite, in which the PbCSP central repeats were replaced by the VK210 PvCSP repeats (Pb/Pv sporozoite). In the present study, the chimeric PvCSP allelic variants (VK210, VK247, and P. vivax-like) were fused with the mumps virus nucleocapsid protein in the absence (NLP-CSPR) or presence of the conserved C-terminal (CT) domain of PvCSP (NLP-CSPCT). To elicit stronger humoral and cellular responses, Pichia pastoris yeast was used to assemble them as nucleocapsid-like particles (NLPs). Mice were immunized with each recombinant protein adjuvanted with Poly (I:C) and presented a high frequency of antigen-specific antibody-secreting cells (ASCs) on days 5 and 30, respectively, in the spleen and bone marrow. Moreover, high IgG titers against all PvCSP variants were detected in the sera. Later, these immunized mice with NLP-CSPCT were challenged with Pb/Pv sporozoites. Sterile protection was observed in 30% of the challenged mice. Therefore, this vaccine formulation use has the potential to be a good candidate for the development of a universal vaccine against P. vivax malaria.

Structural and ultrastructural characteristics of the tongue of the Collared Peccary (Pecari tajacu, Linnaeus, 1758).

The tongue is an important organ in species due to its feeding functions, and its structure is influenced by the habitat and diet. The Collared Peccary (Pecari tajacu, Linnaeus, 1758) is a terrestrial mammal that is distributed on the American continents and has an omnivorous diet. This study aimed to describe the morphological characteristics of the tongue, lingual papillae and the connective tissue cores (CTCs) of the Collared Peccary. Eight tongues were collected from the Wild Animals Multiplication Center. The samples (n = 6) were processed for three-dimensional analysis of their dorsal epithelium, and their CTCs by scanning electron microscopy and the other samples (n = 2) were used to observe ultrastructural characteristics by transmission electron microscopy. Filiform papillae were observed in the lingual apex and body with their conical CTC demonstrating ripples in their extent. Two types of fungiform papillae were observed, the first in the apex and body with a dome-shaped CTC and the second forming a dorsolateral line with a radial pattern CTC. The vallate papillae were bilateral in the caudal region with the CTCs, characterised by numerous projections, and foliate papilla were not observed. We found a new type of papilla anteriorly to the vallate papilla with an irregular groove and a CTC formed by triangular-shaped projections with a duct opening at the top. We concluded that the lingual papillae of the Collared Peccary resemble those papillae of other mammals, however, were revealed another form of their CTCs and a new lingual papillae morphological description.

Effects of physical training on sarcomere lengths and muscle-tendon interface of the cervical region in an experimental model of menopause.

The aim of this study was to describe the structural and ultrastructural aspects of the myotendinous junction (MTJ) and the proximal and distal sarcomeres of the sternomastoid of aged Wistar rats subjected to an experimental model of menopause and swimming training. A total of 20 female elderly rats were divided into the following four groups (n=5 in each group): sedentary/no-menopausal (SNM), trained/no-menopausal (TNM), sedentary/menopausal (SM), and trained/menopausal (TM). The MTJ samples were dissected and analyzed using transmission electron microscopy. We showed that the TNM Group rats exhibited changes in morphological characteristics as a consequence of physical exercise, which included an increase of 36.60% (P<0.001) in the evagination length of the MTJ and a reduction in the length of the distal (77.38%) (P<0.0001) and proximal (68.15%) (P<0.0001) sarcomeres. The SM Group exhibited a reduction of about 275.93% (P<0.001) in the muscle-tendon interface and in the lengths of distal sarcomeres (55.87%) (P<0.0001) compared with SNM Group. Our results suggest that the swimming training under experimental model of menopause promoted tissue reorganization and increased muscle-tendon interaction with a drastic development in the length and thickness of the sarcoplasmatic invaginations and evaginations. In addition, the sarcomeres exhibited different lengths and a reduction in both groups subjected to swimming training.

Application of a Purified Protein From Natural Latex and the Influence of Suture Type on Achilles Tendon Repair in Rats.

BACKGROUND: The prolonged tendon-healing process, the high costs associated with treatment, the increase in the number of injuries over the past decades, and the lack of consensus on the optimal treatment of tendon injuries are a global problem. Restoring the normal tendon anatomy and decreasing the healing time are key factors for treatment advancement. HYPOTHESIS: Application of a purified protein from natural latex (PPNL) accelerates the healing process, increasing collagen synthesis and decreasing metalloproteinases. PPNL associated with a simpler suture technique should decrease the healing time. STUDY DESIGN: Controlled laboratory study. METHODS: Injury, surgery, and treatment with PPNL were conducted with male Sprague-Dawley rats. Two suture techniques were used: U-suture, a simpler and lesser traumatic technique, and Kessler-Tajima, to avoid strangulation of the microcirculation. Achilles tendons were completely sectioned, and 100 µL of 0.1% PPNL was applied on the tendon during surgery. Tendon morphology, distribution, and quantity of collagen types I and III, as well as expression of TIMP-1, TIMP-2, MMP-2, and MMP-9 and ultrastructural aspects of cells and collagen fibrils, were assessed after 2 and 4 weeks. RESULTS: PPNL treatment improved collagen type I synthesis and reduced MMP-2 expression. All groups showed a 6.8-times increase in tendon weight as compared with the control group after 2 weeks and a 5.2-times increase after 4 weeks. All groups showed an increase in diameter after 4 weeks, except for the ones treated with PPNL, which showed a slight reduction in diameter. The peak of concentration of collagen fibrils with a 80-nm diameter was 27.79% in the control group; all other experimental groups presented fibrils between 50 and 60 nm. However, the best results were observed with Kessler-Tajima suture associated with PPNL. CONCLUSION/CLINICAL RELEVANCE: There are no known medicines or substances capable of aiding the tendon healing process besides surgery. The discovery of a substance able to improve this process and decrease its duration represents an important advancement in orthopaedic medicine.