Keratin as an effective coating material for in vitro stem cell culture, induced differentiation and wound healing assays.

The utilization of stem cells in tissue engineering holds great promise as efficient tools for tissue regeneration and in treating numerous musculoskeletal diseases. However, several limiting factors, such as precise delivery and control of differentiation of these stem cells as well as mimicking the microenvironment required to modulate stem cell behaviour in-vivo, have given rise to an urgent need for the development of new biomaterials which could be tailored to enhance cell renewal and/or direct cell fates. Keratin-rich biological materials offer several advantages, such as biocompatibility, tailorable mechanical properties, huge bioavailability, non-toxicity, non-immunogenic, and intrinsic tissue repair and/or regeneration capabilities, which makes them highly valued. In the present work, we report the preparation of keratin-based bio-materials from goat hair waste and its effectiveness as a coating material for in vitro culture and induced differentiation of mesenchymal stem cells (MSC's) and primary goat fibroblast cells. Since no known keratinase enzymes are expressed as such in human and/or animal systems, these keratin biomaterials could be used to slow the rate of degradation and deliver keratin-loaded stem cell scaffolds to induce their directed differentiation in vivo. The generated keratin materials have been characterized for surface morphology, protein structures, size and other properties using SDS-PAGE, LC/MS-MS, SEM, FTIR etc. Also, in vitro cell culture assays such as cell adhesion, viability using MTT, live dead assays, differentiation assays and in vitro scratch/wound healing assays were performed. Our results provide important data supporting tissue engineering applications of these keratinous biomaterials by combining the unique biological characteristics of goat hair-derived keratin material with the regenerative power of stem cells and their combinatorial use in applications such as disease treatment and injury repair as well as their use in the preparation of wound healing products, such as dressings and bandages, for management of clinical care in animals.

A comprehensive study on the arterial vasculature of the brain in water buffalo (Bubalus bubalis): Clinical correlates.

The present study was designed to provide a comprehensive analysis of the anatomical aspects of arterial blood vasculature in the water buffalo brains. Fifty cranial cavities of adult water buffaloes were opened via both the dorsal and ventral approaches and the arteries were exposed and photographed. The buffalo rostral epidural rete mirabile generally resembled that of large ruminants. The oculomotor, abducent and trigeminal nerves were intimately associated with the rostral rete. Similar to the majority of ruminants, the arterial circle of the brain was heart-shaped in buffalos and presented all collateral blood vessels as mentioned in the previous literature. The study further revealed that the cranial nerve roots were closely related to the arterial circle of the brain and could be used as indicators for differentiating various branches of the arterial circle of the brain. In addition to the usual variations of the arterial circle of the brain, a bihemispheric rostral cerebral artery along with an aplastic rostral cerebral artery were reported for the first time. The deviant behaviour of the rostral cerebral artery forms a baseline to study various clinical conditions of the blood vasculature in the buffalo brain. The rostral choroidal, middle cerebral and the caudal cerebral arteries constantly were emerged as single vessels without any variations. In conclusion, the anatomy of the arterial vasculature of the buffalo brain observed in the present study provided evidence of its morphological resemblance to other species of the Bovini tribe.

Biogenic Fabrication of Iron Oxide Nanoparticles from Leptolyngbya sp. L-2 and Multiple In Vitro Pharmacogenetic Properties.

Metallic nanoparticles have received a significant amount of reflection over a period of time, attributed to their electronic, specific surface area, and surface atom properties. The biogenic synthesis of iron oxide nanoparticles (FeONPs) is demonstrated in this study. The green synthesis of metallic nanoparticles (NPs) is acquiring considerable attention due to its environmental and economic superiorities over other methods. Leptolyngbya sp. L-2 extract was employed as a reducing agent, and iron chloride hexahydrate (FeCl3·6H2O) was used as a substrate for the biogenic synthesis of FeONPs. Different spectral methods were used for the characterization of the biosynthesized FeONPs, ultraviolet-visible (UV-Vis) spectroscopy gave a surface plasmon resonance (SPR) peak of FeONPs at 300 nm; Fourier transform infrared (FTIR) spectral analysis was conducted to identify the functional groups responsible for both the stability and synthesis of FeONPs. The morphology of the FeONPs was investigated using scanning electron microscopy (SEM), which shows a nearly spherical shape, and an X-ray diffraction (XRD) study demonstrated their crystalline nature with a calculated crystallinity size of 23 nm. The zeta potential (ZP) and dynamic light scattering (DLS) measurements of FeONPs revealed values of -8.50 mV, suggesting appropriate physical stability. Comprehensive in-vitro pharmacogenetic properties revealed that FeONPs have significant therapeutic potential. FeONPs have been reported to have potential antibacterial and antifungal properties. Dose-dependent cytotoxic activity was shown against Leishmania tropica promastigotes (IC50: 10.73 µg/mL) and amastigotes (IC50: 16.98 µg/mL) using various concentrations of FeONPs. The cytotoxic potential was also investigated using brine shrimps, and their IC50 value was determined to be 34.19 µg/mL. FeONPs showed significant antioxidant results (DPPH: 54.7%, TRP: 49.2%, TAC: 44.5%), protein kinase (IC50: 96.23 µg/mL), and alpha amylase (IC50: 3745 µg/mL). The biosafety of FeONPs was validated by biocompatibility tests using macrophages (IC50: 918.1 µg/mL) and red blood cells (IC50: 2921 µg/mL). In conclusion, biogenic FeONPs have shown potential biomedical properties and should be the focus of more studies to increase their nano-pharmacological significance for biological applications.

Scavenger receptor B1 facilitates the endocytosis of Escherichia coli via TLR4 signaling in mammary gland infection.

SCARB1 belongs to class B of Scavenger receptors (SRs) that are known to be involved in binding and endocytosis of various pathogens. SRs have emerging role in regulating innate immunity and host-pathogen interactions by acting in co-ordination with Toll-like receptors.Query Little is known about the function of SCARB1 in milk-derived mammary epithelial cells (MECs). This study reports the role of SCARB1 in infection and its potential association in TLR4 signaling on bacterial challenge in Goat mammary epithelial cells (GMECs). The novelty in the establishment of MEC culture lies in the method that aims to enhance the viability of the cells with intact characteristics upto a higher passage number. We represent MEC culture to be used as a potential infection model for deeper understanding of animal physiology especially around the mammary gland. On E.coli challenge the expression of SCARB1 was significant in induced GMECs at 6 h. Endoribonuclease-esiRNA based silencing of SCARB1 affects the expression of TLR4 and its pathways i.e. MyD88 and TRIF pathways on infection. Knockdown also affected the endocytosis of E.coli in GMECs demonstrating that E.coli uses SCARB1 function to gain entry in cells. Furthermore, we predict 3 unique protein structures of uncharacterized SCARB1 (Capra hircus) protein. Overall, we highlight SCARB1 as a main participant in host defence and its function in antibacterial advances to check mammary gland infections. Video Abstract.

Tribo-corrosive behavior of additive manufactured parts for orthopaedic applications.

Background: Additive manufacturing (AM) being an integral component of the production offers a wide variety of applications in the production of different components. The medical industry after the introduction of Additive Manufacturing has resulted in several advancements. The production of intricate patient-specific implants is one of such advancements which greatly assist a surgeon during a surgery. Orthopedic implants apart from possessing good mechanical strength are also expected to exhibit good tribological and corrosion behavior. As a result, the development of various orthopaedic implants and tools has become simple with the use of additive manufacturing. Objectives and Rationale: In the current paper an effort has been made to discuss actual scientific knowledge on the tribo-corrosive behavior of additive manufactured parts for orthopedic applications. Different studies dealing with the mechanisms of lubrication and friction in synovial joints have also been considered. A special focus has also been laid down to study the corrosive effect of implants on the human body. A section dedicated to texturing of orthopedic implants has also been provided. The paper further elaborates the different research challenges and issues related to the use of additive manufacturing for the production of optimized orthopedic implants. Conclusion: The study revealed that additive manufacturing has greatly aided in the manufacture of different orthopaedic implants with enhanced properties. However, a detailed study of the effect of processes like friction, wear, lubrication and corrosion in these implants needs to be done. The performance of these implants in the presence of various synovial fluids also needs to be addressed. However, the lack of more biocompatible materials, scalability and cost issues hinder the widespread use of AM in the different orthopaedic applications.

Functionally graded additive manufacturing for orthopedic applications.

Background: Additive Manufacturing due to its benefits in developing parts with complex geometries and shapes, has evolved as an alternate manufacturing process to develop implants with desired properties. The structure of human bones being anisotropic in nature is biologically functionally graded i,e. The structure possesses different properties in different directions. Therefore, various orthopedic implants such as knee, hip and other bone plates, if functionally graded can perform better. In this context, the development of functionally graded (FG) parts for orthopedic application with tailored anisotropic properties has become easier through the use of additive manufacturing (AM). Objectives: and Rationale: The current paper aims to study the various aspects of additively manufactured FG parts for orthopedic applications. It presents the details of various orthopedic implants such as knee, hip and other bone plates in a structured manner. A systematic literature review is conducted to study the various material and functional aspects of functionally graded parts for orthopedic applications. A section is also dedicated to discuss the mechanical properties of functionally graded parts. Conclusion: The literature revealed that additive manufacturing can provide lot of opportunities for development of functionally graded orthopedic implants with improved properties and durability. Further, the effect of various FG parameters on the mechanical behavior of these implants needs to be studied in detail. Also, with the advent of various AM technologies, the functional grading can be achieved by various means e.g. density, porosity, microstructure, composition, etc. By varying the AM parameters. However, the current limitations of cost and material biocompatibility prevent the widespread exploitation of AM technologies for various orthopedic applications.

Successful laparoscope-assisted orchiectomy in three cryptorchid sheep.

Three adult Corriedale cryptorchid sheep were subjected to laparoscope-assisted orchiectomy of the retained testicles. One (n = 2) or both (n = 1) the testicles were missing in their scrotal sac and inguinal regions. Ultrasonography was used to locate the retained testicles and their distance from the abdominal surface. The animals were restrained in dorsal recumbency and Trendlenburg posture under lumbosacral epidural anaesthesia using 2% lignocaine hydrochloride. Two laparoscopic ports were created in the caudal abdomen adjacent to the retained testicles. They were identified by their ovoid shape, white glistening surface (Tunica albuginea) and typical vasculature. Laparoscope-assisted exteriorization of the testicles after enlarging the ports, ligation of their blood supply and resection of the spermatic cord was performed successfully. The scrotal testes in two rams were then subjected to routine Burdizzoo castration. The laparoscopic port sites healed without complications and all the animals continued to do well subsequently. From this case report, it is concluded that in sheep the laparoscopy; a minimally invasive procedure can confirm abdominal retention of testicle/s and may also be used for their retrieval in a single sitting. Although total laparoscopic procedure is expected to reduce the incision size further but requires advanced laparoscopic instruments and expertise.

Effect of intravitreal bevacizumab on macular thickness: exploring serum and vitreous proangiogenic biomarkers in patients with diabetic macular edema

Background/aim: This study evaluates diabetic macular-edema (DME) patients for the effect of intravitreal bevacizumab (IVB) injection on macular thickness and proangiogenic biomarkers in serum and vitreous. Materials and methods: Forty DME patients were analyzed for macular thickness (MT). Twelve proangiogenic biomarkers in serum and vitreous were analyzed before and after IVB. Results: Significant decrease in MT with vitreal vascular endothelial growth factor-A (VEGF-A) was observed as expected after IVB, while serum VEGF-A did not follow a decreasing trend in contrast to VEGF-C, which decreased both in serum and vitreous. Other vitreal factors like bone morphogenetic protein-9 (BMP9) and fibroblast growth factor (FGF) were also significantly decreased, while endothelial growth factor (EGF) increased following IVB. Before IVB, significant negative correlations were vitreous BMP9 with serum FGF, vitreous human growth factor (HGF) and interleukin-8 (IL-8) with serum endothelin, and vitreous and serum FGF and serum placental growth factor (PLGF) with EGF. After IVB, negative correlations in serum vs. vitreous were found for both HGF and PLGF with BMP9, and angiopoietin with FGF. Cube average thickness was negatively correlated with serum FGF and positively correlated with vitreous PLGF and endothelin. Conclusion: Vascular endothelial growth factors are not the only factors that cause macular edema in diabetic patients. The effect of IVB on different proangiogenic biomarkers indicated a complex interplay of other factors in DME.

Advances in genome editing for improved animal breeding: A review.

Since centuries, the traits for production and disease resistance are being targeted while improving the genetic merit of domestic animals, using conventional breeding programs such as inbreeding, outbreeding, or introduction of marker-assisted selection. The arrival of new scientific concepts, such as cloning and genome engineering, has added a new and promising research dimension to the existing animal breeding programs. Development of genome editing technologies such as transcription activator-like effector nuclease, zinc finger nuclease, and clustered regularly interspaced short palindromic repeats systems begun a fresh era of genome editing, through which any change in the genome, including specific DNA sequence or indels, can be made with unprecedented precision and specificity. Furthermore, it offers an opportunity of intensification in the frequency of desirable alleles in an animal population through gene-edited individuals more rapidly than conventional breeding. The specific research is evolving swiftly with a focus on improvement of economically important animal species or their traits all of which form an important subject of this review. It also discusses the hurdles to commercialization of these techniques despite several patent applications owing to the ambiguous legal status of genome-editing methods on account of their disputed classification. Nonetheless, barring ethical concerns gene-editing entailing economically important genes offers a tremendous potential for breeding animals with desirable traits.

Long non-coding RNAs: Mechanism of action and functional utility.

Recent RNA sequencing studies have revealed that most of the human genome is transcribed, but very little of the total transcriptomes has the ability to encode proteins. Long non-coding RNAs (lncRNAs) are non-coding transcripts longer than 200 nucleotides. Members of the non-coding genome include microRNA (miRNA), small regulatory RNAs and other short RNAs. Most of long non-coding RNA (lncRNAs) are poorly annotated. Recent recognition about lncRNAs highlights their effects in many biological and pathological processes. LncRNAs are dysfunctional in a variety of human diseases varying from cancerous to non-cancerous diseases. Characterization of these lncRNA genes and their modes of action may allow their use for diagnosis, monitoring of progression and targeted therapies in various diseases. In this review, we summarize the functional perspectives as well as the mechanism of action of lncRNAs.