Impose of KNDy/GnRH neural circuit in PCOS, ageing, cancer and Alzheimer's disease: StAR actions in prevention of neuroendocrine dysfunction.

The Kisspeptin1 (KISS1)/neurokinin B (NKB)/Dynorphin (Dyn) [KNDy] neurons in the hypothalamus regulate the reproduction stage in human beings and rodents. KNDy neurons co-expressed all KISS1, NKB, and Dyn peptides, and hence commonly regarded as KISS1 neurons. KNDy neurons contribute to the "GnRH pulse generator" and are implicated in the regulation of pulsatile GnRH release. The estradiol (E2)-estrogen receptor (ER) interactions over GnRH neurons in the hypothalamus cause nitric oxide (NO) discharge, in addition to presynaptic GABA and glutamate discharge from respective neurons. The released GABA and glutamate facilitate the activity of GnRH neurons via GABAA-R and AMPA/kainate-R. The KISS1 stimulates MAPK/ERK1/2 signaling and cause the release of Ca2+ from intracellular store, which contribute to neuroendocrine function, increase apoptosis and decrease cell proliferation and metastasis. The ageing in women deteriorates KISS1/KISS1R interaction in the hypothalamus which causes lower levels of GnRH. Because examining the human brain is so challenging, decades of clinical research have failed to find the causes of KNDy/GnRH dysfunction. The KISS1/KISS1R interactions in the brain have a neuroprotective effect against Alzheimer's disease (AD). These findings modulate the pathophysiological role of the KNDy/GnRH neural network in polycystic ovarian syndrome (PCOS) associated with ageing and, its protective role in cancer and AD. This review concludes with protecting effect of the steroid-derived acute regulatory enzyme (StAR) against neurotoxicity in the hippocampus, and hypothalamus, and these measures are fundamental for delaying ageing with PCOS. StAR could serve as novel diagnostic marker and therapeutic target for the most prevalent hormone-sensitive breast cancers (BCs).

Design strategies for composite matrix and multifunctional polymeric scaffolds with enhanced bioactivity for bone tissue engineering.

Over the past few decades, various bioactive material-based scaffolds were investigated and researchers across the globe are actively involved in establishing a potential state-of-the-art for bone tissue engineering applications, wherein several disciplines like clinical medicine, materials science, and biotechnology are involved. The present review article's main aim is to focus on repairing and restoring bone tissue defects by enhancing the bioactivity of fabricated bone tissue scaffolds and providing a suitable microenvironment for the bone cells to fasten the healing process. It deals with the various surface modification strategies and smart composite materials development that are involved in the treatment of bone tissue defects. Orthopaedic researchers and clinicians constantly focus on developing strategies that can naturally imitate not only the bone tissue architecture but also its functional properties to modulate cellular behaviour to facilitate bridging, callus formation and osteogenesis at critical bone defects. This review summarizes the currently available polymeric composite matrices and the methods to improve their bioactivity for bone tissue regeneration effectively.

Generation of hybrid tissue engineered construct through embedding autologous chondrocyte loaded platelet rich plasma/alginate based hydrogel in porous scaffold for cartilage regeneration.

Over the past decades, various attempts have been made to develop suitable tissue-engineered constructs to repair or regenerate the damaged or diseased articular cartilage. In the present study, we embedded Platelet rich plasma (PRP)/Sodium Alginate (SA) based hydrogel in porous 3D scaffold of chitosan (CH)/chondroitin sulfate (CS)/silk fibroin (SF) to develop hybrid scaffold for cartilage tissue construct generation with abilities to support shape recovery potential, facilitate uniform cells distribution and mimic gel like cartilage tissue extracellular matrix.The developed hybrid matrix shows suitable pore size (55-261 µm), porosity (77 ± 4.3%) and compressive strength (0.13 ± 0.04 MPa) for cartilage tissue construct generation and its applications. The developed SA/PRP-based cartilage construct exhibits higher metabolic activity, glycosaminoglycan deposition, expression of collagen type II, and aggrecan in comparison to SA based cell-scaffold construct. In-vivo animal study was also performed to investigate the biocompatibility and cartilage tissue regeneration potential of the developed construct. The obtained gross analysis of knee sample, micro-computed tomography, and histological analysis suggest that implanted tissue construct possess the superior potential to regenerate hyaline cartilage defect of thickness around 1.10 ± 0.36 mm and integrate with surrounding tissue at the defect site. Thus, the proposed strategy for the development of cartilage tissue constructs might be beneficial for the repair of full-thickness knee articular cartilage defects.

Symphony of the DNA flexibility and sequence environment orchestrates p53 binding to its responsive elements.

The p53 tumor suppressor protein maintains the genome fidelity and integrity by modulating several cellular activities. It regulates these events by interacting with a heterogeneous set of response elements (REs) of regulatory genes in the background of chromatin configuration. At the p53-RE interface, both the base readout and torsional-flexibility of DNA account for high-affinity binding. However, DNA structure is an entanglement of a multitude of physicochemical features, both local and global structure should be considered for dealing with DNA-protein interactions. The goal of current research work is to conceptualize and abstract basic principles of p53-RE binding affinity as a function of structural alterations in DNA such as bending, twisting, and stretching flexibility and shape. For this purpose, we have exploited high throughput in-vitro relative affinity information of responsive elements and genome binding events of p53 from HT-Selex and ChIP-Seq experiments respectively. Our results confirm the role of torsional flexibility in p53 binding, and further, we reveal that DNA axial bending, stretching stiffness, propeller twist, and wedge angles are intimately linked to p53 binding affinity when compared to homeodomain, bZIP, and bHLH proteins. Besides, a similar DNA structural environment is observed in the distal sequences encompassing the actual binding sites of p53 cistrome genes. Additionally, we revealed that p53 cistrome target genes have unique promoter architecture, and the DNA flexibility of genomic sequences around REs in cancer and normal cell types display major differences. Altogether, our work provides a keynote on DNA structural features of REs that shape up the in-vitro and in-vivo high-affinity binding of the p53 transcription factor.

Design and evaluation of ciprofloxacin loaded collagen chitosan oxygenating scaffold for skin tissue engineering.

Hypoxia and sepsis are key concerns towards modern regenerative medicine. Oxygen generating biomaterials having antibacterial property aims to answer these concerns. Hypoxia promotes reactive oxygen species at the implant site that delays wound healing. Sepsis in wound also contributes to delay in wound healing. Therefore, scaffold with antibacterial property and oxygen-producing capacities have shown ability to promote wound healing. In the present study oxygen releasing, ciprofloxacin loaded collagen chitosan scaffold was fabricated for sustained oxygen delivery. Calcium peroxide (CPO) acted as a chemical oxygen source. Oxygen release pattern exhibited a sustained release of oxygen with uniform deposition of CPO on the scaffold. The drug release study shows a prolonged, continuous, and sustained release of ciprofloxacin. Cell culture studies depict that scaffold has suitable cell attachment and migration properties for fibroblasts. In vivo studies performed in the skin flip model visually shows better wound healing and less necrosis. Histological studies show the maintenance of tissue architecture and the deposition of collagen. The results demonstrate that the proposed CPO coated ciprofloxacin loaded collagen-chitosan scaffold can be a promising candidate for skin tissue engineering.

Generation of scaffold incorporated with nanobioglass encapsulated in chitosan/chondroitin sulfate complex for bone tissue engineering.

Over the past decade, various composite materials fabricated using natural or synthetic biopolymers incorporated with bioceramic have been widely investigated for the regeneration of segmental bone defect. In the present study, nano-bioglass incorporated osteoconductive composite scaffolds were fabricated through polyelectrolyte complexation/phase separation and resuspension of separated complex in gelatin matrix. Developed scaffold exhibits controlled bioreactivity, minimize abrupt pH rise (~7.8), optimal swelling behavior (2.6+-3.1) and enhances mechanical strength (0.62 ± 0.18 MPa) under wet condition. Moreover, in-vitro cell study shows that the fabricated scaffold provide suitable template for cellular attachment, spreading, biomineralization and collagen based matrix deposition. Also, the developed scaffold was evaluated for biocompatibility and bone tissue regeneration potential through implantation in non-union segmental bone defect created in rabbit animal model. The obtained histological analysis indicates strong potential of the composite scaffold for bone tissue regeneration, vascularization and reconstruction of defects. Thus, the developed composite scaffold might be a suitable biomaterial for bone tissue engineering applications.

Osteogenic differentiation ability of human mesenchymal stem cells on Chitosan/Poly (Caprolactone)/nano beta Tricalcium Phosphate composite scaffolds.

Our work depicts the development and characterization of Chitosan/Poly (caprolactone)/nano beta-Tricalcium phosphate (CS/PCL/ß-TCP) porous composite scaffolds by freeze drying method. Addition of PCL to CS/ß-TCP composite scaffolds had significantly increased the compressive strength besides decelerating the degradation rate. Human mesenchymal stem cells (hMSCs) were chosen to assess in-vitro biocompatibility of the prepared scaffolds in terms of cell viability, cell attachment and proliferation by MTT assay, SEM and DNA Quantification assays respectively. Further, increased osteogenic differentiation assay results (Alkaline Phosphatase assay and Total calcium content) revealed the role of ß-TCP in composite scaffolds. Altogether, results suggest the potentiality of prepared porous freeze dried composite scaffolds in bone tissue engineering applications.

Design and evaluation of chitosan/chondroitin sulfate/nano-bioglass based composite scaffold for bone tissue engineering.

Chitosan, a natural biopolymer with osteoconductive properties is widely investigated to generate scaffolds for bone tissue engineering applications. However, chitosan based scaffolds lacks in mechanical strength and structural stability in hydrated condition and thereby limits its application for bone tissue regeneration. Thus in the present study, to overcome the limitations associated with chitosan based scaffolds, we fabricated polyelectrolyte complexation mediated composite scaffold of chitosan and chondroitin sulfate incorporated with nano-sized bioglass. Developed scaffolds were successfully characterized for various morphological, physico-chemical, mechanical and apatite forming properties using XRD, FT-IR, FE-SEM and TEM. It was observed that polyelectrolyte complexation followed by incorporation of bioglass significantly enhances mechanical strength, reduces excessive swelling behavior and enhances structural stability of the scaffold in hydrated condition. Also, in-vitro cell adhesion, spreading, viability and cytotoxity were investigated to evaluate the cell supportive properties of the developed scaffolds. Furthermore, alkaline phosphatase activity, biomineralization and collagen type I expression were observed to be significantly higher over the composite scaffold indicating its superior osteogenic potential. More importantly, in-vivo iliac crest bone defect study revealed that implanted composite scaffold facilitate tissue regeneration and integration with native bone tissue. Thus, developed composite scaffold might be a suitable biomaterial for bone tissue engineering applications.

Design and evaluation of chitosan/poly(l-lactide)/pectin based composite scaffolds for cartilage tissue regeneration.

Poor regenerative potential of cartilage tissue due to the avascular nature and lack of supplementation of reparative cells impose an important challenge in recent medical practice towards development of artificial extracellular matrix with enhanced neo-cartilage tissue regeneration potential. Chitosan (CH), poly (l-lactide) (PLLA), and pectin (PC) compositions were tailored to generate polyelectrolyte complex based porous scaffolds using freeze drying method and crosslinked by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), N-hydroxysuccinimide (NHS) solution containing chondroitin sulfate (CS) to mimic the composition as well as architecture of the cartilage extracellular matrix (ECM). The physical, chemical, thermal, and mechanical behaviors of developed scaffolds were done. The scaffolds were porous with homogeneous pore structure with pore size 49-170µm and porosities in the range of 79 to 84%. Fourier transform infrared study confirmed the presence of polymers (CH, PLLA and PC) within the scaffolds. The crystallinity of the scaffold was examined by the X-ray diffraction studies. Furthermore, scaffold shows suitable swelling property, moderate biodegradation and hemocompatibility in nature and possess suitable mechanical strength for cartilage tissue regeneration. MTT assay, GAG content, and attachment of chondrocyte confirmed the regenerative potential of the cell seeded scaffold. The histopathological analysis defines the suitability of scaffold for cartilage tissue regeneration.

Strategies on process engineering of chondrocyte culture for cartilage tissue regeneration.

The current work is an attempt to study the strategies for cartilage tissue regeneration using porous scaffold in wavy walled airlift bioreactor (ALBR). Novel chitosan, poly (L-lactide) and hyaluronic acid based composite scaffold were prepared. The scaffolds were cross-linked with 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide, N-hydroxysuccinimide and chondroitin sulfate to obtain interconnected 3D microstructure showing excellent biocompatibility, higher cellular differentiation and increased stability. The surface morphology and porosity of the scaffolds were analyzed using scanning electron microscopy (SEM) and mercury intrusion porosimeter and optimized for chondrocyte regeneration. The study shows that the scaffolds were highly porous with pore size ranging from 48 to 180 µm and the porosities in the range 80-92%. Swelling and in vitro degradation studies were performed for the composite scaffolds; by increasing the chitosan: HA ratio in the composite scaffolds, the swelling property increases and stabilizes after 24 h. There was controlled degradation of composite scaffolds for 4 weeks. The uniform chondrocyte distribution in the scaffold using various growth modes in the shake flask and ALBR was studied by glycosaminoglycans (GAG) quantification, MTT assay and mixing time evaluation. The cell culture studies demonstrated that efficient designing of ALBR increases the cartilage regeneration as compared to using a shake flask. The free chondrocyte microscopy and cell attachment were performed by inverted microscope and SEM, and from the study it was confirmed that the cells uniformly attached to the scaffold. This study focuses on optimizing strategies for the culture of chondrocyte using suitable scaffold for improved cartilage tissue regeneration.