Decellularization of caprine esophagus using fruit pericarp extract of Sapindus mukorossi.

Biological detergents like sodium deoxycholate, sodium dodecyl sulphate and Triton X-100 impairs the collagenous and non-collagenous proteins, glycosaminoglycans and growth factors. Further, certain chemical and enzymes are responsible for residual cytotoxicity in the decellularized extracellular matrix. The main focus of this study was to explore the decellularization property of soap nut pericarp extract (SPE) for development of decellularized tubular esophageal scaffold. For this 2.5, 5.0 and 10% concentrations of SPE were used for decellularization of caprine esophageal tissues. Histological analysis of hematoxylin and eosin and Masson's trichrome stained tissue samples confirmed decellularization with preservation of extracellular matrix microarchitecture. Scanning electron microscopic images of luminal surface of decellularized esophageal matrix showed randomly oriented collagen fibres with large interconnected pores and cells were absent. However, the external surface was more textured with fibrous structures and collagen fibres were well preserved. DAPI stained decellularized tissues revealed complete removal of nuclear components, verified by DNA content measurement and SDS-PAGE. The FTIR spectra of decellularized esophagus shows absorption peaks of amide A, B, I, II and III. Elastic modulus of the decellularized esophagus scaffolds increased (P > 0.05) as compared to native tissues. Histological and scanning electron microscopic evaluation of in vitro seeded scaffolds showed attachment and growth of primary chicken embryo fibroblasts over and within the decellularized scaffolds. It was concluded that 5% SPE is ideal for preparation of cytocompatible decellularized caprine esophageal scaffold with well-preserved extracellular matrix architecture and, may be used as an alternative to biological detergents and other chemicals.

Decellularization of Skin Tissue.

Biomaterials science encompasses elements of medicine, biology, chemistry, materials, and tissue engineering. They are engineered to interact with biological systems to treat, augment, repair, or replace lost tissue function. The choice of biomaterial depends on the procedure being performed, the severity of the patient's condition, and the surgeon's preference. Prostheses made from natural-derived biomaterials are often derived from decellularized extracellular matrix (ECM) of animal (xenograft) or human (allograft) origin. Advantages of using ECM include their resemblance in morphology and three-dimensional structures with that of tissue to be replaced. Due to this, scientists all over are now focusing on naturally derived biomaterials which have been shown to possess several advantages compared to synthetic ones, owing to their biocompatibility, biodegradability, and remodeling properties. Advantages of a naturally derived biomaterial enhance their application for replacement or restoration of damaged organs/tissues. They adequately support cell adhesion, migration, proliferation, and differentiation. Naturally derived biomaterials can induce extracellular matrix formation and tissue repair when implanted into a defect by enhancing attachment and migration of cells from surrounding environment. In the current chapter, we will focus on the natural and synthetic dermal matrix development and all of the progress in this field.

Development of decellularized aortic scaffold for regenerative medicine using Sapindus mukorossi fruit pericarp extract.

The aim of this study is to develop a novel decellularization method using aqueous extract of soap nut pericarp (SPE) and its evaluation using hematoxylin-eosin staining, scanning electron microscopy, diamidino-2-phenylindol (DAPI) staining, mechanical testing, sodium dodecyl sulfate polyacrylamide gel electrophoresis and DNA quantification. The presently available decellularization agent raises some concerns due to the potential for presence of residual cytotoxic agents in the extracellular matrix. Histological analysis of hematoxylin and eosin and masson's trichrome stained processed aortic samples shows complete decellularization with preservation of extracellular matrix microarchitecture at 120 h. Further, staining of tissue samples with DAPI demonstrates complete removal of DNA fragments. Quantitative evaluation of DNA in the decellularized aorta tissues demonstrated a significant (P < 0.01) decrease in DNA content as compared to native tissues. Collagen quantification assay indicate no significant (P> 0.05) difference in its content between native and decellularized caprine aorta. Tensile strength of the decellularized scaffolds decreased non-significantly (P > 0.05) when compared to native tissues. There was no significant (P > 0.05) difference in young's modulus of elasticity, stiffness and stretch ratio between native aortic tissues and decellularized aortic scaffolds. Histological and scanning electron microscopic examination of in vitro cultured scaffold demonstrated the cell viability and proliferation of primary chicken embryo fibroblasts. SPE treatment is thus capable of producing cytocompatible decellularized caprine aorta scaffold with preservation of extracellular matrix architecture for vascular tissue engineering and could be applied widely as one of the decellularization agent.

Sleep quality and insulin resistance in adolescent subjects with different circadian preference: A cross-sectional study.

BACKGROUND: Studies have shown that alterations in the sleep cycle can predispose to several disorders. Most of the previous studies were done on the adults. Hence, the aim of the study was to see the effect of circadian disruption on the health of adolescent population. MATERIALS AND METHODS: In this cross-sectional study, 203 subjects were enrolled. Study subjects were divided into three groups: definite evening chronotype, intermediate chronotype, and definite morning chronotype. Sleep quality was measured by Pittsburgh Sleep Quality Index (PSQI). Daytime sleepiness and chronotype were measured by Epworth Sleepiness Score and Morningness-Eveningness Questionnaire Self-Assessment version, respectively. Two hours postprandial glucose was measured after oral glucose tolerance test. Fasting blood glucose and fasting insulin were measured. Homeostasis model of assessment for insulin resistance (HOMA-IR) was calculated. Data were summarized as mean ± standard deviation. Crude odds ratios and Karl Pearson's correlation coefficient of metabolic parameters with poor sleep were calculated. RESULTS: Statistically significant difference was found in the mean value of poor sleep quality, 2 h postprandial blood glucose level, and insulin resistance among subjects of three groups. Subjects of evening chronotype have more significant positive correlation of 2 h postprandial blood glucose level and HOMA-IR value with poor sleep quality when compared with subjects of intermediate and morning chronotypes. CONCLUSION: Subjects with evening chronotype are more prone for development of metabolic syndrome compared with subjects of intermediate and morning chronotypes if proper health policies are not adopted for adolescents.

Role of Vitamin-D in the prevention and treatment of Alzheimer's disease.

Alzheimer's disease is the most common form of age related cognitive impairment. Aim of the present study was to see the effect of vitamin D on cognitive function in elderly. The study was conducted in Department of Physiology, King George's Medical University (KGMU). A total of 80 subjects were enrolled based on Mini Mental State Examination (MMSE) score < 24 and vitamin D deficiency. They were divided into two groups as Group A (case) and Group B (control), each group having 40 subjects. Intervention (Vitamin D supplementation) was given in Group A. The assessment of dementia was done by Mini Mental State Examination (MMSE) score. Every subject was clinically evaluated and estimation of vitamin D was done by direct ELISA kit. Gender, weight, height, BMI, residence and education were also similar between two groups. A significant (p=0.0001) change in MMSE score was observed in both Group A and Group B from baseline to 3 & 6 months and from 3 to 6 months, however, mean change was higher in Group A than Group B. In conclusion, vitamin D supplementation caused significant improvement in the cognitive performance in subjects with senile dementia.

Primary chicken embryo fibroblasts seeded acellular dermal matrix (3-D ADM) improve regeneration of full thickness skin wounds in rats.

Rat skins were deepithelialized and decellularized by hypertonic saline and sodium deoxycholate (SDC), respectively. Primary chicken embryo fibroblasts (P-CEF) were cultured and seeded on prepared acellular dermal matrix (ADM). A full thickness skin defect (20×20 mm(2)) was created in thirty-six rats and randomly divided into three equal groups. Defect was left open, repaired with ADM and ADM seeded with P-CEF (3-D ADM) in groups 1, 2 and 3, respectively. By day 28, the treated wounds healed completely without scar. By day 7 hydroxyproline contents was higher in group 3 as compared to groups 1 and 2. There was slightly more B cell response in animals implanted with ADM and 3-D ADM. At day 21, stimulation index was lower with acellular dermis antigen as compared to 3-D ADM antigen. In group 1 on day 3, the granulation tissue showed more inflammatory reaction, fibroplasia and neovascularization as compared to group 2 and 3. By day 28, there was complete epithelization was observed in all groups over. However, a large scar was observed in group 1. The graft was completely absorbed and replaced with densely thick and best arranged collagen fibers. On day 7, malonyldialdehyde and superoxide dismutase levels were significantly (P<0.05) increased in group 1. Reduced glutathione values increased and reached to near normal in groups 2 and 3. Catalase values were significantly (P<0.05) higher in group 1 at different time intervals. SEM samples of group 2 showed ingrowth of fibroblasts into acellular matrix at host graft junction. However, in group 3 fibroblasts were infiltrated within the pores of graft. It was concluded that P-CEF cells seeded ADM facilitated early and better healing.